Objective To construct the over-expressed lentivirus vector of PRDM5 gene and establish the Neuro-2a cells stably transfected PRDM5， and to provide the basis evidence for exploring the effect of PRDM5 in pathogenesis of ischemic stroke（IS）. Methods The sequence of PRDM5 was searched and designed based on NCBI. The PRDM5 gene was amplified by PCR and ligated with the lentiviral vector GV492 digested by BamHⅠ and AgeⅠ restriction enzymes to form the GV492-PRDM5 over-expression recombinant plasmid. The positive clones with similar length and size to the target gene fragment were screened by PCR and sent to Shenggong Bioengineering （Shanghai） Co. Ltd. for identification. The correctly-sequenced GV492-control plasmid and GV492-PRDM5 over-expression recombinant plasmid were transfected into the HEK293T cells， respectively. After 48 h of transfection， the lentiviruses were collected by centrifugation， and they were GV492-control lentivirus and GV492-PRDMS over-expression lentivirus； the titers of these two lentiviruses were determined by lentiviral titer assay. The Neuro-2a cells were divided into GV492-control group and GV492-PRDM5 group， and then infected with GV492-control lentivirus and GV492-PRDM5 over-expression lentivirus， respectively， with a lentivirus multiplicity of infection （MOI） of 100. The Neuro-2a cells successfully infected with GV492-control lentivirus and GV492-PRDM5 over-expression lentivirus were screened with puromycin （10 mg·L^-1） after 72 h of infection. The growth status and the expression of green fluorescence protein of Neuro-2a cells in GV492-control group and GV492-PRDM5 group were observed by fluorescence microscope. The expression levels of PRDM5 mRNA and PRDM5 protein in the Neuro-2a cells in two groups were detected by real-time fluorescence quantitative RCR（RT-qPCR） and Western blotting methods. Results The PCR results showed that the length of the positive transformant of GV492-PRDM5 recombinant plasmid was about 684 bp， and the gene sequence of GV492-PRDM5 over-expression recombinant plasmid was consistent with the designed and synthesized PRDM5 over-expression sequence. The titers of GV492-control lentivirus and GV492-PRDM5 over-expression lentivirus were both 2.5×10⁸ TU·mL^-1. The Neuro-2a cells in GV492-control group and GV492-PRDM5 group grew well， and the expressions of green fluorescence protein were found under fluorescence microscope.The RT-qPCR results showed that the expression level of PRDM5 mRNA in the Neuro-2a cells in GV492-PRDM5 group was significantly increased compared with GV492-control group（P<0.01）. The Western blotting results showed that the specific bands appeared in the Neuro-2a cells in GV492-control group and GV492-PRDM5 group with a relative molecular weight of 75 000； compared with GV492-control group， the expression level of PRDM5 protein in the Neuro-2a cells in GV492-PRDM5 group was increased（P<0.01）. Conclusion The over-expression lentivirus vector of PRDM5 gene is successfully constructed， and the stably transfected GV492-PRDM5-Neuro-2a cells are established.

Objective To discuss the anti-tumor effect of low dose of methotrexate（MTX） combined with sorafenib（SFN） on the human osteosarcoma（OS）, and to clarify the possible mechanism. Methods Four types of human OS cells（143B cells, HOS cells, U2OS cells, and MG63 cells） were cultured in vitro. Western blotting method was used to detect the expression levels of vascular endothelial growth factor（VEGF） and vascular endothelial growth factor receptor 2（VEGFR2） proteins in the above four kinds of cells. The human OS xenograft model was established in the nude mice, and 20 successfully modeled BALB/C nude mice were randomly divided into control group（given 2% dimethyl sulfoxide+98% corn oil）, low dose of MTX group（given 2 mg·kg^-1 MTX）, SFN group （given 15 mg·kg^-1 SFN）, and combined drug group（given 2 mg·kg^-1 MTX+15 mg·kg^-1 SFN）； there were 5 mice in each group. The tumor volumes of the mice in various groups were detected and tumor growth curves were plotted； HE staining was used to observe the morphology of tumor tissue of the mice in various groups； immunohistochemistry was used to detect the positive expression rates of VEGFR2， proliferation marker Ki-67, and hypoxia-inducible factor-1（HIF-1） proteins in tumor tissue of the mice in various groups.The human OS 143B cells were divided into 0, 0.125, 0.250, 0.500, 1.000, 2.000, and 4.000 μmol·L^-1 MTX groups（given 0, 0.125, 0.250, 0.500, 1.000, 2.000, and 4.000 μmol·L^-1 MTX， respectively）. CCK-8 method was used to detect the proliferation rates of the 143B cells in various groups, and half inhibityory concentration （IC₅₀） was calculated； the concentration of MTX that had no effect on 143B cell survival was selected as low dose of MTX. The human OS 143B cells were divided into control and low dose of MTX groups (given 0 and 0.250 μmol·L^-1 MTX). ELISA method was used to detect the levels of VEGF in the 143B cells in various groups. Results Compared with 143B cells， the expression levels of VEGF and VEGFR2 proteins in the HOS cells， U2OS cells， and MG63 cells were significantly increased （P<0.001）. In the xenograft model， compared with control group， the tumor volumes of the mice in SFN group， and combined drug group were decreased （P<0.001）； compared with low dose of MTX group and SFN group， the tumor volume of the mice in combined drug group was decreased （P<0.01）. The immunohistochemical results showed that compared with control group， the positive expression rates of Ki-67， VEGFR2， and HIF-1 proteins in tumor tissue of the mice in combined drug group were significantly decreased（P<0.05）. The CCK-8 results showed that there was no change in the proliferation of the 143B cells treated with 0.25 μmol·L^-1 MTX. The ELISA results showed that compared with control group， the level of VEGF in the 143B cells in MTX group was signyicantly decreased （P<0.05）. Conclusion Low dose of MTX enhances the anti-tumor effect of SFN on the human OS， which may be due to the inhibition of VEGF secretion by the OS cells， thereby enhancing the anti-tumor effect of SFN on the human OS.

Objective To discuss the effect of prostaglandin E2 （PGE₂）， a pyrogenic mediator， on the discharge activity of warm-sensitive neurons （WSNs） in median preoptic nucleus （MnPO） of hypothalamus in the female mice， and to clarify its mechanism. Methods Coronal brain slices of MnPO were prepared from the female mice. The slices were then perfused with artificial cerebrospinal fluid （ACSF） containing synaptic transmission blockers （STBs）. The discharge frequency was monitored using the patch clamp technique while changing the temperature of the perfusate to identify the WSNs. A total of 32 MnPO WSNs were divided into base line group（n=32） and PEG₂ （n=32）. The patch clamp technique was employed to monitor the discharge frequencies of MnPO WSNs following the perfusion of ACSF and 1 μmol·L^-1 PGE₂， respectively. The MnPO WSNs with good activity and significant change in discharge frequency after PGE₂ perfusion were divided into PGE₂ receptor E-series prostaglandin receptrol （EP）1 antagonist （EP1 ant）+PGE₂ group （n=7）， EP3 ant+PGE₂ group （n=7）， and EP4 ant+PGE₂ group （n=7）. The patch clamp technique was used to monitor the discharge frequencies of MnPO WSNs following the perfusion of 3 μmol·L^-1 EP1 ant and 1 μmol·L^-1 PGE₂ mixture， 10 μmol·L^-1 EP3 ant and 1 μmol·L^-1 PGE₂ mixture， and 10 μmol·L^-1 EP4 ant and 1 μmol·L^-1 PGE₂ mixture， respectively. Resluts： After perfuson of the ACSF containing STBs， a total of 188 MnPO neurons from the female mice with an intrinsic temperature sensitivity coefficient （m value） were identified； out of these， 32 neurons had an m value of ≥0.8 and were identified as MnPO WSNs， accounting for approximately 17% of all recorded neurons. Compared with baseline discharge frequency， the discharge frequency of MnPO WSNs after addition of PGE2 was decreased （P<0.05）. Compared with PGE₂ group， the percentage change in discharge frequency of MnPO WSNs in EP3 ant+PGE₂ group was significantly decreased （P<0.05）. Compared with PGE₂ group， the percentage change in discharge frequency of MnPO WSNs in EP1 ant+PGE₂ group had no significant difference （P>0.05）. Compared with PGE₂ group， the percentage change in discharge frequency of MnPO WSNs in EP4 ant+PGE₂ group had no significant difference（P>0.05）. Conclusion In the female mice， WSNs make up approximately 17% of the total neurons in MnPO. PGE₂ can directly inhibit the discharge activity of MnPO WSNs in the female mice through postsynaptic mechanism involving EP3 receptors.

Objective To discuss the effect of angiotensin （1-7） ［Ang（1-7）］ on high-turnover bone disease in the uremic rats， and to clarify its possible mechanism. Methods Thirty SD rats were randomly divided into sham operation group（n=6） and experimental group（n=24）. The rats in experimental group underwent 5/6 nephrectomy（Platt method） + high-phosphorus（P） diet［1.2% P， 1.0% calcium（Ca）］ to establish the model of uremic high-turnover bone disease. The successfully modeled rats were then randomly divided into model group， Ang（1-7） group， angiotensin-converting enzyme 2（ACE2） activator dimethylacetamide amidoxime（DIZE） group， and Mas receptor antagonist group （A779 group）， with 6 rats in each group. The levels of Ca， P， blood creatinine （Scr）， blood urea nitrogen （BUN）， and 24 h urinary protein （UP） in serum of the rats in various groups were detected by automatic biochemical analyzer at 12 and 18 weeks after operation； immunofluorescence staining was used to detect the levels of intact parathyroid hormone （iPTH） in the rats in various groups； ELISA method was used to detect the levels of osteocalcin（OC）， type Ⅰ collagen N-terminal peptide （NTX）， and tartrate-resistant acid phosphatase （TRAP）-5b in serum of the rats in various groups； high-resolution micro-CT scan was used to detect the bone density （BMD）， tissue mineral density （TMD）， trabecular thickness （Tb.Th）， and trabecular separation （Tb.Sp） in femur tissue of the rats in various groups； Von Kossa staining and Giemsa staining were used to observe the pathomorphology of cortical bone and trabecular bone of the rats in various groups， and the trabecular bone volume （TBV） was calculated； fluorescence microscope was used to detect the mineral apposition rates （MAR） of the rats in vanious groups， and the osteoblast index （OBI） and osteoclast index （OCI） of the rats in various groups were calculated. Results At 12 and 18 weeks after operation， compared with sham operation group， the weights of the rats in model group， Ang（1-7） group， DIZE group， and A779 group were decreased （P<0.05）. At 12 and 18 weeks after operation， compared with sham operation group， the levels of 24 h UP， Scr， and BUN in serum of the rats in model group， Ang（1-7） group， DIZE group， and A779 group were increased （P<0.05）； at 18 weeks after operation， compared with model group， the levels of 24 h UP and Scr in serum of the rats in Ang（1-7） group and DIZZ group were decreased （P<0.05）， and the levels of 24 h UP， Scr and BUN in serum of the rats in A779 group were increased （P<0.05）. The successful establishment of the uremic high-turnover bone disease model was confirmed. At 12 and 18 weeks after operation， compared with sham operation group， the serum iPTH， P， OC， NTX， and TRAP-5b levels of the rats in model group， Ang（1-7） group， DIZE group， and A779 group were all significantly increased （P<0.05）； at 18 weeks after operation， compared with model group， the serum NTX and TRAP-5b levels of the rats in Ang（1-7） group and DIZE group were decreased（P<0.05）， while the serum iPTH， P， NTX and TRAP-5b levels in A779 group were increased（P<0.05）. The high-resolution micro-CT scan results showed that compared with sham operation group， the values of femur BMD and TMD of the rats in model group， Ang（1-7） group， DIZE group， and A779 group were all significantly decreased （P<0.05）； compared with model group， the values of femur BMD and TMD of the rats in Ang（1-7） group and DIZE group were increased （P<0.05）， while the values of BMD and TMD of the rats in A779 group were decreased （P<0.05）. Compared with sham operation group， the femur Tb.Th of the rats in model group was decreased （P<0.05）， and the Tb.Sp was increased （P<0.05）； the femur Tb.Th of the rats in Ang（1-7） group and DIZE group were increased （P<0.05）， and the Tb.Sp was decreased （P<0.05）. Compared with model group， the femur Tb.Th of the rats in A779 group was decreased （P<0.05）， and the Tb.Sp was increased （P<0.05）. The bone pathological examination results showed that compared with sham operation group， the femur TBV of the rats in model group， Ang（1-7） group， DIZZ group and A779 group were decreased （P<0.05）， and MAR， OBI and OCI were increased （P<0.05）； compared with model group， the OBI and OCI of the rats in Ang（1-7） group and DIZE group were decreased （P<0.05）， and TBV was increased （P<0.05）， while the OBI and OCI of the rats in A779 group were increased （P<0.05）， and TBV was decreased （P<0.05）. Conclusion The ACE2/Ang（1-7）/Mas axis improves high-turnover bone disease in the uremic rats.

Objective To investigate the expressions of cystathionine-β-synthase （CBS） and cystathionine-γ-lyase （CSE） and their effects on the malignant biological behaviours of breast cancer cells， and to elucidate their mechanisms. Methods The breast cancer tissue and paracancerous normal tissue from 15 cases of patients were selected， and RT-qPCR and Western blotting methods were used to detect the mRNA and protein expression levels of CBS and CSE in breast cancer tissue， paracancerous normal tissue， MCF-7 cells， and MDA-MB-231 cells. The MCF-7 cells were divided into siNC group （transfected with siNC） and siCBS group （transfected with siCBS）， and the MDA-MB-231 cells were divided into ovNC group （transfected with CSE over-expression empty plasmid） and ovCSE group （transfected with CSE over-expression plasmid）. CCK8 assay was used to detect the proliferation activities of breast cancer cells in various groups， Transwell assay was used to detect the numbers of migration and invasion cells in various groups， and Western blotting method was used to detect the protein expression levels of E-cadherin， N-cadherin and Vimentin proteins in the breast cancer cells in various groups. Results Compared with paracancerous normal tissue， the expression levels of CBS and CSE mRNA and proteins in breast cancer tissue were increased （P<0.05 or P<0.01）. Compared with MDA-MB-231 cells， the CBS mRNA expression level in the MCF-7 cells was increased （P<0.05）； compared with MCF-7 cells， the expression level of CSE protein in the MDA-MB-231 cells was decreased （P<0.05）. Compared with siNC group， the proliferation activity， the numbers of migration and invasion cells， the expression levels of N-cadherin and Vimentin proteins in the MCF-7 cells in siCBS group were significantly decreased（P<0.05）， and the expression level of E-cadherin protein was increased（P<0.05）. Compared with ovNC group， the proliferation activity， the numbers of migratoin and invasion cells， and the expression levels of N-cadherin and Vimentin proteins in the MDA-MB-231 cells in ovCSE group were increased （P<0.05）， while the expression level of E-cadherin protein was significantly decreased （P<0.05）. Conclusion The expressions of CBS and CSE are upregulated in breast cancer tissue， and high levels of CBS and CSE promote proliferation， migration， invasion and epithelial-mesenchymal transition（EMT） of breast cancer cells.

Objective To discuss the inhibitory effect of schisandrin B （SchB） on the migration and invasion of the pancreatic cancer Pan02 cells， and to clarify its mechanism. Methods The pancreatic cancer Pan02 cells were treated with different concentrations of SchB （0， 0.78， 1.56， 3.12， 6.25， 12.50， and 25.00 mg·L^-1） for 24， 48， and 72 h. CCK-8 method was used to detect the survival rates of the cells in various groups， and the concentration of SchB for the subsequent experiments was confirmed. The Pan02 cells were divided into control group， 2.5 mg·L^-1 SchB group， 5.0 mg·L^-1 SchB group， and 10.0 mg·L^-1 SchB group. Wound healing assay was used to detect the wound healing rates of the Pan02 cells in various groups； Transwell chamber assay was used to detect the numbers of migration and invasion Pan02 cells in various groups； Western blotting method was used to detect the expression levels of Vimentin and N-cadherin proteins in the Pan02 cells in various groups.The mouse models of subcutameous transplanted tumor of pancreatic cancer cells were established.Ten successfully modeling mice were randomly divided into control group and SchB group （n=5）. After 28 d of treatment， the weights of tumor of the mice were determined； immunohistochemistry method was used to detect the expressions of Vimentin and N-cadherin proteins in tumor tissue of the mice in various groups. Results The CCK-8 results showed that compared with control group， the survival rates of the pancreatic cancer Pan02 cells in different concentrations of SchB groups were decreased （P<0.05 or P<0.01）. The wound healing results showed that compared with control group， the wound healing rates of the cells in 2.5， 5.0， and 10.0 mg·L^-1 SchB groups were decreased （P<0.05 or P<0.01）. The Transwell chamber results showed that compared with control group， the numbers of migration and invasion Pan02 cells in 2.5， 5.0， and 10.0 mg·L^-1 SchB groups were decreased （P<0.05 or P<0.01）. The Western blotting results showed that compared with control group， the expression levels of Vimentin and N-cadherin proteins in the Pan02 cells in 2.5， 5.0， and 10.0 mg·L^-1 SchB groups were decreased （P<0.05 or P<0.01）. Compared with control group， the tumor volume and weight of the mice in SchB group were significantly decreased（P<0.01）. The immunohistochemistry results showed that compared with control group， the positive expression rates of Vimentin and N-cadherin proteins in tumor tissue of the mice in SchB group were significantly decreased （P<0.01）. Conclusion SchB can inhibit the proliferation， migration， and invasion of the pancreatic cancer Pan02 cells， and its mechanism is related to the reduction of expressions of Vimentin and N-cadherin proteins.

Objective To construct an eukaryotic expression vector pcDNA3.1-MYC-SGK1-mcherry which containing mouse serum and glucocorticoid-induced kinase （SGK）1 gene， and to observe its expression in the transfected HEK293 cells. Methods The SGK1 target gene segments were amplified by PCR method， and the segments were ligated to the pcDNA3.1-MYC-C-mcherry vector which was doubly-digested with Hind Ⅲ and Sbf Ⅰ. After successful verification by enzyme digestion and sequencing， the pcDNA3.1-MYC-SGK1-mcherry expression vector was transfected into the HEK293 cells by liposome transfection. Western blotting method was used to determine the expression level of eukaryotic expression vector in the cells. Results The vector band was located at 5 200 bp and the target gene band was located at 3 100 bp， which was consistent with the expected results. The sequencing results were also consistent when compared with the expected sequence by Snap Gene software， which indicated that the eukaryotic expression vector pcDNA3.1-MYC-SGK1-mcherry was successfully constructed. Successful expression of the eukaryotic expression vector pcDNA3.1-MYC-SGK1-mcherry was observed by Western blotting method， in which the transfected cells showed well-defined bands near the relative molecular mass of 49 000. Conclusion The eukaryotic expression vector pcDNA3.1-MYC-SGK1-mcherry is successfully constructed， laying a solid foundation for the subsequent study on the transition mechanism of SGK1 gene in the early development of mouse fertilized egg cells.

Objective To investigate the effect of nuclear receptor subfamily 2 group F member 2 （NR2F2） on the biological behaviors of human ovarian cancer SKOV3 cells， and to clarify its molecular mechauism and provide the new idea for treatment of ovarian cancer. Methods Gene Expression Profiling Interactive Analysis（GEPIA） Database analyse the expression level of NR2F2 gene in ovarian tissue， and analyse its correlation with clinical prognosis of ovarian cancer patients. The human ovarian cancer SKOV3 cells were divided into control group and NR2F2 over-expression （NR2F2 OE） group， which were transfected with mCherry control virus and NR2F2 OE over-expression virus， respectively， when the cell deusity reached 70%， and the stable transfection SKOV3 cell lines were screened with puromycin（puro） 48 h lafter. Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the transfection efficiencies of the cells； RT-qPCR method was used to detect the expression levels of NR2F2 and sex-determining region Y-box 2 （SOX2） mRNA in the cells in two groups； Western blotting method was used to detect the expression levels of NR2F2， ATP-binding cassette superfamily G member 2 （ABCG2）， and programmed cell death 1-ligand 1 （PD-L1） protcins in the cells in two groups. CCK-8 assay was used to detect the proliferation activities of the cells in two groups； Wound assay was used to detect the migration rates of the cells in two groups； Transwell chamber assay was used to detect the number of transmembrane cells； Spheroidization assay was used to detect the numbers of spheroids in the cells； peripheral blood mononuclear cells （PBMCs）-mediated tumor cell killing assay was used to detect the relative densities of surviving tumor cells； CCK-8 assay was used to detect the half maximal inhibitory concentration （IC₅₀） of paclitaxel （PTX） and carboplatin （CBP）. Results Compared with normal ovarian tissue， the expression level of NR2F2 gene in ovarian tumor tissue was decreased （P<0.05）， and decreased with the improvement of clinical pathological grading of ovarian tumor. The patients with higher expression level of NR2F2 gene had better clincal prognosis. The SKOV3 cells with NR2F2 over-expresson were successfully constructed， and the expression levels of NR2F2 mRNA and protein in the cells in NR2F2 OE group were increased compared with control group （P<0.001）. The CCK-8 assay results showed that compared with control group， the proliferation activities of the cells in NR2F2 OE group were decreased at different time points （1， 2， 3， and 4 d） （P<0.05 or P<0.01）. The cell wound assay results showed that compared with control group， the migration rate of the cells in NR2F2 OE group was decreased （P<0.001）. The Transwell assay results showed that compared with control group， the number of transmembrane cells in NR2F2 OE group was decreased （P<0.01）. Compared with control group， the number of the spheroids in NR2F2 OE group was decreased （P<0.05）， and the expression levels of SOX2 mRNA（P<0.01） and protein （P<0.001） were increased. Compared with control group， the relative density of surviving tumor cells in NR2F2 OE group was decreased， but the difference was not significant （P<0.05）， and the expression level of PD-L1 protein was decreased （P<0.05）. Compared with control group， the proliferation activities of cells in NR2F2 OE group were decreased （P<0.05）， and the drug sensitivities of the cells to PTX and CBP were enhanced （P<0.05）； the IC₅₀ of PTX was significantly reduced， while the IC₅₀ of CBP could not be calculated due to excessively high drug concentration； the expression level of ABCG2 protein was decreased （P<0.05）. Conclusion The over-expression of NR2F2 may inhibit the proliferation， migration， and invasion of the human ovarian cancer SKOV3 cells， decrease the expression levels of SOX2， PD-L1 and ABCG2 proteins， suppress the stemness and immune evasion ability of the SKOV3 cells， and enhance the sensitivities of the SKOV3 cells to PTX and CBP.

Objective To discuss the effect of Yes-associated protein （YAP） silencing on the proliferation， migration， and invasion capabilities of the human cervical cancer （CC） SiHa cells. Methods The human CC SiHa cells were cultured in vitro， and the lentiviral YAP shRNA was transfected into the SiHa cells to establish stably transfected YAP-shRNA experimental group （sh-YAP group） and empty plasmid control group （control group）. Western blotting method was used to detect the silencing effect of YAP； immunofluorescence method was used to detect the microfilament number and morphology of actin filaments （F-actin） in the cells in both groups； CCK-8 method was used to detect the survival rates of the cells in two groups； Transwell chamber assay and wound healing assay were used to detect the numbers of migration and invasion cells and scratch healing rates of the cells in two groups； Western blotting method was used to detect the expression levels of epithelial-mesenchymal transition （EMT） markers （E-cadherin and Snail）， DNA damage repair-related proteins （γ-H2AX）， and apoptosis-related proteins ［c-MYC and B-cell lymphoma-2 （Bcl-2）］ in the cells in two groups. Results The results of lentiviral YAP shRNA transfection into SiHa cells showed that the expression level of YAP protein in the SiHa cells was significantly decreased （P<0.05）. The immunofluorescence results showed that after YAP silencing， the F-actin in SiHa cells was sparse and regularly arranged， with a reduced number of cells and a shriveled appearance. The CCK-8 results showed that compared with control group， the survival rate of the SiHa cells in sh-YAP group was significantly decreased cultured for 24 and 48 h （P<0.01）. The results of Transwell chamber assay and the wound healing assay showed that compared with control group， the numbers of migration and invasion SiHa cells in sh-YAP group were significantly decreased （P<0.01）， and the cell scratch healing rates were signifiantly decreased（P<0.05）. The Western blotting results showed that compared with control group， the expression level of E-cadherin protein in the cells in sh-YAP group was increased （P<0.05）， and the expression levels of c-MYC， Bcl-2， and γ-H2AX proteins were decreased （P<0.05 or P<0.01）. Conclusion YAP gene silencing leads to the depolymerization of F-actin in the human CC SiHa cells and regulates the apoptosis and DNA damage repair， potentially reversing the EMT process， thereby inhibiting the proliferation and migration of the tumor cells.

Objective To investigate the protective effect of Dendrobium officinale polysaccharides （DOP） on intestinal mucosal barrier damage， and to elucidate the possible mechanism. Methods Ten female C57BL/6 mice， aged 5 months， were selected as young group； twenty femal C57BL/6 mice， aged 15 months， were randomly divided into aged group and DOP treatment group （200 mg·kg^-1， DOP group）， with 10 mice in each group. The mice in DOP group were administrated with DOP by gavage. The body mass， food intakes and hanging time of the mice in various groups were detected. HE staining was used to observe the pathomorphology of intestinal and spleen tissues of the mice in various groups. Immunohistochemical staining was used to detect the expressions of intestinal atresin 1 （ZO-1） and Mucin 2（MUC2） in intestinal tissue of the mice in various groups. The intestinal baterial metabolite medium（IBMM） were prepared to intervene the Caenorhabditis elegans （C.elegans）， and the C.elegans were randomly divided into Young-IBMM group， Aged-IBMM group， and DOP-IBMM group.Immuno-fluorescence method was used to analyze the intestinal lipofuscin accumulation levels on the 1st day and the 12th day of the C. elegans in various groups. Brilliant blue staining was used to assess the intestinal leakage on the 1st day and the 12th day of C. elegans in various groups.The Caco-2 cells were randomly divided into Young-IBMM， Aged-IBMM and DOP-IBMM groups， and Western blotting method was used to detect the expression levels of ZO-1，Occludin，tumor necrosis factor-α（TNF-α），interleukin-6 （IL-6），phosphorylated myosin light chain（p-MLC），myosin light chain kinase（MLCK）proteins in the Caco-2 cells in various groups. Results Compared with young group， the body mass of the mice in aged group was increased （P<0.05）， the amount of food intake was decreased （P<0.05）， and the hanging time was decreased （P<0.05）； compared with aged group， the body mass of the mice in DOP group was significantly decreased （P<0.01）， the amount of food intake was increased （P<0.05）， and the hanging time was significantly extended （P<0.01）. The HE staining results showed that compared with young group， the thickness of intestinal mucosa of the mice in aged group became thinner， the goblet cells were reduced， the intestinal villi were disordered with different lengths， a large amount of hemosiderin was found on the surface of the spleen， the cell components in the red medullary were reduced， and the lymphatic sheath and lymphatic nodes around the intra-white pulp artery remained or almost disappeared； compared with aged group， the thickness of the intestinal mucosa of the mice in DOP group was increased， the goblet cells were increased， the length of the intestinal villi was consistent and neatly arranged， the overall function of the red pulp of the spleen was improved， and the components of the white pulp were increased. The immunohistochemical staining results showed that compared with young group， the expression levels of ZO-1 and MUC2 proteins in intestinal tissue of the mice in aged group were significantly decreased （P<0.05 or P<0.001）； compared with aged group， the expression levels of ZO-1 and MUC2 proteins in the intestinal tissue of the mice in DOP group were significantly increased （P<0.05 or P<0.001）. The immuno-fluorescence analysis showed that compared with Young-IBMM group， the intestinal lipofuscin accumulation level of C.elegans in Aged-IBMM group was significantly increased （P<0.001）；compared with Aged-IBMM group，the intestinal lipofuscin accumulation level of C.elegans in DOP-IBMM group was significantly reduced （P<0.001）. The brilliant blue staining showed that compared with Young-IBMM group， the bright blue dye leaked into the whole body of C.elegans from intestinal tissue in Aged-IBMM group， and the intestinal structure became blurred and was difficulted to be observed；compared with Aged-IBMM group， the leakage of bright blue dye of C.elegans in DOP-IBMM was reduced. The Western blotting results showed that compared with Young-IBMM group， the expression levels of TNF-α， IL-6， p-MLC， and MLCK proteins in the Caco-2 cells in Aged-IBMM group were significantly increased （P<0.01 or P<0.001）， and the expression levels of ZO-1 and Occludin proteins were significantly decreased （P<0.05 or P<0.01）； compared with Aged-IBMM， the expression levels of TNF-α， IL-6， p-MLC and MLCK proteins in the Caco-2 cells in DOP-IBMM group were significantly decreased （P<0.01）， and the expression levels of ZO-1 and Occludin proteins were significantly increased （P<0.05 or P<0.01）. Conclusion DOP has an ameliorating effect on intestinal mucosal barrier damage in the aged mice，and its mechanism may be related to the improvement of intestinal barrier damage by regulating intestinal bacterial metabolites， inhibiting the p-MLC/MLCK signal pathway， restoring the expression of tight junction complexes， and reducing the level of intestinal inflammation.

Objective To discuss the effect of asiatic acid （AA） on the inflammation and oxidative stress damage induced by lipopolysaccharide （LPS） in the primary cultured hippocampus neurons， and to clarify its mechanism. Methods The primary cultured rat hippocampus neurons （cell purity identified by immunofluorescence staining） were divided into control group， LPS （10 mg·L^-1） group， and LPS+AA group （10 mg·L^-1 LPS+10， 20， and 40 μmol·L^-1 AA）， AA group （20 μmol·L^-1 AA）， ML385 group ［10 μmol·L^-1 nuclear factor erythroid 2-related factor （Nrf2） inhibitor］， and LPS+ML385+AA group （10 mg·L^-1 LPS+10 μmol·L^-1 ML385+20 μmol·L^-1 AA）. After drug treatment， methylthiazolyldiphenyl-tetrazolium bromide （MTT） method was used to detect the survival rates of the hippocampus neurons in various groups； lactate dehydrogenase （LDH） kit was used to detect the LDH leakage rates of the hippocampus neurons in various groups； enzyme linked immunosorbent assay （ELISA） kit was used to detect the expression levels of inflammatory factors ［interleukin （IL）-1β and tumor necrosis factor （TNF）-α］ and the activities of superoxide dismutase （SOD） and malondialdehyde （MDA） levels in the hippocampus neurons in various groups； Griess method was used to detect the nitric oxide （NO） levels in supernatant of the hippocampus neurons in various groups； immunofluorescence staining was used to detect the expressions of Nrf2 and heme oxygenase-1 （HO-1） proteins in the hippocampus neurons in various groups； Western blotting method was used to detect the expression levels of Nrf2， HO-1， nuclear factor-kappa B（NF-κB）， and B-cell lymphoma 2 （Bcl-2） proteins in the hippocampus neurons in various groups. Results Compared with control group， the survival rate of the hippocampus neurons， SOD activity， and Bcl-2 expression level in the cells in LPS group were significantly decreased （P<0.01）， while the LDH leakage rate， expression levels of IL-1β and TNF-α， MDA level， and NO level， as well as the expression level of NF-κB protein， were significantly increased （P<0.01）； the fluorescence intensities and expression levels of Nrf2 and HO-1 proteins in hippocampus neurons were significantly decreased （P<0.01）. Compared with LPS group， the survival rates of hippocampus neurons， SOD activities， and expression levels of Bcl-2 in the cells in LPS+10 μmol·L^-1 AA group and 20 μmol·L^-1 AA group were significantly increased （P<0.01）， while the LDH leakage rates， expression levels of IL-1β and TNF-α， MDA levels， and NO levels， as well as expression levels of NF-κB protein， were significantly decreased （P<0.05 or P<0.01）， and the fluorescence intensities and protein expression levels of Nrf2 and HO-1 in the cells were significantly increased （P<0.01）. Compared with LPS+20 μmol·L^-1 AA group， the fluorescence intensities of Nrf2 and HO-1 in the cells in LPS+ML385+AA group were significantly decreased （P<0.01）， and the expression levels of Nrf2 protein in the nucleus and cytoplasm， the expression levels of HO-1 and Bcl-2 proteins in the cells were significantly decreased （P<0.01）， while the expression level of NF-κB protein was significantly increased （P<0.01）. Conclusion AA can improve LPS-induced inflammation and oxidative stress damage in the primary cultured rat hippocampus neurons， and its mechanism may be related to the activation of the Nrf2/HO-1 signaling pathway.

Objective To investigate the effect of betaine in oxygen-glucose deprivation injury of rat brain microvascular endothelial cells（BMECs）， and to clarify the regulatory effect of betaine on phosphoinositide 3-kinase（PI3K）/protein kinase B（AKT） pathway. Methods Five SD rats aged 7 d were selected and the rat BMSEs were obtained. The oxygen-glucose deprivation model of rat BMECs was prepared under hypoxic and hypoglycemic conditions； the experiment was divdided into model group， and low dose， medium dose， and high dose of betaine groups and positive control group， at the same time， blank control group （without modeling） was set up. The BMECs in blank control group and model group were treated with fresh medium， the BMECs in positive control group were given a final concentration of 10 μmol·L^-1 nimodipine， and the BMECs in low， medium and high doses of betaine groups were treated with betaine at final concentration of 0.5， 1.0 and 2.0 mmol·L^-1， respectively. The survival rates of BMECs in various groups were determined by CCK-8 method at 12， 24 and 48 h after culture； the activities of lactate dehydrogenase （LDH） and the levels of adenine ribonucleoside triphosphate （ATP） in the rat BMECs in various groups were determined using kits， and the levels of tumor necrosis factor-α （TNF-α）， interleukin（IL）-6， IL-1β， and IL-18 in supernatants of the BMECs in various groups were determined by enzyme-linked immunosorbent assay （ELISA）； the activities of superoxide dismutase （SOD） and levels of malondialdehyde （MDA） in the BMECs in various groups were determined by kits； the transendothelial resistance （TEER） values of rat BMSCs in various groups were determined by TEER analyzer， and the horseradish peroxidase （HRP） permeabilities of BMECs in various groups were determined by an insertion cell culture apparatus. TUNEL staining was used to determine the apoptotic rates of rat BMECs in vaisous groups， and Western blotting method was used to determine the ratios of phosphory lated PI3K（p-PI3K）/PI3K and phosphorylated AKT（p-AKT）/AKT in the rat BMECs in various groups. Results Compared with blank control group， the survival rate of BMECs， activity of SOD， and level of ATP， value of TEER， and ratios of p-PI3K/PI3K and p-AKT/AKT of the rat BMECs in model group were significantly decreased （P<0.05）， while the activity of LDH， the levels of TNF-α， IL-6， IL-1β， IL-18， and MDA， the apoptotic rate of the BMECs， and HRP permeability were significantly increased（P<0.05）. Compared with model group， the survival rates of the BMECs， activities of SOD， and levels of ATP， values of TEER， and ratios of p-PI3K /PI3K and p-AKT /AKT of the BMECs in low， medium， and high doses of betaine groups and positive control group were significantly increased（P<0.05）， while the activities of LDH， the levels of TNF-α， IL-6， IL-1β， IL-18， and MDA， the apoptotic rates of BMECs and HRP permeabilities were significantly decreased（P<0.05）. Conclusion Betaine can significantly repair the oxygen-glucose deprivation/reperfusion injury in the rat BMECs， inhibit the oxidative damage and apoptosis of BMECs， and improve the permeability of the cells； its mechanism may be related to the regulation of the PI3K/AKT pathway.

Objective To study the protective effect of Pien-Tze-Huang on acetaminophen-induced liver injury，and to clarify the possible mechanism. Methods The human normal hepatocytes（L02 cells） were divided into control group， APAP group （10 mmol·L^-1 APAP），APAP+PZH group （10 mmol·L^-1 APAP and 0.4 mg·mL^-1 PZH）， and PZH group （0.4 mg·mL^-1 PZH）. The survival rates of the cells in various groups were determined by MTT method， the morphology was observed by inverted microscope， and the apoptotic rates were detected by flow cytometry. The activities of superoxide dismutase （SOD） and lactate dehydrogenase （LDH） and the levels of malondialdehyde （MDA） in the cell supernant in various groups were detected by the kits， the reactive oxygen species（ROS） levels and the mitochondrial membrane potential （MMP） in the hepatocytes in various groups were detected by fluorescence probe. Western blotting method was used to examine the expression levels of apoptosis-related proteins， phosphatidylinositol 3 kinase （PI3K）/protein kinase B（AKT） signaling pathway proteins， nuclear factor-κB（NF-κB） signaling pathway proteins and inflammatory factors in the cells in various groups. Results The results of MTT showed that compared with control group， the survival rate of the cells in APAP group was markedly decreased （P<0.05）； compared with APAP group， the survival rate of the cells in APAP+PZH group was significantly increased （P<0.05）. Compared with control group， the number of the L02 cells in APAP group showed a decreasing and loosely arranged trend； compared with APAP group， the number and arrangement of the L02 cells in APAP+PZH group were notably improved. The results of flow cytometry showed that compared with control group， the apoptotic rate of the L02 cells in APAP group was significantly increased（P<0.05）； compared with APAP group， the apoptotic rate of the cells in APAP+PZH group was significantly decreased（P<0.05）. Compared with control group，the activity of LDH and level of MDA in the cells in APAP group were significantly increased （P<0.05）， while the activity of SOD was significantly decreased （P<0.05）； compared with APAP group，the activity of LDH and level of MDA in the cells in APAP+PZH group were significantly decreased（P<0.05）， while the activity of SOD was significantly increased（P<0.05）. Compared with control group， the fluorescence intensity of ROS in the cells in APAP group was significantly increased；compared with APAP group，the fluorescence intensity of ROS in APAP+PZH group was significantly decreased. Compared with control group， the MMP of the L02 cells in APAP group was significantly decreased； compared with APAP group，the MMP of the L02 cells in APAP+PZH group was significantly increased. The results of Western blotting showed that compared with control group， the expression levels of caspase-9， B-cell lymphoma 2（Bcl-2） associated X protein（BAX）， phosphorylated PI3K （p-PI3K）， phosphorylated AKT （p-AKT）， phosphorylated NF-κB inhibitor alpha （p-IKBα）， p-P65， phosphorylated inhibitor of kappaB kinaseβ （p-IKKβ）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α（TNF-α） proteins in the cells in APAP group were significantly increased （P<0.05）， while the level of Bcl-2 proteins in the cells in APAP group was significantly decreased （P<0.05）； compared with APAP group， the levels of caspase-3， BAX， p-PI3K， p-AKT， p-IKBα， p-P65， p-IKKβ， IL-1β， IL-6， and TNF-α proteins in the cells in APAP+PZH group were significantly decreased， while the level of Bcl-2 protein was significantly increased（P<0.05）. Conclusion PZH may reduce the oxidative stress and inflammatory response in the cells by regulating the PI3K/AKT and NF-κB signaling pathway， therefore attenuate the L02 cell injury induced by APAP.

Objective To discuss the effect of DEAD-box RNA helicase 39A（DDX39A） gene silencing on the proliferation，migration and invasion of the esophageal cancer TE-1 cells，and to clarify its possible mechanism. Methods For bioinformatics analysis， GSE63941， GSE77861， GSE20347， and GSE16153 chip data were downloaded from the GEO database. The esophagel cancer-related data were selected from the TCGA Database.R software was used to analyze the differentially expressed genes.STRING Database was used to construct the protein-protein interaction （PPI） network.Identification of key genes of high relevance was achieved using the MCODE plugin in Cytoscape.The expression of key genes in normal esophageal tissue and esophageal cancer tissue were analyzed with the GEPIA 2 database. Kaplan-Meier Plotter was used to perform survived analysis and plotting for the screened key genes.Cytological experiments were carried out on esophageal cancer TE-1 cells， and small interfering RNA （siRNA）technology was used to silence the expression of DDX39A gene.The TE-1 cells in the logarithmic growth phase were selected and divided into blank （MOCK） group， negative control （si-NC） group， and silencing （si-DDX39A） group. Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of DDX39A mRNA and protein in the TE-1 cells in various groups；CCK-8 assay was conducted to detect the proliferation activity of cells in various groups， and the cell scratch assay was used to measure the migration rate of cells in various groups； Transwell chamber assay was used to detect the number of invasion cells in various groups；Western blotting method was used to detect the expression levels of β-catenin， glycogen synthase kinase-3β（GSK3β）， phosphorylated glycogen synthase kinase-3β（p-GSK3β）， c-MYC， Cyclin D1 and nuclear β-catenin proteins in the cells in various groups. Results Analyses using TCGA database combined with the GEO Database yielded a total of 56 differentially expressed genes. MCODE plugin in Cytoscape software identified 41 key genes of high relevance； DDX39A was screened by analyzing 41 genes through the GEPIA 2 and Kaplan-Meier plotter Databases. The results of RT-qPCR and Western blotting methods showed that compared with si-NC group， the expression levels of DDX39A mRNA and protein in the cells in si-DDX39A group were decreased （P<0.05）. The CCK-8 results showed that the proliferation activity of the cells in si-DDX39A group was lower than that in si-NC group （P<0.05）. The cell scratch assay results showed that the cell migration rate in si-DDX39A group after 24 h was lower than that in si-NC group （P<0.05）.The results of Transwell chamber assay showed that the number of invasion cells in si-DDX39A group was lower than that in si-NC group （P<0.05）. Compared with si-NC group， the expression levels of β-catenin， p-GSK3β， c-MYC， Cyclin D1， and nuclear β-catenin in the TE-1 cells in si-DDX39A-1 group and si-DDX39A-3 group were decreased （P<0.01）， but the expression levels of GSK3β protein had no significant differences （P>0.05）. Conclusion Silencing of DDX39A gene could inhibit the proliferation， migration and invasion of TE-1 cells， and the mechanism may be related to the regulation of Wnt/β-catenin signaling pathway.

Objective To explore the correlations between the cerebral infarction area and cytokines and immune status in patients with acute ischemic stroke， and to provide the theoretical basis for immunotherapy of the patients with different degrees of cerebral infarction. Methods Sixty-seven patients with acute ischemic stroke within 72 h of the onset were randomly selected according to the inclusion and exclusion criteria， and were divided into large-area cerebral infarction group （n=34） and non-large-area cerebral infarction group（n=33） on the basis of the biggest infarction area in the sequences of magnetic resonance diffusion-weighted imaging（CDWI）. Clinical baseline characteristics such as gender， age， and medical history were collected from the patients in two groups， the serum levels of interleukin （IL）-2， IL-6， IL-10， and IL-17A， tumor necrosis factor-α （TNF-α）， and interferon-γ （IFN-γ） were measured using flow cytometry； the absolute values of lymphocytes （LYM#）， lymphocyte percentages （LYM%）， and neutrophil/lymphocy ratios（NLR） in peripheral blood of the patients caiculated， and the ratios of IFN-γ/IL-4， TNF-α/IL-4， and TNF-α/IL-10 rations were also calculated. The values of National Institutes of Health Stroke Scale （NIHSS） scores of the patients were evaluatd on the basis of the assessment of clinical neurological signs. The correlations of the cerebral infarction area and NIHSS score， cytokines and immune status groups of the patients in two were tested by rank correlation analysis. Results Compared with non-large-area cerebral infarction group， the serum levels of IL-2， IL-6， IL-10， IL-17A， TNF-α， and IFN-γ as well as the NLR in the peripheral blood of the patients in large-area cerebral infarction group were significantly increased （P<0.01）， while the LYM#， LYM% and TNF-α/IL-4 were significantly decreased （P<0.01）. There was a positive correlation between cerebral infarction area and NIHSS score in the patients in large-area cerebral infarction group （r_s =0.521， P<0.05）， and there was a significantly positive correlation between cerebral infarct area and NIHSS score in the patients in non-large-area cerebral infarction group （r_s =0.721， P<0.001）. The NIHSS scores were positively correlated with serum IL-6 （r_s =0.306， P=0.005）， IL-4 （r_s =0.252， P<0.001）， IL-2 （r_s =0.109， P=0.025）， IL-17A （r_s =0.405， P<0.001）， and IFN-γ （r_s =0.146， P<0.001） levels in two groups； no correlations were found between NIHSS scores and TNF-α （r_s =0.039， P=0.726） and IL-10 （r_s =0.121， P=0.192） levels. NIHSS scores of the patients in two groups had negative correlatious with the serum level of LYM# （r_s =-0.026， P=0.036） and LYM% （r_s =-0.008， P=0.002） ，and had positive correlated with NLR （r_s =0.315， P=0.009）. Conclusion The infarction area of the patients with actue cerebral infarction is correlated with the NIHSS score， the inflammatory response， the degree of adaptive immune injury， and the immune status. The have positive correlation with cytokines and immune markers and the overall size of the infarction area. Compared with the patients with non-large-acea cerebral infarction， the immunosuppression of the patients with large-area infarcted areas is more likely to occure.

Objective To explore the promotion effect on remineralization of demineralized dentin of urushiol primer applicated in acid-etch-rinse adhesives， and to clarify its impact on the longevity of dentin adhesion. Methods Ninety-six freshly extracted，caries-free third molars were selected to prepare the dentin specimens. Following acid etching with 37% phosphoric acid gel， the specimens were randomly divided into blank control group， 0.3%， 0.7%， 1.0%， and 1.5% urushiol groups， and positive control group（acetone solvent）. The treated samples were placed in modified simulated body fluids for remineralization for 14 and 28 d. Attenuated total reflection Fourier transform infrared spectroscopy（ATR-FTIR） was used to detect the relative mineralization mass of minerals in the dentinal tubules in various groups， and X-ray Diffractometery and Energy Dispersive spectrometer were used to analyze the dentin surface material compositions in various groups. Scanning electron microscope（SEM） was used to observe the surface morphology of the specimens in various groups， Vickers hardness tester was used to measure the microhardness of the dentin surface in various groups， and microtensile strength （μTBS） was used to examine the effect of the bond strengthes in various groups. Results Compared with blank control group， the conversion rate of adhesive double bonds by primer in positive control group was decreased， but the difference was not significant （P>0.05）； but the conversion rates of adhesive double bonds by primer in 0.3%， 0.7%， 1.0%， and 1.5% urushiol groups were increased （P<0.05）. The SEM results revealed that at 14 and 28 d， compared with bland control group， a minimal membranous deposit in dentinal tubules was seen in positive control group， minimal mineralization displayed in 0.3 % urushiol group， significant deposition of loose mineral particles with blocking the tubule orifices was found in 0.7% urushiol group， noticeable mineral precipitates exhibited in 1.0% urushiol group， and relatively empty dentinal tubules were seen in 1.5% urushiol group. The microhardness results showed that at 14 d after remineralization， compared with blank control group， the microhardness in positive control group showed no significant improvement（P>0.05）， while the differences in 0.3%， 0.7%， 1.0% and 1.5% urushiol groups were statistically significant （P<0.05）； at 14 d after remineralization compared with positive control group， the microhardness of dertin in 0.7 % and 1.0% urushiol groups were increased（P<0.05）； at 28 d after remineralization， compared with blank control and postive control groups， the microhardness in urushiol groups were significantly increased （P<0.05）， espectially in 0.7% to 1.5% urushiol groups （P<0.05）. In the μTBS test， at 14 d after remineralization， compared with postive control group， the μTBS in 0.3%， 0.7%， 1.0% and 1.5% urushiol groups were increased（P<0.05）； at 28 d after remineralization， the μTBS in blank control group was the lowest； compared with blank control group， there was no significant difference in the μTBS in postive control group（P>0.05）； compared with positive control group， the μTBS in 0.3%， 0.7%， 1.0%， and 1.5% urushiol groups were increased （P<0.05）， espectially in 0.7%，1.0， and 1.5% urushiol groups （P<0.05）. Conclusion Natural-derived urushiol， as a novel primer， can pretreat the demineralized dentin substrates， and facilitate collagen cross-linking within the dentin matrix； moreover， it leverages the phenolic hydroxyl groups within its structure to attact calcium and phosphate ions， envelope dentin collagen fibers to promote remineralization， in order to enhance the strength of the resin-dentin bonding interface.

Objective To investigate the efficacy and safety of fospropofol disodium （FP） in the induction and maintenance of general anesthesia in the adult patients graded Ⅰ or Ⅱ by the American Society of Anesthesiologists （ASA） undergoing elective surgery， and to provide the theoretical basis for application of EP in the induction and maintenance of general anesthesia. Methods Adult patients of ASA grade Ⅰ or Ⅱ undergoing elective surgery were selected with a total of 100 patients recruited sequentially according to the time of visit， and they were randomly divided into FP group （50 cases） and propofol group （50 cases）. All patients were prepared preoperatively， and received a slow injection of midazolam （2 to 3 mg） and sufentanil （0.3 μg·kg^-1）， followed by induction of anaesthesia 1 to 2 min later. The patients in FP group were given FP （10.0-12.5 mg·kg^-1） intravenously， and the patients in propofol group were given propofol （1.5-2.0 mg·kg^-1） intravenously. After the Modified Obserational Assessment Alertness/Sedation （MOAA/S） score dropped to 1， muscle relaxant was administrated and the induction was completed. During the maintenance of anaesthesia， the patients in FP group received a continuous intravenous infusion of FP at a rate of 12.5-15.0 mg·kg^-1·h^-1， and the patients in propofol group received a continuous infusion of propofol at a starting rate of 6 mg·kg^-1·h^-1. The patients in two groups additionally received remifentanil （0.1-0.4 μg·kg^-1·min^-1） for co-analgesia， and the rate of administration was adjusted according to the patient’s status. Systolic blood pressure （SBP）， diastolic blood pressure （DBP）， mean arterial pressure （MAP）， heart rate （HR） and bispectral index （BIS） values of the patients in two groups were recorded at different time points： before induction （T₁）， immediately after tracheal intubation （T₂）， 5 min after induction （T₃）， 10 min after induction （T₄）， 20 min after induction （T₅）， 30 min after induction （T₆）， 40 min after induction （T₇） and at the end of the procedure （T₈）. The time to onset of sedation/anaesthesia （MOAA/S≤1）， the time to eye opening， and the time to awakening （MOAA/S=5） of the patients in two groups were recorded. The lowest intraoperative SBP and BIS values and the time required of the patients in two groups were observed. The incidence of adverse reactions related to agitation， choking， nausea， vomiting and cardiovascular system or respiratory system were compared between two groups. Results There were no statistically differences in the general informations and the duration of surgery of patients between two groups （P>0.05）. The induction time of the patients in FP group （2.39 min） was significantly longer than that in propofol group （0.70 min） （P<0.05）. In the recovery period of general anesthesia， the eye opening time and recovery time of the patients in FP group were significantly longer than those in propofol group （P<0.05）. There were no significant differences in MAP of the patients between two groups at different time points （P>0.05）. The HR at T₄， T₅， T₆， and T₇ time points of the patients in FP group were lower than those in propofol group （P<0.05）. The lowest value of BIS of the patients in FP group was significantly smaller than that in propofol group， and the time taken to reach the lowest value of BIS in FP group was significantly longer than that in propofol group （P<0.05）. The time taken to reach the lowest value of SBP of the patients in FP group was longer than that in propofol group （P<0.05）. However， the lowest value of SBP of the patients and the incidence of adverse reations of the patients in two groups showed no statistical differences （P>0.05）. Conclusion Compared with propofol， FP injection is safe and effective in the induction and maintenance of general anesthesia in adult patients with ASA class Ⅰ or Ⅱ undergoing elective surgery， with a low incidence of adverse reactions， which is a new anesthesia option.

Objective To observe the effectiveness and safety of different doses of esketamine combined with propofol for intravenous administration in the child patients undergoing enhanced computed tomography （CT） examination， and to clarify the optimal clinical dose of esketamine in combination with propofol for sedation. Methods This study is a randomized， controlled， double-blind （blinded to subjects and evaluators）， and single-center clinical trial. A total of 120 preschool children undergoing enhanced CT examination were randomly divided into propofol group （P group）， propofol+0.3 mg·kg^-1 esketamine group （P+K3 group）， and propofol+0.5 mg·kg^-1 esketamine group （P+K5 group）， and there were 40 cases in each group. All the children were given 2 mg·kg^-1 propofol， and additional propofol was administered in increments of 1 mg·kg^-1 until the sedation criteria for entering the CT room were met ［Modified Observer’s Assessment of Alertness/Sedation （MOAA/S） score ≤3］. The vital signs of the children were observed and recorded at four time points： before sedation （T₀）， when sedation was satisfactory （T₁）， during contrast agent injection （T₂）， and upon awakening （T₃）. The examination time， time to satisfactory sedation （from the start of sedation to MOAA/S score≤3）， and awakening time （from the end of the examination to MOAA/S score>4） of the children in various groups were recorded. The total dose of propofol and the proportion of cases requiring additional propofol were compared among various groups. Adverse reactions during induction， examination， and after awakening were also compared among various groups. Results There were no significant differences in general conditions of the children in three groups （P>0.05）. Hemodynamic parameters： at T₂， compared with P group， the SpO₂ levels of the children in P+K3 group and P+K5 group were increased （P<0.05）； at T₁， compared with P group， the SBP levels of the children in P+K3 group and P+K5 group were increased （P<0.05）. There were no significant differences in examination time of the children in three groups （P>0.05）. Compared with P group and P+K3 group， the time to satisfactory sedation of the children in P+K5 group was shorter （P<0.05）. Compared with P group， the awakening time of the children in P+K3 group and P+K5 group was shorter （P<0.05）. Compared with P group and P+K3 group， the total dose of propofol of the children in P+K5 group was decreased （P<0.05）， and the proportion of cases requiring additional propofol was lower （P<0.05）. Adverse reaction indicators： compared with P group， the incidence of respiratory depression of the children in P+K3 group and P+K5 group was lower （P<0.05）， and the incidence of nausea and vomiting was lower （P<0.05）. Compared with P group and P+K3 group， the incidence of movement during the examination of the children in P+K5 group was lower （P<0.05）， and the incidence of dizziness was higher （P<0.05）. There were no significant differences in the incidence of increased airway secretions of the children in three groups （P>0.05）. Conclusion The use of 0.5 mg·kg^-1 esketamine combined with 2 mg·kg^-1 propofol for intravenous administration in the child patients for enhanced CT examination sedation can improve the efficiency of such examinations and offers high safety and effectiveness.

Objective To explore the differential flora between depression group and control group Based on the GMrepo database of intestinal flora， after excluding the factors such as gender， age， body mass index（BMI）， and country， and to further clarify the characteristics of differential intestinal flora in the depression patients with different ages and genders. Methods The subjects were selected from the GMrepo database with phenotypes of “depression” and “health”， and the relevant microbial abundance datasets of the screened research subjects were downloaded based on inclusion and exclusion criteria. Using the case matching function of SPSS 27.0 statistical software， 95 control subjects and 95 depression patients were matched into two groups based on gender （1∶1）， age （±5 years）， BMI （±1.5 kg·m^-2）， and country （1∶1）； univariate analysis on intestinal flora using non-parametric tests was conducted to screen the differential intestinal flora with a P<0.05 under hypothesis testing； Wald’s forward stepwise selection method was used to construct a binary Logistic regression model， stratified analysis was conducted based on gender （male， female） and age （≤65 years， >65 years） and the significantly differential flora between the subjects in control group and the patients in depression group were determined based on odds ratio （OR） and P-value within different subpopulatious. Results Compared with control group， Paraprevotella ［OR=0.661，95% confidence interval （CI）=0.489-0.893， P=0.007］ and Prevotella （OR=0.946， 95% CI=0.903-0.992， P=0.022）showed significantly lower abundance in the patients in depression group， which were the protective factors for the occurrence of depression. Paraprevotella（OR=0.358， 95% CI=0.146-0.883， P=0.026） was identified as the differential flora in the male population between depression group and control group， while Faecalibacterium （OR=0.565， 95% CI=0.322-0.990， P=0.046） and Alistipes （OR=0.513， 95% CI=0.289-0.911， P=0.023） were the differential flora in the female population. Prevotella （OR=0.654， 95% CI=0.476-0.899， P=0.009） was the differential flora among the individuals’ age≤65 years between depression group and control group. Conclusion Paraprevotella， Prevotella， Faecalibacterium， and Alistipes are the characteristic intestinal flora associated with depression， and the changes in their abundances may have significant impacts on the occurrence and development of depression.

Objective To detect the levels of sex hormones and insulin in follicular fluid（FF） and the expression level of phosphatase and tensin homolog deleted on chromosome ten（PTEN） in granulosa cells in the infertile patients with polycystic ovary syndrome（PCOS）， and to preliminarily explain the correlations between the insulin level and the expression level of PTEN mRNA. Methods Seventy infertile patients were selected as the subjects and divided into PCOS group and control group （tubal obstruction or infertility due to male factors） according to infertility factors. All patients received in vitro fertilization-embryo transfer（IVF-ET） treatment. FF and ovarian granulosa cells were collected on the day of ovulation. The expression levels of PTEN mRNA in ovarian granulosa cells of the patients in two groups were detected by real-time fluorescence quantitative PCR（RT-qPCR） method. The levels of sex hormone and insulin in FF were measured by electrochemiluminescence. The correlations of the PTEN mRNA expression level in ovarian granulosa cells and testosterone（T） in FF with the level of insulin in FF were analyzed by Spearman correlation analysis method. Results There were no significant differences in age， infertility years， body mass index （BMI）， basic sex hormone， total dose of gonadotropin（Gn） and days of ovulation induction in two groups（P>0.05）. Compared with control group， the anti-Mullerian hormone（AMH） and antral follicle counting（AFC） of the patients in PCOS group were significantly increased（P<0.05）. The RT-qPCR results showed that the PTEN mRNA expression level in ovarian granulosa cells of the patients in the PCOS group was higher than that in control group（P<0.001）. The electrochemiluminescence results showed that the levels of T and insulin in FF of the patients in PCOS group were higher than those in control group（P<0.05）， whereas the estrogen and progesterone levels were lower than those in control group（P<0.05）. The Spearman correlation analysis showed that that T level in FF was positively correlated with the insulin level of the patients in PCOS group（r=0.577， P<0.001）， and the PTEN mRNA expression level in ovarian granulosa cells was positively correlated with the insulin levels in FF （r=0.616， P<0.001）； in control group， there was no correlation between T level and insulin level in FF（r=0.266， P=0.123）， and there was no correlation between the expression level ofPTEN mRNA in granulosa cells and the insulin level in FF in control group （r=-0.214， P=0.216）. Conclusion The high expression of PTEN in granulosa cells of the infertile patients with PCOS may be related to the local hyperinsulin level in the ovary， and PTEN participates in the occurrence and development of PCOS.

Objective To discuss the associations of mismatch repair （MMR） in gastric cancer with preoperative inflammatory indicators and clinicopathological features in the gastric cancer patients， and to construct a gastric cancer MMR predictive model based on preoperative inflammatory indicators and clinicopathological features of the gastric cancer patients， and to provide new ideas for evaluation of MMR status in gastric cancer. Methods The data of 254 gastric cancer patients who underwent surgical treatment from September 2020 to October 2023 were included. According to the expression of MMR protein， the patients were divided into MMR normal （proficiout MMR， pMMR） group and MMR deficient （dMMR） group. The preoperative inflammatory indicators and clinicopathological features data of the gastric cancer patients in two groups were collected. The associations between inflammatory indicators， clinicopathological features， and MMR in dMMR group and pMMR group were analyzed usingChi-square test. The independent predictive factors for dMMR were selected to construct the nomogram. Receiver operating characteristic （ROC） curve and calibration curve were used to evaluate the predictive efficacy， and decision curve was used to evaluate the practicality of the predication model. Results A total of 254 gastric cancer patients were included in the study， with 221 patients （87%） in pMMR group and 33 patients （13%） in dMMR group. There were statistically significant differences （P<0.05） in age， tumor location， tumor differentiation degree， maximum tumor diameter， platelet-to-lymphocyte ratio （PLR）， alkaline phosphatase （AKP）， alkaline phosphatase-to-albumin ratio （AAR）， fibrinogen（FB）-to-lymphocyte （FLR）， FB-to-albumin（AL） （FAR）， D-dimer （D-D）， and FB of the gastric cancer patients between dMMR group and pMMR group. Univariate and multivariate Logistic regression analysis revealed maximum tumor diameter ［odd ratio（OR）=2.958， 95% confidence interval （CI）：1.196-7.314， P=0.019］， tumor location （OR=4.013，95%CI：1.596-10.089， P=0.003）， tumor differentiation （OR=3.006， 95%CI： 1.250-7.230， P=0.014）， FAR （OR=2.793， 95%CI：1.179-6.616， P=0.020）， and carbohydrate antigen 199（CA199） （OR=0.279， 95%CI：0.084-0.929， P=0.038） were the independent predictors of dMMR. The area under the ROC curve （AUC） value of the gastric cancer MMR prediction model constructed based on inflammatory indicators and clinical pathological characteristics was 0.800 with the sensitivity of 0.851 and the specificity of 0.606. The calibration curve of the nomogram was found to fit the ideal curve well，and in Hosmer-Lemeshow test P=0.412， the clinical decision curve showed a better net benefit. Conclusion The preoperative inflammatory indicators and clinicopathological features are associated with MMR in gastric cancer； maximum tumor diameter， tumor location， tumor differentiation， CA199， and FAR are the independent predictors of dMMR. The prediction model based on the above predictors could predict the MMR status of the dMMR gastric cancer patients.

Objective To analyze the changes in the levels of inflammatory factors in gingival crevicular fluid of the patients with periodontal disease underwent invisible appliance， and to provide the reference for clinical practice. Methods PubMed， Embase， Cochrane Library， China National Knowledge Infrastructure（CNKI）， Chinese Biology Medicine（CBM）， Wanfang and VIP Databases were used for literature collection about the treatment using invisible appliance and fixed appliance for periodontal disease. The retrieval time was limited from January 1997 to November 2023. Two researchers independently screened the literatures and extracted data based on the inclusion and exclusion criteria，and conducted quality evaluation. Review Manager 5.4 software was used to analyze the changes in the levels of inflammatory cytokines including tumor necrosis factor-α（TNF-α），C-reaction protein（CRP）， interleukin-6（IL-6），interleukin-1β（IL-1β） and interleukin-2（IL-2） before and 6 months after appliance placement. Results Six randomized controlled trials were included in this study with a total sample size of 601 cases. Subgroup analysis was conducted based on the type of brackets. The results of Meta-analysis showed that compared with fixed appliance group，the changes in the levels of TNF-α（MD=-1.32， 95%CI：-1.87--0.77，P<0.001），IL-6（MD=-0.78， 95%CI：-1.22--0.35， P<0.001），CRP（MD=-1.03， 95%CI：-1.30--0.76， P<0.001） and IL-1β （MD=-1.45， 95%CI：-2.21--0.70， P<0.001） in invisible appliance group were significantly decreased， while the change in IL-2 level（MD=0.74，95%CI：0.61-0.87， P<0.001） was significantly increased. Conclusion Compared with fixed appliance， the invisible appliance can better control the levels of inflammatory factors in gingival crevicular fluid of the patients with periodontal disease，which is beneficial to the periodontal tissue health of these patients.

Objective To discuss the expressions of autotaxin （ATX） and lysophosphatidic acid receptor 3 （LPA3） in serum and lung tissue of the chronic obstructive pulmonary disease （COPD） patients， and to clarify the role of ATX and LPA3 in the occurrence and development of COPD. Methods A total of 40 COPD patients were collected and brought into acute exacerbation of COPD group （AECOPD group）； after treatment， those stabilized patients were included in COPD stable group （n=40）； additionally， 40 healthy individuals were recruited as control group. Enzyme-linked immunosorbent assay （ELISA） was used to detect the ATX levels in the serum of the subjects in various groups. Another 80 patients who underwent lobectomy were divided into COPD smoking group （CS group， n=20）， COPD non-smoking group （CNS group， n=20）， non-COPD smoking group （HS group， n=20）， and non-COPD non-smoking group （HNS group， n=20）. The general informations of the subjects in various groups were collected. HE staining was used to observe the pathomorphology of lung tissue of the patients in various groups；immunohistochemical staining was used to detect the expression levels of ATX and LPA3 proteins in lung tissue of the patients in various groups；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of ATX and LPA3 mRNA in lung tissue of the patients in various groups；Pearson correlation analysis was used to evaluate the correlations of continuous variables with normal distribution， and Spearman correlation analysis was used to evaluate the correlations of other variables. Results Compared with control group， the percentage of forced expiratory volume in the first second to predicted value （FEV1%pred） and the percentage of forced expiratory volume in the first second to forced vital capacity （FEV1/FVC） of the patients in COPD stable group were significantly decreased （P<0.05）. Compared with COPD stable group and control group， the level of ATX in serum of the patients in AECOPD group was increased （P<0.05）； compared with control group， the level of ATX in serum of the patients in COPD stable group was increased （P<0.05）. The serum ATX level of the patients in AECOPD group was positively correlated with COPD Assessment Test （CAT） scores （r=0.581， P<0.001） and showed no correlations with smoking history， white blood cells （WBC）， percentage of neutrophils （NEUT%）， neutrophil to lymphocyte ratio （NLR）， and body mass index （BMI） （P>0.05）. The serum ATX level of the patients in COPD stable group was positively correlated with WBC and CAT score （r=0.384， P=0.014； r=0.463， P=0.003） and negatively correlated with FEV1%pred and FEV1/FVC （r=-0.393， P=0.012； r=-0.353， P=0.025）. Compared with CS group and CNS group， the FEV1%pred and FEV1/FVC of the patients in HS group and HNS group were increased （P<0.05）. The immunohistochemical staining results showed that compared with CS group， the expression levels of ATX and LPA3 proteins in lung tissue of the patients in HS group and HNS group were decreased （P<0.05）. Compared with CNS group， the expression levels of ATX and LPA3 proteins in lung tissue of the patients in HS group and HNS group were decreased （P<0.05）. The RT-qPCR results showed that compared with CS group， the expression levels of ATX and LPA3 mRNAs in lung tissue of the patients in HS group and HNS group were decreased （P<0.05）； compared with CNS group， the expression levels of ATX and LPA3 mRNAs in lung tissue of the patients in HS group and HNS group were decreased （P<0.05）. The correlation analysis results showed that the expression level of ATX protein in lung tissue of the COPD patients was positively correlated with the expression level of LPA3 protein （r=0.723， P<0.001）. Conclusion The expression levels of ATX and LPA3 mRNA and proteins in lung tissue of the COPD patients are increased. The serum ATX levels in both AECOPD group and COPD stable group are increased， suggesting that these factors may be involved in the inflammatory response of COPD， promoting its occurrence and development.

The Streptococcus suis meningitis patients often accompany with visual and hearing impairment， and their life quality is affected seriously after recovery. Our department admitted a 59-year-old male patient. The patient presented headache accompanied by fever， followed by decreased vision acuity in both eyes and reduced hearing in the right ear. The patient was alerted and oriented， with only light sensation in both eyes and reduced hearing in both ears （more pronounced on the right side）. The neck strength was 3 horizontal fingers， and Kernig sign was positive. Magnetic resonance imaging （MRI） head scan showed patchy abnormal signals in the white matter of the right frontal lobe， while further enhanced MRI scan did not reveal any abnormal signals. The cerebrospinal fluid appeared slightly yellow and turbid color with blood droplets at the initial drip， and the total number of leukocytes and protein level were significantly increased.The glucose and chloride levels were decreased with the multinucleated cells accounting for 79% of the cellular components. The cerebrospinal fluid IgG level was 82.25 mg·L^-1. Blood culture yielded Streptococcus suis as the causative pathogen. Metagenomic next generation sequencing （mNGS） in cerebrospinal fluid indicated Gram-positive bacteria， with 7 001 Streptococcus suis sequences detected. The patient was diagnosed with Streptococcus suis meningitis and treated with anti-infective therapy complemented by hormone and symptomatic treatment. Follow-up interviews at 6 and 18 months post-discharge revealed a satisfactory recovery of visual acuity， while the hearing did not improve significantly compared to at the time of discharge. In the patients with a history of exposure with sick or dead pigs who develop signs of meningitis accompanied by simultaneous involvement of other organs such as eyes and ears， Streptococcus suis meningitis should be considered and empirical anti-infective treatment should be provided as soon as possible. Blood culture and mNGS for definitive diagnosis should be conducted promptly. Combined application of Streptococcus suis-sensitive antibiotics and dexamethasone may help improve hearing impairments in these patients.

Application of invisible orthodontic appliances in the treatment of adult skeletal malocclusions， especially in the cases requiring tooth extraction， has always been one of the challenges in invisible orthodontic technology. Our department received a 30-year-old female patient in March 2019； the patient presented with complaints of “crooked teeth and protruding mouth，” seeking orthodontic treatment to improve her facial profile. The patient had bilateral distal molar relationship， proclined maxillary and mandibular anterior teeth， and deep overjet； radiographic examination revealed an ANB angle of 6.2°， indicating a skeletal Class Ⅱ malocclusion. By extracting four first premolars and using four miniscrews， after 20 months of treatment， the extraction spaces were fully closed， the dental arch was aligned， and the occlusal relationship was favorable； the anterior teeth were retracted， the mandible showed a reverse rotation， and the soft tissue profile was significantly improved. Application of invisible orthodontic appliances in conjunction with miniscrews anchorage can achieve good three-dimensional control of tooth movement in adult extraction cases， providing a reference for clinical practitioners treating such patients.

The author of this paper repored the diagnostic and treatment process of one patient with secondary primary tracheal adenoid cystic carcinoma（TACC）. From the perspective of disease occurrence， the patient was successively diagnosed with tracheal basal cell adenocarcinoma and tracheal adenoid cystic carcinoma， which was extremely rare in clinical practice， providing a reference for studying the correlation and differences in the incidence of different types of tracheal cancer. At the same time， during the treatment process， massive bleeding from the tracheostomy site occurred during radiotherapy， deepening the understanding of radiotherapy complications in TACC. The patient， a 61-year-old female， underwent surgical treatment for tracheal basal cell adenocarcinoma five years ago. One year ago， the patient experienced exertional dyspnea， which gradually worsened， severely affecting her daily life， leading to her hospital admission for further diagnosis and treatment. The physical examination results showed a 2 cm×1 cm irregular mass in the right neck， with normal skin temperature and color， no tenderness or pain on pressure， and good mobility. Enhanced computed tomography（CT） of the larynx indicated cauliflower-like soft tissue masses on the right and posterior walls of the trachea and a nodule on the anterior margin of the sternocleidomastoid muscle in the right supraclavicular region， suggesting recurrence of an intratracheal tumor.The differential diagnosis included tracheal squamous cell carcinoma， which often forms keratin pearls and exhibits more significant cellular atypia. The immunohistochemical markers are also different， and the results of pathology and immunohistochemistry examinations can effectively distinguish them. The patient underwent resection of the mass along with the tracheal wall and excision of the right supraclavicular mass. The postoperative pathology confirmed adenoid cystic carcinoma of the trachea with local neural involvement. Given the patient’s symptoms of tracheal obstruction and the possibility of cervical lymph node metastasis of TACC， surgery was the primary treatment choice. Postoperative radiotherapy further controlled residual tumor cells， reduced the risk of recurrence， and improved the local control rates. After 12 months of follow-up post-radiotherapy， no signs of tumor recurrence were observed. The clinicians should reinforce diagnostic thinking and be highly vigilant for the possibility of secondary primary tumors in the trachea. They should comprehensively assess using various examination methods to improve the early diagnosis accuracy and avoid the misdiagnosis and missed diagnosis， thereby providing the best treatment opportunity for the patients.

Giant cell-rich osteosarcoma（GCRO） is a special subtype of osteosarcoma， and is rare. This paper reported the clinical and imaging data of the patient with GCRO who had special imaging manifestations， and to provide the reference for clinical diagnosis and treatment of GCRO. The patient， a 69-year-old female，was admitted to the hospital due to the discovery of a mass in the right lower leg for 1 month. The X-ray imaging manifestations showed that the cortical bones on the opposite edges of the tibia and fibula presented mild moth-eaten changes. The plain CT scan imaging manifestations showed that there was a mass within the muscle， and the cortical bones on the opposite edges of the tibia and fibula were destroyed. The tumor only involved the superficial cortical bones and did not invade the deep cortical bones and the medullary cavity. The MRI imaging manifestations showed that the local bones of the tibia and fibula adjacent to the tumor were destroyed， and the medullary cavity was not invaded. An extended resection of the mass was performed， and the postoperative pathological diagnosis was GCRO. The patient received chemotherapy after the operation. However， a tumor in the right lower leg recurred 15 months after the operation. The patient passed away 21 months after the operation. Giant cell-rich osteosarcoma often leads to misdiagnosis or missed diagnosis due to its non-specific clinical manifestations. This paper explored the clinical manifestations and imaging data of the patient with GCRO in order to improve the clinical understanding level and the level of diagnosis and treatment of the clinicians about this disease.

Objective To discuss the clinical application value of echocardiography in evaluating the early outcomes of transcatheter aortic valve replacement （TAVR） via the transapical approach， and to clarify the role of echocardiography in assessing the efficacy of the surgery. Methods The clinical data of 85 patients who received J-Valve prosthetic valves via the transapical TAVR were retrospectively analyzed. The patients were divided into AS group （simple aortic stenosis， n=20）， AR group （simple aortic regurgitation， n=37）， and AS&AR group （aortic stenosis with regurgitation， n=28）. Echocardiographic examination was performed on all the patients before operation， 1 week after operation， 3 months after operation， and 6 months after operation. The parameters including left ventricular end-diastolic volume （LVEDV）， left ventricular end-systolic volume （LVESV）， left ventricular ejection fraction （LVEF）， left ventricular fractional shortening （LVFS）， interventricular septal thickness （IVST）， left ventricular posterior wall thickness （LVPWT）， aortic valve peak flow velocity （AV V_max）， aortic valve mean transvalvular pressure gradient （AV PG_mean）， and paravalvular leak （PVL） width were measured to evaluate the cardiac function and the function of the prosthetic valve； the occurrence of postoperative complications of the patients in various groups was also analyzed. Results J-Valve prosthetic valves were successfully implanted in all 85 patients. There were no significant differences in age， gender， New York Heart Association （NYHA） heart function classification， history of hypertension， history of diabetes， history of hyperlipidemia， and history of coronary artery disease among various groups befor operation（P>0.05）， ensuring comparability. Compared with before operation， 1 week after operation， the AV V_max and AV PG_mean of the patients in AS group and AS&AR group were decreased （P<0.05）； there were no significant differences in various parameters of the patients in AR group （P>0.05）. Compared with before operation， 3 months after operation， the LVEF and LVFS of the patients in AS group were increased （P<0.05）， while the AV V_max and AV PG_mean were decreased （P<0.05）； the LVEDV and LVESV of the patients in AR group were decreased （P<0.05）， while the LVEF and LVFS were increased （P<0.05）； the LVEDV， LVESV， AV V_max， and AV PG_mean of the patients in AS&AR group were decreased （P<0.05）， while the LVEF and LVFS were increased （P<0.05）. Compared with before operation， LVEDV， LVESV， IVST， and LVPWT of the patients in all three groups 6 months after operation were decreased （P<0.05）， while LVEF and LVFS were increased （P<0.05）； the AV V_max and AV PG_mean of the patients in AS group and AS&AR group were decreased （P<0.05）； the AV PG_mean of the patients in AR group was decreased （P<0.05）. The postoperative complications included 3 cases of permanent pacemaker implantation （2 cases in AS group， 1 case in AR group）， 1 case of stroke （in AS group）， and 13 cases of PVL （4 cases in AS group， 5 cases in AR group， 4 cases in AS&AR group）. No deaths occurred during follow-up. Conclusion Echocardiography can accurately and quantitatively evaluate early changes in cardiac function and the functional state of prosthetic valves after transapical TAVR， providing objective evidence for evaluating surgical outcomes and postoperative complications.

Objective To prepare a polyvinyl butyral （PVB） nanofiber membrane loaded with propolis， and to clarify its physicochemical properties， drug release behavior， antibacterial activity， and biocompatibility. Methods Propolis with a mass fraction of 0.12% was dissolved in 18% （mass fraction） PVB methanol solution. PVB and PVB-propolis （PVB-P） nanofiber membranes were prepared using electrospinning from PVB and PVB-P solutions， respectively. Scanning electron microscope （SEM） was used to observe the microscopic morphology of PVB-P nanofibers， and the Nano Measurer software was used to analyze the fiber diameter distribution. Fourier transform infrared spectroscopy （FTIR） was used to analyze the chemical composition of PVB-P. Water contact angle measurements were used to evaluate the hydrophilicity of PVB-P. UV spectrophotometer was used to detect the cumulative release of propolis at different time points. PVB and PVB-P nanofiber membranes were co-cultured with Staphylococcus aureus （S.aureus）， Escherichia coli （E.coli）， Candida albicans （C.albicans）， Salmonella， and Pseudomonas aeruginosa （P.aeruginosa） and divided into PVB group and PVB-P group. The absorbance method was used to calculate the antibacterial values of PVB and PVB-P against the five types of bacteria. The NIH-3T3 cells were seeded on PVB and PVB-P nanofiber membranes and divided into PVB group and PVB-P group. The CCK-8 method was used to detect the survival rates of NIH-3T3 cells on the nanofiber membranes in PVB group and PVB-P group at 1， 3， and 7 d. Results The SEM results showed that PVB and PVB-P nanofibers were interconnected in a mesh-like porous structure， with uniform thickness and no beads. The Nano Measurer software measurement results showed that the diameter of PVB nanofibers was （0.50±0.10） μm， and the diameter of PVB-P nanofibers was （0.54±0.16） μm. FTIR analysis showed that PVB-P nanofiber membranes exhibited characteristic peaks of PVB （1 140 and 1 002 cm^-1） and propolis （1 161 cm^-1）. The water contact angle measurement results showed that the contact angle of PVB membrane was （144.26°±2.90°） and that of PVB-P membrane was （128.13°± 1.36°）. The UV spectrophotometer results showed that the cumulative release of propolis was 0.04 mg at 1 d， 0.07 mg at 3 d， and reached a steady state of 0.79 mg at 7 d. The absorbance method results showed that the antibacterial values of PVB-P nanofiber membranes against S.aureus， E.coli， C.albicans， Salmonella， and P. aeruginosa were 4.39， 1.27， 5.68， 3.16， and 1.87， respectively. The CCK-8 method results showed that the survival rates of NIH-3T3 cells on PVB and PVB-P nanofiber membranes was>90% at 1， 3， and 7 d， and there were no significant differences between various groups （P>0.05）. Conclusion The diameter of PVB nanofiber membranes loaded with propolis is increased， and these membranes exhibit antibacterial effects against S.aureus and C.albicans， while also achieving sustained release of propolis.

The F-Box protein family is one kind of proteins containing the F-Box domain， which are together with S-phase kinase associated protein 1（SKP1）， Cullin1， and ring box protein 1（RBX1） to form the SKP1-CUL1-F-Box（SCF） E3 ubiquitin ligase complex； this complex mediates substrate ubiquitination modification and subsequent degradation via the proteasome pathway. The host F-Box protein plays an important role during the whole viral infection. The F-Box protein can regulate the replication of various RNA virus and DNA virus such as human immunodeficiency virus（HIV） and epstein-barrvirus（EB）， and the regulation mechanisms are different. After the virus enters the host cells， the host F-Box protein can regulate the stability and degradation of the key proteins during virus replication， enhance the immune response of host cells after virus infection， and inhibit the virus replication. Some F-Box proteins can assist the virus in completing the replication cycle by degrading the host restriction factors and inhibiting the virus activation and the interferon signaling pathway. The viruses also degradade the host factors and promote their own replication by encoding proteins containing the F-Box domain to bind to host SKP1， Cullin1， and RBX1 proteins. There are significant differences in the role of F-Box protein regulation during virus infection； one F-Box protein can regulate the replication of multiple viruses， and one virus can also be regulated by multiple F-Box proteins. This study systematically reviews the role of F-Box proteins in the process of viral infection from the perspectives of host F-Box proteins and viral F-Box proteins， and further explores the significance and potential value of F-Box proteins as targets for developing novel antiviral drugs.

Microglia， as intrinsic immune cells of the central nervous system， play an immune response function in Alzheimer’s disease （AD），which prevents further damages by eliminating abnormal proteins and apoptotic neurons while also leads pathological progression via inducing chronic neuroinflammation.The previous studies have suggested that the functional changes of microglia activation are initiated by AD pathologies. However，the recent genomic studies have challenged this understanding. Large-scale genome-wide association analysis （GWAS） and whole genome/exome sequencing studies have identified more than 70 risk loci of AD. Among these risk loci， most gene variants are involved in encoding microglia function-related molecules or affecting the transcriptional activity of genes associated with the microglial biofunctions.The functional and pathway analysis results have revealed that these risk loci are mainly enriched in signaling pathways regulating microglia phagocytosis， lipid metabolism， and immune response， suggesting that microglia not only act as a “responder” to AD pathologies， but also a “participant” in the development of AD pathogenesis.In-depth studies of these susceptibility genes may further expand our understanding of the regulatory mechanisms and functional spectrum of microglia in AD. This review is based on genetic studies and summarizes the current knowledge of microglial-related susceptibility genes related to AD and their regulatory mechanisms.

Mid-to-late stage ankle arthritis is a chronic degenerative disease that is extremely common in clinical practice. It is characterized by significant cartilage degeneration and subchondral bone sclerosis， accompanied by the formation of osteophytes around the joint， often leading to joint deformity. This condition causes severe pain in the patients during walking， severely restricts their activities， and affects their qualities of life. In recent years， with the continuous improvement of medical standards， the treatment methods for mid-to-late stage ankle arthritis have shown a diversified development trend. Non-surgical treatments primarily include activity restriction， orthotic devices， oral non-steroidal anti-inflammatory drugs （NSAIDs）， and intra-articular injections of the talocrural joint.The surgical treatments primarily include joint distraction arthroplasty， periacetabular osteotomy， total ankle arthroplasty， and ankle arthrodesis. Tissue engineering therapy， as an emerging method， has also received considerable attention. This article systematically reviewed the selection principles and research progress of various treatment options for mid-to-late stage ankle arthritis， including traditional treatments， non-surgical treatments， surgical treatments， and tissue engineering treatments. By deeply analyzing the basic principles and advantages and disadvantages of each treatment method， and combining the latest research findings on clinical outcomes， a scientific and comprehensive clinical decision-making reference system was constructed to provide clearer and more comprehensive treatment choices for both doctors and patients， thereby effectively improving treatment outcomes and enhancing the quality of life for the patients.