Objective To discuss the effect of NOD-like receptor protein 3 （NLRP3） inflammasome on the renal interstitial fibrosis in the unilateral ureteral obstruction （UUO） model rats， and to clarify its potential mechanism. Methods Thirty healthy male Wistar rats were randomly divided into sham operation group （n=6） and UUO group （n=24）. The rats in sham operation group underwent the dissection of the ureter without ligation， while the rats in UUO group were sacrificed on the 3rd， 7th， and 14th days after operation， and based on the treatment duration，the rats were divided into UUO 3 d group （n=8）， UUO 7 d group （n=8）， and UUO 14 d group （n=8）. HE staining and Masson staining were used to observe the pathomorphology of kidney tissue of the rats in various groups； reagent kits were used to detect the levels of malondialdehyde （MDA）， activities of superoxide dismutase （SOD）， and levels of hydroxyproline （HYP） in kidney tissue of the rats in various groups；immunohistochemistry method was used to detect the expression levels of α-smooth muscle actin （α-SMA） and transforming growth factor-β1 （TGF-β1） proteins in kidney tissue of the rats in various groups； Western blotting method was used to detect the expression levels of NLRP3 protein in kidney tissue of the rats in various groups. Results The HE staining results showed significant tubular dilation， interstitial edema， and widening， with increased infiltration of inflammatory cells， and shedding of epithelial cells was seen in parts of the tubular lumina of the rats in UUO group. Compared with sham operation group， the interstitial fibrosis scores of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）； compared with UUO 3 d group and UUO 7 d group， the interstitial fibrosis score of the rats in UUO 14 d group was significantly decreased （P<0.05）. The Masson staining results showed that in UUO group， there was evident infiltration of inflammatory cells in the renal interstitium and a noticeable increase in fibrotic tissue proliferation； with the increasing of duration of UUO， some tubular structures disappeared， and the interstitial widened further with gradually increasing collagen deposition， particularly at the corticomedullary junction. Compared with sham operation group， the interstitial fibrosis scores of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）； and compared with UUO 3 d and UUO 7 d groups， the interstitial fibrosis score of the rats in UUO 14 d group was significantly decreased （P<0.05）. Compared with sham operation group， the levels of MDA in obstructed kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）， and the SOD activities were significantly decreased （P<0.05）. Compared with sham operation group， the levels of HYP in obstructed kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were also significantly increased （P<0.05）；compared with UUO 3 d group， the level of HYP in obstructed kidney tissue of the rats in UUO 14 d group was significantly increased （P<0.05）. The immunohistochemistry results showed that compared with sham operation group， the expression levels of α-SMA protein in kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significant increased （P<0.05）； compared with UUO 3 d and UUO 7 d groups， the expression levels of α-SMA protein in kidney tissue of the rats in UUO 14 d group was significantly increased （P<0.05）； compared with sham operation group，the expression levels of TGF-β1 protein in renal tubular epithelial cells and renal tubule interstitial tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were also significantly increased （P<0.05）； compared with UUO 3 d group， the expression levels of TGF-β1 protein in the tubular epithelial cells and renal tubule interstitial tissue of the rats in UUO 14 d group were significantly decreased（P<0.05）. The Western blotting results showed that compared with sham operation group， the expression levels of NLRP3 protein in kidney tissue of the rats in UUO 7 d and UUO 14 d groups were significantly increased （P<0.05）. Conclusion The NLRP3 inflammasome plays a critical role in renal fibrosis of the UUO rats，and its mechanism may be related to the increasing of oxidative stress and the increasing of expression level of TGF-β1 protein.

Objective To discuss the improvement effect of uric acid （UA） on the postoperative cognitive dysfunction （POCD） in the mice anesthetized with isoflurane for a long duration， and to clarify its possible mechanism. Methods Twenty-four healthy male C57BL/6 mice were randomly divided into blank control group，anesthesia group， and UA group, and there were eight mice in each group. The mice in UA group were injected intraperitoneally with 200 μL UA solution daily for 2 d before anesthesia. The mice in blank control group and anesthesia group were given the same volume of saline; the mice in anesthesia group and UA group were used to prepare the models of long-duration isoflurane anesthesia， while the mice in blank control group were untreated. Y-maze tests was used to detect the alternation success rate， movement distances， and movement speeds of the mice in various groups； situational fear experiment was used to detect the percentages of freezing time； Western blotting method was used to detect the expression levels of interleukin (IL)-1β， IL-10， and mature brain-derived neurotrophic factor (mBDNF) proteins in hippocampus tissue of the mice in various groups. Results The Y-maze test results showed that compared with blank control group， the alternation success rate of the mice in anesthesia group was significantly decreased (P<0.01)； compared with anesthesia group， the alternation success rate of the mice in UA group was significantly increased (P<0.01). The situational fear experiment results showed that compared with blank control group, the percentage of freezing time of the mice in anesthesia group was significantly decreased (P<0.01)； compared with anesthesia group， the percentage of freezing time of the mice in UA group was significantly increased (P<0.05). The cued memory experiment resutls showed that there were no significant differences of the percentage of freezing time of the mice between various groups (P>0.05). The Western blotting results showed that compared with blank control group， the expression level of IL-1β protein in hippocampus tissue of the mice in anesthesia group was increased (P<0.01)， while the expression levels of IL-10 and mBDNF proteins were decreased (P<0.01)； compared with anesthesia group, the expression level of IL-1β protein in hippocampus tissue of the mice in UA group was decreased (P<0.05), and the expression levels of IL-10 and mBDNF proteins were increased (P<0.05 or P<0.01). Conclusion UA can improve the POCD in the mice, and its mechnasim may be related with its anti-inflammatory activity inhibiting the central inflammation and upregulating the mBDNF protein expression.

Objective To observe the development process of the neuronal synapse in cerebral cortex and basal ganglionic eminence （GE） regions of the mice， and to clarify the differences in the development of excitatory and inhibitory synapses in different brain regions in vivo and in vitro. Methods The female C57BL/6 mice were euthanized by cervical dislocation from the 13.5th day to the 15.5th day during the pregnancy， and the embryos were collected under the sterile conditions. The cortex and GE regions of brain tissue of the embryonic mice were gradually isolated under microscope. The primary neurons from the embryonic mice were cultured in vitro， and the cell samples were collected on the 3rd， 7th， 14th， and 21th days， respectively， and regarded as culture 3 d， 7 d， 14 d， and 21 d groups. The expression levels of postsynaptic density 95 （PSD95） and Gephyrin mRNA in the primary neurons from the cortex and GE regions of the mice in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR）method. Immunofluorescence method was used to detect the expression levels of vesicular glutamate transporter 1 （vGLUT1）， PSD95， vesicular GABA transporter （vGAT）， and Gephyrin proteins in the neurons from the cortex and GE regions of the mice in various groups. Immunofluorescence method was also used to detect the expression levels of vGLUT1 and vGAT proteins in the neurons from the cortical and GE regions in brain tissue of the embryonic mice. Results Compared with culture 3 d group， the expression levels of PSD95 and Gephyrin mRNA in cortex and GE regions of the mice in culture 14 d and 21 d groups were significantly increased （P<0.01）. Compared with cortex area， the expression level of Gephyrin mRNA in the neurons from GE region of the mice in culture 14 d group was significantly decreased （P<0.01）. The microscope observation results showed that the excitatory and inhibitory synapses in the neurons from cortex and GE regions of the mice in culture 14 d group showed preliminary development， with positive expression of relevant proteins； among them， the excitatory synaptic proteins showed more distinct positive expression in the cortex neurons， and the presynaptic vGLUT1 and postsynaptic PSD95 molecules exhibited co-localization in the cell bodies and protrusions of the cortical neurons； the inhibitory presynaptic vGAT protein and postsynaptic Gephyrin protein in the neurons from GE region also exhibited co-localization in the cell bodies and protrusions， and there were more distinct expressions of the presynaptic molecule proteins than postsynaptic molecule proteins. Compared with cortex region， the levels of vGLUT1 and PSD95 proteins in the neurons from GE region of the mice in culture 14 d group were significantly decreased （P<0.01）， while the levels of vGAT and gephyrin proteins were significantly increased （P<0.01）. In culture 21 d group， the positive expressions of synaptic protein in the neurons from cortex and GE regions were increased， and the excitatory and inhibitory synapses further matured and enhanced. In the neurons from cortex and GE regions， rich patterns of corresponding pre- and postsynaptic expression were formed in the cell bodies and protrusions， and synapse structures showed gradual， positive development， with more apparent expression of presynaptic molecules compared wih postsynaptic proteins. Compared with cortex region， the levels of vGLUT1 and PSD95 proteins in the neurons from GE region of the mice in culture 21 d group were significantly decreased （P<0.01）， and the levels of vGAT and Gephyrin proteins were significantly increased （P<0.01）. Compared with cortex region， the expression level of vGLUT1 protein in the neurons from GE region in brain tissue of the embryonic mice was significantly decreased （P<0.01）， while the expression level of vGAT protein was significantly increased （P<0.05）. Conclusion There are distinct differences in synaptic development between the neurons from cortex and GE regions, the excitatory synapses develope earlier in the cortical region and the inhibitory synapses develope earlier in the GE region. The region-specific development of synapses suggests that different types of neural diseases with different cell types might originate from different developmental processes.

Objective To discuss the effects of leucovorin on tumor growth and angiogenesis in the mice with lung cancer transplantation tumor， and to elucidate its mechanism. Methods Thirty-two healthy C57BL/6 mice were selected to prepare the Lewis lung cancer transplantation tumor models. The mice were randomly divided into model group （0.9% NaCl）， low dose of leucovrin group （25 mg·kg^－1 leucovrin）， high dose of leucovrin group （50 mg·kg^－1 leucovrin） and positive control group （60 mg·kg^－1 cyclophosphamide）， and there were 8 mice in each group. The levels of interleukin-2 （IL-2）， interferon-γ （INF-γ） and tumor necrosis factor-α （TNF-α） in serum of the mice in various groups were detected by enzyme linked immunosorbent assay（ELISA） method； the expression of vascular endothelial growth factor （VEGF） in tumor tissue of the mice in various groups were detected by immunohistochemistry， and the microvascular density （MVD） and VEGF protein expression scores were calculated； the expression levels of nuclear factor-κB（NF-κB） and hypoxia inducible factor-1α（HIF-1α） in tumor tissue of the mice in various groups were detected by Western blotting method. Results Compared with model group， the transplantation tumor masses of the mice in low and high doses of leucovorin groups and positive control group were decreased （P<0.05）； compared with low dose of leucovorin group， the transplantation tumor masses of the mice in high dose of leucovorin group and positive control group were decreased （P<0.05）， and the tumor inhibitory rates were increased （P<0.05）； compared with high dose of leucovorin group， the transplantation tumor mass of the mice in positive control group was decreased （P<0.05）， and the tumor inhibitory rate was increased （P<0.05）. Compared with model group， the spleen indexes and thymus indexes of the mice in low and high doses of leucovorin groups and positive control group were increased （P<0.05）； compared with low dose of leucovorin group， the spleen indexes and thymus indexes of the mice in high dose of leucovorin group and positive control group were increased （P<0.05）； compared with high dose of leucovorin group， the spleen index and thymus index of the mice in positive control group were increased （P<0.05）.The ELISA method results showed that compared with model group， the levels of IL-2， INF-γ and TNF-α in serum of the mice in low and high doses of leucovorin groups and positive control group were increased （P<0.05）； compared with low dose of leucovorin group， the levels of IL-2， INF-γ and TNF-α in serum of the mice in high dose of leucovorin group and positive control group were increased （P<0.05）； compared with high dose of leucovorin group， the levels of IL-2， INF-γ and TNF-α in serum of the mice in positive control group were increased（P<0.05）. Compared with model group， the MVD in tumor tissue of the mice in low and high dose of leucovorin groups and positive control group was decreased （P<0.05）； compared with low dose of leucovorin group， the MVD in tumor tissue of the mice in high dose of leucovorin group and positive control group was decreased （P<0.05）； compared with high dose of leucovorin group， the MVD in tumor tissue of the mice in positive control group was decreased （P<0.05）. Compared with model group， the expression amounts of VEGF protein in tumor tissue of the mice in low and high dose of leucovorin groups and positive control group were decreased； compared with low dose of leucovorin group， the expression amounts of VEGF protein in tumor tissue of the mice in high dose of leucovorin group and positive control group were decreased； compared with high dose of leucovorin group， the expression amount of VEGF protein in tumor tissue of the mice in positive control group was decreased. The differences in VEGF protein expression scores between model group， low dose of leucovorin group， high dose of leucovorin group and positive control group were statistically significant （P<0.05）. Compared with model group， the expression levels of NF-κB and HIF-1α in tumor tissue of the mice in low and high doses of leucovorin groups and positive control group were decreased（P<0.05）； compared with low dose of leucovorin group， the expression levels of NF-κB and HIF-1α in tumor tissue of the mice in high dose of leucovorin group and positive control group were decreased （P<0.05）； compared with high dose of leucovorin group， the expression levels of NF-κB and HIF-1α in tumor tissue of the mice in positive control group were decreased （P<0.05）. Conclusion Leucolorin can inhibit the tumor growth， protect the immune organs and suppress the tumor angiogenesis in the Lewis lung cancer transplantation tumor mice， and may exert the therapeutic effect by targeting NF-κB/HIF-1α signaling pathway and down-regulating the expression levels of NF-κB and HIF-1α proteins.

Objective To screen the differentially expressed genes （DEGs） under high phosphate-induced calcification in the vascular smooth muscle cells （VSMCs） by mRNA high-throughput sequencing technology， and to analyze the key genes and signaling pathways involved in the VSMCs calcification. Methods The human VSMCs were divided into control group and model group. The cells in model group was exposed to the high-phosphate medium， while the cells in control group were cultured in DMEM supplemented with 10% fetal bovine serum under the same conditions. The VSMCs in two groups， stably transfected， were cultured for 12 d. The morphology of the cells in two groups were observed and photographed under inverted microscope. The DEGs were selected by Hisat2 software， and Gene Ontology （GO） functional and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were performed by Stringtie software from three aspects， such as biological processes （BP）， molecular functions （MF）， and cellular components （CC）. The calcification of the cells in two groups was observed by Von Kossa staining method. Real-time fluorescence quantitative PCR （RT-qPCR） method was used to analyze the expression levels of alkaline phosphatase （ALP）， bone morphogenetic protein 2 （BMP2）， alpha-smooth muscle actin （α-SMA）， tumor protein 53 （Tp53）， glutathione peroxidase 4 （GPX4）， ferritin light chain 1 （Ftl1）， and glycosylphosphatidylinositol-specific phospholipase D1 （GPLD1） mRNA in the cells in two groups. Results Compared with control group， there were 2 524 DEGs in the cells in model group， and there were 1 368 upregulated DEGs and 1 156 downregulated DEGs. Clustering of DEGs between the cells in two groups was distinct. The GO functional and KEGG pathway enrichment analysis results showed that the upregulated DEGs were primarily involved in regulating the microtubule cytoskeleton， cell polarity， protein localization， and cell cycle regulation among BPs； in constructing cell membrane， microtubule organization， chromosomes， and kinetochore among CCs； and functioning in phosphatidylinositol phosphate， Rho GTPase protein binding， transmembrane transport， and protein kinase regulatory activity among MFs. Downregulated DEGs were mainly involved in cytoplasmic translation， protein membrane localization， mRNA metabolism， and protein endoplasmic reticulum localization among BPs； in forming ribosome subunits， cell membrane， and autophagy among CCs； and functioning in single-stranded DNA， ribonucleoprotein complex， growth factor binding， regulating protein kinase activity， and catalytic activity among MFs. Seven signaling pathways were significantly enriched in upregulated genes， most notably in the biosynthesis of glycosylphosphatidylinositol （GPI） anchors； whereas 18 signaling pathways were significantly enriched in the downregulated genes， most notably in ferroptosis.The RT-qPCR results showed that compared with control group， the expression levels of GPX4， Ftl1， and Tp53 mRNA in the cells in model group were significantly decreased （P<0.01）， while the expression level of GPLD1 mRNA was significantly increased （P<0.01）； compared with control group， the expression level of α-SMA mRNA in the cells in model group was significantly decreased （P<0.01）， and the expression levels of ALP and BMP2 mRNA were significantly increased （P<0.01）. Conclusion The VSMCs underwent calcification and normal cells exhibit the DEGs.The key signaling pathways in the calcification induced by high phosphate in the VSMCs include ferroptosis and GPI anchor biosynthesis， mediated primarily through GPX4， Ftl1， Tp53， and GPLD1.

Objective To discuss the regulatory effect of serum and glucocorticoid-induced protein kinase 1 （SGK1） in the early development of fertilized eggs at G₁ phase of the mice， and to clarify the related mechanism. Methods Some female mice aged 4-6 weeks and weighed about 20 g， and several male mice aged over 8 weeks and weighed about 30 g were selected. The female mice were intraperitoneally injected with 10 IU of pregnant mare serum gonadotropin （PMSG）， followed by 10 IU of human chorionic gonadotropin （HCG） after 48 h. After HCG injection， the female mice were caged overnight with the male mice at a ratio of 1∶1.The fertilized eggs at G₁， S， G₂， and M phases were collected at 12-21 h， 21-26 h， 26-28 h， and 28-30 h after injected with HCG， and their cellular morphology at different cell cycles were observed under light microscope. The mouse fertilized eggs at G₁ phase after superovulation were collected， the mRNA was synthesized in vitro， and divided into no injection group， Tris-EDTA buffer injection group （TE injection group）， and SGK1-mRNA injection group.The SGK1 antibodies were mixed with KSOM culture medium with the concentrations of 1∶25， 1∶50， 1∶100， 1∶200， and 0 to culture the mouse fertilized eggs at G₁ phase. Western blotting method was used to detect the expression levels of SGK1 protein in fertilized eggs of the mice in various groups and the dephosphorylation for phosphorylated SGK1-Threonine 256 site tyrosine15 site of cell diusion cyclin 2（Cdc2）（Cdc2-pTyr15） in the fertilized eggs of the mice in various groups and different concentrations of SGK1 antibody groups and the developmental states of the fertilized eggs in the fertilized eggs of the mice in various groups and different concentrations of SGK1 antibody groups were observed under phase contrast microscope； the expression levels of phosphorylated SGK1-Thr256 （SGK1-pThr256） and Cdc2-pTyr15 proteins in fertilized eggs at different post-HCG injection times were detected by Western blotting method. Results Compared with no injection and TE injection groups， the expression level of SGK1 protein in the cells in SGK1-mRNA injection group was significantly increased （P<0.01）. 27-28 h after injected with HCG， the phosphorylation signaling of Cdc2-pTyr15 in fertilized eggs of the mice in SGK1-mRNA injection group was gradually disappeared， and there was no phosphorylation signaling 29 h after injected with HCG. At 28-29 h after injected with HCG，the phosphorylation signaling of Cdc2-pTyr15 in fertilized eggs of the mice in no injection and TE injection groups gradually disappeared， completely disappeared at 30 h after injected with HCG. With the increasing of the concentration of SGK1 antibody， the disappearing time of the Cdc2-pTyr15 phosphorylation signaling was increased. At 27 h after injected with HCG，the fertilized eggs of the mice in SGK1-mRNA injection group was initiated cleavage； at 31 h after injected with HCG， nearly all the fertilized eggs turned into G₂ phase； at 33 h after injected with HCG， all the fertilized eggs in 0 and 1∶200 SGK1 antibody groups underwent cleavage. However， with the increasing of SGK1 antibody concentration， the cleavage of the fertilized eggs in 1∶25， 1∶50， and 1∶100 SGK1 antibody groups was gradually decreased， particularly at 1∶25 SGK1 antibody group. Compared with no injection and TE injection groups， the death rate of the fertilized eggs of the mice in SGK1-mRNA injection group was significantly decreased at 31 h after injected with HCG （P<0.05）， and the cleavage rate was increased（P<0.05）. With the increasing of the SGK1 antibody concentration， the death rates of the fertilized eggs in different concentrations of SGK1 antibody group were increased （P<0.05）， with the extending of cleavage time was increased， and the cleavage rate of the fertilized eggs was decreased in a dose-dependent manner， and the cleavage rate of fertilized eggs in 1∶25 SGK1 antibody group was the lowest.The expression level of SGK1-pThr256 protein in fertilized eggs of the mice was gradually increased from 27 h after injected with HCG （P<0.05 or P<0.01） in a time-dependent manner； at 28 to 29 h after injected with HCG， the expression levels of Cdc2-Tyr15 protein were gradually decreased （P<0.05） in a time-dependent manner， and had completely disappeared at 30 h after injected with HCG. Conclusion Both the over-expression and inhibition of SGK1 can affect the time for the fertilized eggs at G₁ phase to entry into M phase， suggesting that SGK1 protein may be one of the regulatory factors in the early development of fertilized eggs at G₁ phase of the mice， and it may regulate the development of the fertilized eggs at G₁ phase through regulation of Cdc2.

Objective To discuss the inhibitory effect of Schisandrin B on the proliferation of pancreatic cancer Pan02 cells， and to clarify the mechanism. Methods CCK-8 method was used to detect the proliferation rates of the Pan02 cells after treated with different concentrations（0， 0.78， 1.56， 3.12， 6.25， 12.50， and 25.00 mg·L^－1 ） of Schisandrin B to select the optimal concentration and treatment time of Schisandrin B. The mouse pancreatic cancer Pan02 cells were divided into control group （0 mg·L^－1 Schisandrin B）， 2.5 mg·L^－1 Schisandrin B group， 5.0 mg·L^－1 Schisandrin B group， and 10.0 mg·L^－1 Schisandrin B group. The morpholoy of Pan02 cells invarious groups was observed with light microscope； 5-ethynyl-2'-deoxyuridine （EdU） staining assay was used to detect the positive expression rates of the Pan02 cells in various groups； flow cytometry was used to detect the percentages of the Pan02 cells at different cell cycles and the apoptotic rates of the cells in various groups； Western blotting method was used to detect the expression levels of cell cycle and apoptosis-related proteins in the cells in various groups. Results The CCK-8 method results showed that after treated with Schisandrin B for 48 and 72 h， compared with 0 mg·L^－1 Schisandrin B， the proliferation rates of the Pan02 cells after treated with different concentrations of Schisandrin B were decreased （P<0.01）， especially at 72 h. 0.25， 5.0， and 10.0 mg·L^－1 Schisandrin B were selected to treat the Pan02 cells， and 72 h was the treatment time. In control group， the Pan02 cells had a spindle shape， with good condition， and grew closely adhered to the wall with normal organelles and cytoplasm， in 2.5 and 5.0 mg·L^－1 Schisandrin B groups， the cell volume was decreased， the intercellular adhesion was disappeared， and the cell membrane was intact but more permeable； the cytoplasm shrank and vacuolar structures appeared inside the cells， with some fragmented and floating on the surface of the solution； in 10.0 mg·L^－1 Schisandrin B group， the Pan02 cells exhibited notable apoptotic bodies， indicating an apoptotic state. The EdU staining results showed that compared with control group， the rates of EdU positive cells in 2.5， 5.0， and 10.0 mg·L^－1 Schisandrin B groups were significantly decreased （P<0.01）. The flow cytometry results showed that compared with control group， the percentages of the cells at S phase in 2.5， 5.0， and 10.0 mg·L^－1 Schisandrin B groups were significantly increased （P<0.01）， while the percentages of the cells at G₂/M phase were significantly decreased （P<0.01）， and the percentages of the cells at G₀/G₁ phase in 5.0 amd 1.0 mg·L^－1 Schisandrin groups were decreased （P<0.01）； compared with control group， the apoptotic rates of the cells in 2.5， 5.0， and 10.0 mg·L^－1 Schisandrin B groups were significantly increased （P<0.01）. The Western blotting results showed that compared with control group， the expression levels of p27， B-cell lymphoma 2 （Bcl-2） associated X protein （Bax）， cleaved cysteine aspartic acid protease-3 （cleaved Caspase-3）， and cleaved poly adenosine diphosphate（ADP） ribose polymerase （cleaved PARP） proteins in the cells in 2.5 mg·L^－1 Schisandrin B group were significantly increased （P<0.05 or P<0.01）， the expression levels of cyclin A2， cyclin E2， and Bcl-2 proteins in the cells in 5.0 and 10.0 mg·L^－1 Schisandrin B groups were significantly decreased （P<0.05 or P<0.01）， while the expression levels of p27， Bax， cleaved Caspase-3， and cleaved PARP proteins in the cells in 5.0 and 10.0 mg·L^－1 Schisandrin B groups were significantly increased （P<0.01）. Conclusion Schisandrin B has an inhibitory effect on proliferation of the pancreatic cancer Pan02 cells， and its mechanism may be related to the activation of the cysteine aspartic acid protease-3 （Caspase-3） pathway to induce the apoptosis and activating p27 protein to induce the arrest of cell cycle at S phase.

Objective To discuss the effect of D-limonene on the proliferation and apoptosis of the glioblastoma （GBM） cells，and to clarify its possible mechanism. Methods The GBM cells were divided into control group （0 mmol·L^-1 D-limonene） and 0.2， 0.4， 0.6， 0.8， and 1.0 mmol·L^-1 D-limonene groups. CCK-8 method was used to detect the inhibitory rates of proliferation of the cells in various groups； clone formation assay was used to detect the clone formation rates of the cells in various groups； Annexin Ⅴ-FITC/PI method was used to detect the apoptotic rates of the cells in various groups； Western blotting method was used to detect the expression levels of protein kinase B （AKT）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and poly adenosine diphosphate（ADP）-ribose polymerase （PARP） proteins in the cells in various groups； imunofluorescence method was used to detect the expression levels of cleaved Caspase-3 protein in the cells in various groups. Fifteen model mice with subcutaneous tumor xenografts were randomly divided into blank group （0 mg·kg^-1·d^-1 D-limonene）， low dose of D-limonene group （200 mg·kg^-1·d^-1 D-limonene）， and high dose of D-limonene group （400 mg·kg^-1·d^-1 D-limonene）， and there were 5 mice in each group.The inhibitory rates of the tumor in vitro in various groups were calculated； HE staining and immunohistochemical staining were used to observe the morphology of subcutaneous tumor tissue of the mice in various groups and the growth curves of the tumor were drawn； immunohistochemical assay was used to detect the positive expression rates of Ki67 protein in subcutaneous tumor tissue of the mice in various groups； TUNEL staining was used to detect the apoptosis of the tumor cells in various groups. Results In control group， the cells were spindle-shaped， in good condition， growing closely and adherently， with normal organelles and cytoplasm. After treated for 48 h， the cells in 0.6 mmol·L^-1 D-limonene group showed reduced volume， intact but more permeable cell membranes， shrunken cytoplasm， internal vacuole structures， and some fragments floating in the solution. The cells in 0.8 and 1.0 mmol·L^-1 D-limonene groups exhibited significant apoptotic bodies and were in an apoptotic state. The CCK-8 results showed that compared with control group， the inhibitory rates of proliferation of the U87， LN229， and GL261 cells in 0.6， 0.8， and 1.0 mmol·L^-1 D-limonene groups were significantly increased （P<0.01）， the inhibitory rates of proliferation of the U87 and GL261 cells were significantly increased （P<0.01）. The clone formation assay results showed that compared with control group， the clone formation rates of the U87， LN229， and GL261 cells in 0.4， 0.6， and 0.8 mmol·L^-1 D-limonene groups were significantly decreased （P<0.05 or P<0.01）. The Annexin Ⅴ-FITC/PI results showed that compared with control group， after treated with D-limonene for 48 h， the apoptotic rates of the LN229 cells in 0.6， 0.8， and 1.0 mmol·L^-1 D-limonene groups were significantly increased （P<0.01）. The Western blotting results showed that compared with control group， the expression levels of Bax proteins in the LN229 cells in 0.6， 0.8， and 1.0 mmol·L^-1 D-limonene groups were significantly increased （P<0.01）， while the expression levels of AKT and Bcl-2 proteins were significantly decreased （P<0.01）， the expression level of PARP protein in the LN229 cells in 0.8 and 1.0 mmol·L^-1 D-limonene group was significanthy increased（P<0.01）.The immunofluorescence results showed that compared with control group， the expression levels of cleaved Caspase-3 protein in the LN229 cells in 0.6， 0.8， and 1.0 mmol·L^-1 D-limonene groups were significantly increased （P<0.01）. Compared with blank group， the tumor volumes of the mice in low and high doses of D-limonene groups were significantly decreased （P<0.01）. Compared with blank group， the tumor weights of the mice in low and high doses of D-limonene groups were significantly decreased （P<0.05）， and the inhitory rates of tumor were significantly increased （P<0.05）. The tumor cells in blank group were diffusely distributed， with deepened nuclear staining and increased nucleocytoplasmic ratio； a large number of degenerated and necrotic tumor cells were observed in tumor tissue of the mice in low and high doses of D-limonene groups. Compared with blank group， the positive expression rates of Ki67 protein in tumor tissue of the mice in low and high doses of D-limonene groups were significantly decreased （P<0.01）. Compared with blank group， the apoptotic rates of tumor cells of the mice in low and high doses of D-limonene groups were significantly increased （P<0.01）. Conclusion D-limonene has the inhibitory effect on the proliferation of the GBM cells； its mechanism may be related to the regulation of AKT protein expression and the activation of the Caspase-3 pathway to induce the apoptosis.

Objective To discuss the improvement effect of Shenshuai Yingyang Capsule in the renal osteodystrophy model rats and its effect on the bone morphogenetic protein （BMP）-7/Smads signaling pathway，and to clarify the relationship between renal function and bone metabolism in the renal osteodystrophy model rats. Methods Fifty SPF SD rats were selected， and 10 of which were randomly chosen as control group， and the remaining 40 rats were used to establish the renal osteodystrophy model. Thirty successful modeling rats were divided into model group， positive control group， and Shenshuai Yingyang Capsule group， and there were 10 rats in various groups. The rats in control and model groups were given saline， the rats in positive control group were given 0.01 mg·kg^-1 calcitriol， and the rats in Shenshuai Yingyang Capsule group were given 1.2 g·kg^-1 Shenshuai Yingyang Capsule， all the rats were treated for 12 weeks. HE staining was used to observe the pathomorphology of femur tissue of the rats in various groups； automatic biochemical analyzer and chemiluminescence were used to detect the kidney function and calcium-phosphorus metabolism indicators in serum of the rats in various groups； real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of BMP-7 and Smad1/5/8 mRNA in femur tissue of the rats in various groups； Western blotting method was used to detect the expression levels of BMP-7， phosphorylated BMP-7（p-BMP-7）， Smad1/5/8， and phosphorylated-Smad1/5/8 （p-Smad1/5/8） proteins in femur tissue of the rats in various groups. Results The HE staining results showed that there was normal trabecular arrangement in control group with unchanged osteoblasts and osteoid area. In model group， the trabecular width and average osteoid area were increased， while the number of osteoblasts was decreased. In positive control and Shenshuai Yingyang Capsule groups， the trabecular width and average osteoid area were decreased， and the number of osteoblasts was increased. Compared with control group，the levels of blood urea nitrogen （BUN） and serum creatinine （Scr） of the rats in model group were increased （P<0.05）； compared with model group， the levels of BUN and Scr of the rats in positive control and Shenshuai Yingyang Capsule groups were decreased （P<0.05）； compared with positive control group， the levels of BUN and Scr of the rats in Shenshuai Yingyang Capsule group were decreased （P<0.05）. Compared with control group， the level of calcium ion （Ca²⁺） in serum of the rats in model group was decreased（P<0.05）， and the levels of phosphorus ion （P^3-）， parathyroid hormone （PTH）， and alkaline phosphatase （ALP） were increased （P<0.05）； compared with model group， the levels of Ca²⁺ in serum of the rats in positive control and Shenshuai Yingyang Capsule groups were increased（P<0.05）， and the levels of P^3-， PTH， and ALP were decreased （P<0.05）； compared with positive control group， the level of Ca²⁺ in serum of the rats in Shenshuai Yingyang Capsule group was increased， and the levels of P^3-， PTH， and ALP were decreased （P<0.05）. The RT-qPCR results showed that compared with control group，the expression levels of BMP-7 and Smad1/5/8 mRNA in femur tissue of the rats in model group were decreased （P<0.05）； compared with model group， the expression levels of BMP-7 and Smad1/5/8 mRNA of the rats in positive control drug group and Shenshuai Yingyang Capsule group were increased （P<0.05）； compared with positive control group， the expression levels of BMP-7 and Smad1/5/8 mRNA in femur tissue of the rats in Shenshuai Yingyang Capsule group were increased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of BMP-7， p-BMP-7， Smad1/5/8， and p-Smad1/5/8 proteins in femur tissue of the rats in model group were decreased （P<0.05）； compared with model group， the expression levels of BMP-7， p-BMP-7， Smad1/5/8， and p-Smad1/5/8 proteins in femur tissue of the rats in positive control group and Shenshuai Yingyang Capsule group were increased （P<0.05）； compared with positive control group， the expression levels of BMP-7， p-BMP-7， Smad1/5/8， and p-Smad1/5/8 proteins in femur tissue of the rats in Shenshuai Yingyang Capsule group were increased （P<0.05）. Conclusion Shenshuai Yingyang Capsule can improve the kidney function and calcium-phosphorus metabolism disorders in the renal osteodystrophy model rats， promote the proliferation of the bone marrow stromal cells， and enhances the bone metabolism； its mechanism may be related to the activation of BMP-7/Smads signaling pathway.

Objective To discuss the effects of monocyte chemoattractant protein-1 （MCP-1） on the migration and invasion of lung cancer A549 cells， and to clarify the mechanisms. Methods Immunohistochemistry method was used to detect the expression of MCP-1 protein in 80 cases of non-small cell lung cancer （NSCLC） and adjacent normal lung tissues. The human lung cancer A549 cells were cultured in vitro. The MCP-1-small interfering RNA （siRNA） experiment was divided into blank group， negative control group （si-NC group）， MCP-1-siRNA-1 group， and MCP-1-siRNA-2 group. The MCP-1 over-expression experiment was divided into control group， empty vector control group （OE-NC， transfected with MCP-1 over-expression empty vector）， over-expression MCP-1 group （OE-MCP-1 group， transfected with MCP-1 over-expression plasmid）， over-expression MCP-1+extracellular regulated protein kinase （ERK） pathway inhibitor PD98059 group （OE-MCP-1+PD98059 group， co-transfected with MCP-1 over-expression plasmid and PD98059）， and PD98059 group （transfected with PD98059）.The MCP-1 siRNA and plasmids were transfected into the lung cancer A549 cells； Western blotting method was used to verify the transfection efficiencies of the cells in various groups； the migration rate and the number of invasion cells in various groups were observed by wound healing assay and Transwell chamber assay， respectively； Western blotting method was also used to detect the expression levels of phosphorylated ERK （p-ERK）， total ERK （t-ERK）， and epithelial-mesenchymal transition （EMT）-related proteins in the A549 cells in various groups. Results Compared with adjacent tissue， the positive expression rate of MCP-1 protein in NSCLC tissue was significantly increased （P<0.05）， and the expression level of MCP-1 protein was related to TNM stage and lymph node metastasis （P<0.05）. Compared with si-NC group， the expression level of MCP-1 protein in the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups was significantly decreased （P<0.01）. Compared with control group and OE-NC group， the expression level of MCP-1 protein in the cells in OE-MCP-1 group was significantly increased （P<0.01）. The wound healing assay results showed that compared with si-NC group， the migration rate of the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups were significantly decreased （P<0.01）. Compared with OE-NC group， the migration rate of the cells in OE-MCP-1 group was significantly increased （P<0.01）； compared with OE-MCP-1 group， the migration rate of the cells in OE-MCP-1+PD98059 group was significantly decreased （P<0.01）. Compared with OE-MCP-1+PD98059 group， the migration rate of the cells in PD98059 group was significantly decreased （P<0.01）. The Transwell chamber assay results showed that compared with si-NC group， the number of invasion cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups was significantly decreased （P<0.01）. Compared with OE-NC group， the number of invasion cells in OE-MCP-1 group was significantly increased （P<0.01）； compared with OE-MCP-1 group， the number of invasion cells in OE-MCP-1+PD98059 group was significantly decreased （P<0.01）； compared with OE-MCP-1+PD98059 group， the number of invasion cells in PD98059 group was significantly decreased （P<0.01）.The Western blotting results showed that compared with si-NC group， the expression levels of p-ERK， Vimentin， and N-cadherin protein in the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups were significantly decreased （P<0.05 or P<0.01）， and the expression level of E-cadherin proteins was significantly increased （P<0.01）. Compared with OE-NC group， the expression levels of p-ERK， Vimentin， and N-cadherin proteins in the cells in OE-MCP-1 group were significantly increased （P<0.01）， and the expression level of E-cadherin protein was significantly decreased （P<0.01）. Compared with OE-MCP-1 group， the expression levels of p-ERK， Vimentin， and N-cadherins proteins in the OE-MCP-1+PD98059 group were significantly decreased （P<0.01）， and the expression level of E-cadherin protein was significantly increased （P<0.05）. Compared with OE-MCP-1+PD98059 group， the expression levels of p-ERK， Vimentin， and N-cadherin proteins in the cells in PD98059 group were significantly decreased （P<0.05 or P<0.01）， and the expression level of E-cadherin protein was increased （P<0.01）. Conclusion MCP-1 protein can upregulate the expression of EMT-related proteins in the lung cancer A549 cells， and promote the migration and invasion of the lung cancer A549 cells； its mechanism may be related to the activation of the ERK signaling pathway.

Objective To discuss the effect of long-term use of chlorhexidine on the resistance of Enterococcusfaecalis （E.faecalis）， and to clarify its mechanism. Methods The standard strain of E.faecalis was repeatedly exposed to chlorhexidine for 10 generations， and the minimum inhibitory concentration （MIC） was recorded at each passage. The bacteria collected from the 10th generation with increased MIC values were designated as the E.faecalis chlorhexidine-resistant strains （E.faecalis-Cs）. The growth curves of two strains were drawn； the morphology of two strains were observed by transmission electron microscope；the number of biofilm formation of two strains was detected by crystal violet staining；the bacterial hydrophobicities of two strains were detected by microbial adhesion to hydrocarbons （MATH） method； the expression levels of S-ribosylhomocysteine lyase （LuxS） mRNA in the bacterial biofilms of two strains were detected by real-time fluorescence quantitative PCR （RT-qPCR） method. Results From the 0th to the 10th generation， the MIC values of E.faecalis were gradually increased. The growth curves of E.faecalis and E.faecalis-Cs showed no significant differences. The transmission electron microscope observation results showed that both E.faecalis and E.faecalis-Cs appeared oval or diplococcal， with intact cell wall structures， smooth edges， and evenly distributed cytoplasm. There were no significant differences in the morphology， size， cell wall thickness， or integrity between two types of bacteria.The crystal violet staining results showed that compared with E.faecalis， the number of biofilm formation of E.faecalis-Cs was significantly increased （P<0.05）. The MATH results showed tha the hydrophobicity of E.faecalis-Cs was significantly higher than that of E.faecalis （P<0.05）. The RT-qPCR results showed that the expression level of LuxS mRNA in the biofilms of E.faecalis-Cs was significantly higher than that of E.faecalis （P<0.05）. Conclusion E.faecalis develops the resistance after repeated exposure to the chlorhexidine，and the pathogenicity of the resistant strain is enhanced. The high expressin of quorum sensing （QS） system LuxS gene and stronger biofilm forming ability of bacteria may be the potential mechanism for E.faecalis to tolerate the chlorhexidine.

Objective To discuss the effects of different doses of ganoderic acid A （GAA） on the biological activities， glycolysis， and the expression of rate-limiting enzymes of non-small cell lung cancer （NSCLC） PC9 cells， and to clarify the mechanism. Methods The NSCLC PC9 cells were cultured in vitro and divided into blank control group， low dose （25 μmol·L^-1） of GAA group， medium dose （50 μmol·L^-1） of GAA group， and high dose （100 μmol·L^-1） of GAA group. The methylthiazolydiphenyltetrazolium（MTT） assay was used to detect the survival rates of the PC9 cells in various groups；Transwell chamber assay was used to detect the number of migration cells of the PC9 cells in various groups； the glucose uptakes of the PC9 cells in various groups were detected by glucose assay kit；the levels of ATP in the PC9 cells in various groups were detected by ATP assay kit； the levels of lactic acid in the PC9 cells in various groups were detected by lactate assay kit； the expression levels of hexokinase 2 （HK2） and pyruvate kinase M2 （PKM2） mRNA in the PC9 cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method； the expression levels of HK2 and PKM2 proteins in the PC9 cells in various groups were detected by Western blotting method. Results The MTT assay results showed that， at 24， 48， and 72 h of culture， compared with blank control group， the survival rates of the cells in medium and high doses of GAA groups were significantly decreased （P<0.05）. At 72 h of culture， compared with low dose of GAA group， the survival rate of the cells in high dose of GAA group was significantly decreased （P<0.05）. The Transwell chamber assay results showed that compared with blank control group， the numbers of migration cells in medium and high doses of GAA groups were significantly decreased （P<0.05）； compared with low doses of GAA group， the number of migration cells in high dose of GAA group was significantly decreased （P<0.05）. Compared with blank control group， the glucose uptakes and levels of lactic acid in the cells in medium and high dose of GAA groups were significantly decreased （P<0.05）， and the level of ATP in the cells in high dose of GAA group was significantly decreased （P<0.05）. The RT-qPCR results showed that compared with blank control group， the expression levels of HK2 and PKM2 mRNA in the cells in medium and high doses of GAA groups were significantly decreased （P<0.05）. The Western blotting results showed that compared with blank control group， the expression levels of HK2 and PKM2 proteins in the cells in medium and high doses of GAA groups were significantly decreased （P<0.05）. Conclusion Medium and high doses of GAA can inhibit the biological activities of proliferation and migration of the PC9 cells， reduce the glycolysis， and its mechanism may be related to the inhibition of the expressions of the key rate-limiting enzymes HK2 and PKM2.

Objective To discuss the anti-fatigue effect of Wujia Shengmai Yin，and to clarify its mechanism. Methods Thirty-six male ICR mice were randomly devided into control group （equivalent volume of distilled water）， Shengmai Yin group （500 mg·kg?1 of Shengmai Yin）， and Wujia Shengmai Yin group （600 mg·kg?1 of Wujia Shengmai Yin）. The body weights of the mice in various groups were detected every 7 d， and the mental states were observed. The rotating rod test and exhaustive swimming test were used to detect the duration on the rod and the swimming time to exhaustion of the mice in various groups， respectively；the levels of urea nitrogen （BUN） and lactate （LA）， and the activities of lactate dehydrogenase （LDH） in serum， the levels of liver glycogen （LG）， muscle glycogen （MG）， and malondialdehyde （MDA）， and activities of glutathione peroxidase （GSH-Px） and superoxide dismutase （SOD） in muscle tissue of the mice in various groups were detected by kits； the expression levels of glucose metabolism-related proteins in liver tissue of the mice in various groups were detected by Western blotting method. Results Compared with before experiment， the body weights after experiment of the mice in various groups showed a increasing trend but the differences were not statistically significant （P>0.05）. The rotating rod test results showed that compared with control group， the duration on the rod of the mice in Wujia Shengmai Yin group was significantly increased （P<0.01）. The exhaustive swimming test results showed that compared with control group， the swimming time to exhaustion of the mice in Shengmai Yin group and Wujia Shengmai Yin group was significantly increased （P<0.01）. Compared with control group， the levels of BUN in serum of the mice in Shengmai Yin group and Wujia Shengmai Yin group were significantly decreased （P<0.01）， and the activities of LDH were significantly increased （P<0.01）； the level of LA of the mice in Wujia Shengmai Yin group was significantly decreased（P<0.01）. Compared with Shengmai Yin group，the levels of BUN and LA in the serum and LDH activity of the mice in Wujia Shengmai Yin group were significantly decreased （P<0.01）. Compared with control group， the levels of LG in liver tissue and the levels of MG in muscle tissue of the mice in Shengmai Yin group and Wujia Shengmai Yin group were significantly increased （P<0.01）； compared with Shengmai Yin group， the level of LG in liver tissue and the level of MG in muscle tissue of the mice in Wujia Shengmai Yin group were increased （P<0.01）. Compared with control group， the activities of GSH-Px and SOD in muscle tissue of the mice in Shengmai Yin group and Wujia Shengmai Yin group were significantly increased， and the levels of MDA in Shengmai Yin group and Wujia Shengmai Yin group was significantly decreased （P<0.01）； compared with Shengmai Yin group， the activities of GSH-Px and SOD in muscle tissue of the mice in Wujia Shengmai Yin group were significantly increased and the level of MDA was decreased （P<0.01）. The Western blotting results showed that compared with control group， the expression levels of phosphorylated phosphoinositide 3-kinase （p-PI3K）， phosphorylated protein kinase B （p-AKT）， phosphorylated glycogen synthase kinase 3 beta （p-GSK3β）， and glycogen synthase （GS） proteins in liver tissue of the mice in Shengmai Yin group and Wujia Shengmai Yin group were significantly increased （P<0.05 or P<0.01）； compared with Shengmai Yin group， the expression levels of p-PI3K， p-AKT， p-GSK3β， and GS proteins in liver tissue of the mice in Wujia Shengmai Yin group were significantly increased（P<0.01）. Conclusion Wujia Shengmai Yin enhances the anti-fatigue effect by activating the phosphoinositide 3-kinase （PI3K）/protein kinase B （AKT）/glycogen synthase kinase 3 beta （GSK3β） signaling pathways， and improves the body’s antioxidant capacity， and increases the glycogen synthesis.

Objective To discuss the effect of Xuebijing on brain tissue damage and immune imbalance of helper T lymphocyte 17 （Th17）/regulatory T lymphocyte （Treg） in cerebrospinal fluid （CSF） of the N-methyl-D-aspartate （NMDA） receptor encephalitis model mice， and to clarify its therapeutic effect. Methods Sixty healthy male C57BL/6J mice were randomly divided into control group， model group， low dose of Xuebijing group， and high dose of Xuebijing group， and there were 15 mice in each group. Except for control group， the mice in the other three groups were injected with the antigen combined with immunostimulation to establish the NMDA receptor encephalitis models. The mice in low and high doses of Xuebijing groups were injected intraperitoneally with 5 and 10 mL·kg^-1 of Xuebijing injection， respectively. HE staining was used to observe the pathomorphology of brain tissue of the mice in various groups； TUNEL assay was used to detect the apoptotic rates of the neurons in hippocampus CA1 region of brain tissue of the mice in various groups；enzyme-linked immunosorbent assay （ELISA） method was used to detect the levels of interleukin （IL）-6， IL-10， IL-17， and transforming growth factor β （TGF-β） in serum of the mice in various groups； flow cytometry was used to detect the percentages of Th17 and Treg cells in CSF of the mice in various groups； Western blotting method was used to detect the expression levels of retinoic acid-related orphan receptor γt （RORγt）， forkhead box protein 3 （Foxp3）， IL-10， and IL-17 proteins in brain tissue of the mice in various groups；immunohistochemistry method was used to detect the rates of IL-17 and Foxp3 positive cells in brain tissue of the mice in various groups. Results The HE staining results showed that the hippocampus CA1 region of brain tissue of the mice in control group had a clear structure without obvious lesions； compared with control group， the mice in model group showed partial pyramidal cell shrinkage， elongation of apical dendrites， loss of a few neurons， and sparse tissue in the hippocampus CA1 region of brain tissue； compared with model group， the mice in low and high doses of Xuebijing groups showed that the damage of the cells in the hippocampus CA1 region of brain tissue was decreased， and the morphological recovery， more orderly arrangement， and more significant improvement could be seen in hippocampus CA1 region of the mice in high dose of Xuebijing group. The TUNEL assay results showed that compared with control group， the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in model group was significantly increased （P<0.05）； compared with model group， the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased （P<0.05）； compared with low dose of Xuebijing group， the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in high dose of Xuebijing group was significantly decreased （P<0.05）. The ELISA results showed that compared with control group， the levels of IL-6 and IL-17 in serum of the mice in model group were significantly increased （P<0.05）， while the levels of IL-10 and TGF-β were significantly decreased （P<0.05）； compared with model group， the levels of IL-6 and IL-17 in serum of the mice in low and high doses of Xuebijing groups were significantly decreased （P<0.05）， while the levels of IL-10 and TGF-β were significantly increased （P<0.05）； compared with low dose of Xuebijing group， the levels of IL-6 and IL-17 in serum of the mice in high dose of Xuebijing group were significantly decreased （P<0.05）， while the levels of IL-10 and TGF-β were significantly increased （P<0.05）. The flow cytometry results showed that compared with control group， the percentage of CD4+IL-17A+ Th17 cells in CSF of the mice in model group was significantly increased （P<0.05）， while the percentage of CD25+Foxp3+ Treg cells was significantly decreased （P<0.05）； compared with model group， the percentages of CD4+IL-17A+ Th17 cells in CSF of the mice in low and high doses of Xuebijing groups were significantly decreased （P<0.05）， while the percentage of CD25+Foxp3+ Treg cells was significantly increased （P<0.05）； compared with low dose of Xuebijing group， the percentage of CD4+IL-17A+ Th17 cells in CSF of the mice in high dose of Xuebijing group was significantly decreased （P<0.05）， while the percentage of CD25+Foxp3+ Treg cells was significantly increased （P<0.05）. The Western blotting results showed that compared with control group， the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in model group were significantly increased （P<0.05）， while the expression levels of Foxp3 and IL-10 proteins were significantly decreased （P<0.05）； compared with model group， the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased （P<0.05）， while the expression levels of Foxp3 and IL-10 proteins were significantly increased （P<0.05）； compared with low dose of Xuebijing group， the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in high dose of Xuebijing group were significantly decreased （P<0.05）， while the expression levels of Foxp3 and IL-10 proteins were significantly increased （P<0.05）. The immunohistochemistry results showed that compared with control group， the rate of IL-17 positive cells in brain tissue of the mice in model group was significantly increased （P<0.05）， while the rate of Foxp3 positive cells was significantly decreased （P<0.05）； compared with model group， the rates of IL-17 positive cells in brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased （P<0.05）， while the rates of Foxp3 positive cells were significantly increased （P<0.05）； compared with low dose of Xuebijing group， the rate of IL-17 positive cells in brain tissue of the mice in high dose of Xuebijing group was significantly decreased （P<0.05）， while the rate of Foxp3 positive cells was significantly increased （P<0.05）. Conclusion Xuebijing can effectively ameliorate the brain tissue injury， regulate the cytokine levels， and intervene in immune imbalance of Th17/Treg in the mice with anti-NMDA receptor encephalitis.

Objective To discuss the effect of N-myc downstream-regulated gene 1 （NDRG1） on the enzalutamide （ENZA） resistance in the castration-resistant prostate cancer （CRPC）， and to clarify its mechanism. Methods The human CRPC C4-2 cells and ENZA-resistant strain C4-2/ENZA cells were cultured in vitro. The expression levels of NDRG1 mRNA in the C4-2/ENZA cells and their parental C4-2 cells were detected by real-time fluorescence quantitative PCR （RT-qPCR） method. The expression levels of NDRG1， androgen receptor （AR）， and prostate-specific antigen （PSA） proteins in the cells were detected by Western blotting method to verify the transfection efficiency of the cells. The C4-2/ENZA cells were divided into blank group （normally cultured without treatment）， negative control lentivirus （Lv-NC） group （transfected with Lv-NC）， Lv-NDRG1 group （transfected with Lv-NDRG1）， Lv-NC+ENZA group （transfected with Lv-NC followed by ENZA treatment）， Lv-NDRG1+ENZA group （transfected with Lv-NDRG1 followed by ENZA treatment）， Lv-NDRG1+epidermal growth factor （EGF） group （transfected with Lv-NDRG1 followed by EGF treatment）， and Lv-NDRG1+EGF+ENZA group （transfected with Lv-NDRG1 followed by EGF and ENZA treatment）. The half-maximal inhibitory concentration （IC₅₀）， resistance index （RI）， and proliferation activity of the cells were detected by MTT assay；the apoptotic rates of the cells in various groups were detected by flow cytometry； RT-qPCR method was used to detect the expression levels of NDRG1 mRNA in the cells in various groups； Western blotting method was used to detect the expression levels of NDRG1， AR， phosphorylated AR at serine²¹³ （p-ARSer²¹³）， phosphorylated AR at serine⁸¹ （p-AR^Ser81）， and PSA proteins in the cells in various groups. Results Compared with C4-2 cells， the expression levels of NDRG1 mRNA and protein in the C4-2/ENZA cells were significantly decreased （P<0.01） and the expression levels of AR and PSA proteins were increased （P<0.01）， indicating low expression of NDRG1 in the ENZA-resistant C4-2/ENZA strain. Compared with Lv-NC group， the expression levels of NDRG1 mRNA and protein in the cells in Lv-NDRG1 group were significantly increased （P<0.01）， indicating the successful construction of an NDRG1 gene over-expression strain of C4-2/ENZA resistant cells. The MTT assay results showed that compared with the C4-2 cells，the IC₅₀ of the C4-2/ENZA cells was increased （P<0.01） and the RI was 17.78； compared with Lv-NC group， the IC₅₀ of the C4-2/ENZA cells in Lv-NDRG1 group was decreased （P<0.01）. After 24 h of EGF treatment， compared with Lv-NC group， the IC₅₀ of the C4-2/ENZA cells in Lv-NC+EGF group was significantly increased （P<0.01）； compared with Lv-NDRG1 group， the IC₅₀ of the C4-2/ENZA cells in Lv-NDRG1+EGF group was increased （P<0.01）. Compared with before ENZA treatment， after 24 h of ENZA treatment， the proliferation activities of C4-2 and C4-2/ENZA cells were gradually decreased （F=223.80， P<0.01； F=81.46， P<0.01）. Compared with Lv-NC group， the proliferation activity in the C4-2/ENZA cells in Lv-NDRG1 group after 24 h of ENZA treatment was significantly decreased （P<0.01）. After 24 h of EGF treatment， compared with Lv-NC group， the proliferation activity of the C4-2/ENZA cells in Lv-NC+EGF group was significantly increased （P<0.01）， while the the proliferation activity of the C4-2/ENZA cells in Lv-NDRG1+EGF group was significantly decreased （P<0.01）. The chosen concentration and treatment duration for further testing were 10 000 μmol·L^-1 ENZA and the intervention time was 24 h. The flow cytometry results showed that after 24 h of ENZA treatment， compared with Lv-NC group， the apoptotic rate of the cells in Lv-NDRG1 group was significantly increased （P<0.01）； compared with Lv-NC+ENZA group，the apoptotic rate of the cells in Lv-NDRG1+ENZA group was significantly increased （P<0.01）. After 24 h of EGF treatment， compared with Lv-NDRG1 group， the apoptotic rate of the cells in Lv-NDRG1+EGF group was significantly decreased （P<0.01）， while the apoptotic rate of the cells in Lv-NDRG1+ENZA group was significantly increased （P<0.01）； compared with Lv-NDRG1+ENZA group， the apoptotic rate of the cells in Lv-NDRG1+EGF+ENZA group was significantly decreased （P<0.01）. The Western blotting results showed that after 24 h of ENZA treatment， compared with Lv-NC group， the expression levels of AR and PSA proteins and the ratio of p-ARSer²¹³/AR and p-ARSer⁸¹/AR in the cells in Lv-NDRG1 group were significantly decreased （P<0.05 or P<0.01）. After 24 h of EGF treatment， compared with Lv-NC group， the expression levels of AR and PSA proteins and the ratio of p-ARSer²¹³/AR and p-ARSer⁸¹/AR in the cells in Lv-NC+EGF group were significantly increased （P<0.05 or P<0.01）； compared with Lv-NDRG1 group， the expression levels of AR and PSA proteins and the ratio of p-ARSer²¹³/AR and p-ARSer⁸¹/AR in the cells in Lv-NDRG1+EGF group were significantly increased （P<0.01）. Conclusion Over-expression of NDRG1 can reduce the resistance of CRPC to ENZA， and its mechanism may be related to the inhibition of AR signaling.

Objective To discuss the effect of adipose-derived stem cell-derived exosomes （ADSC-Exos） on the migration ability of the macrophages RAW264.7， and to clarify its role in promoting function of the macrophages. Methods The adipose tissue adjacent to epididymis of the SD rats was isolated to perform primary culture of the adipose-derived stem cells （ADSCs）. The adipogenic and osteogenic differentiation induction was conducted， and the multidirectional differentiation potential of the ADSCs was detected by oil Red O and Alizarin red staining. Western blotting and immunofluorescence methods were used to detect the positive expressions of the ADSCs markers CD29 and CD44； the ADSC-Exos were extracted by Exos isolation kit， and the morphology， size， and distribution of particle size of the ADSC-Exos were examined by transmission electron microscope and nanoparticle tracking analyzer； the expression levels of exosome-specific markers CD9 and TSG101 proteins in the ADSC-Exos were detected by Western blotting method； the uptake of ADSC-Exos by the macrophages was observed by tracing method. The macrophages RAW264.7 were divided into control group， 10 mg·L^-1 ADSC-Exos group， 20 mg·L^-1 ADSC-Exos group， and 40 mg·L^-1 ADSC-Exos group. The activities of the macrophages in various groups were detected by 5-ethynyl-2'-deoxyuridine （EdU） staining； the number of migration macrophages in various groups was detected by Transwell chamber assay； the adhesion of macrophages in various groups was observed by fluorescence microscope. Results After 24 h of primary culture， the ADSCs adhered to the wall and exhibited scattered， elongated shapes； after 7 d of culture， the adherent cells showed a comb-like， vortex-like orderly arrangement， resembling fibroblasts； after 10 passages， the irregular morphology of the ADSCs and decreased proliferation rate were found. The isolated ADSCs showed potential for the osteogenic and adipogenic differentiation， and the expressions of CD29 and CD44 proteins were positive.The transmission electron microscope observation resuls showed that the ADSC-Exos appeared disc-shaped， and the main peak of particle size distribution was around 132 nm. The CD9 and TSG101 proteins were positively expressed in the ADSC-Exos， indicating successful extraction. The fluorescence microscope results showed red fluorescence signals around the nuclei of the RAW264.7 cells， indicating the uptake of ADSC-Exos by the macrophages. Compared with control group， the rates of EdU positive cells in 10，20， and 40 mg·L^-1 ADSC-Exos groups were significantly increased（P<0.05）； compared with 10 mg·L^-1 ADSC-Exos group， the rate of EdU positive cells in 20 mg·L^-1 ADSC-Exos group was significantly increased （P<0.05）. Compared with control group， the numbers of migration cells in 10， 20， and 40 mg·L^-1 ADSC-Exos groups were significantly increased （P<0.05）； compared with 10 mg·L^-1 ADSC-Exos group， the numbers of migration cells in 20 and 40 mg·L^-1 ADSC-Exos groups were significantly increased （P<0.05）. Compared with control group， the numbers of the adherent macrophages in 10， 20， and 40 mg·L^-1 ADSC-Exos groups were significantly increased （P<0.05）； compared with 10 mg·L^-1 ADSC-Exos group， the number of adherent macrophages in 20 mg·L^-1 ADSC-Exos group was significantly increased （P<0.05）. Conclusion The ADSC-Exos can be internalized by the macrophages and they can enhance the migration ability of the macrophages by affecting the cell adhesion.

Objective To discuss the inhibitory effect of microRNA-487a （miR-487a） on the M2 polarization of tumor-associated macrophages （TAMs） in gastric cancer， and to clarify its effect on the proliferation， invasion， and migration of the gastric cancer AGS cells. Methods The TAMs from gastric cancer tissue and adjacent normal tissue macrophages （NTMs） from adjacent tissue of the primary gastric cancer patients were isolated and cultured. The human monocyte THP-1 cells were induced in vitro to differentiate into TAMs， and the differentiated M0， M1， and M2 macrophages were cultured for 24 h by conditioned medium （CM） to obtain the TAMs， M1-TAMs， and M2-TAMs respectively. The TAMs were transfected and then divided into blank group， inhibitor-NC group， miR-487a inhibitor group， miR-487a inhibitor+si-NC group， and miR-487a inhibitor+si-TIA1 group. The transfection efficiencies of the cells in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods. The M2-TAMs were co-cultured with the AGS cells， and divided into AGS group， AGS+inhibitor-NC group， AGS+miR-487a inhibitor group， AGS+miR-487a inhibitor+si-NC group， and AGS+miR-487a inhibitor+si-TIA1 group. RT-qPCR method was used to detect the expression levels of miR-487a and lymphocyte intracytoplasmic antigen-1 （TIA1） mRNA in TAMs from gastric cancer tissue and NTMs from adjacent normal tissue in various groups； Western blotting method was used to detect the expression level of TIA1 protein in TAMs from gastric cancer tissue and NTMs from adjacent normal tissue and TAMs in various groups； flow cytometry was used to detect the levels of CD206 and CD163 in TAMs in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-10 （IL-10）， transforming growth factor-beta （TGF-β）， vascular endothelial growth factor A （VEGF-A）， and arginase-1 （Arg-1） in culture supernatant of the TAMs cells； CCK-8 assay was used to detect the proliferative activity of the AGS cells in various groups； wound healing assay was used to detect the migration rates of the AGS cells in various groups； Transwell assay was used to detect the number of invasion AGS cells in various groups. Results The RT-qPCR results shoued that compared with NTMs from adjacent tissue， the expression level of miR-487a in the TAMs from gastric cancer tissue was significantly increased （P<0.01） and the expression level of TIA1 mRNA was significantly decreased （P<0.01）. Compared with TAMs， the expression level of miR-487a in M1-TAMs was significantly decreased （P<0.01）， and the expression level of TIA1 mRNA was increased （P<0.01）； the expression level of miR-487a in M2-TAMs was significantly increased （P<0.01）， and the expression level of TIA1 mRNA was decreased （P<0.01）. After transfection， compared with blank group and inhibitor-NC group， the expression level of miR-487a in the cells in miR-487a inhibitor group was significantly decreased （P<0.01）， indicating successful transfection. The Western blotting results showed that compared with NTMs from adjacent normal tissue， the expression level of TIA1 protein in TAMs from gastric cancer tissue was decreased （P<0.01）； compared with TAMs， the expression level of T1A1 protein in M1-TAMs was significantly increased （P<0.01）， and the expression of TIA1 protein in M2-TAMs was significantly decreased （P<0.01）； after co-transfection， compared with inhibitor-NC group， the expression level of TIA1 protein in the cells in miR-487a inhibitor group was significantly increased （P<0.01）； compared with miR-487a inhibitor+si-NC group， the expression level of TIA1 protein in the cells in miR-487a inhibitor+si-TIA1 group was significantly decreased （P<0.01）. The flow cytometry results showed that compared with blank group and inhibitor-NC group， the levels of CD206 and CD163 in the cells in miR-487a inhibitor group were significantly decreased （P<0.01）； after co-transfection， compared with inhibitor-NC group， the levels of CD206 and CD163 in the cells in miR-487a inhibitor group were significantly decreased （P<0.01）； compared with miR-487a inhibitor+si-NC group， the levels of CD206 and CD163 in the cells in miR-487a inhibitor+si-TIA1 group were significantly increased （P<0.01）. The ELISA results showed that compared with blank group and inhibitor-NC group， the levels of IL-10， TGF-β， VEGF-A， and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor group were significantly decreased （P<0.01）； after co-transfection， compared with inhibitor-NC group， the levels of IL-10， TGF-β， VEGF-A， and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor group were significantly decreased （P<0.01）； compared with miR-487a inhibitor+si-NC group， the levels of IL-10， TGF-β， VEGF-A， and Arg-1 in culture supernatant of the TAMs in miR-487a inhibitor+si-TIA1 group were significantly increased （P<0.01）. The CCK-8 assay results showed that compared with AGS group， the proliferation activity of the cells in AGS+inhibitor-NC group was significantly increased （P<0.01）； compared with AGS+inhibitor-NC group， the proliferation activity of the cells in AGS+miR-487a inhibitor group was significantly decreased （P<0.01）； compared with AGS+miR-487a inhibitor+si-NC group， the proliferation activity of the cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased （P<0.01）. The wound healing assay results showed that compared with AGS group， the migration rate of the cells in AGS+inhibitor-NC group was significantly （P<0.05）； compared with AGS+inhibitor-NC group， the migration rate of the cells in AGS+miR-487a inhibitor group was significantly decreased （P<0.01）； compared with AGS+miR-487a inhibitor+si-NC group， the migration rate of the cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased （P<0.05）. The Transwell assay results showed that compared with AGS group， the number of invasion AGS cells in AGS + inhibitor-NC group was significantly increased （P<0.01）； compared with AGS + inhibitor-NC group， the number of invasion AGS cells in AGS+miR-487a inhibitor group was significantly decreased （P<0.01）； compared with AGS+miR-487a inhibitor+si-NC group， the number of invasion AGS cells in AGS+miR-487a inhibitor+si-TIA1 group was significantly increased （P<0.01）. Conclusion Silencing the miR-487a expression can inhibit the M2 polarization of the gastric cancer-associated macrophages by targeted upregulation of TIA1， and suppress the proliferation， migration， and invasion of the gastric cancer cells.

Objective To clone the Helicobacter pylori （Hp） hp0169 gene and conduct the crystallographic study， and to clarify its secondary and tertiary structures. Methods The hp0169 gene and its encoded protein sequence of the Hp NCTC26695 strain were retrieved from the UniProt database. Bioinformatics method was used to analyze the physicochemical properties of the Hp recombinant protease （HpPrtC） protein； SOPMA and DNAStrar softwares were used to predict the secondary structure characteristics of HpPrtC protein； SWISS-MODEL software was used to construct the tertiary structure of the HpPrtC protein； IEDB and ABCpred softwares were used to predict the antigenic epitopes of the B lymphocytes HpPrtC protein； SYFPEITMI website was used to predict the antigenic epitopes of the T lymphocytes of HpPrtC protein； the expert pool （EP） and random forest （RF） algorithms were used to predict the crystallizability of the HpPrtC protein；the HpPrtC recombinant protein was expressed in the prokaryotic system； the HpPrtC recombinant protein was purified by Ni²⁺ affinity chromatography and size-exclusion chromatography；the crystallization conditions for HpPrtC were screened by crystallization kit. Results The hp0169 gene contained 1 269 base pairs and encoded the protein of 422 amino acids， the theoretical isoelectric point was 7.64 and the relative molecular weight was 47 300. HpPrtC was a hydrophilic and soluble protein. The number of amino acids of alpha helices of HpPrtC accounted for 35.78%， beta sheets 18.72%， beta turns 6.87%， and random coils 38.63%. The antigen epitope analysis results showed that HpPrtC contained five dominant linear epitopes of B lymphocytes， three conformational epitopes， and multiple potential dominant epitopes of T lymphocytes. The homology modeling results showed that HpPrtC formed a dimer， and each monomer displayed a barrel structure surrounded by β sheets， alpha helices， and random coils. HpPrtC was predicted to have moderate crystallizability without signal peptides and transmembrane helices. Small clustered needle-like crystals of HpPrtC were obtained under the conditions of 0.2 mol·L^-1 magnesium chloride， 0.1 mol·L^-1 tris （hydroxymethyl） amino methane（Tris）， 3.4 mol·L^-1 hexanediol， and pH=8.5. Conclusion HpPrtC is a hydrophilic protein that forms a dimeric structure and crystallizes into small clustered needle-like crystals under suitable conditions. HpPrtC contains dominant antigenic epitopes of the T lymphocytes and B lymphocytes and can serve as an antigen for the design of Hp vaccines to establish the multivalent fusion vaccines or multi-epitope vaccines； the results provide an experimental basis for the prevention and control of Hp.

Objective To screen the differentially expressed immune-related genes （DEIRGs） in in-stent restenosis （ISR）， and to analyze the immune cell infiltration in ISR， and to clarify the mechanism of occurrence and development of ISR. Methods The mRNA gene expression data of GSE46560 dataset samples were downloaded from the Gene Expression Omnibus （GEO），and divided into ISR group and non-ISR group. The “Limma” package in R software was used to identify the differentially expressed genes （DEGs） which were then intersected with immune-related genes （IRGs） to identify the DEIRGs in ISR； R software was used for Gene Ontology （GO） functional enrichment andalysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis on DEIRGs；the STRING database was used to construct the protein-protein interaction （PPI） network， which was visualized and analyzed for Hub genes by Cytoscape software； the receiver operating characteristic （ROC） curve of the Hub genes were plotted， and the area under the curve （AUC） was calculated and the diagnostic value was evaluated； CIBERSORT software was used to analyze the immune cell infiltration in ISR； Pearson correlation analysis was used to analyze the relationships between the immune cells and the relationships between the immune cells and key genes. Results A total of 331 DEGs were identified （P<0.05， | log₂FC| >1）， including 176 upregulated genes and 155 downregulated genes， and 38 DEIRGs were obstained. The GO functional enrichment analysis results showed that the DEIRGs were mainly enriched in biological processes （BP） such as defense response， immune response， and immune system； in cellular components （CC），the DEIRGs were located primarily in the extracellular region and cytoplasmic membrane； and in molecular functions （MF）， the DEIRGs were mainly involved in regulating signaling receptor binding and cytokine receptor activity.The KEGG signaling pathway enrichment analysis results indicated that the DEIRGs in ISR were primarily enriched in the phosphatidylinositol 3-kinase/protein kinase B （PI3K-AKT） and transforming growth factor-β （TGF-β） signaling pathways. In the PPI network， CD19 had the highest node among the top 10 Hub genes. Compared with non-ISR group， the expression level of the CD19 gene in the samples in ISR group was increased （P<0.05）. The AUC value in the ROC curve of CD19 gene expression was 0.92 （P<0.05）. The immune cell infiltration analysis results showed that compared with non-ISR group， the infiltration level of T lymphocyte follicular helper （Tfh） cells in the patients in ISR group were increased （P<0.05）， the infiltration levels of immature B lymphocytes， CD8+T lymphocytes， naive CD4+T lymphocytes， and M0 macrophages were increased， but the differences were not statistically significant （P>0.05）， while the infiltration levels of memory B lymphocytes， activated memory CD4+ T lymphocytes， regulatory T cells， resting natural killer （NK） cells， activated NK cells， monocytes， resting mast cells， and neutrophils were decreased， but the differences were not statistically significant （P>0.05）. There were positive correlations between Tfh cells and M0 macrophages and resting mast cells （r=0.88，P<0.05；r=0.68，P<0.05）， and there were negative correlations between Tfh cells and monocytes and neutrophils（r=-0.49，P<0.05；r=-0.42，P<0.05）. Conclusion CD19 may influence the occurrence and development of ISR by regulating the activation of the PI3K-AKT signaling pathway to affect the Tfh and B lymphocytes. CD19 can serve as a biomarker for the diagnosis of ISR.

Objective To discuss the mechanism of salidroside in the treatment of triple negative breast cancer （TNBC） by using the bioinformatics and network pharmacology methods， and to clarify the main targets and signaling pathways involved in the therapeutic effect. Methods The dataset GSE45827 was obtained from the Gene Expression Omnibus （GEO） database； the gene set enrichment analysis （GSEA） was performed by using the R software package GSEABase；the differentially expressed genes （DEGs） between the adjacent normal tissue and TNBC tissue were identified by limma R software package；the Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway enrichment analysis were performed on the DEGs， and the DEGs were integrated with the drug targets to import into gene/protein interaction retrieval tool String database， and the protein-protein interaction （PPI） networks were constructed；the functional module screening of the PPI network was conducted by MCODE plugin， and the top 2 modules ranked by SCORE value were further subjected to GO functional enrichment analysis and KEGG signaling pathway analysis. The pathways obtained from the two rounds of KEGG enrichment analysis were intersected with the results of GSEA enrichment analysis to identify the pathways involved in the therapeutic effect of salidroside on TNBC. The top 10 key node genes in the highest scoring module determined by the maximum clique centrality （MCC） score caculated by CytoHubba plugi were considered as the core genes； the molecular docking was performed by AutoDock Vina1.1.2 and PyMOL2.3.0 Software. Results The intersection of KEGG and GSEA enrichment analysis results showed 13 singaling pathways， including the cell cycle， cellular senescence， and p53 signaling pathways，and so on. The biological processes involved in the GO functional analysis， such as mitosis， nuclear division， and sister chromatid separation， were closely related to the cell cycle and consistented with the results of the KEGG signaling pathway enrichment analysis. The top ranked module based on the SCORE value contained 5 drug target genes of Rhodiola glycoside，such as cyclin A2 （CCNA2）， checkpoint kinase 1 （CHEK1）， kinesin family member 11 （KIF11）， DNA topoisomerase 2-alpha （TOP2A）， and thymidylate synthase （TYMS）. The molecular docking results demonstrated strong binding affinities between the above proteins and Rhodiola glycoside （binding energy<－7.0 kcal·mol^－1）. Conclusion The tightly binding target of salidroside is located in the key functional modules of DEGs of TNBC， which can directly regulate by binding with CCNA2 and protein， and indirectly regulate the key differentially genes of TNBC by binding with KIF11， TOPA2， CHEK1 and TYMS proteins. Therefore， salidroside may be a potential clinical therapeutic drug for TNBC.

Objective To discuss the predictive value of lung ultrasound score （LUS） for the use of mechanical ventilation （MV） and pulmonary surfactant （PS） in the preterm infants with late-onset respiratory distress syndrome （RDS）. Methods The prospective analysis was conducted on the late-onset preterm infants （gestational age 34^0/7-36^6/7 weeks） complicated with RDS； in total， 67 late-onset infants complicated with RDS were included. The infants were divided into MV group（n=36）， non-MV group（n=31）， PS group（n=30）， and non-PS group（n=37） based on the necessity to use MV and PS within 48 h after birth. Lung ultrasound examination was performed on all the infants 2 h after admission， and before the application of PS， and the LUS for 6-zone， 10-zone， and 12-zone partitions were calculated. Receiver operating characteristic （ROC） curve for the prediction of MV and PS application in the infants with late-onset RDS were drawn by LUS with different partitions， and the predictive values of different partition methods were compared by DeLong method. Results Compared with non-PS group， the birth weight， LUS， positive end expiratory pressure （PEEP）， mean airway pressure （MAP）， MAP×fraction of inspired oxygen （FiO₂）/PaO₂ value， duration of mechanical ventilation， and hospital stay of the infants in PS group were increased （P<0.05）， and the ratio of PaO₂/FiO₂ was decreased （P<0.01）. Compared with non-MV group， the birth weight， LUS， PEEP， MAP， MAP × FiO₂/PaO₂ value， duration of mechanical ventilation and hospital stay of the infants in MV group were increased （P<0.05）， and the ratio of PaO₂/FiO₂ was decreased （P<0.01）. PEEP， MAP， and LUS were identified as the influencing factors for application of PS in the late-onset preterm infants complicated with RDS when employing 6-zone LUS to predict the application of PS［odds ratio（OR）>1， P<0.05］. When employing 10-zone and 12-zone LUS for the use of PS， MAP × FiO₂/PaO₂ and LUS were the influencing factors （OR>1， P<0.05）. The area under curve （AUC） for predicting the application of PS in the late-onset infants complicated with RDS by 6-zone， 10-zone， and 12-zone LUS were 0.909， 0.904， and 0.915， respectively， all showing good predictive values； the AUCs for predicting the application of MV by 6-zone， 10-zone， and 12-zone LUS were 0.868， 0.872， and 0.887， respectively， all showing good predictive values as well. Conclusion LUS can effectively predict the necessity for whether or not applying MV and PS in the late-onset infants complicated with RDS， and MAP combined with LUS can enhance the capability to predict the application of MV.

Objective To analyze the causal relationship between lifestyle-based factors and the occurrence and development of hepatobiliary malignancies by Mendelian randomization study method， and to provide the potential clinical evidence for the prevention and treatment of hepatobiliary malignancies. Methods The data from large-scale， independent genome-wide association studies （GWAS） were selected， and seven-step inclusion criteria for the instrumental variable screening were set up. The exposure lifestyles included the percentage of carbohydrate intake， percentage of fat intake， percentage of protein intake in the diet， coffee intake， weekly alcohol consumption times， leisure electronic screen exposure time， moderate to vigorous intensity physical activity （MVPA） during leisure time， sedentary behavior at work， age at first smoking， daily smoking quantity， current smoking status， and past smoking status， totaling 12 phenotypes. The primary analysis method used was the random effect model of the inverse variance weighted （IVW） method， and the heterogeneity was detected by Cochrane’s Q test and the horizontal pleiotropy was detected by MR-Egger intercept method. Results The current smoking status was significantly positively correlated with the increasing risk of extrahepatic cholangiocarcinoma （OR=1.607， 95% CI： 1.113-2.322， P=0.011）. Higher coffee intake was causally linked to a higher risk of liver cancer and intrahepatic cholangiocarcinoma （OR=1.000， 95% CI： 0.999-1.000， P=0.012）. In the physical activity， more MVPA was associated with the lower risk of liver cancer and intrahepatic cholangiocarcinoma （OR=0.998， 95% CI： 0.996-0.999， P=0.002）. The Cochrane’s Q test results showed that there was mild heterogeneity between MVPA and extrahepatic cholangiocarcinoma（Q=18.354，P=0.049） as well as the percentage of protein intake and intraphepatic cholangiocarainoma（Q=12.715，P=0.026）， and the MR-Egger intercept method results showed there was no horizontal pleiotropy. Conclusion There is a causal relationship between current smoking status and extrahepatic cholangiocarcinoma， and there is a causal relationship between more MVPA and the lower risk of liver cancer and intrahepatic cholangiocarcinoma. Education on smoking and physical activity for the patients may offer potential benefits for the prevention of hepatobiliary malignancies.

Objective To discuss the correlation between homocysteine （Hcy） and inflammatory responses as well as oxidative stress， and to analyze its role in the occurrence and development of acute ischemic stroke （AIS）. Methods Thirty-eight patients with their first incidence of AIS were selected as AIS group， and according to the principles of case-control study， 45 healthy individuals undergwent routine health examination during the same period were selected as control group. The levels of homocysteine and inflammatory cytokine interleukin （IL）-6， and tumor necrosis factor α （TNF-α） in serum of the subjects in two groups were detected by enzyme-linked immunosorbent assay （ELISA） method； the level of highly sensitive C-reactive protein （hs-CRP） of the subjects in two groups was detected by immunoturbidimetry； the levels of malondialdehyde （MDA） and activities of superoxide dismutase （SOD） in serum of the subjects in two groups were detected by chemical colorimetry； Pearson’s correlation analysis was used to analyze the correlation between serum Hcy level and levels of inflammatory markers and oxidative stress indicators of the AIS patients. Results Compared with control group， the levels of Hcy， hs-CRP， TNF-α， IL-6， and MDA in serum of the patients in AIS group were significantly increased （P<0.05）， while the activity of SOD was significantly decreased （P<0.05）. The level of Hcy in serum of the AIS patients was positively correlated with the levels of hs-CRP， TNF-α， IL-6， and MDA （r=0.615， P<0.05； r=0.632， P<0.05； r=0.598， P<0.05； r=0.612， P<0.05）， and negatively correlated with the activity of SOD （r=-0.325， P<0.05）. Conclusion The Hcy level in serum of the AIS patients is closely associated with the inflammatory factors and oxidative damage. Hcy can promote the production of oxidative free radicals and inflammatory factors and cause damage to the endothelial cells and play a significant clinical role in the occurrence and development of AIS.

Objective To analyze the application effect of narrow elastic disposable tourniquet in liposuction reduction surgery for the lower extremity lymphedema，and to provide the basis for its clinical application. Methods The retrospective analysis was conducted on the clinical data of 309 patients who underwent liposuction reduction surgery for the lower extremity lymphedema. The patients were divided into non-narrow elastic disposable tourniquet group （n=163） and narrow elastic disposable tourniquet group （n=146）. The intraoperative blood loss， rates of allogenic blood transfusion， incidence of adverse reactions related to dilation fluid into blood， incidence of blood pressure fluctuations， and preoperative and postoperative 24 h levels of hemoglobin （Hb） and albumin （Alb） of the patients in two groups were compared. Results Compared with non-narrow elastic disposable tourniquet group， the intraoperative blood loss， allogenic blood transfusion rate， and incidence of adverse reactions related to dilation fluid into blood of the patients in narrow elastic disposable tourniquet group were decreased （P<0.05）， while the incidence of intraoperative blood pressure fluctuations was increased （P<0.05）. Compared with non-narrow elastic disposable tourniquet group， the ΔHb level （preoperative Hb level-postoperative 24 h Hb level） and ΔAlb level（preoperative Alb level-postoperative 24 h Alb level） of the patients in narrow elastic disposable tourniquet group were decreased （P<0.05 or P<0.01）. Conclusion The application of narrow elastic disposable tourniquet in liposuction reduction surgery for the lower extremity lymphedema can effectively reduce the intraoperative bleeding and the need for allogenic transfusions， decrease the level of ΔAlb， and prevent the occurrence of adverse reactions related to dilation fluid into blood. However， attention should be given to the potential adverse reactions related to intraoperative circulatory fluctuations.

Objective To discuss the changes in the levels of chemokine ligand 19 （CCL19） and soluble CD163 （sCD163） in serum of the patients with systemic lupus erythematosus （SLE） during pregnancy， and to clarify their effects on the maternal and infant outcomes. Methods A total of 180 pregnant SLE patients were selected as SLE group and then divided into successful pregnancy group （n=132） and pregnancy failure group （n=48） based on the maternal and infant outcomes. A total of 180 healthy pregnant women underwent prenatal checks during the same period were randomly selected as control group. The general data of the patients in two groups were collected， and the serum levels of CCL19 and sCD163， along with related serum factors， were detected by kits. Multivariate Logistic regression analysis was used to detect the risk factors for pregnancy failure in the SLE patients， and receiver operating characteristic （ROC） curve was used to evaluate the effectiveness of serum CCL19 and sCD163 levels in predicting the pregnancy outcomes of the patients in SLE group. Results Compared with control group， the levels of complements C3 and C4 in the serum of the patients in SLE group were significantly decreased （P<0.05）， and the levels of erythrocyte sedimentation rate （ESR）， creatinine （CR）， anti-cardiolipin antibody （ACA）-IgG， anti-β2 glycoprotein Ⅰ （anti-β2GPⅠ）， CCL19， and sCD163 of the patients were significantly increased （P<0.01）. Compared with successful pregnancy group， the levels of complement C3 and C4 pregnancy of the patients in failure group were significantly decreased （P<0.01）， and the levels of ESR， CR， ACA-IgG， anti-β2GPⅠ， CCL19， and sCD163 were significantly increased （P<0.01）. The serum levels of CCL19， sCD163， ESR， CR， ACA-IgG， and anti-β2GPⅠ were the risk factors for pregnancy failure of the SLE patients （P<0.05 or P<0.01）， while the levels of complement C3 and C4 were the protective factors （P<0.01）. The area under the ROC curve （AUC） of the serum CCL19 level for predicting the pregnancy failure of the SLE patients was 0.726， and the AUC of serum SCD163 level for predicting the pregnancy failure of the SLE patients was 0.789； the AUC of combination of both markers for predicting the pregnancy failure of the SLE patients was 0.835. The predictive performance of CCL19 and sCD163 for pregnancy outcomes of the SLE patients was superior to either marker alone （Z_{combined-CCL19}=3.066， P=0.002； Z_{combined-sCD163}=2.087， P=0.037）. Conclusion The serum levels of CCL19 and sCD163 in the SLE patients during pregnancy are significantly increased， which may cause the poor outcomes in the patients.

Objective To discuss the changes of serum levels of Klotho and fibroblast growth factor 23 （FGF23） in the small for gestational age （SGA） infants after birth， and to clarify their relationships with growth and development. Methods A total of 35 SGA and 53 appropriate for the gestational age（AGA） infants were selected and divided into SGA group（n=35） and AGA group（n=53）， including 51 infants in premature group， among them 20 infants in preterm SGA group and 31 infants in preterm AGA group； among them 37 infants in full-term group， 15 infants in full-term SGA group and 22 infants in full-term AGA group. The clinical materials of the infants in various groups were collected. The levels of Klotho and FGF23 in serum and clinical biochemical markers of the infants on the 7th and 14th days after birth were detected. The relationships between the levels of Klotho and FGF23 in serum on the 7th and 14th days postnatally and newborn growth indicators such as body weight， body length， head circumference， chest circumference， and Kopu index， as well as their correlations with calcium and phosphorus metabolism were analyzed. Results Compared with AGA group， the body weight， body length， head circumference， chest circumference， and Kopu index of the infants in SGA group were decreased （P<0.05）. On the 7th and 14th days after birth， compared with preterm group， the serum levels of Klotho and FGF23 of the infants in full-term group were significantly increased （P<0.01）. Compared with the 7th day after birth，the levels of serum Klotho of the infants in preterm and full-term groups on the 14th day were significantly increased （P<0.01）， and the levels of FGF23 in serum were decreased （P<0.01）. Compared with AGA group， the levels of serum Klotho and FGF23 of the infants in SGA group on the 7th and 14th days after birth were significantly decreased （P<0.05 or P<0.01）. Compared with the 7th day after birth， the levels of serum Klotho of the infants in both AGA and SGA groups on the 14th days after birth were significantly increased （P<0.01）， and the FGF23 levels were decreased （P<0.05 or P<0.01）. Compared with preterm AGA group， the levels of Klotho and FGF23 in serum of the infants in preterm SGA group on the 7th and 14th days after birth were significantly decreased （P<0.05 or P<0.01）. Compared with full-term AGA group， the levels of Klotho and FGF23 in serum of the infants in full-term SGA group on the 7th and 14th days after birth were significantly decreased （P<0.05 or P<0.01）. In SGA group，the serum levels of Klotho and FGF23 on the 7th day after birth were positively correlated with the gestational age， body weight， body length， head circumference， chest circumference， and Kopu index （P<0.05 or P<0.01）； there was a positive correlation between the serum level of Klotho and the serum level of FGF23 （P<0.05）. In terms of calcium-phosphorus metabolism， in SGA group，the serum level of Klotho on the 7th day after birth was positively correlated with serum phosphorus level （P<0.01）， and the level of serum FGF23 on the 7th day after birth was positively correlated with serum calcium and phosphorus levels （P<0.05 or P<0.01）. Conclusion Klotho and FGF23 proteins are closely associated with growth and development and phosphate metabolism of the infants. The expression levels of Klotho and FGF23 in serum of the SGA infants postnatally are lower， but the secretion of Klotho is increased with the gradul improvement of each organ， and the decrease of FGF23 may be the adaptive response.

Objective To analyze the levels of serum soluble intercellular adhesion molecule-1 （sICAM-1）， soluble vascular cell adhesion molecule-1 （sVCAM-1）， and superoxide dismutase （SOD） activity in the patients with maintenance hemodialysis （MHD）， and to discuss their relationships with coronary artery calcification （CAC）. Methods The clinical materials from 102 MHD patients （MHD group） were retrospectively analyzed. Additionally， 74 volunteers underwent routine health examination at the same time （health examination group） were selected. The CAC scores （CACs） of the patients in MHD group were detected by multi-slice computed tomography （MSCT）， and the patients were categorized into non-calcification group， mild calcification group， moderate calcification group， and severe calcification group. The general data and serum levels of sICAM-1， sVCAM-1， and SOD activities of the subjects in two groups were compared. The levels of calcium （Ca）， phosphorus （P）， parathyroid hormone （PTH）， sICAM-1， sVCAM-1， and SOD activities in serum of the patients with different degrees of calcification were analyzed. Pearson’s correlation analysis was used to analyze the correlations between the levels of sICAM-1， sVCAM-1， and SOD activity in serum of the MHD patients and CACs. Results Compared with health examination group， the levels of sICAM-1 and sVCAM-1 in serum of the patients in MHD group were significantly increased （P<0.01）， and the SOD activity was significantly decreased （P<0.01）. Compared with non-calcification group， the levels of PTH， sICAM-1， and sVCAM-1 in serum of the MHD patients in mild， moderate， and severe calcification groups were significantly increased （P<0.05）， and the SOD activities were significantly decreased （P<0.05）； the levels of P in serum of the MHD patients in moderate and severe calcification groups were significantly increased （P<0.05）. Compared with mild calcification group， the levels of P， PTH， sICAM-1， and sVCAM-1 in serum of the MHD patients in moderate and severe calcification groups were significantly increased （P<0.05）， and the SOD activities significantly decreased （P<0.05）. Compared with moderate calcification group， the levels of sICAM-1， sVCAM-1， and P in serum of the MHD patients in severe calcification group were significantly increased （P<0.05）， and the SOD activity was significantly decreased （P<0.05）. The SOD activity in serum of the MHD patients was negatively correlated with CACs （r =-0.484， P<0.01）， while the levels of sICAM-1 and sVCAM-1 were positively correlated with CACs （r =0.441， P<0.01； r = 0.561， P<0.01）. Conclusion The levels of sICAM-1 and sVCAM-1 and activity of SOD in serum of the MHD patients are abnormal. With the decreasing of the SOD activity and increasing of the levels of sICAM-1 and sVCAM-1， the degree of CAC in the MHD patients is aggravated.

Objective To discuss the clinical characteristics， diagnosis， and treatment of Shwachman-Diamond syndrome （SDS）， and to enhance the clinicians’ awareness of the disease. Methods The clinical materials of one patient diagnosed with SDS， primarily presented with neutropenia and elevated transaminase levels， confirmed by genetic testing were retrospectively analyzed. The clinical manifestations， genetic features， diagnosis， and treatment methods of SDS were analyzed complemented with the relevant literatures. Results This patient was a male child， aged 27 months. His initial clinical presentations were neutropenia and elevated transaminase levels. The patient had previously experienced diarrhea when the patient was 3 months old， which improved after treated with oral pancreatic enzyme dispersion. Over the past six months， the patient had recurrent respiratory infections. Upon admission， the examination results showed there was dental enamel hypoplasia， and the imaging results showed the abnormal bone density in the long bones of the limbs.The genetic sequencing results showed a homozygous mutation in the Shwachman-Bodian-Diamond syndrome（SBDS） gene （c.258+2T>C）. During hospitalization， the patient received the hepatoprotective care and granulocyte augmentation supportive treatment， leading to an improvement in his condition， and the patient was discharged. During a one-year follow-up， the patient’s condition was stable. Conclusion The typical presentation of the SDS patient includes diarrhea， liver function abnormalities， hematologic abnormalities， and skeletal anomalies， particularly neutropenia； there may also be developmental delays and involvement of the heart， liver， central nervous system， skeleton， and immune system. The genetic testing of suspected children is crucial， and it can aid in the early diagnosis and treatment of SDS patients.

Objective To discuss the pathological diagnostic process of one case of invasive adenocarcinoma of lung complicated with metastatic nuclear protein of testis （NUT） midline carcinoma of mediastinal lymph node，and to provide the basis for the clinical diagnosis of this disease. Methods The clinical materials of one patient with invasive adenocarcinoma of lung complicated with metastatic NUT midline carcinoma of mediastinal lymph node were collected. Intraoperative frozen section pathological diagnosis of the lung tumor was performed to determine the nature of the lesion， and postoperative mediastinal tumor was sent for slow pathological examination. Both lung and mediastinal tumors underwent routine pathological examination and immunohistochemical staining， the pathological diagnostic process was analyzed combined with the relevant literatures， and the pathological differentiation was performed. Results The patient， a 59-year-old man， underwent a CT scan at an external hospital， which revealed a soft tissue density shadow in the anterior mediastinum and a ground-glass nodule in the lower lobe of the right lung， both were considered to be neoplastic lesions. The intraoperative rapid pathological diagnosis of the lung tumor suggested pulmonary invasive adenocarcinoma， and the postoperative immunohistochemical staining results confirmed it as primary pulmonary invasive adenocarcinoma. The postoperative mediastinal tumor was confirmed as lymph node metastatic NUT midline carcinoma through immunohistochemical staining， external consultation， and genetic testing. Conclusion NUT midline carcinoma is a rare poorly differentiated squamous cell carcinoma that often occurs in the midline structures and may involve other organs and lymph node metastasis； its diagnosis requires a combination of histological morphology， imaging data， and genetic testing results.

Objective To construct the eukaryotic cell translation initiation factor 4A3 （EIF4A3） short hairpin RNA （shRNA） lentiviral vector， and to establish the Neuro-2a-EIF4A3-shRNA stable transfection cell line. Methods The EIF4A3 gene sequence was retrieved from the National Center for Biotechnology Information （NCBI） database； the PCR identification primers were designed and synthesized， and connected to the lentiviral GV493 vector digested with EcoR I and Age I enzymes to construct the GV493-EIF4A3-shRNA lentiviral plasmid； PCR method was used to screen the positive clones， which were sequenced for the identification； the GV493 empty plasmid and GV493-EIF4A3-shRNA recombinant plasmid were transfected into the HEK293T cells， regarded as GV493 control lentivirus and GV493-EIF4A3-shRNA lentivirus， respectively. After 48 h of transfection， the lentiviruses were collected for packaging and the viral titer was determined. The Neuro-2a cells were divided into blank group， GV493 control group， and GV493-EIF4A3 shRNA group. The Neuro-2a cells in blank group were untreated， and the Neuro-2a cells in GV493 control group and GV493-EIF4A3 shRNA group were infected with the respective lentiviruses at a multiplicity of infection （MOI） of 100.The infected Neuro-2a cells were selected by 10 mg·L^-1 puromycin， and the growth status and green fluorescence expression of the Neuro-2a cells in various groups were observed under fluorescence microscope； real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of EIF4A3 mRNA and protein in the Neuro-2a cells in various groups. Results The PCR sequencing results showed that the gene sequence of the GV493-EIF4A3-shRNA recombinant plasmid was consistent with the designed EIF4A3-shRNA sequence， indicating successful construction of the GV493-EIF4A3 lentiviral vector. The fluorescence microscope observation results showed that there was strong fluorescence expression and good growth status in the HEK293T cells， confirming successful lentiviral packaging. The viral titers for GV493 control lentivirus and GV493-EIF4A3-shRNA lentivirus both were 2×10⁸ TU·mL^-1. The growth status of the Neuro-2a cells in GV493 control group and GV493-EIF4A3 shRNA group was good， and they expressed green fluorescence， indicating successful construction of the stable transfection cell line. The RT-qPCR results showed that compared with blank group and GV493 control group， the expression level of EIF4A3 mRNA in the cells in GV493-EIF4A3 shRNA group was significantly decreased （P<0.01）. The Western blotting results showed that the specific bands was at a relative molecular mass of 49 000， indicating successful EIF4A3 protein expression in the Neuro-2a cells. Compared with blank group and GV493 control group， the expression level of EIF4A3 protein in the cells in GV493-EIF4A3 shRNA group was significantly decreased （P<0.01）. Conclusion The GV493-EIF4A3-shRNA lentiviral vector is succfssfully constructed， and the Neuro-2a-EIF4A3-shRNA stable transfection cell line is established； the results provide the reference for the study of the effect of EIF4A3 on the intracranial atherosclerosis.

Objective To discuss the method for establishing the rat models of irritable bowel syndrome （IBS） by chronic water avoidance stress （WAS） method， and to evaluate its feasibility. Methods Thirty male Wistar rats were randomly divided into control group （n=10） and model group （n=20）. The rats in model group were induced by WAS method for 1 h everyday， lasting for 10 consecutive days； the rats in control group underwent no interventions. After modeling， the general conditions and body weights of the rats in two groups were observed and recorded. The elevated plus maze （EPM） test was used to detect the percentages of the number of open arm entries （OE） and the time spent in open arms （OT） of the rats in two groups；the abdominal withdrawal reflex （AWR） test was used to assess the visceral sensitivity of the rats in two groups； electrocardiography was used to detect the heart rate variability （HRV） of the rats in two groups； electromyography （EMG） of the external oblique muscle was used to detect the colorectal pain sensitivity thresholds of the rats in two groups； multi-channel physiological signal recorder was used to monitor the slow wave frequency of the colon of the rats in two groups. Results There were no death rats in both groups during the modeling period. After modeling， the rats in model group exhibited poor mental status， reduced spontaneous activity， hypoactivity， disordered and dull fur， irritability， and unclean anal areas； whereas， the rats in control group showed no significant changes in the mental state， spontaneous activity， fur， and perianal area. Compared with control group， the body weight of the rats in model group was significantly decreased （P<0.05）. The EPM test results showed that compared with control group， the OE percentage and OT percentage of the rats in model group were significantly decreased （P<0.01）. The AWR test results showed that 12 rats in model group scored ≥3 points， indicating that the successful rate in creating the visceral pain models was 60%. Compared with control group， the low frequency （LF） signals and the ratio of LF/high frequency （HF） of the rats in model group were significantly increased （P<0.01）， and the HF was significantly decreased （P<0.05）. The EMG results showed that compared with control group， the coloretal pain sensitivity threshold of the colon of the rats in model group was significantly decreased （P<0.01）， and the slow wave frequency of the colon was significantly increased （P<0.01）. Conclusion The WAS method for establishing the rat model of IBS effectively demonstrates the changes in behavior and mental state， increased the visceral sensitivity， accelerated colonic slow wave frequency， and autonomic nervous system imbalance； the WAS method can serve as an effective modeling approach for observing and evaluating the related drugs and interventions on treatment of IBS.

Carpal tunnel syndrome （CTS） is one of the most common peripheral nerve entrapment disorders， the elevated pressure in the carpal tunnel， high-intensity activities and obesity are the main causes， and the patients with mild to moderate CTS are more prevalent. The main pathogenesis of CTS involves the increasing of carpal tunnel pressure and impaired local blood oxygen supply leading to reduced nerve conduction. Currently， the clinical treatment methods for mild to moderate CTS mainly include surgical and nonsurgical treatments. Nonsurgical treatment is the preferable choice for the patients with mild to moderate CTS. The western medical treatment primarily rely on oral medications， but their long-term use is limited due to the certain adverse effects； the local blockade and extracorporeal shock wave therapies show better efficacy for the patients with frequent activities and severe symptoms； the traditional Chinese medicine treatment also becomes a choice for some CTS patients due to their advantages of less pain， lower medical costs， and significant effectiveness. This study reviews the recent advancements in the pathogenesis and treatment of mild to moderate CTS， in order to design the personalized treatment methods for the mild to moderate CTS patients based on their specific conditions in clinical settings and provide the references for precise treatment of the mild to moderate CTS patients.

Cancer therapy-related cardiovascular toxicity（CTR-CVT） is gradually becoming a critical factor affecting the prognosis of cancer survivors. Vascular endothelial growth factor （VEGF） and its receptor inhibitors （VEGFIs）， developed as novel anti-cancer drugs targeting VEGF， are now widely used in clinical practice. They can extend the survival period of the cancer patients and improve the prognosis of the patients. However， the hypertension induced by VEGFIs， as the most common CTR-CVT， may limit and impact their use and leads to severe cardiovascular diseases （CVD）. It is essential to closely monitor blood pressure in the cancer patients treated with VEGFIs， conduct early assessments， and optimize the management to achieve the best anti-cancer efficacy and minimize the risk of CTR-CVT. This review discusses the clinical manifestations， pathogenesis， diagnosis， and treatment strategies of VEGFIs-related hypertension， in order to provide better guidances for managing and addressing VEGFIs-related hypertension for the clinicians.

The macrophages， as a crucial component of the body’s immune system， can be polarized into M1 and M2 types by different cellular molecules in various environments， and contribute to the progression of various diseases. In inflammatory responses， the M1 macrophages are primarily regarded as the pro-inflammatory cells， facilitate the inflammation progression， tissue destruction， and bone resorption， while M2 macrophages， as inflammatory cells， participate in tissue healing and bone repairment. In the tumor microenvironment， the roles of M1 and M2 macrophages are reversed. The periodontitis， pulpitis， and oral squamous cell carcinoma （OSCC） are the most prevalent inflammation and tumor in the oral cavity. Therefore， this article summarizes the relevant researches from home and abroad on the polarization of macrophages in oral inflammatory responses such as periodontitis， peri-implantitis， and pulpitis， bone remodeling during orthodontic treatment， and OSCC， and elucidate the metabolic activities of macrophages in infiammation，tumor，and bone remodeling and the mechanism of regulating the onset and development of diseases by the macrophage polarization， and provides new perspectives for the clinical treatment.

Postherpetic neuralgia （PHN） is a typical chronic neuropathic pain syndrome. Both peripheral and central nervous system mechanisms are believed to be involved in PHN， but the central nervous system-related brain network structure and function are not yet fully elucidated， limiting the study on the clinical analgesic drugs and other intervention strategies. In recent years， the research on pain matrix-related brain networks has helped to reveal the central nervous system regulation mechanism of pain， but there are few reports on the PHN pain matrix. This review summarizes the recent studies on the PHN pain matrix， retrospectively analyzes the functional and structural changes in specific pain-related brain regions， in order to provide the new insights for exploring the effective targeted analgesic treatments.