Objective: To study the recovery effect of Siwutang on the anemia induced by chemotherapy in the mice, and to clarify its mechanism.Methods: A total of 24 mice were randomly divided into control group(n=8,not given any drugs),model group(n=8,given 5-fluorouracil),Siwutang group(n=8,given 5-fluorouracil +Siwutang). After administration for 15 d,the peripheral blood of mice was taken,and the morphology and the number of red cells were observed under microscope. The number of peripheral blood red cells and the levels of hemoglobin of the mice in various groups were calculated by full-automatic hemocytometer; Double staining was used to mark the surface antigens of red cells,then antibody binding and staining were performed.Flow cytometry was used to detect the number of peripheral blood-labelled and spleen-labelled red cells.Results: Compared with control group, the number of red cells in peripheral blood of the mice in model group was significantly decreased(P<0.05),and the cells shrinkaged. Compared with model group, the number of red cells in peripheral blood of the mice in Siwutang group was significantly increased(P<0.05) and the cells showed rounding. Compared with control group, the level of hemoglobin in peripheral blood of the mice in model group was significantly decreased (P<0.05); compared with model group, the level of hemoglobin in peripheral blood of the mice in Siwutang group was significantly increased (P<0.01). The flow cytometry results showed that compared with control group, the expression level of CD71/Ter119 in peripheral blood of the mice in model group was significantly decreased (P<0.01),and the number of red cells in various quadrants was decreased(P<0.01); compared with model group, the expression level of CD71/Ter119 in peripheral blood of the mice in Siwutang group was significantly increased (P<0.01),and the number of red cells in various quadrants was increased(P<0.01);compared with control group, the expression level of CD71/Ter119 in spleen blood of the mice in model group was significantly decreased (P<0.01),and the number of cells in various quadrants was decreased(P<0.01); compared with model group, the expression level of CD71/Ter119 in spleen blood of the mice in Siwutang group was significantly increased (P<0.01),the number of red cells in UL quadrant was decreased (P<0.01),and the number of red cells in UR and LR quadrants was increased(P<0.01).Conclusion: Siwutang can promote the recovery of anemia induced by chemotherapy in the mice, increase the number of red cells,improve the morphology of red blood cells,increase the level of hemoglobin in peripheral blood, and increase the expression levels of marker antigens on the surface of red cells in the peripheral blood and spleen.

Objective: To discuss the effects of extremely low frequency electromagnetic fields(ELF-EMF) exposure on the function and morphology of liver,kidney and spleen of the SD rats, and to clarify the mechanism of radiation damage of ELF-EMF.Methods: Twenty male SD rats were randomly divided into exposure group and control group (n=10). The rats in exposure group were exposed in an ELF-EMF device with a magnetic induction of 0.1T and a frequency of 50 Hz; the exposure was conducted for 8 h per day for 30 d. The rats in control group didn't receive any treatment. After exposure for 30 d, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (UN), creatinine (Cr) of the rats in two groups were detected. HE staining was used to detect the morphology of the liver, kidney, spleen tissues of the rats in two groups.Results: Compared with control group, the serum levels of ALT, AST, UN, and Cr of the rats in exposure group were increased(P<0.05). In exposure group,the dilatation and congestion were found in the central veios of liver and hepatic sinusoids, glomeruli and interstitial capillaries of kidney, and insinusoids of spleen of the rats; part of the hepatocytes showed necrosis. There was no abnormality morphology in the liver, kidney,and spleen tissues of the rats in control group.Conclusion: Exposure to ELF-EMF(0.1 T, 50 Hz) can affect the function and structures of liver, kidney and spleen tissues of the SD rats, and its mechanism may be related to oxidative stress induced by ELF-EMF.

Objective: To investigate the expression of glucose regulating protein 78(GRP78) in the sciatic nerve cells of the rats with diabetic peripheral neuropathy (DPN), and to elucidate the clinical significance and mechanism of Adjust Zang Dredge Meridianelectroacupuncture in the treatment of DPN.Methods: A total of 86 adult male SD rats were divided into normal control group(n=5)and model group (n=81). The diabetic rat models were built through one-time injection of streptozocin(STZ), and then the diabetic rats were randomly divided into model control group(n=27),medicine intervention group(n=26), and electroacupuncture intervention group(n=26).The rats in normal control group and model control group were fed with the maintenance feed.During the course of experiment, no special treatment was done for the rats in normal control group and model control group.The rats in drug intervention group were treated with intragastric administration of mecobalamin,20 mg·kg^-1·d^-1,once per day. The rats in electroacupuncture intervention group were treated with Adjust Zang Dredge Meridian electroacupuncture in the bilateral Feishu, Pishu, Shenshu, Hegu, Zusanli,Sanyinjiao and Taichong, once per day, 30 min per time.The thermal sensitivity value, motor nerve conduction velocity(MCV)and sensory nerve conduction velocity(SCV)of the tibial nerve of the rats in various groups were measured, the ultrastructures of sciatic nerve of the rats in various groups were observed under transmission electron microscope, the apoptotic rates of sciatic nerve cells of the rats in various groups were detected by TUNEL method, the expressions of GRP78 in sciatic nerve cells of the rats in various groups were detected by immumofluorescence,and the expression levels of GRP78 in sciatic nerve of the rats in various groups were detected by Western blotting method.Results: Compared with medicine intervention group, the thermal sensitivity value of the rats in electroacupuncture intervention group was significantly decresed (P<0.01). Compared with model control group, the MCV and SCV of tibial nerve of the rats in medicine intervention group and electroacupuncture intervention group were increased after 12-week-intervention (P<0.01); compared with before intervention, the MCV and SCV of tibial nerve of the rats in various groups were decreased after 12-week-intervention (P<0.01).Under transmission electron microscope, some of the myelin sheath lamellae of sciatic nerve of the rats in model control group showed irregular membrane-like mass, and most of the thicker myelinated nerve fibers were seriously involved; the above changes were alleviated in medicine intervention group and electroacupuncture intervention group.The TUNEL assay results showed that compared with model control group, the apoptotic rates of sciatic nerve cells of the rats in medicine intervention group and electroacupuncture intervention group were significantly decreased (P<0.01). The Western blotting results showed that compared with model control group,the expression levels of GRP78 in sciatic nerve cells of the rats in medicine intervention group and electroacupuncture intervention group were significantly decreased (P<0.05).Conclusion: Adjust Zang Dredge Meridian electroacupuncture can reduce the expression level of GRP78 in sciatic nerve cells of the DPN rats, inhibit the occurrence of endoplasmic reticulum stress(ERS), decrease the apoptotic rate of cells, and protect the nerves.

Objective: To analyze the characteristics of clustered regularly interspaced short palindromic repeats (CRISPR) in Shigella from the database and clinic, and to explore the characteristics of CRISPR-Q1 and the correlations among CRISPRs.Methods: According to the genome database of Shigella, the primer sequences and PCR amplication were used to obtain the CRISPR sequences of Shigella, and the multiple sequence alignment was used to analyze the CRISPR distribution, the number and features of spacer; BLAST was used to analyze the repeats and spacers in CRISPR-Q1;the distribution characteristics of the repeats and spacers in Shigella and the location and features of CRISPR-Q1 in the strain were detected; the relationships between the spacer number of CRISPR-Q1 and CRISPR1/CRISPR2 in the database and laboratory were analyzed.Results: Among 107 Shigella strains in the database, 106 Shigella strains contained CRISPR1 or CRISPR2, 8 Shigella strains contained severial spacers; 106 Shigella strains contained CRISPR-Q1, and 13 Shigella strains contained severial spacers.A total of 6 among 12 Shigella strains in the laboratory contained CRISPR1 or CRISPR2, all of the 12 strains contained CRISPR-Q1. A portion of the CRISPR-Q1 spacer (about 35 bp) widely distributed in the genome of the same strain of chromosome, as was the repeat sequence; CRISPR-Q1 contained the repetitive extragenic palindromic elements (REP). The spacer number of CRISPR-Q1 was related to the CRISPR1 or CRISPR2 of Shigella in the database(χ²=61.60,P<0.01) and in the laboratory (P=0.015).Conclusion: CRISPR-Q1 widely distributes in Shigella and the REP elements can be found in CRISPR-Q1.There are correlations between CRISPR1 or CRISPR2 and CRISPR-Q1 in Shigella.

Objective: To observe the changes of dopamine (DA) levels in the hippocampal dentate gyrus (DG) region of the rats during the establishment and extinction process of active avoidance conditioned reflex, and to investigate the effect of D1 receptor in the active avoidance learning of the rats and its mechanism.Methods: A total of 24 male SD rats were randomly divided into non-training group, training group, control group and SCH group (n=6). The rats in training group were trained for active avoidance whereas the rats in non-training group were only put into the shuttle box without training, and then the DA levels in extracellular fluid in DG region of the rats in two groups were measured. In control and SCH groups, the saline or SCH-23390 was injected into the DG region of the rats before the active avoidance training, and then the glutamate (Glu) levels and the field excitatory postsynaptic potential (fEPSP) amplitudes in extracellular fluid in DG region of the rats in two groups were examined. The rates of active avoidance of the rats were recorded by behavioral analysis system of the shuttle box. The levels of DA and Glu in DG region of the rats were measured by microdialysis and HPLC techniques, and the amplitude of fEPSP in DG region of the rats was examined by electrophysiological recording.Results: The DA level in DG region of the rats in training group was gradually increased during the establishment process and was gradually decreased during the extinction process of conditioned reflex; compared with the 1st day, the DA level in DG region of the rats on the 5th day was markedly increased (P<0.05); the DA level in DG region of the rats in non-training group did not significantly change at different time points during the experimental process (P>0.05). In control group, the rats reached the establishment criterion on the 5th day (active avoidance rate >65%) and reached the criterion of extinction on the 7th day (active avoidance rate <35%); compared with the 1st day, the Glu level and fEPSP amplitude in DG region of the rats on the 5th day were significantly increased (P<0.05). In SCH group, the rats did not acquire the conditioned reflex, and the Glu level and fEPSP amplitude in DG region of the rats were not significantly changed during the whole behavioral training process (P>0.05). Compared with control group, the Glu level and fEPSP amplitude of the rats in SCH group on the 5th day were markedly decreased (P<0.05).Conclusion: DA in hippocampal DG region of the rats can facilitate the active avoidance learning via activation of D1 receptors, and its mechanism is associated with the enhancement of Glu level and synaptic transmission efficiency.

Objective: To investigate the expression of microRNA-155(miRNA-155) in the cerebral cortex tissue of the rats with cerebral ischemia-reperfusion injury, and to clarify the effect of miRNA-155 on cerebral ischemia-reperfusion injury.Methods: A total of 48 healthy adult male SD rats were randomly divided into sham operation group(n=24) and cerebral ischemia reperfusion group(I/R group,n=24).The cerebral ischemia-reperfusion models in I/R group were established by Longa modified suture occlusion in the right middle cerebral artery of the rats.The rats in sham operation group only received blood vessel isolation.TTC staining was used to calculate the ischemic infarction volume of the rats in various groups.qRT-PCR method was used to detect the expression levels of miR-155 in cerebral cortex tissue of the rats in various groups.Results: Compared with sham operation group, the ischemic infarction volume of the rats in I/R group was significantly increased at 24,48,and 72 h after reperfusion(P<0.05);the ischemic infarction volumes of the rats in I/R group were decreased gradually with the prolongation of reperfusion time.Compared with sham operation group,the expression levels of miR-155 in cerebral cortex tissue of the rats in I/R group were significantly increased at 24, 48, and 72 h after reperfusion (P<0.05); the expression levels of miR-155 were gradually decreased with the prolongation of reperfusion time.Conclusion: The expression levels of miR-155 in cerebral cortex tissue of the rats with cerebral ischemia-reperfusion injury are high, and miR-155 may participate in the process of cerebral ischemia-reperfusion injury.

Objective: To explore the inhibitory effect of 18β-glycyrrhetinic acid (18β-GA) on the inflammation-related gastric cancer, and to clarify its mechanism.Methods: A total of 72 K19-Wnt/C2mE transgenic mice with gastric cancer were randomly divided into 18β-GA treatment group (drinking water contain 0.1% 18β-GA, n=36) and control group (drinking normal water, n=36). After 52 weeks, the incidence of gastric cancer and morphology of gastric mucosa of the mice in two groups were detected. Immunohistochemistry staining was used to detect the histochemistry scores(H-score) of Ki-67, F4/80, ATP4a, KCNE2, pepsinogen C (PGC), Wnt-1, β-catenin, and cyclooxygenase-2 (COX-2) in gastric mucosa epithelial cells of the mice in two groups.Results: The gastric mucosa of the mice in control group showed protruded lesions, atypical hyperplasia and chronic gastritis. Compared with control group,the incidence(P=0.019) and the volume of gastric cancer of the mice in 18β-GA treatment group were significantly decreased (P<0.01);the structural heterozygosities of gastric cells and tissue were decreased,and the inflammation reactions of the mice in 18β-GA treatment group were alleviated. Compared with control group, the H-scores of Ki-67, F4/80, Wnt-1, β-catenin, and COX-2 in gastric mucosa epithelial cells of the mice in 18β-GA treatment group were decreased (P<0.05), and the H-scores of ATP4a, KCNE2,and PGC were increased (P<0.05).Conclusion: 18β-GA can significantly inhibit the occurrence of gastric cancer in the K19-Wnt/C2mE transgenic mice. The inhibitory effect may be related to alleviating the inflammation reaction and promoting the differentiation of gastric mucosa epithelial cells and inhibiting the occurrence of gastric cancer.

Objective: To investigate the effect of niacinamide mononucleotide (NMN) on the fibrosis of renal cells in the rats with diabetic nephropathy(DN), and to elucidate the mechanism of NMN in regulating the fibrosis of renal parenchymal cells through silent information regulator 1(Sirt1) and AKT pathways.Methods: The rat models of type 2 diabetes mellitus were induced by streptozotocin (STZ) and the model rats were randomly divided into experiment group(n=30) and control group(n=10).The rats in experiment group were divided into diabetes+NMN group(n=15) and diabetes+PBS group(n=15).The rats in diabetes+ NMN group were given subcutaneous injection of NMN for 20 d and the rats in diabetes+PBS group were given 200 μL sterile PBS in the same way.Then the rats were decapitated and the kidney tissues were taken for section and protein extraction. The expression levels of Sirt1, AKT, p-FoxO3a and Cav-1 proteins in kidney tissue of the rats in various groups were detected by Western blotting method and immuno-confocal focusing. The glomerular mesangial HBZY-1 cells were treated with high concentration of glucose (200 mmol·L^-1) for 3-6 d, and then the cells were further randomly divided into 4 groups (treated with 0,50,100,and 200 μmol·L^-1 NMN)and the cells only treated with 5.6 mmol·L^-1 glucose were regareded as control group. After 24 h culture, the cells were collected and the expression levels of Sirt1, AKT, and p-FoxO3a proteins in the HBZY-1 cells in various groups were detected by Western blotting method.Results: Compared with diabetes +PBS group,the expression levels of Sirt1 and AKT proteins in the renal parenchyma cells of the rats in diabetes+ NMN group were significantly increased (P<0.01) and the expression levels of p-FoxO3a and Cav-1 proteins in the renal parenchyma cells of the rats in diabetes + NMN group were also increased(P<0.01). Compared with control group, the expression levels of Sirt1 and AKT proteins in the HBZY-1 cells of the rats in 50 μmol·L^-1 NMN group were significantly increased (P<0.01), and the expression levels of Sirt1, AKT,and p-FoxO3a proteins in the HBZY-1 cells in 100 and 200 μmol·L^-1 NMN groups were increased significantly (P<0.05 or P<0.01).Conclusion: NMN can increase the expression levels of endogenous p-FoxO3a and Cav-1 proteins in the glomerular cells of the DN rats by regulating the expression levels of Sirt1 and AKT proteins, indicating that NMN and its analogues may play an important role in the prevention and treatment of the renal fibrosis of the DN rats.

Objective: To detecet the effect of herbicide atrazine(ATR) on immunologic function of the rats, and to clarify the immunotoxicity of ATR.Methods: A total of 32 Wistar rats were randomly divided into control group, low, middle and high doses of ATR groups (n=8); the rats in various groups were given 0,5, 25, and 125 mg·kg^-1 ATR for 28 d, respectively. The body weights and spleen indexes of the rats in various groups were detected; HE staining was used to observe the morphology of spleen tissue of the rats in various groups;lymphocyte transformation assay was used to determine the lymphocyte transformation rates in spleen cells of the rats in various groups; the killing activities of NK cells in spleen cells of the rats in various groups were detected;flow cytometry was used to detect the percentages of CD3⁺ and CD8⁺ T cells in spleen cells of the rats in various groups; ELISA assay was used to detect the serum levels of interleukin-1 (IL-1) and interleukin-6(IL-6) of the rats in various groups.Results: Compared with control group, the body weights of the rats in different doses of ATR groups had no statistically significant differences(P>0.05).Compared with control group, the spleen indexes of the rats in low and middle doses of ATR groups had no statistically significant differences(P>0.05),and the spleen index of the rats in high dose of ATR group was decreased(P<0.05).The HE staining results showed that compared with control group, the morphology of spleen tissue of the rats in low dose of ATR group had no significant changes; the germinal centers of spleen tissue of the rats in middle dose of ATR group were slightly decreased; the morphology of spleen tissue of the rats in high dose of ATR group had obviously degenerative changes, which were characterized by disappearance of germinal centers, diminution of white pulp, and congestion of red pulp. The results of killing activity detection of NK cells showed that compared with control group, the killing activities of NK cells of the rats in different doses of ATR groups had no significant differences(P>0.05).The spleen lymphocyte transformation assay results showed that compared with control group, the lymphocyte transformation rates of spleen cells of the rats in low and middle doses of ATR groups had no significant differences(P>0.05);the lymphocyte transformation rate of spleen cells of the rats in high dose of ATR group was decreased(P<0.05).The flow cytometry results showed that compared with control group, the percentages of CD3⁺ and CD8⁺ T cells of the rats in low and middle doses of ATR groups had no significant differences(P>0.05),and the percentages of CD3⁺ and CD8⁺ T cells of the rats in high dose of ATR group were decreased(P<0.05).The cytokine detection results showed that compared with control group, the serum levels of IL-1 and IL-6 of the rats in low and middle doses of ATR groups had no signifiant differences(P>0.05),and the serum levels of IL-1 and IL-6 of the rats in high dose of ATR group were increased(P<0.05).Conclusion: High dose of ART can induce the obvious immunotoxicity.

Objective: To analyze the changes of gene profile of over-expression WWOX gene in the human oral squamous cell carcinoma(OSCC) Tca8113 cells, and to explore the tumor inhibition mechanism of WWOX gene.Methods: The Tca8113 cells were divided into WWOX over-expression group and control group. In WWOX over-expression group, the lentiviral plasmid carrying the full-length cDNA fragment of WWOX gene was infected into the Tca8113 cells, while in control group, the lentiviral plasmid carrying the empty pGV287-LV vector was infected into the Tca8113 cells. The quantitative real time PCR (qRT-PCR)and Western blotting methods were used to detect the expressions of WWOX mRNA and protein. The gene chip technique was used to screen the differientially expressed genes and biological analysis was performed. The quantitative PCR was used to detect the relative mRNA expression levels of differientially expressed genes involving in the mitogen-activated protein kinase(MAPK) signaling pathway.Results: After infection, the relative mRNA expression level of WWOX gene in the Tca8113 cells in WWOX over-expression group was higher than that in control group(P<0.05), and the expression of WWOX protein in the Tca8113 cells in WWOX over-expression group was higher. A total of 347 differientially expressed genes with a 1.5-fold-change(FC) were screened by gene chip, in which 171 genes showed up-regulated expression, and 176 genes showed down-regulated expression. The Bioinformation analysis results showed that these different genes distributed in several pathways, which were related to cancer, p53, MAPK, and interleukin-2(IL-2) pathways and so on. Compared with control group,the relative expression levels of MAP2K,NR4A1,DUSP5,and DUSP6 mRNA in MAPK signaling pathway in WWOX over-expression group were increased(P<0.05), and the relative expression level of fibroblast growth factor receptor 2(FGFR2) mRNA was decresed(P<0.05).Conclusion: The over-expression of WWOX gene in the Tca8113 cells can induce the changes of gene profile of Tca8113 cells. The tumor inhibition mechanism of WWOX gene may be related to regulating the expressions of some genes involving in MAPK signaling pathway.

Objective: To observe the recovery effect of polylactide-glycolic acid (PLGA) microsphere-loaded velvet antler peptide(VAP) combined with bone marrow mesenchymal stem cells(BMMSCs) on the sciatic nerve injury of the rats, and to explore the related mechanisms.Methods: The sciatic nerve injury models were made by the cross-sectional method.A total of 60 rats were randomly divided into sciatic nerve transection control group(control group),VAP group,BMMSCs transplantation (BMC) group and VAP microsphere combined with BMMSCs transplantation(VAP-BMC) group(n=15).Seven days after modeling, the rats in control group were injected with PBS solution in the transecting sciatic nerve region,the rats in VAP group were injected with PBS solution containing PLGA microspheres loading drug, the rats in BMC group were injected with PBS solution containing BMMSCs,and the rats in VAP-BMC group were injected with PBS solution containing PLGA microspheres loading drug and BMMSCs.After three months,the neurological function of the rats in various groups were evaluated by Basso,Beattie,Bresnahan(BBB) functional sports rating scale,and the morphology of sciatic nerves of the rats in various groups were detected by HE staining.The expression levels of protein disulfide isomerase(PDI),glucose regulated protein-78(GRP-78) and Caspase-12 in sciatic nerve of the rats in various groups were detected by Western blotting method.Results: The results of BBB functional sports rating scale showed that compared with control group, the BBB scores of the rats in VAP group,BMC group and VAP-BMC group were increased (P<0.01),especially in VAP-BMC group.The results of HE staining showed that compared with control group, the sciatic nerve fibers of the rats in VAP group,BMC group and VAP-BMC group got thicker and the diameters of axon were longer (P<0.01).The results of Western blotting showed that compared with control group, the expression levels of PDI and Caspase-12 in the sciatic nerve of the rats in VAP group,BMC group and VAP-BMC group were significantly decreased(P<0.01),and the expression level of GRP-78 protein in the sciatic nerve of the rats in VAP-BMC group was significantly increased (P<0.01).Conclusion: PLGA microsphere-loaded VAP combined with BMMSCs can significantly promote the repairment of injured sciatic nerves, and its mechanism may be related to the inhibition of endoplasmic reticulum stress.

Objective: To investigate the expression levels of matrix metalloproteinase 2(MMP-2) and matrix metalloproteinase 9(MMP-9) in dentin of healthy adults under different adhesion conditions, and to provide the basis for improving the binding effect of matrix metalloproteinases(MMPs) inhibitors in clinic.Methods: A total of 20 adult freshlyextracted molars were collected and ground into dentin powder under liquid nitrogen cooling.The oral condition was simulated.The dentin was treated with self-etching bonding (Single Bond Universal) and total-etching bonding (35% phosphate gel +Adpter Single Bond 2). The dentin without any treatment was regarded as negative control group, the dentin treated with 10% phosphoric acid(self-etching bonding)or 10% phosphoric acid+ 35% phosphate gel (total-etching bonding) was regarded as positive control group, the dentin treated with Single Bond Universal(self-etching bonding) or 35% phosphate gel+Adper Single Bond 2 (total-etching bonding) was regarded as blank control group; the dentin pretreated with the MMPs inhibitors chlorhexidine(CHX) and minocycline(MI) during the bonding process respectively was regarded as CHX group and MI group,and the dentin treated with 10% sodium hypochlorite was regarded as aging group; on the basis of aging group, the dentin treated with CHX and MI was regarded as CHX aging group and MI aging group.Gelatin zymography was used to perform the polyacrylamide gel electrophoresis; after incubating, staining and decoloring, the bands were analyzed by gel image analysis system, and the expression levels of MMP-2 and MMP-9 in dentin were calculated.Results: Under the condition of self-etching bonding, compared with blank control group, the expression levels of MMP-2 and MMP-9 in dentin in CHX group were decreased (P<0.05), and the expression levels of MMP-2 and MMP-9 in the dentin powder in MI group were decreased (P<0.05);compared with aging blank control group, the expression levels of MMP-2 and MMP-9 in dentin in CHX aging group were decreased (P<0.05), and the expression levels of MMP-2 and MMP-9 in dentin in MI aging group were decreased (P<0.05). Under the condition of total-etching bonding, compared with blank control group, the expression levels of MMP-2 in dentin in CHX group and MI group were decreased (P<0.05); compared with aging blank control group, the expression levels of MMP-2 in dentin in CHX aging group and MI aging group were decreased (P<0.05).Conclusion: In the process of dentin bonding, CHX and MI could slow down the enzymatic reaction and improve the bonding strength by decreasing the expression levels of MMP-2 and MMP-9.

Objective: To investigate the stem cell-like properties of glioma U87 cells undergoing epithelial to mesenchymal transition (EMT), and to clarify the inhibitory effect of resveratrol (Res) on their stem cell-like properties.Methods: The glioma U87 cells were treated with transforming growth factor-β1(TGF-β1) to induce EMT, and the morphology of U87 cells was observed under inverted microscope. Western blotting method was used to detect the expression levels of mesenchymal markers (N-cadherin, Vimentin) and EMT-inducing transcription factors(Snail, Slug, Zeb1, Twist1). After the EMT of glioma cells was confirmed, the experiment was divided into control group (U87 cells didn't receive any treatment), EMT group (the glioma cells presented EMT induced by TGF-β1) and Res treatment group (the glioma cells with EMT were treated with Res). The number of second-generation gliospheres and the expression levels of stem cell markers (Bmi1 and Sox2) in glioma U87 cells in various groups were detected.Results: The glioma U87 cells treated with TGF-β1 for 48 h showed dispersed,elongated, and similar to the appearance of fibroblasts. The expression levels of N-cadherin, Vimentin, Snail, Slug, Zeb1 and Twist1 in glioma cells in different concentrations of TGF-β1 groups were significantly increased, suggesting the presence of EMT in glioma U87 cells. Compared with control group, the number of second-generation gliospheres in the U87 cells in EMT group was increased (P<0.01), and the expression levels of stem cell markers (Bmi1 and Sox2) in the glioma U87 cells in EMT group were markedly increased (P<0.05 or P<0.01). Compared with EMT group, the number of second-generation gliospheres in the glioma U87 cells in Res treatment group was significantly decreased(P<0.01), and the expression levels of Bmi1 and Sox2 in the glioma U87 cells in Res treatment group were decreased (P<0.01).Conclusion: The glioma U87 cells undergoing EMT exhibit the stem cell-like properties, and Res can inhibit their stem-like properties.

Objective: To discuss the protective effect of velvet antler polypeptides(VAP)combined with Schwann cells(SCs) modified by glial cell line derived neurotrophic factor(GDNF)gene on the apoptosis of spinal cord neurons induced by beta-amyloid 25-35(Aβ_25-35).Methods: The spinal cord cells of fetal mice were prepared and the spinal cord neurons in logarithmic growth phase were taken; the apoptosis of spinal cord neurons was induced by Aβ_25-35.The spinal cord neurons were divided into normal cell group (the normal spinal cord neurons), induced apoptosis group (the apoptotic spinal cord neurons induced by Aβ_25-35, SCs group (the apoptotic spinal cord neurons induced by Aβ_25-35+SCs), GDNF group (the apoptotic spinal cord neurons induced by Aβ_25-35+GDNF), SCs + GDNF group (the apoptotic spinal cord neurons induced by Aβ_25-35+GDNF-transfected SCs) and VAP combination group (the apoptotic spinal cord neurons induced by Aβ_25-35+VAP combined with GDNF-transfected SCs).Flow cytometry was used to detect the apoptotic rates of spinal cord neurons in various groups; immunohistochemical staining was used to detect the number of caspase-3 positive cells in the spinal cord neurons in various groups.Results: After suspension inoculation of fetal spinal cord neurons, most of them were round at the initial stage.The flow cytometry results showed that compared with induced apoptosis group, the apoptotic rates of spinal cord neurons in SCs,GDNF,SCs+GDNF,and VAP combination groups were decreased (P<0.05). Compared with SCs+GDNF group,the apoptotic rate of spinal cord neurons in VAP combination group was decreased (P<0.05).The immunohistochemistry results showed that the expression of caspase-3 in spinal cord neurons in various groups could be found. There were no significant differences in the number of caspase-3 positive cells and cell staining between SCs group, GDNF group and SCs+GDNF group; but the number of caspase-3 positive cells in SCs group, GDNF group and SCs+GDNF group were significantly higher than that in induced apoptosis group (P<0.05).Conclusion: VAP combined with GDNF-transfected SCs has the protective effect on the apoptosis of spinal cord cells by reducing the expression of caspase-3 in spinal cord neurons.

Objective: To establish the acute lung injury (ALI) mouse models induced by lipopolysaccharide (LPS),and to investigate the protective effect of berberine on ALI and its mechanism in vivo.Methods: A total of 40 C57BL/6 mice were randomly divided into control group, LPS group, berberine +LPS group and berberine group(n=10). The mice in control group were treated with saline;the mice in LPS group were treated with 5 mg·kg^-1 LPS for 6 h; the mice in berberine +LPS group were pretreated with 50 mg·kg^-1 berberine for 5 d before LPS treatment, then treated by 5 mg·kg^-1 LPS for 6 h;the mice in berberine group were pretreated with 50 mg·kg^-1 berberine for 5 d.The morphology of lung tissue of the mice in various groups were detected by HE staining. The ratios of wet to dry weights of lung tissue of the mice in various groups were detected. The concentrations of total protein and the number of effusion cells in bronchoalveolar lavage fluid(BALF) of the mice in various groups were detected; the levels of tumor necrosis factor(TNF-α) and interleukin-6(IL-6) in BALF of the mice in various groups were detected by ELISA method; the reactive oxygen species (ROS) levels in lung tissue of the mice in various groups were detected by dihydreethidium(DHE) method. The mitochondrial membrane potential in lung tissue of the mice in various groups were detected by JC-1 method.The expression levels of Bcl-2 and Bax proteins in lung tissue of the mice in various groups were detected by Western blotting method,and the ratios of Bcl-2/Bax were caculated.Results: Compared with control group, the inflammatory cell infiltration of lung interstitium and the alveolar space of the mice in LPS group were significantly increased, and the alveolar wall was thickened, the ratio of wet to dry weight of lung tissue was significantly increased (P<0.01), the level of total protein (P<0.05) and the number of exudation cells in BALF were increased (P<0.01),the levels of TNF-α and IL-6 in BALF were significantly increased (P<0.01), the ROS level in lung tissue was increased, the mitochondrial membrane potential in lung tissue was decreased (P<0.01),and the ratio of Bax/Bcl-2 in lung tissue was increased (P<0.01). Compared with LPS group, the inflammatory changes of lung tissue of the mice in berberine+LPS group were improved, the ratio of wet to dry weight of lung tissue was decreased (P<0.05), the level of total protein and the number of exudation cells in BALF were increased (P<0.05),the levels of TNF-α and IL-6 in BALF were decreased (P<0.01), the ROS level in lung tissue was decreased, the mitochondrial membrane potential in lung tissue was increased (P<0.05),and the ratio of Bax/Bcl-2 in lung tissue was decreased (P<0.05).Conclusion: Berberine could improve the degrees of LPS-induced ALI and inflammation of the mice,and the mechanism might be related to reduction of the ROS level and protection of mitochondrial function in lung tissue.

Objective: To investigate the protective effects of hyperbaric oxygen (HBO) in the exercise-induced fatigue mice, and to elucidate its mechanism.Methods: A total of 30 healthy male Kunming mice were randomly divided into control group, fatigue group and HBO intervention group. The mice in control group accepted normal exercise and swimming training without weight-bearing everyday. The mice in fatigue group and HBO intervention group accepted weight-bearing swimming everyday; after each exhaustive training, the mice in fatigue group were placed in the cage and the mice in HBO intervention group were immediately placed into the hyperbaric chamber with fresh sodium lime. The exhaustive swimming time of the mice in various groups were detected. The levels of malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), glutathione-peroxidase (GSH-Px) and total antioxidant capacity (T-AOC) in liver tissue of the mice in various groups were measured by ELISA method. The morphology of liver tissue of the mice in various groups were observed by electron microscope. The activities of ATPase in liver tissue of the mice in various groups were detected.Results: At the last exhaustive training, the exhaustive swimming time of the mice in HBO intervention group was significantly longer than that in fatigue group (P=0.002). Compared with control group, the levels of MDA in liver tissue of the mice in fatigue group and HBO intervention group were increased (P<0.05); while the levels of CAT, SOD, GSH-Px and T-AOC were decreased (P<0.05). Compared with fatigue group, the level of MDA in liver tissue of the mice in HBO intervention group was decreased (P<0.05), while the levels of CAT, SOD, GSH-Px and T-AOC were increased (P<0.05). There were no obvious abnormal changes in liver tissue of the mice in control group; the livercells of the mice in fatigue group showed concentration and endoplasmic reticulosis, the mitochondria were swollen or concentrated, and the structure of the iliac crest was broken, deformed and fused; but in HBO intervention group, the cytomembrane of liver tissue was intact, the mitochondrial changes were less than that in fatigue group, the structure of fistula was intact, and the changes of endoplasmic reticulum were also alleviated. Compared with control group, the activities of Na⁺-K⁺-ATPase, Ca²⁺-Mg²⁺-ATPase and total ATPase in liver tissue of the mice in fatigue group and HBO intervention group were decreased (P<0.05);compared with fatigue group, the activities of Na⁺-K⁺-ATPase, Ca²⁺-Mg²⁺-ATPase and total ATPase in liver tissue of the mice in HBO intervention group were increased (P<0.05).Conclusion: HBO intervention could effectively prolong the exhaustive swimming time of the mice and protect the function of liver,and its mechanism may be related to improving the antioxidant capacity and ATPase activity of liver tissue.

Objective: To investigate the effects of miR-26a targeting high mobility group protein 1(HMGA1) gene on the growth, invasion and migration of colon cancer cells,and to clarify whether the HMGA1 gene was the target gene of miR-26a.Methods: The miR-NC(miR-NC group), miR-26a mimics(miR-26a mimics group) and miR-26a inhibitor (miR-26a inhibitor group) were transfected into the human colon cancer SW480 cells. RT-PCR and Western blotting methods were used to detect the mRNA and protein expression levels of HMGA1. Luciferase reporter gene detection kit was used to detect the double luciferase activity in SW480 cells and to determine whether HMGA1 was the target gene of miR-26a.The cell proliferation activities of cells in various groups were detected by CCK8 assay; the number of invasion and migration cells was detected by Transwell chamber method.Results: HMGA1 gene was the target gene of miR-26a.Compared with miR-NC group, the proliferation activities of colon cancer SW480 cells in miR-26a mimics group at after 24, 48 and 72 h after transfection were significantly decreased(P<0.01), while the proliferation activities of colon cancer SW480 cells in miR-26a inhibitor group were significantly increased (P<0.01). Compared with miR-NC group, the proliferation activities of colon cancer SW480 cells,the number of invasion and migration cells in miR-26a mimics group and miR-26a mimics+pcDNA3.1-HMGA1 group at different time points were significantly decreased(P<0.01); the proliferation activities of colon cancer SW480 cells,the number of invasion and migration cells in miR-NC+pcDNA3.1-HMGA1 group were increased (P<0.01).Compared with miR-26a mimics group, the proliferation activities of colon cancer SW480 cells,the number of invasion and migration cells in miR-26a mimics+pcDNA3.1-HMGA1 group at different time points were increased (P<0.01);compared with miR-NC+pcDNA3.1-HMGA1 group, the proliferation activities of colon cancer SW480 cells,the number of invasion and migration cells in miR-26 mimics+pcDNA3.1-HMGA1 group at different time points were decreased(P<0.01).Conclusion: HMGA1 gene is the target gene of miR-26a.Up-regulation of the expression of miR-26a can inhibit the proliferation of colon cancer SW480 cells and down-regulation of the expression of miR-26a can promote the proliferation of colon cancer SW480 cells.

Objective: To investigate the effects of WNT4 gene silecing on the migration and invasion of renal tubular epithelial cells of the rats stably transfected with paired box gene 2(PAX2) gene,and to clarify whether the PAX2 gene regulates the biological activities of renal tubular epithelial cells through WNT4 gene or not.Methods: The renal tubular epithelial cells transfected with PAX2 gene in the logarithmic phase were divided into stably transfected group and control group. The cells in stably transfected group were divided into stably transfected control group(without WNT4 siRNA silencing) and silencing 72 h group(with WNT4 siRNA silencing for 72 h). The relative expression amounts of PAX2 and WNT4 proteins in renal tubular epithelial cells of the rats in stably transfected group and control group were detected by Western blotting method. The relative expression amounts of WNT4 mRNA in renal tubular epithelial cells of the rats in stably transfected control group and silencing 72 h group were detected by Real time PCR method. The migration rates of renal tubular epithelial cells of the rats in stably transfected control group and silencing 72 h group were detected by cell wound scratch assay. The number of transmembrane renal tubular epithelial cells of the rats in stably transfected control group and silencing 72 h group was detected by Transwell experiment.Results: The Western blotting results showed that the relative expression amounts of PAX2 and WNT4 proteins in renal tubular epithelial cells of the rats in stably transfected group were higher than those in control group(P<0.05). The Real time PCR results showed that the relative expression amount of WNT4 mRNA in renal tubular epithelial cells of the rats in silencing 72 h group was lower than that in stably transfected control group(P<0.05).The cell wound scratch assay results showed that the migration rate of renal tubular epithelial cells in silencing 72 h group after 10 h was lower than that in stably transfected control group,but there was no significant difference (P>0.05); the migration rate of renal tubular epithelial cells in silencing 72 h group after 18 h was lower than that in stably transfected control group(P<0.05).The Transwell experiment results showed that the number of transmembrane renal tubular epithelial cells in silencing 72 h group was lower than that in stably transfected control group(P<0.05).Conclusion: PAX2 gene can regulate the biological activities of renal tubular epithelial cells through WNT4 gene.

Objective: To investigate the effects of fingolimod (FTY720) on the migration and proliferation of neural stem cells (NSCs) and astrocytes of the mice, and to elucidate the protective effect of FTY720 on spinal cord injury(SCI) of the mice and its mechanism.Methods: The NSCs and astrocytes were cultured in vitro and divided into control group(treated with PBS) and experiment group(treated with 0.1 μmol·L^-1 FTY720). The SCI mice were divided into control group (given PBS by oral,n=10) and expriment group (given 1 mg·kg^-1·d^-1 FTY720 by oral,n=10). The number of migration cells of NSCs and astrocytes in two groups was detected by Transwell experiment, and the number of migration cells and the number of proliferative neurospheres of NSCs were detected by immunofluorescence staining.The SCI void areas of the mice in two groups were detected by HE staining, and the dynamic recovery of motor function of the mice was assessed by Basso Mouse Scale(BMS) behavioral score.Results: The Transwell detection results showed that compared with control group, the number of migration cells of NSCs in expriment group was increased(P<0.05), and the number of migration cells of astrocytes in expriment group was decreased(P<0.05). The immunofluorescence detection results showed that compared with control group, the number of proliferative neurospheres of NSCs in expriment group was increased(P<0.05),and the number of astrocytes in damaged SCI in expriment group was decreased(P<0.05).The HE staining results showed that compared with control group, the SCI void area was decreased(P<0.05). The BMS score results showed that the BMS score of the mice in expriment group was higher than that in control group(P<0.05).Conclusion: FTY720 can promote the proliferation and migration of NSCs, inhibit the migration of astrocytes, decrease the formation of glial scar, and promote the motor function recovery of the mice after SCI.

Objective: To explore the effects of plumbagin (PLB) on the proliferation and apoptosis of hepatocellular carcinoma hepG2R cells resistant to sorafenib,and to clarify its mechanism.Methods: The hepatocellular carcinoma cells hepG2 were cultured in vitro and the models of sorafenib-resistant HepG2R cells were set up. MTT assay was used to identify the resistance factor and screen the concentration and time of drug. According to the results, the concentrations of sorafenib and PLB were confirmed as 5 μmol·L^-1 and 2 μmol·L ^-1. The action time in various groups was 48 h. The HepG2R cells were randomly divided into control group, sorafenib (5 μmol·L^-1) group, PLB(2 μmol·L^-1) group, sorafenib (5 μmol·L^-1) combined PLB(2 μmol·L^-1) group (combination group). MTT assay was used to detect the cell vitality in various groups. The clone formation rates of cells in various groups were detected by clone formation assay. The apoptotic rates of cells in various groups were determined with Hoechst33342 assay and flow cytometry(FCM). The expression levels of cleaved Caspase-3,Bax and Bcl-2 proteins in the cells in various groups were examined by Western blotting method,and the Bax/Bcl-2 ratio was calculated. The reactive oxygen (ROS) levels in the cells in various groups were detected by ROS detector kit.Results: As the increasing of concentrations of sorafenib, the cell vitalities of HepG2 and HepG2R cells were gradually decreased;the IC₅₀ of sorafenib-resistant HepG2R cells was 4.5 times as much as HepG2 cells (P<0.05). The sensitivities of sorafenib resistant HepG2R cells were increased with the increasing of PLB concentrations and prolongation of time. Compared with control group, sorafenib group and PLB group, the clone formation rate of the cells in combination group was decreased(P<0.05 or P<0.01). The Hoechst 33342 assay results showed the nuclei were lightly stained; a few of the nuclei in sorafenib group and PLB group were strongly stained and bright; while in combination group, the nuclei were mostly stained and chromatin was condensed and bright.The FCM results showed that compared with control group, sorafenib group and PLB group, the apoptotic rate of cells in combination group was significantly increased (P< 0.05 or P< 0.01). The Western blotting results showed that the expression level of cleaved Caspase-3 in the cells and the Bax/Bcl-2 ratio in combination group were increased significantly (P<0.05 or P<0.01). The ROS level in the cells in combination group was higher than those in the other groups(P< 0.05 or P<0.01).Conclusion: PLB can improve the resistance of hepartocellular carcinoma to sorafenib and the mechanism may be related to increasing the ROS level.

Objective: To investigate the effects of acetaminophen on the hemodynamics, left ventricular function and plasma level of brain natriuretic peptide(BNP) in the premature young rats,and to clarify its mechanism in the treatment of patent ductus arteriosus(PDA) in the premature infants.Methods: The pregnant rats were injected intraperitoneally with lipopolysaccharide(LPS) to prepare the premature young rat models. The premature young rats were divided into model control group (n=18)and administration group (n=19);another 10 young rats with normal gestratonal age were selected and used as blank control group. The young rats in blank control group didn't receive any treatment, the young rats in model control group were not given any drug, and the premature young rats in administration group were continunously administrated with acetaminophen for 3 d. Color Doppler ultrasonography was used to detect the hemodynamic parameters of the young rats in various groups, including left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVEDd), left ventricular end systolic diameter (LVESd), fractional shortening (FS), left ventricular end-systolic volume (ESV), and left ventricular end-diastolic volume (EDV).The plasma BNP levels of young rats were detected.Results: The body weights of the young rats in administration group and model control group were lower than that in blank control group (P<0.05). After administration of acetaminophen for 3 d, compared with model control group, the body weight of the young rats in administration group was increased (P<0.05). Compared with model control group,the LVEF, FS and EDV of the young rats in administration group were increased (P<0.05),and the LVEDd, LVESd and ESV of the young rats were decreased(P<0.05). Compared with blank control group, the LVEF and FS of the young rats in administration group were decreased (P<0.05); the LVEDd, LVESd, EDV and ESV of the young rats were increased, but the differences were not statistically significant (P>0.05).Compared with blank control group, the plasma BNP level of the young rats in model control group was significantly increased(P=0.004);compared with model control group, the plasma BNP level of the young rats in administration group was significantly decreased(P=0.009).Conclusion: Acetaminophen can protect the left ventricular function of the young rats by improving the hemodynamic indicators and reducing the plasma BNP level.

Objective: To evaluate the efficacy and safety of saxagliptin combined with insulin and insulin used in the treatment of the patients with type 2 diabetes systematically.Methods: CNKI database, VIP database,Wanfang database,CBM database,PubMed and Cochrane Library were searched.The time was from the begining of the databases to Dec 31,2017. The randomized controlled trials (RCT) were selected according to the inclusion and exclusion criteria. Then the qualities of included studies were evaluated and the data was extracted. The Meta-analysis was performed by Review Manager 5.3 and Stata 12.0 softwares.Results: A total of 647 articles were selected initially and 22 articles involving 2 243 patients were included in this Meta-analysis. The results of Meta-analysis of effectiveness indicators showed that compared with insulin used alone(control) group, the HbA1c level(MD=-0.86, 95%CI:-1.03——0.68,P<0.01), FBG level (MD=-0.95, 95%CI:-1.22——0.67,P<0.01), PBG level (MD=-1.28, 95%CI:-1.61——0.94,P<0.01) and level of daily insulin dosage(MD=-16.17, 95%CI:-17.40——14.95,P<0.01) of the patients in saxagliptin combined with insulin(case) group were decreased. The safety analysis results showed that the total incidences of adverse reactions (OR=0.39, 95%CI:0.28-0.52,P<0.01) and the incidence of hypoglycemia (OR=0.31, 95%CI:0.22-0.45,P<0.01) of the patients in case group were lower than those in control group.The funnel-plot of these indicators displayed a symmetrical figure, and there was no publication bias; the sensitivity analysis results showed that there was no difference between all the indicators, and the results were stable.Conclusion: Compared with insulin used alone, saxagliptin combined with insulin therapy has certain advantages to reduce the levels of HbA1c, FPG, PBG of the type 2 diabetes patients and the daily insulin dosage, has high safety and dosen't increase the risk of hypoglycemia and other adverse reactions.

Objective: To explore the changes of incidence of major vascular lesions after intensive intervention of multiple controllable risk factors in the type 2 diabetes mellitus (T2DM) patients in Han population in Northeast China, and to establish an effective, standardized and cost-effective intervention scheme for the primary prevention of major vascular lesions of the T2DM patients.Methods: A total of 300 T2DM patients newly diagnosed or diagnosed in one year without major vascular lesions were randomly divided into conventional treatment group(n=150) and intensive intervention group (n=150). The clinical study method of parallel control was used to intervene in the some controllable risk factors for 2 years to varying degrees.The body mass index (BMI), waist/hip ratio (WHR), systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting blood glucose (FPG), 2 h postprandial blood glucose (2 h PBG), glycosylated hemoglobin (HbA1c), serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-c), high density lipoprotein (HDL-c), intima media thickness (IMT) and incidence of majoy vascular lesions of atherosclerosis(AS) of the patients in two groups were measured,and the medical expenses of the patients were analyzed statistically.Results: After 2-year-follow-up,the DBP, FBG, 2 h PBG, HbA1c, TG, TC and HDL-c of the patients in conventional treatment group were significantly lower than those before treatment (P<0.05).After 2-year-follow-up, the BMI, SBP, FBG, 2 h PBG, HbA1c, TG, TC, HDL-c and LDL-c of the patients in intensive intervention group were significantly lower than those before treatment (P<0.05). After 2-year-follow-up,the BMI, TC and IMT of the patients in intensive intervention group were significantly lower than those in conventional treatment group (P<0.05); compared with before treatment, the IMT of the patients in conventional treatment group after 2-year-follow-up was significantly decreased (P<0.05);compared with intensive intervention group, the IMT of the patients in conventional treatment group after 2-year-follow-up was significantly increased (P<0.05).The incidence of AS of the patients in intensive intervention group was lower than that in conventional treatment group (P<0.05). The medical expenses of the patients in both two groups were increased year by year; the 2-year total medical expenses, per capita annual medical expense in the first year and second year of the patients in intensive treatment group were lower than those in conventional treatment group (P<0.05).Conclusion: Intensive intervention of multivessel risk factors of is an effective way to prevent and treat the major vasular lesions of the T2DM patients.

Objective: To construct a three-dimensional finite element model of "2×4" appliance system simulating the clinical reality, and to investigate the effect of variation of bending location of tip-forward bending arch wires on the mechanical behavior in "2×4" appliance system.Methods: The 3-dimensional finite element model of "2×4" appliance system including maxillary dentition and brackets of tip-forward arch wires were constructed by reverse engineering method and the tip-forward bending arch wires with different bending locations[arch wire length from the point of arch wire to the central point of central incisor bracket/arch wire length from the central point of the buccal tube to the central point of central incisor bracket (A/L) were 0.8,0.7,0.6,0.5,0.4,and 0.3]were designed. The forces and moments of teeth under different tip-forward arch wires and the initial displacement of teeth under molar tip-forward arch wires were calculated by finite analysis software.Results: The force system changed as the bending locations of tip-forward arch wires varied. There were no vertical forces existed at A/L=0.41 for the first molar and incisors. There were no neutral points existed in the vertical force system(make the vertical forces=0 N in vertical direction). The moment of the first molar reversed when A/L=0.38 and the moment of lateral incisor reversed when A/L=0.59.The central incisor was affected under extrusion force without force and moment reverse as the bending location varied. The initial displacement of tooth was not conducive to the correction of anterior tooth apertognathia when A/L=0.8.Conclusion: The force system of three-dimensional finite element model of "2×4" appliance system changes as the varying of bending location of tip-forward arch wires. The molar tip-forward arch wire is unfavorable when correcting the anterior tooth apertognathia and the auxiliaries should be added.

Objective: To analyze the effects of serum hepatitis B virus covalently closed circular DNA(HBV-cccDNA)level on the liver and kidney functions, detection of HBV antigens in kidney tissue, pathological grading and staging of liver tissue in the children with hepatitis B virus associated glomerulonephritis (HBV-GN), and to evaluate the clinical value of HBV-cccDNA level detection in the diagnosis of the children with HBV-GN.Methods: A total of 39 HBV-GN children (observation group) were selected and all of them underwent the liver and kidney biopsy. A total of 40 HBV-carried children with normal liver function were selected as control group. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) of the children were detected.Molecular beacons PCR method was used to detect the serum HBV-cccDNA level of the children. The morphology of liver and kidney tissues of the children was detected with HE staining.The detection rates of HBsAg, HBeAg and HBcAg in kidney tissue of the children were detected by immunofluorescence. Receiver operating characteristic(ROC) curve and area under the ROC curve (AUC) were used to evaluate the value of HBV-cccDNA level detection in the diagnosis of HBV-GN. The Ax fluorescence value of HBV-cccDNA > 21 was determined as positive, and the Ax fluorescence value of HBV-cc cDNA < 21 was determined as negative; the children with HBV-GN were divided into HBV-cccDNA positive group(n=24) and HBV-cccDNA negative group(n=15). The serum levels of HBV DNA and HBV-cccDNA were measured at the 2nd, 4th, 8th and 12th weeks after antiviral therapy.Results: The abnormal rates of ALT and AST levels,positive rates of HBeAg and urine protein of the children in HBV-cccDNA positive group were higher than those in HBV-cccDNA negative group(χ²=4.454,P=0.035;χ²=5.912,P=0.022;χ²=8.770,P=0.007).There were no significant differences in the degrees of liver inflammation and fibrosis in the children between HBV-cccDNA positive and negative groups (P>0.05). Membranous nephropathy (MN) was the main pathological type of kidney biopsy in the HBV-GN.HBeAg and HBcAg were the main components of HBV antigens. The detection rates of HBeAg and HBcAg of the children in HBV-cccDNA positive group were significantly higher than those in HBV-cccDNA negative group(χ²=5.652, P=0.027; χ²=12.523,P=0.001). The ROC curve analysis results showed that the HBV-cccDNA level detection could effectively distinguish the HBV-GN children in observation group from those in control group, AUC=0.804 (95% CI 0.709-0.883).The levels of HBV-cccDNA of 10 cases of HBV-GN children underwent standardized treatment with complete follow-up treatment data were decreased significantly at the 2nd week after treatment. At the 12th week after treatment, in 7 cases the HBeAg turned negative, without proteinuria and hematuria symptoms, and the AST and ALT levels were normal. The HBV-cccDNA levels of 3 cases with ineffective treatment were higher than that those of the remaining 7 cases.Conclusion: The high expression of HBV-cccDNA is closely related to the liver function, proteinuria and HBV antigen detection in kindney tissue of the HBV-GN children. The detection of HBV-cccDNA level has potential clinical value for the auxiliaty diagnosis and evaluation on the curative effect of the HBV-GN.

Objective: To investigate the associations between the levels of human plasma anti-gliadin antibodies and occurrence of schizophrenia in the patients with different ages in Chinese Han population,and to clarify the effects of human plasma anti-gliadin antibodies in the occurrence of schizophrenia.Methods: The plasma samples were collected from 313 schizophrenia patients (schizophrenia group) and 408 healthy controls (healthy control group) in Chinese Han population. The levels of human anti-gliadin IgA and IgG antibodies of the subjects in various groups were detected by enzyme-linked immnosorbent assay(ELISA) method. The levels of human plasma anti-gliadin IgA and IgG antibodies in the subjects in schizophrenia group and healthy control group were analyzed by Pearson's correlation analysis and compared by Mann-Whitney U test; receiver operating characteristic (ROC) curve analysis was performed.Results: The level of plasma anti-gliadin IgA antibody showed a significantly positive correlation with the age and age ranges of the subjects(r=0.177,P<0.01;r=0.171,P<0.01); the level of plasma anti-gliadin IgG antibody showed significantly negative correlation with the age and age ranges of the subjects (r=-0.104, P<0.01;r=-0.127, P<0.01).Compared with healthy control group, the levels of plasma anti-gliadin IgA antibodies of the patients aged 20-30 years old and 50-60 years old in schizophrenia group were increased(Z=-3.746,P<0.01;Z=-2.186,P<0.05). Compared with healthy control group, the level of plasma anti-gliadin IgG antibody of the patients aged 20-30 years old in schizophrenia group was increased, but the difference was not statistically significant(P>0.05). The ROC curve analysis results showed that the area under ROC curve(AUC) of anti-gliadin IgA and IgG antibodies were 0.573 (SE=0.022, 95%CI:0.53-0.62) and 0.520 (SE=0.022, 95%CI:0.48-0.56). The sensitivities of anti-gliadin IgA and IgG antibodies were 13.7% and 12.8% when the specificity was 92.2%.Conclusion: The increasing of plasma anti-gliadin antibodies level is associated with the occurrence of schizophrenia in the patients with different age ranges in Chinese Han population, and the human plasma anti-gliadin antibodies maybe play an important role in the young schizophrenia patients.

Objective: To explore the serum levels and the positive expression rates of human mammaglobin(hMAM) and extracellular matrix metalloproteinase inducer(CD147) of the patients with breast cancer,and to clarify their clinical significnaces in the diagnosis of breast cancer.Methods: A total of 122 patients with breast cancer(breast cancer group), 21 patients with breast fibroadenoma (breast fibroadenoma group)and 16 healthy controls (healthy control group)were selected as the subjects. The serum samples of subjects in various groups were collected.The serum levels and the positive expression rates of hMAM and CD147 of the subjects in various groups were measured by ELISA method. The serum levels and the positive expression rates of hMAM and CD147 of the breast cancer patients with different clinicopathological features were analyzed.The cut-off values of serum levels of hMAM and CD147 of the patients with breast cancer and their sensitivities and specificities in the dignosis of breast cancer were comfirmed by receiver operating characteristic (ROC) curve.Results: The serum levels of hMAM and CD147 of the patients in breast cancer group were higher than those in healthy control group and breast fibroadenoma group(P<0.05). The positive expression rates of hMAM and CD147 of the patients in breast cancer group were higher than those in healthy control group and breast fibroadenoma group(P<0.01); while the positive expression rates of serum hMAM combined with CD147 of the patients in breast cancer group were higher than those in healthy control group and breast fibroadenoma group(P<0.01).There were significant differences in the positive expression rates of serum hMAM and CD147 of breast cancer patients with lymph node metastasis or not (χ²=10.375,P<0.01; χ²=15.556, P<0.01). There were significant differences in the positive expression rates of serum CD147 of the breast cancer patients with different TNM stages, ER and Her-2 (χ²=8157, P<0.05;χ²=6.035,P<0.05; χ²=5.385,P<0.05).With the increasing of TNM stages, the positive expression rates of serum hMAM and CD147 were increased.The area under ROC curve(AUC) of serum level of hMAM was 0.809, 95%CI:0.703-0.916;the AUC of serum level of CD147 was 0.721, 95% CI:0.582-0.861. When the cut-off value of hMAM was 6.51 μg·L^-1,its sensitivity and specificity were 61.1% and 87.5%, respectively. The sensitivity and specificity were 73.6% and 68.7% respectively when the cut-off value of CD147 was 144.92 ng·L^-1. The AUC of detection of hMAM combined with CD147 was 0.880, 95% CI:0.798-0.962; the sensitivity and specificity were 80.6% and 81.2%, respectively.Conclusion: The serum level of hMAM has the high sensitivity and specificity in the diagnosis of breast cancer. Combined detection of hMAM and CD147 can improve the positive rate of diagnosis.

Objective: To investigate the association between single nucleotide polymorphism(SNP) of integrin α-subunit 1 (ITGA1) gene and Helicobacter pylori(H. pylori) infection, and to provide the basis for the prevention and treatment of H. pylori infection-related diseases.Methods: The blood samples of 281 examinees were detected. ELISA method was used to confirm the status of H. pylori infection in the blood of the examinees.The samples were divided into H. pylori negative group(n=159) and H. pylori positive group(n=122)according to the antibody titers. The genotypes of rs1862610, rs2432143, and rs2447867 sites in ITGA1 gene were detected by TaqMan method.The haplotype was constructed by Haploview software 4.0 and unconditional Logistic regression was used to evaluate the associations between the SNPs of alleles, genotypes and haplotypes and H. pylori infection.Results: The SNPs of rs1862610 and rs2432143 sites in ITGA1 gene were not associated with H. pylori infection(P>0.05). The SNPs of rs2447867 site in ITGA1 gene was significantly associated with H. pylori infection(P<0.05). Carrying CT genotype decreased the risk of H. pylori infection (CT vs CC:OR=0.52, 95% CI:0.31-0.86). The SNPs between rs1862610 and rs2432143 sites in ITGA1 gene had a strong linkage disequilibrium (D'=1) and formed a block. None of the haplotypes was associated with H. pylori infection(P>0.05).Conclusion: The SNP of rs2447867 site in ITGA1 gene is associated with H. pylori infection,and carrying CT genotype can decrease the risk of H. pylori infection.

Objective: To compare the effects of different methods on the glucose treat-to-target time,glucose fluctuation,hypoglycemia and insulin doses in the type 2 diabetic patients treated by insulin bump, and to find the best method to make the glucose to reach the standand level safely,fast and effectively in the type 2 diabetic patients treated by insulin bump.Methods: Sixty hospitalized type 2 diabetic patients inadequatly controlled by premix insulin treatment were randomly divided into convention group (n=20),Bolus Wizard group (n=20),and Bolus Wizard combined with monitoring(combination) group (n=20) according to the random number grouping method.The insulin doses of the patients in convention group were adjusted according to the glucose monitoring of fingertip and the doctor's experiences; the insulin doses of the patients in Bolus Wizard group were adjusted according to the Bolus Wizard software in insulin bump, and the glucose monitoring of fingertip; the insulin doses of the patients in combination group were adjusted according to the Bolus Wizard software combined with real time continuous glucose monitoring system(RTCGM). The level of fingertip glucose was tested.The standard deviation of blood glucose(SDBG)and largest amplitude of glycemic excursion(LAGE)were used to evaluate the glucose fluctuation of the patients in various groups. The treat-to-target time,glucose fluctuation,hypoglycemia and daily insulin doses of the patients in various groups were recorded.Results: Compared with convention group,the treat-to-target time of the patients in Bolus Wizard group was decreased(t=2.30,P<0.05);compared with Bolus Wizard group,the treat-to-target time of the patients in combination group was decreased(t=3.50,P<0.05).On the 3rd day of treatment, compared with convention group,the SDBG and LAGE of the patients in Bolus Wizard group were decreased(t_SDBG=3.11,t_LAGE=2.54,P<0.05); compared with Bolus Wizard group, the LAGE of the patients in combination group was decreased(t_LAGE=2.47,P<0.05).There were no significant differences in the incidence of total hypoglycemia events (χ²=2.192, P=0.532), significant hypoglycemia events (χ²=2.765, P=0.322) and nocturnal hypoglycemia events (χ²=2.192, P=0.532) among the patients in various groups; there were no significant differences in the average insulin dosage (F=2.102, P=0.131), the non-basic insulin dosage (χ²=2.328, P=0.107) and the percentage of non-basic insulin (χ²=2.104, P=0.131) among the patients in various groups.Conclusion: Bolus Wizard software combined with real-time dynamic RTCGM has better effect in the treatment of type 2 diabetes without increasing the risk of hypoglycemia and insulin dosage.

Objective: To investigate the effect of chemotherapy combined with sodium cantharidinate vitamin B6 injection in the patients with non-small cell lung cancer (NSCLC) and its effect on the patients' quality of life, and to clarify its mechanism.Methods: A total of 110 NSCLC patients were divided into chemotherapy group and combined group according to the different treatment methods adopted by the patients. A total of 55 patients were treated with chemotherapy alone (chemotherapy group) and 55 patients treated with chemotherapy combined with sodium cantharidate vitamin B6 injection (combined group). The total effective rate of the patients in two groups were compared; EORTC QLO-C43 scale was used to evaluate the quality of life of the patients in two groups before and after treatment; the serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-α), and interleukin-12 (IL-12) of the patients in two groups before and after treatment were detected by double-antibody sandwich enzyme-linked immunoassay method.Results: After treatment, the total effective rate of the patients in combined group was higher than that in chemotherapy group, but the difference was not statistically significant (χ²=0.806, P=0.369). Before treatment, there were no significant differences in the scores of functional scale, overall health status, symptom subscale and lung cancer specific subscale of the patients between combined group and chemotherapy group (P>0.05). After treatment, the scores of functional scale and overall health status of the patients in combined group were significantly higher than those in chemotherapy group (P<0.05); the scores of symptom subscale and lung cancer specific subscale of the patients in combined group were significantly lower than those in chemotherapy group (P<0.05). Before treatment, there were no significant differences in the serum levels of VEGF, TNF-α, and IL-12 of the patients between combined group and chemotherapy group (P>0.05). After treatment, the serum level of VEGF of the patients in combined group was significantly lower than that in chemotherapy group (t=3.608, P<0.05), and the serum levels of TNF-α and IL-12 were significantly higher than those in chemotherapy group (t=3.426, P<0.05; t=2.849, P<0.05).Conclusion: The effect of chemotherapy combined with sodium cantharidate vitamin B6 injection in the treatment of the NSCLC patients is good, and it can significantly improve the quality of life of the patients;its mechanism may be related to decreasing the serum VEGF level and increasing the serum TNF-α and IL-12 levels.

Objective: To explore the clinical efficacy and safety of beraprost sodium (BPS) combined with glucocorticoid (GC) and(or) immunosuppressive agents in the treatment of the patients with primary nephrotic syndrome (PNS),and to provide evidence for its application in the treatment of PNS.Methods: Eighty-six patients diagnosed as PNS definitely were selected.They were treated with GC and(or) immunosuppressive agents and were divided into BPS group (administrated with BPS, n=42) and control group (administrated with dipyridamole or aspirin,n=44) according to their willing to the acceptance of different anti-platelet treatment regimens. The relevant laboratory indexes of the patients in two groups before and after treatment were analyzed, and the effective rate,incidence of complications and drug adverse reactions of the patients in two groups were compared.Results: Compared with control group, the urinary protein levels of the patients in BPS group at the 1st and 6th months after treatment were significantly decresed(P<0.05) and the serum albumin (ALB) levels of the patients in BPS group were significantly increased (P<0.05); the levels of fibrinogen (FIB) and D-dimer (DD) of the patients in BPS group at the 3rd, 6th and 12th months after treatment were significantly decreased (P<0.05 or P<0.01). Compared with control group, the systolic blood pressure(SBP)and diastolic blood pressure (DBP) of the patients in BPS group at the 1st month after treatment were decreased significantly (P<0.05).The cholesterol (TC) level and total effective rate of the patients in BPS group at the 6th month after treatment were higher than those in control group (P<0.05). Compared with control group, the incidence of elevation of blood pressure of the patients with normal basal blood pressure in BPS group was significantly decreased(P<0.05);the incidence of headache and dizziness was significantly increased(P<0.05).Conclusion: The safety of BPS combined with GC and(or)immunosuppressive agents in treating the PNS patients is higher and superior to the conventional antiplatelet agents.

Objective: To analyze the clinical manifestion and treatment method of one patient with severe periodontitis complicated with oral lichen planus, and to clarify the effect of combined treatment method and the importance of regular follow-up.Methods: The clinical manifestion of the patient was that the bilateral moral teeth had become looseness and the gum had bled for six months,and the patients had a history of oral lichen planus in gum for 2 years old. The probing depth(PD) of all the teeth was 5-6 mm and the attachment loss(AL)was 8-9 mm,with Ⅱ°-Ⅲ°mobility and bleeding on probing (BOP)(+).The teeth was treated with the regular basis periodontal treatment combined with gingival triamcinolone acetonide injection treatment.When the inflammation situation remained stable, the delayed implantation was performed and the crown restoration treatment was done for 26,27,36 and 37.The splinting treatment was done for 31 and 41 Ⅱ° mobile teeth. The regular follow-up periods were 1 week,2 weeks,1 month,3 months,and 6 months after treatment during periodontal maintenance.Results: After combined treatment and regular follow-up,the PD of both anterior and premolar were 4 mm,and the BOP was negative. The clinical symptom of oral lichen planus of the patient was relieved, and there were no symptoms of congestion and erosion on the gum. After the inflammation situation was controlled, the chewing function of the left molar region was restored by implantation rehabilitation. A 3-month-follow-up cycle and excellent treatment compliance were beneficial for the control of disease. The optimal follow-up period of gingival condition maintained best was once per 2 weeks.Conclusion: The combined treatment can relieve the clinical symptoms of the patient with severe periodontitis complicated with oral lichen planus. Regular follow-up and excellent treatment compliance are conducive to controlling the condition of disease.

Objective: To analyze the clinical features of Sj gren syndrome (SS) patient by investigating the diagnosis and treatment of one SS patient and perform the review of relative literatures,and to improve the understanding of the clinicians for the rare clinical symptoms and imaging features of SS.Methods: The patient was a 23-year-old woman with dyspnea associated with cough and expectoration for 1 month,and went to the hospital after hemoptysis for 3 d.The physical examination results showed pale conjunctiva and there were no other obvious positive signs.The chest CT results showed there were multiple cystic changes in both lungs.Further bronchoscopic biopsy,labial gland biopsy and rheumatism examinations and other assistant examinations were performed.The patient received related treatment.Results: The patient was initially diagnosed as lymphangioleiomyomatosis (LAM) and eventually diagnosed as SS by related examinations.The patient was treated with oral glucocorticoids and immunosuppressive agents.The symptoms of the patient were improved after treatment.The dyspnea was relieved, hemoptysis was not found,and the lung diffusion function was improved significantly.There were no significant changes in chest CT examination of the patient 2 months after treatment.Conclusion: There is rarely patients with hemoptysis and multiple cystic lesions in both lungs simultaneously among the SS patients,SS mainly occur in the women with childbearing age and it should be differentiated from LAM.

Objective: To explore the method to construct the progesterone receptor(PR)-superparamagnetic iron oxide (SPIO)-poly 1actic-co-glycolic acid (PLGA) and perfluorohexane(PFP) molecular probe(PR-s-PFP/PLGA), and to investigate its targeted possibility of binding the breast cancer cells in vitro.Methods: The s-PFP/PLGA nanoparticles were prepared and the diameter and average potential of s-PFP/PLGA nanoparticles were detected. Cytotoxicity experiment was used to compare the relative proliferation rates (RGR) of cells with different concentrations of s-PFP/PLGA. The magnetic resonance imaging (MRI), photoacoustic imaging (PAI) and ultrasound imaging of s-PFP/PLGA nanoparticles were detected,and the echo intensity of s-PFP/PLGA nanoparticles was observed by high intensity focused ultrasound (HIFU) in vitro.The PR-s-PFP/PLGA probe was prepared by carbodiimide method. The breast cancer cell line T-47d with high expression of PR was cultured in vitro. The T-47d cells with DiO green fluorescence were divided into targeted imaging agent group, non-targeted group and blank control group. The binding of the probe to the cells in various groups was observed.Results: The s-PFP/PLGA nanoparticles were spherical under transmission electron microscope (TEM).The SPIO particles were evenly distributed on the shell, the average particle size was (738.5±181.2) nm and the average potential was (-15.8±5.7) mV, and the obvious photoacoustic signals were found. The ultrasound imaging of s-PFP/PLGA nanoparticles in vitro showed a punctate high-level echo, and the echo intensity was enhanced after HIFU irradiation. With the increasing of iron concentrations, the MRI signals showed a increasing trend in T1W1. Under laser scanning confocal microscope, the s-PFP/PLGA nanoparticles, which was phagocytized by T-47d cells, emitted red fluorescence.Conclusion: PR-SPIO-PLGA has excellent physicochemical properties, good stability and strong targeting effect on the cancer cells.