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Absteact Objective To investigate the immunopathological changes of B lymphocyte activation in MRL/lpr lupus-like mice. Methods 20-week-old female MRL/lpr mice were sacrificed by cervical dislocation. The index of thymus and spleen of mice were calculated. The ratio of T lymphocyte subsets, total B cells and plasma cells in spleen lymphocyte suspension were analyzed with flow cytometry. Kidney and spleen pathology were observed with HE staining. The level of immune complex C3 expression in kidney and IL-21 mRNA in spleen were detected by immunofluorescence and q-PCR respectively. Serum IgG1, IgG2 a, ANA, anti-dsDNA, B lymphocyte stimulating factor(BLyS) and IL-10 levels were examinated by ELISA kits. The 20-week-age female BALB/c mice were used as control. Results Compared with the control group, the spleen and thymus index of MRL/lpr mice increased significantly, the proportion of TFH cells, Th17 cells, activated T cells and plasma cells increased while total B cells and immature T cells decreased obviously, and the differences were statistically significant. Furthermore, the C3 immune complex in glomerulus in kidney and the levels of serum IgG1, IgG2 a, anti-dsDNA, ANA, BLyS and IL-10 in MRL/lpr mice increased remarkably, displaying an interstitial nephritis. The spleen HE staining result of model mice displayed diffuse proliferation of white pulp, and the expression of IL-21 in spleen distinctly increased. The differences were statistically significant. Conclusion Similar with SLE, MRL/lpr mice display distinct interstitial nephritis which includes B lymphocyte hyperactivation, a mass of autoantibodies, immune complex deposition and high levels of BLyS, IL-10, IL-21.

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Objective To investigate the changes of age-related hippocampal glutamate receptor 1 level and its correlation with spatial learning and memory ability in CD-1 mice exposed to inflammation during late pregnancy.Methods Maternal mice were intraperitoneally injected with lipopolysaccharide(LPS, 50 μg/kg) or normal saline at days 15～17 of pregnancy. Their offspring were treated as LPS group and normal saline group, and completed Morris water maze at the age of 3 months and 15 months respectively. The level of GluR1 was detected by Western blotting.Results The 15-month normal saline group(compared to the 3-month normal saline group) and the 15-month LPS group(compared to 15-month normal saline group) had longer swimming distanceand lower swimming distance percentage in the target quadrant(all P<0.05) in the Morris water maze, with statistical significance and lower hippocampal GluR1 levels( all P<0. 01). The difference had statistical significance. Correlation analysis showed that only the hippocamppal level of GluR1 in the 15-month mice negatively correlated with swimming distance and positively correlated with the percentage of swimming distance in target quadrant. Conclusion The results suggest that the middle-aged CD-1 mice decrease GluR1 content of hippocampus which correlates with impaired ability of spatial learning and memory,and maternal exposure to inflammation in the late pregnancy accelerate this change.

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Objective To investigate the role of triptolide-induced down-regulation of cytochrome P450 3 A4(CYP3 A4)expression in causing liver injury. Methods L-02 cells and C57 BL/6 male mice were randomly divided into 4 groups respectively, including normal group(C), triptolide(TP) group, triptolide(TP)+carbamazepine(CBZ) group and triptolide(TP)+amlodipine(AML) group. After 24 h, the medium was collected, and C57 BL/6 male mice were euthanized and taken blood and liver 2 weeks later. The apoptosis rate of each group was detected by flow cytometry, while the damage of mice liver cells was observed by hematoxylin-eosin staining. The kit was uesd to detectethe level of ALT and AST in cell supernatant and mouse serum. The mRNA and protein expression levels of CYP3 A4 in cells and mouse liver were tested by Western blot and RT-PCR. The content of triptolide in cell culture medium and mouse serum was detected by HPLC. Results The results showed that the TP group significantly had more apoptosis and tepatocyte injury than the normal group. However, compared to the TP group, the apoptosis and liver cell injure of the TP+CBZ group significantly relieved, while the TP+AML group aggravated. The levels of ALT and ASTin vitroandin vivoof the TP group were significantly higher than the normal group, and the lower level was observed in the TP+CBZ group and higher level was observed in the TP+AML group. The expression of CYP3 A4 in TP group and TP+AML group significantly declined conapared with group C, while the expression of in the TP+CBZ group significantly higher. The results of HPLC showed that the TP content in the TP+CBZ group was lower than that in the TP group, while it was opposite in the TP+AML group. Conclusion TP can slow down cells metabolism by inhibiting its main metabolic enzyme CYP3 A4, which may be one of the underlying mechanisms of liver damage.

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Objective To study effects of low-frequency repetitive transcranial magnetic stimulation(rTMS) on relapse behavior, memory function and hippocampus of rats with methamphetamine(MA) addiction.Method 60 SD rats were randomly divided into control group, model group and experimental group. Fixed ratio 1 was applied in model group and experimental group to establish self-administration models of MA rats. In the process from model establishment to successful natural withdrawal, rats in experimental group were treated with rTMS for 7 d, while rats in model group and control group were only treated with sham stimulation. After completion, learning and memory ability of rats in each group was determined by wilderness experiment and Morris water maze test. After rats in model group and experimental group were given conditional clue induction, presence or absence of relapse behaviors was observed. After the end of the experiment, rates were sacrificed. The cerebral hippocampus was taken for slicing up. After HE staining,Nissl staining and uranyl acetate-lead citrate staining,changes of hippocampus were observed. Results The MA addiction rat models were successfully prepared. The learning and memory ability in model group was reduced. The r TMS treatment could effectively improve learning and memory treatment of rats with MA addiction. After natural withdrawal,there were relapse behaviors in rats of model group given conditional clue induction. After r TMS treatment,number of effective nasal touching under conditional clue induction was significantly decreased. In hippocampus of MA addiction rat models,number of Nissl bodies and average optical density decreased. The number of synapses in CA3 area reduced. There was fusion in anterior and posterior membranes.The number of synaptosomes reduced. The r TMS treatment could alleviate pathological changes to some extent.Conclusion The r TMS treatment can improve memory function and hippocampus tissue damage in MA addiction rats,and inhibit MA relapse behavior under conditional clue induction.

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Objective To explore the effect of interferon regulatory factor IRF9 gene silencing on apoptosis of cerulein induced exocrine pancreas AR42 J cells and elucidate the role of IRF9 gene in the development of acute pancreatitis. Methods The AR42 J cells at logarithmic growth stage were divided into blank control group, negative control group and IRF9 silencing group. After successful transfection of siRNA, the model of acute pancreatitisin vitrowas constructed by cerulean treatment. The mRNA and protein expression levels of IRF9 gene, Caspase 3 and Caspase 9 in AR42 J cells were detected by qRT-PCR and Western blot. The apoptosis rate of AR42 J cells in each group was detected by Annexin V-FITC/PI double staining assay. Results ① The mRNA and protein levels of IRF9 gene in each group increased after cerulean treated 24 hours, and the mRNA and protein levels of Caspase 3 and Caspase 9 also significantly increased, indicating that the expression levels of IRF9 and apoptosis-related genes were up-regulated when acute pancreatitis occurred.② Compared with the blank control group, the apoptosis rate of AR42 J cells in IRF9 silencing group significantly reduced after cerulean treatment(P<0.05). Conclusions The expression levels of IRF9, Caspase 3 and Caspase 9 were up-regulated and the apoptosis rate increased when acute pancreatitis occured. The silencing of IRF9 gene reduced the apoptosis of cerulean induced acute pancreatitisin vitro.

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Objective To examine the effect of sevoflurane exposure at the neonatal period on memory and synaptic plasticity at puberty in mice. Methods 30 neonatal mice(6～7 days old) were randomly divided into control group and sevoflurane exposure group, 15 mice in each group. The neonatal mice in the control group were not exposed to sevoflurane. The neonatal mice in the sevoflurane exposure group were anesthetized with 3.5% sevoflurane for 30 min, then with 3% sevoflurane for 30 min, and finally with 2.5% sevoflurane for 30 min. At 2-month-old, spatial memory, fear memory and social interaction memory were evaluated by morris water maze test, contextual fear and cue fear test and triple chambered social behavioral assay, respectively. The levels of Caspase-3 and BDNF in hippocampus were detected by immunohistochemical staining. The frequency and amplitude of mEPSC in GABA-ergic neurons were evaluated by whole cell patch clamp. Results Behavioral experiments show that compared with control group, mice in sevoflurane exposure group showed impairment in spatial memory, fear memory and social interaction memory at 2-month-old(allP<0.001). Immunohistochemistry showed that compared with the control group, the number of BDNF positive cells significantly reduced(P<0.001) while the number of Caspase-3 positive cells significantly increased(P<0.001) in hippocampus of mice in sevoflurane exposure group at 2-month-old. Whole cell patch clamp showed that compared with the control group, the frequency and amplitude of mEPSC significantly reduced(allP<0.01) in GABA-ergic neurons of mice in sevoflurane exposure group at 2-month-old. Conclusion Sevoflurane exposure at the neonatal period in mice can lead to neurodegeneration, memory impairment and synaptic plasticity decrease at puberty.

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Objective To establish a stable and efficient method for the culture of mouse bone marrow mesenchymal stem cells and the extraction of exosomes. Methods The whole bone marrow, differential attachment and hypoxia environment were used to culture bone marrow mesenchymal stem cells(BM-MSCs).Then the cells were collected, the surface markers of stem cells were identified, and osteogenesis, adipogenesis and chondrogenesis were induced. The supernatant was collected, and the exosomes were extracted by overspeed separation and identified. Result The growth of mouse BMSCs was great, and the purity of the third generation cells could reach more than 95% and stable passage to about 15 generations. The cells expressed the markers of mesenchymal stem cells, and could be successfully induced to differentiate into osteogenesis,adipogenesis and chondrogenesis. The extracellular vesicles obtained by ultracentrifugation had typical exosome-like structure, and Western blot identification showed that they had characteristic protein markers of exosomes. Conclusion The whole bone marrow culture method makes the loss of BMSCs in mice minimum, and the differential attachment method under 5% oxygen environment makes the proliferation of BMSCs in mice fast and high purity. Moreover, the mouse BMSCs cultured by this method can secrete exosomes vesicles steadily. This provides a great method for the culture of mouse mesenchymal stem cells and the acquisition of their exosomes.

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Objective To investigate the effect of MAST3 on proliferation and migration in SGC-7901 cells.Methods Western blot and qRT-PCR were used to detect MAST3 expression in gastric cancer tissues. MAST3-RNAi and p Ex-3-MAST3 were further transfected into SGC-7901 cells,and the expressions of CyclinD1,C-myc,MMP9 and MMP3 were detected by qRT-PCR and Western blot,respectively,and the effect of MAST3 on the proliferation and migration of SGC-7901 cells was observed. Cell cycle was detected by flow cytometry. Results Western blot results showed that MAST3 was significantly higher in gastric cancer tissues than that in adjacent tissues( P<0. 01),and the expression increased significantly after TGF-β1 stimulation. After MAST3-RNAi inhibited the expression of MAST3,the expression of CyclinD1 and C-myc protein was significantly lower than that of the control group( P<0. 01),and the cells in S phase and G2/M phase significantly decreased,while the expression of MMP-9 and MMP-3 protein was also significantly lower than that of the control. The expression of CyclinD1 and Cmyc protein was significantly higher than that of the control group after p Ex-3-MAST3 over-expression of MAST3( P<0. 01),and the cells in S phase and G2/M phase significantly increased,MMP-9 and MMP-3 protein expression was also significantly higher than of the control group. Conclusion MAST3 can regulate the proliferation and migration of gastric cancer SGC-7901 cells.

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Objective To study the expression levels of miR-486-5 p in colorectal cancer(CRC) cell lines and CRC stem cells(CSCs), and to explore its effect on CRC CSCs stemess and the targeted regulation of miR-486-5 p on Forkhead box class O1(FOXO1). Methods qRT-PCR was used to detect the expression levels of miR-486-5 p in CRC cell lines SW480, HCT116, SW620 and HT29 and normal colon cell line NCM460. CSCs were cultured in HCT116 cells. Western blot was used to detect the expressions of Nanog, SOX2, OCT4 stemess marker protein and miR-486-5 p in CSCs. After transfected with miR-486-5 p mimic and control(NC) 48 h, MTS was used to detect the proliferation ability of CSCs, Transwell assay was used to detect the metastasis ability of CSCs, and Boyden assay was used to detect the invasive ability of CSCs. CSCs stably overexpressing miR-486-5 p were screened by geneticin(G418) and injected into BALB/c nude mice to observe the effect of miR-486-5 p on the tumorigenic ability of CSCsin vivo. The targeting effect of mir-486-5 p on FoxO1 gene was verified by targetscan 7.1 software prediction and double luciferase reporter gene test. qRT-PCR was used to detect the effect of miR-486-5 p on FOXO1 mRNA expression in CSCs. Results qRT-PCR results showed that the expression of miR-486-5 p in CRC cell lines was lower than that in normal colon cell line(P<0.05). The expressions of OCT4, Nanog and SOX2 stem cell markers were up-regulated in CSCs, and the expression of miR-486-5 p decreased in CSCs. The proliferation, metastasis and invasion of CSCs decreased after miR-486-5 p mimic transfected(P<0.05). The tumorigenic ability of CSCsmiR-486-5 pnude mice decreased(P<0.05). TargetScan7.1 prediction software and double luciferase reporter gene assay confirmed that FOXO1 was a target gene of miR-486-5 p, and the expression of FOXO1 mRNA in miR-486-5 p mimic group decreased(P<0.05). Conclusion miR-486-5 p has low expression in CRC cell lines and CSCs, and miR-486-5 p may inhibit the stemness of CSCs by negative regulation of FOXO1.

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Objective To investigate the improvement effect of troxerutin on myocardial lesions in type 2 diabetic cardiomyopathy model rats and its possible mechanism. Methods The mice were divided into 4 groups: control group(20 rats, normal saline), model group(20 rats, normal saline), low-dose group [20 rats, troxerutin 50 mg/(kg·d)] and high-dose group [20 rats, troxerutin 100 mg/(kg·d)]. Cardiac function was detected by cardiac color Doppler ultrasound. Plasma biochemical indicators were detected by automatic biochemical analyzer.Histopathological changes were observed by Masson,HE and TUNEL staining. Apoptosis index was detected by TUNEL staining. Immunohistochemistry and Western blot were used to detect related protein expression. Results The results of cardiac color Doppler ultrasound showed that troxerutin significantly improved cardiac function. Masson,HE and TUNEL staining showed that compared with the model group,the cardiomyocytes in the low-dose group and the high-dose group were arranged neatly,the membrane was intact,the tissue fibrosis was reduced,and the collagen fibrosis was alleviated more effectively in the high-dose group( P<0. 05). TUNEL results showed that troxerutin significantly reduced myocardial apoptosis index( P<0. 01). Immunohistochemistry and Western blot showed that troxerutin could inhibit the expression of Collagen I and Caspase-3( P<0. 05) and enhance the expression of p-AKT and p-XIAP( P<0. 05). Conclusion Triclopine has a certain improvement effect on myocardial lesions in type 2 diabetic rats,and its mechanism may be related to activation of AKT/XIAP signaling pathway,inhibition of myocardial apoptosis and myocardial fibrosis.

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Absrtact Objective To evaluate the identification effect, homology analysis ability and self-library effect of MALDI-TOF MS technology for Acinetobacter baumannii. Methods 107 strains of Acinetobacter baumannii were identified by MALDI-TOF MS technology, and then drug sensitivity analysis was carried out. Tigacycline-resistant and tigacycline-susceptible Acinetobacter baumannii were screened out. Databases of tigacycline-resistant and tigacycline-susceptible Acinetobacter baumannii were established using the screened Acinetobacter baumannii. Finally, self-built databases were verified. Results MALDI-TOF MS identified 107 strains of bacteria as 103 strains of Acinetobacter baumannii, 3 strains of Acinetobacter junii and 1 Acinetobacter nosocomium. K-B and broth dilution methods were used to detect the sensitivity of 103 strains of Acinetobacter baumannii to tegacycline. 37 strains of Acinetobacter baumannii resistant to tegacycline and 39 strains of Acinetobacter baumannii susceptible to tegacycline were obtained. The consistency rate of the methods was 68.82%(64/93). Homology analysis for 76 strains of Acinetobacter baumannii revealed the distribution of pan-drug-resistant Acinetobacter baumannii in hospital. For self-built databases, the correct detection rate of tigacycline-resistant Acinetobacter baumannii and tigacycline-sensitive Acinetobacter baumannii was 87.4% and 77.5% respectively. Conclusion MALDI-TOF MS can quickly identify Acinetobacter baumannii, Acinetobacter junii and Acinetobacter nosocomium, and it can carry out Homology analysis of Acinetobacter baumannii. The resistance of Acinetobacter baumannii to tegacycline can be predicted by self-built databases.

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Objective To observe the effects of losartan on rat myocardial microvascular endothelial cells and Janus protein tyrosine protein kinase 2/signal transducer and transcriptional activator 3(JAK2/STAT3) signaling pathway, and explore protective mechanism of losartan on diabetes. Methods Rats were divided into a blank control group,(advanced glycation end products) AGEs group, AGEs plus losartan treatment group, and losartan treatment group for advanced glycation end products and losartan intervention. The activity and apoptosis rate of myocardial microvascular endothelial cells in rats were detected by MTT assay and TUNEL assay. The expression levels of related proteins in JAK2/STAT3 signaling pathway were detected by Western blot and RT-PCR. The levels of IL-6 and tumor necrosis factor-β(TNF-β) were detected by ELISA. Results Compared with the blank control group, the viability of myocardial microvascular endothelial cells in the AGEs group was significantly weakened and the apoptotic rate was increased. The expression of related proteins and the expression of related inflammatory factors IL-6 and TNF-β in the JAK2/STAT3 signaling pathway were also significantly elevated(P<0.05). Compared with the AGEs group, the AGEs plus losartan treatment group showed an increase in cell viability and a decrease in apoptotic rate. The expression of related proteins in the JAK2/STAT3 signaling pathway and the expression of related inflammatory factors IL-6 and TNF-β also decreased(P<0.05). There was no significant difference in the expression of related proteins and inflammatory mediators in the losartan group compared with the blank control group(P>0.05). Conclusion Losartan can effectively inhibit the advanced glycation end products through the JAK2/STAT3 signaling pathway, thereby alleviating the damage to myocardial microvascular endothelial cells and protecting them.

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Objective To explore the expressions of TSP-1 in the gingiva tissues of rat periodontitis model, and to verify the effect of TSP-1 on the expressions of interleukin-6(IL-6) and cyclooxygenase-2(COX-2) in primary cultured human periodontal ligament fibroblasts(hPDLFs). Methods 12 female SD rats were randomized into 2 groups, periodontitis group(ligature induced,n=6), control group(left unligated,n=6). Western blot was used to detect the protein level of TSP-1, and qRT-PCR was used to detect the mRNA level of TSP-1, IL-6 and COX-2 in the gingiva tissues in each group. After primary culturing hPDLFs with tissue block method, recombinant adenovirus(Ad-TSP-1-GFP) carrying TSP-1 gene was constructed and cells were transfected with no-load adenovirus(Ad-GFP) as a negative control, or add recombinant tsp-1 protein(rtsp-1) to the supernatant of hPDLFs culture. Il-6 expression level of hPDLFs was detected by ELISA. The expression of COX-2 in hPDLFs was detected by Western blot. mRNA expression levels of IL-6 and COX-2 were detected by qRT-PCR. Results Compared to the normal group, TSP-1 expressions in gingiva tissues were markedly increased in periodontitis group, and mRNA levels of IL-6 and COX-2 were synchronously elevated. Compared with Ad-GFP group, the level of TSP-1 was significantly higher in Ad-TSP-1-GFP group, meantime, the expression of IL-6 and COX-2 at both protein and mRNA levels in Ad-TSP-1-GFP group were also promoted.The results were confirmed by adding rTSP-1 into cell culture, since the Western blot and qRT-PCR analyses revealved a prominent upregulation of IL-6 and COX-2 after being induced by rTSP-1. Conclusion TSP-1 can significantly upregulate the expressions of IL-6 and COX-2 in hPDLFs, and then aggravate the symptoms of periodontitis.

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Objective To investigate the effect of morphine preconditioning in rostral ventrolateral medullar(RVLM) on myocardial ischemia-reperfusion(I/R) injury in rats and the changes of β-endorphin(β-EP) contents in T2-5 spinal cord and left ventricular muscle during the process. Methods 72 adult male SD rats were randomly divided into 4 groups(18 in each group):sham-operated group(Sham group), ischemia-reperfusion group(IR group), ischemic preconditioning group(IPC group), and RVLM morphine preconditioning group(MPC group). The ischemia reperfusion(I/R) model was treated with coronary artery left anterior descending branch ligation for 30 min and then reperfusion for 120 min. The ischemic preconditioning(IPC) was myocardial ischemia for 5 min, reperfusion for 5 min, and repeated the above operations for 3 times. Sham group, IR group and IPC group were injected 0.9% saline through the catheter. Then I/R was performed in the IR group, and IPC was performed before I/R in the IPC group. In the MPC group,morphine was injected before I/R. The numbers of ventricular arrhythmias [premature ventricular contraction( PVCs) and ventricular tachycardia/ventricular fibrillation( VT/VF) ]were recorded within 30 min of reinfusion by electrocardiogram monitoring. Cardiac specimens were stained with TTC.Left ventricle( LV),right ventricle( RV),infarct area( IS),ischemic risk area( AAR) volume were measured,and the IS/AAR ratio was calculated. Western blot assay was used for RVLM nucleus c-fos protein expression detection. ELISA assay and immunohistochemical staining assay were used for quantitative and qualitative detection the levels of β-EP expression in T2-5 spinal cord and left ventricular muscle,respectively. Results Compared with the Sham group,the IS volume,IS/AAR,the numbers of PVCs,VT/VF,and expression of c-Fos in RVLM and β-EP in spinal cord and myocardium were increased in IR group( P<0. 01). Compared with. IR group,the IS volume,IS/AAR,the numbers of PVCs,VT/VF,and the levels of c-Fos expression were significantly decreased( P<0. 01),but the levels of β-EP expression in myocardium and spinal cord were significantly increased in IPC group and MPC group( P<0. 01). Conclusion Morphine preconditioning in RVLM can alleviate myocardial I/R injury in rats,which may be related to the decrease of c-Fos expression in RVLM nucleus after I/R． β-endorphin may be involved as a transmitter in the myocardial protective effect of morphine preconditioning in RVLM.

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Objective To investigate the killing effect of Cold atmoshperic plasma on human osteosarcoma cell line MG63 and its possible mechanism. Methods The proliferation inhibition effect of Cold atmoshperic plasma on MG63 cells was determined by MTT assay and colony formation assay. The effect of Cold atmoshperic plasma on the morphology of MG63 cells was observed under a microscope. Apoptosis was detected by flow cytometry, and the changes in expression of apoptosis related proteins was detected by Western blot. Results Microscopic observation showed that Cold atmoshperic plasma could induce the shrinkage of MG63 cells. In both MTT and colony formation experiments, Cold atmoshperic plasma showed significant proliferation inhibition effect on MG63 cells in a dose-dependent manner. The cellular analysis showed that Cold atmoshperic plasma caused inhibition of cell proliferation by inducing apoptosis. Western blot showed that Cold atmoshperic plasma could induce the expression of apoptotic proteins in a dose-dependent manner. Conclusion Cold atmoshperic plasma has a significant killing effect on MG63 cells, which may be related to the induction of apoptosis.

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Objective To investigate the mechanism of miR-21 in anti-tuberculosis drug-induced hepatocyte injury. Methods Human normal hepatocytes(HL-7702) were cultured to construct the hepatic injury cell model at the optimum concentration of three drugs. Cells were divided into six groups: control group, drug group, drug+miR-21 inhibitor group, inhibitor negative control group, drug+miR-21 mimics group and mimics negative control group. HE staining was used to observe the morphological changes of the cells in each group. The alanine aminotransferase(ALT) and aspartate aminotransferase(AST) contents in each group were detected by the Lai method. The mRNA expressions of miR-21 and nuclear factor κB(NF-κB), tumor necrosis factor α(TNF-α) and interleukin-6( IL-6) genes were detected by RT-PCR. The expression of NF-κB protein in each group was detected by Western blot. The expression levels of TNF-α and IL-6 in each group were detected by ELISA. Results Compared with the control group, the expression of miR-21 was up-regulated in the drug group, and the mRNA and protein levels of NF-κB, TNF-α and IL-6 were elevated. Overexpression of miR-21 upregulated the mRNA and protein expression of the inflammatory regulatory factor NF-κB, while the mRNA and protein levels of inflammatory factors TNF-α and IL-6 further increased. Interfering with miR-21 reduced the mRNA and protein levels of NF-κB, TNF-α, and IL-6(allP<0.05). Conclusion The expression of miR-21 is up-regulated in anti-tuberculosis drug-induced liver injury cells. Down-regulation of miR-21 can reduce the expression of inflammatory factors to alleviate hepatocyte injury.

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Objective To investigate immunoglobulin D(IgD) can activate the function of T cells through immunoglobulin D receptor(IgDR)-lymphocyte-specific protein tyrosine kinase(Lck) signaling on human T cells and its effect on B cell activation.Methods Peripheral blood from healthy controls and RA patients were collected and peripheral blood mononuclear cells(PBMCs) were separated. CD4+T cells and CD19+B cells were sorted by flow cell sorting. Different concentration of IgD-stimul-ated CD4+T cells were co-cultured with CD19+B cells in transwell chamber. Laser confocal microscopy was used to observe Lck and IgDR co-localization on CD4+T cells.Western blot was used to detect the protein expression of P-Lck and CD40 L on CD4+T cells and the protein expression of CD40 on CD19+B cells. CCK-8 method was used to detect the effect of IgD-stimulated T cells on B cell activation. Results IgD significantly up-regulated the co-expression of Lck and IgDR proteins and the expression of P-Lck protein on CD4+T cells of healthy control. Both in healthy controls and RA patients,IgD-stimulated T cells could induce B cell activation. IgD could significantly up-regulate the expression of CD40 L protein on CD4+T cells and the expression of CD40 protein on CD19+B cells. Conclusion IgD could activate T cells via the IgDR-Lck signaling and stimulate B cell activation by up-regulating protein expression of T-B co-stimulatory molecules CD40 L and CD40.

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Objective To investigate the effect of hypoxia on miR-486 expression in human embryonic lung fibroblast MRC5 cells and identify mechanism of miR-486 in pulmonary fibrosis induced by hypoxia. Methods MRC5 cells were treated with hypoxia. Quantitative real-time PCR(qRT-PCR) was applied to detect the expression of miR-486,fibronectin(FN) and alpha smooth muscle actin(α-SMA),and Western blot was used to measure the expression of p-SMAD2 and GAPDH in MRC5. Lentivirus was used to mediate the overexpression or inhibition of miR-486 in MRC5 cells. Then,qQRT-PCR was used to detect the expression of miR-486,FN,α-SMA,Collagen Ⅰ and Collagen Ⅲ,and Western blot was used to measure the p-SMAD2 and GAPDH expression. Results Hypoxia inhibited the expression of miR-486 and promoted the FN,α-SMA and p-SMAD2 expression. MiR-486 overexpression successfully increased miR-486 expression and inhibited FN,α-SMA,Collagen Ⅰ,Collagen Ⅲ and p-SMAD2 expressions. MiR-486 knockdown upregulated the expressions of FN,α-SMA,Collagen Ⅰ,Collagen Ⅲ and p-SMAD2. Conclusion MiR-486 may regulate the pathogenesis of pulmonary fibrosis induced by hypoxia through targeting SMAD2.

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Objective To clarify the etiological association between different delivery modes and developmental behavior of preschool children. Methods A cross-sectional survey was conducted. A questionnaire survey was conducted on 27 987 preschoolers aged 3 to 6 years old. Questionnaires collected parents' basic demographic characteristics, basic information of children, their lifestyle and so on. The Age and Stages Questionnaire-third edition(ASQ-C) was used to evaluate developmental behaviors of preschool children, excluding children whose parents did not agree to do developmental behavior assessment and did not complete ASQ-C assessment. Finally, a total of 14 634 children completed developmental behavior assessment. Results The detection rates of communication(CM), gross motor(GM), fine motor(FM), problem solving(CG) and person-social(PS) function retardation in preschool children were 22.1%, 19.4%, 18.3%, 23.8% and 24.1%, respectively. After adjusting for confounding factors, compared to women with vaginal delivery, cesarean section with medical indications was positively correlated with gross motor retardation of preschool children(OR=1.14, 95%CI:1.04～1.25) and person-social retardation(OR=1.10, 95%CI:1.01～1.20). There was a positive correlation between cesarean section without medical indications and gross motor(OR=1.16, 95%CI:1.03～1.30), fine motor(OR=1.22, 95%CI:1.08～1.37) and problem solving(OR=1.20, 95%CI:1.07～1.34) retardation in preschool children. Conclusion Compared with vaginal delivery, cesarean section is associated with developmental behavioral delay in preschool children.

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Objective To investigate the association of M-type phospholipase A2 receptor 1(PLA2 R1) gene rs3749117 and rs4664308 with idiopathic membranous nephropathy(IMN). Methods Eighty-eight patients with IMN diagnosed by renal biopsy were selected as the case group,and 122 healthy controls in the physical examination center of the Autonomous Region People's Hospital were collected. The rs3749117 and rs4664308 polymorphism were detected by competitive allele-specific polymerase chain reaction(KASP) and the relationship between PLA2 R1 gene polymorphism and IMN were analyzed. Results The rs4664308 did not meet the Hardy-Weinberg equilibrium. The genotype and allele frequencies of rs3749117 had statistically significant difference between the IMN group and the HC group(χ2=23.415,P<0.001,χ2=25.792,P<0.001). There was no significant difference in the age,gender,systolic blood pressure,diastolic blood pressure,urea nitrogen,creatinine,uric acid,triglyceride,total cholesterol,low density lipoprotein,albumin,urine protein and anti-PLA2 R antibody positive rate between rs3749117 TT genotype group and TC+CC genotype group(P>0.05). The stratified chi-square test showed that there were differences in the distribution of susceptible genotype(TT genotype) and non-susceptible genotype(TC+CC genotype) between IMN and healthy controls in the 18～<40 and 40～<60 age groups(P<0.05),and there was no difference in the distribution of rs3749117 susceptibility genotype(TT genotype) and non-susceptible genotype(TC+CC genotype) between the IMN and the healthy control in the elderly group(P>0.05). Conclusion The PLA2 R1 rs3749117 gene polymorphism may be associated with susceptibility to IMN in the 18～<40 and 40～<60 age groups in Xinjiang,and the gene polymorphism of rs3749117 may be unrelated to the occurrence of IMN in elderly patients.

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Objective To explore the impact of pre-and post-set adaptive statistical iterative reconstruction Veo(ASiR-V) on imaging quality in the chest phantom. Methods The chest phantom had been scanned by GE revolution CT with the different weight of pre-set ASiR-V, which was set to 0%,20%,30%,40%,50%,60%,70%,80%,90%,and 100% successively. Then on the basis of pre-set ASiR-V set as 0%, 30% and 60%, images were reconstructed with post-set ASiR-V weight interval of 10%(from 0% to 100%) consecutively. A standard algorithm was used for original image reconstruction. The slice was 1.25 mm. The CT and SD value of various tissues at different level were recorded. Simultaneously, assessing subjective scores of lung and mediastinal windows under different proportions were evaluated to evaluate the discrepancy of image quality. Results With the increase of pre-ASiR-V weight, subjective scores of mediastinal and pulmonary window reached the best both at 30%. The subjective score of mediastinal window could ≥3 before 70%, while pulmonary window before 90%. There was no obvious change in the CT and SD values of various tissues. On the basic condition of pre-ASiR-V set to 0%, 30% and 60%, with the increase of post-ASiR-V, SD values of different tissues(except lung) showed a clear decreasing trend, while no significant difference was observed in CT values. The subjective scores of mediastinal and pulmonary window could meet the diagnostic requirements while post-set ASiR-V was no higher than 80% in the group of pre-set ASiR-V on 0% and 30%. The subjective scores of mediastinal and pulmonary window displayed fairly well and could meet the diagnostic requirement while post-set ASiR-V had been set on 60%～80% in the group of pre-set ASiR-V on 60%. Conclusion Excessive high pre-ASiR-V will generate the decline of image subjective scores. And excessive high or low post-ASiR-V weight will both do so. 60% pre-position combined with 60%～80% post-position ASiR-V is recommended for Chest CT scan while the default NI value is 14.

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Objective To study the effect of the circulating tumor cells(CTCs) and the CTCs dopamine receptor 1( DRD1) expression level in peripheral blood of breast cancer patients on breast cancer progression and curative effect monitoring. Methods Peripheral blood was collected from 20 healthy people and 20 untreated breast cancer patients, and CTCs and CTCs DRD1 expression levels of them were detected through the differential phase concentration-immune fluorescence in situ hybridization technique(SE-iFISH) in combination with the fluorescent quantitative PCR(Q-PCR). Then 6 cases of breast cancer were selected for operation, and their peripheral blood was collected at 3 and 6 months after operation, to detect CTCs and CTCs DRD1 expression levels and analyze their relationship with disease progression and curative effect. Results CTCs were not detected in peripheral blood of 20 healthy persons while detected in 18 of 20(90.0%) untreated breast cancer patients, which presented high sensitivity and specificity. CTCs and CTCs DRD1 levels evidently decreased in 3 T2 stage patients 3 months and 6 months after surgery(they did not appear recurrence and metastasis). On the contrary, CTCs and CTCs DRD1 levels evidently increased in 3 T3 stage patients 3 months and 6 months after surgery(they appeared prominent recurrence and metastasis). Conclusion Breast cancer patients with high CTCs detection rate and CTCs DRD1 expression often have poor prognosis. On the contrary, patients with low CTCs detection rate of CTCs and expression of CTCs DRD1 show better disease control and good prognosis. Hence, noninvasive CTCs and CTCs DRD1 detection may have high clinical application value in monitoring the efficacy of breast cancer.

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Objective To evaluate the clinical outcome of131Ⅰ therapy in papillary thyroid Cancer(PTC) with preablative stimulated thyroglobulin(ps-Tg)below 10 μg/L and explore correlation between ps-Tg level and the clinical outcome. Methods Ninety eight PTC patients with ps-Tg below 10 μg/L were selected and divided into four groups [excellent response(ER),indeterminate response(IDR),biochemical incomplete response(BIR) and structural incomplete response(SIR)] according to the response to initial131Ⅰ therapy. BIR and SIR were classified as incomplete response(IR).The clinicopathological features and ps-Tg level were compared among the three groups. Three groups of patients were further divided into ER and non-ER groups and the ps-Tg level was compared between them. Lastly,the optimal cut-off point was obtained by analyzing ROC curve to explore the value of ps-Tg for predicting excellent response(ER). Results There was no significant difference in age,sex,neck ultrasound,recurrence risk,number of lesions,cervical lymph node metastasis and tumor size among the three groups(all P>0.05),but there was significant difference in ps-Tg level among them(P<0.001). Two comparisons further made between groups in ps-Tg level,and there was significant difference between ER and IDR(adjusted P=0.001),IR(adjusted P=0.015),but there was no significant difference between IDR and IR(adjusted P=0.869). There was also significant difference in ps-Tg level between ER and non-ER(P<0.001). Area under the ROC curve was 0.777. The diagnostic optimal cut-off point was 1.95 μg/L,with a higher sensitivity and specificity(80.00% and 70.00%,respectively). Conclusion Ps-Tg of no more than 1.95 μg/L can be used to indicate satisfactory clinical outcome in postoperative PTC patients whose ps-Tg was below 10 μg/L.

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Objective The clinical features of children with abdominal type Henoch-Schönlein purpura(HSP) and the changes of gastrointestinal mucosa under endoscopy were analyzed, and the relationship between them was further discussed. Methods The clinical manifestations and endoscopy characteristics of 116 children with abdominal type HSP were analyzed retrospectively, according to the degree of mucosal damage under endoscopy, the children were divided into two groups. The disease course, the symptom score of purpura, the rate of acute pancreatitis and the rate of Henoch-Schönlein purpura nephritis(HSPN) were compared, and the high risk factors of severe mucosal damage in children with HSP were analyzed. The difference of the degree of mucosal damage under endoscopy between the two groups with digestive tract as the first symptom and rash as the first symptom was analyzed. Results ① Abdominal HSP often manifeste as abdominal pain, vomiting, haematemesis, melena and hematochezia, while under endoscopy, it was characterized by purplish red mucous rash with different degrees,of density slightly higher than the surface of mucous membrane, and could be distributed in flakes, and the appearance of interrash mucosa could be normal, which was similar to that of skin purpura. ② There was no significant difference in symptom score and acute pancreatitis complications between the two groups(P>0.05). ③ The total time of recombinant abdominal pain, the time of complete relief of abdominal pain after admission, the length of hospitalization and the incidence of HSPN during hospitalization were significantly higher than those in mild mucosal lesion group(P<0.05). ④ The increase of serum CRP was an independent risk factor for severe mucosal damage in children with abdominal HSP, and its level was positively correlated with the degree of gastrointestinal mucosal injury in children with HSP. ⑤ The damage of mucous membrane in HSP children with digestive tract as the first symptom was more serious than those with rash under endoscopy(P<0.05). Conclusion Endoscopy can be used for early diagnosis of HSP with abdominal pain as the first symptom.The increase of serum CRP in abdominal HSP can be used as an index to predict the degree of gastrointestinal damage. Serious injury of digestive tract mucosa may be a risk factor for renal damage in children with HSP.

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Objective To investigate the expression level of Interleukin-26(IL-26) in hepatitis B virus-associated acute-on-chronic liver failure(HBV-ACLF) and its clinical significance.Methods Serum IL-26 levels were measured by enzyme-linked immunosorbent assay(ELISA) in 53 patients with HBV-ACLF, 53 patients with chronic hepatitis B(CHB) and 18 healthy controls. General data were statistically analyzed, and relevant clinical indicators were analyzed for their correlations with IL-26. Multivariate logistic regression analysis was used to assess independent risk factors for prognosis of patients. ROC curve and survival curve were used to evaluate the prognosis performance of patients. Results The level of IL-26 in HBV-ACLF patients was significantly higher than that in non-aclf patients. The expression of IL-26 was positively correlated with creatinine( CREA) and activated partial thromboplastin time( APTT). Multivariate logistic analysis showed that total bilirubin( TBIL) and IL-26 were independent risk factors. The prognostic model AUC( 0. 933) established by IL-26 combined with MELD was more sensitive and accurate than single index. Serum IL-26>1 019. 68 pg/ml had reliable prognostic accuracy in predicting 90-day mortality in patients with HBV-ACLF. Conclusion The level of IL-26 significantly increased in patients with HBVACLF,which has important clinical significance and application value in monitoring the condition and prognosis of patients with HBV-ACLF.

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Objective To study the expression of CYFIP1 gene in HCC tissue and its biological mechanism in the development of hepatocarcinoma(HCC). Methods The expression of CYFIP1 in 80 pairs HCC tissues was evaluated by staining of tissue microarray slides(TMA), and its clinical relevance was evaluated. The apoptosis assay was conducted with Annexin V-FITC/PI staining. The expression of CYFIP1 in HepG2 cells was performed with Western blot. Results The expression of CYFIP1 in HCC tissues was lower than that in adjacent normal liver tissues(P=0.031), which was clinically relevant with tumor grades, tumor size and prognosis of HCC patients. Knock down of CYFIP1 by shRNA in HepG2 cells increased the proliferation(P=0.019) and decreased the apoptosis(P=0.021) of HepG2 cells. Conclusion CYFIP1 is a suppressor gene in HCC, which may be a potential prognostic marker in HCC.

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To verify the coincidence rate and evaluate the cut-off value of six kinds of domestic enzyme linked immunosorbent assay(ELISA) kits of hepatitis B virus surface antibody(HBsAb), and also to discuss the feasibility of using cut-off serum. HBsAb in clinical samples and the HBsAb standard substances were detected by electro chemiluminescence immunoassay(ECLIA) and six kinds of domestic ELISA kits of HBsAb respectively. Using the cut-off value of each kit to determine the results, the positive rates of reagent E and F were 100% when testing 10 mIU/ml of HBsAb standard substances. Reagent C and E had the highest precision. With 10 mIU/ml of HBsAb standard substances as cut-off serum, the cut-off value and grey zone of the six reagents were different. Reagent D had the lowest cut-off value and the minimum grey zone, while Reagent F had the highest cut-off value and the largest grey zone. When the HBsAb concentration of clinical samples was lower than 10 mIU/ml, 10～<50 mIU/ml and 50～<100 mIU/ml, the positive coincidence rates of the cut-off value determined by the ELISA kit of most reagent manufacturers was different from that of the cut-off serum. When clinical specimens with HBsAb concentration from 100～1 000 mIU/ml were tested, the positive coincidence rates of the two results determined by the two cut-off values of reagent A, B, C, D, E and F were all 100%. There are some differences in the detection performance of the six domestic ELISA kits of HbsAb, and there are deficiencies in the way that the cut-off value was recommended to be calculated. The use of cut-off serum will change the positive and negative coincidence rate of the ELISA kits, which needs to be fully verified by clinical samples before use.

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To study whether Kolliphor®P188 could inhibit the inflammation during autologous fat transplantation,from the perspective of inflammation to analyze the effect of P188 on transplantation. The animal models of autologous fat transplantation were divided into the control group(normal saline group) and the experimental group(P188 group). Weight and volume were measured after fat tissue was removed at time points 1, 2 and 4 weeks. The degree of fat inflammatory infiltration after transplantation was observed by HE staining. Finally, the mRNA and protein levels of inflammation-related factors IL-1α,1 L-1β,IL-6,TNF-α were detected by RT-PCR and ELISA. After 4 weeks of fat transplantation, there were differences in fat weight and volume between the control group and the experimental group [weight:(0.13±0.02) gvs(0.19±0.02) g,P<0.01], volume:(0.15±0.03) mlvs(0.19±0.01) ml,P<0.05]. HE staining results showed that the inflammatory infiltration area of P188 group was lower than that of normal saline group 4 weeks later. RT-PCR and ELISA results showed that 4 weeks after fat transplantation, P188 inhibited the mRNA levels of inflammatory cytokines IL-1α,1 L-1β,IL-6,TNF-α and the protein expressions of IL-1α,1 L-1β,IL-6,TNF-α. It's proved that P188 can inhibit inflammatory infiltration after autologous fat transplantation in rats.

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An imiquimod(IMQ)-induced psoriasis mouse model was established to evaluate the model and explore the effect of gender on IMQ-induced mouse psoriasis model establishment. Half of male and female BALB/c mice were randomly divided into normal control group,model group 1(male) and model group 2(female),and IMQ was applied to the back skin of mice to establish a mouse psoriasis model. The psoriasis area and severity index(PASI) was scored daily for mouse skin. The pathological changes of skin were observed by HE staining. The thymus index and spleen index were calculated. The expression of proliferating cell nuclear antigen(PCNA) was detected by immunohistochemistry and the expression of involucrin was detected by immunofluorescence. Compared with the control mice,IMQ-induced male and female mice showed scaly,erythema and thickening and continued to deteriorate,and the PASI score also showed the same results. HE staining found that compared with control mice,the epidermis of IMQ-induced male and female mice were thickened(P<0.001),telangiectasia and inflammatory cell infiltration were observed in the superficial epidermis. Parakeratosis was observed in the stratum corneum and the granular layer became thinner or even disappeared. The acanthosis was thick,and epidermis extended downward. The spleen index of IMQ-induced mice increased(P<0.001),and there was no significant difference in spleen index between male and female mice. The expression of keratinocytes in IMQ-induced male and female mice increased(P<0.05) and there was abnormal differentiation. There was no significant difference in the expression of PCNA and involucrin between male and female mice. The IMQ-induced mouse psoriasis model has many similarities with human psoriasis,and gender has little effect on the establishment of IMQ-induced mouse psoriasis model.

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In order to look for the plasma differential proteins(DEPs) and the possible plasma protein markers, we analyzed the proteomics in plasma of patients with primary chronic angle closure glaucoma. The plasma was collected, and iTRAQ and LC-MS/MS were performed to find the different proteins(DEPs). The DEPs were analyzed by GO and KEGG to find out the main pathways and key proteins. 143 statistically significant proteins were screened by mass spectrometry(P<0.05), of which 49 were up-regulated and 94 were down-regulated. The DEPs were analyzed from three parts such as the molecular function, biological process and cellular component by GO analysis. The main pathways are metabolic pathway and complement coagulation cascades,the key proteins are divided into the following three kinds according to their different functions:(1) regulating the glaucoma optic ganglion cells: HGFA;(2) oxidative stress related: glutathione peroxidase 3,SOD,Peroxiredoxin-2;(3) immune response related: C4,C4 BP. These differential proteins provide new plasma markers for clinical diagnosis. It helps us to understand the pathological mechanism of primary angle closure glaucoma and provides a new idea for clinical diagnosis.

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To explore the correlation between the serum level of 25 hydroxy vitamin D3[25-(OH)D3] with ovarian responsibility and the pregnancy outcomes of frozen-thawed embryo transfer(FET). 157 patients with tubal infertile factors who received detection of vitamin D before in vitro fertilization and embryos transfer(IVF) were enrolled this study, and assigned into two groups according to the serum level of 25-(OH)D3:25-(OH)VD3<20 μg/L group and 25-(OH)D3≥20 μg/L group. The outcomes of ovarian stimulation and FET were compared between two groups. No statistically significant difference was found in patients' retrieved oocytes, MⅡ, fertilization rate, total dosage duration of gonadotropin(Gn), the total day of Gn injection, high-quality embryonic rate and E2in HCG day(P>0.05). Whereas, clinical pregnancy rate in the 25-(OH)D3sufficiency group(64.3%)was significantly higher than that in 25-(OH)D3deficiency group(45.5%). Although serum 25-(OH)D3has less effect on ovarian responsibility, serum 25-(OH)D3deficiency may have adverse effect on the pregnancy rate of FET.