Aim To investigate the effect of p38 MAPK AS-ODNs on the expression of GLT-1 during the induction of brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP).Methods Eighty healthy male Wistar rats with permanent occlusion of the bilateral vertebral arteries were randomly divided into 6 groups: ①Sham group (n=10);②CIP group (n=10);③ischemic insult (Ⅱ) group (n=10);④CIP+Ⅱ group (n=10);⑤p38 MAPK AS-ODNs+CIP+Ⅱ group (n=30);⑥p38 MAPK S-ODNs+CIP+Ⅱ group (n=10).Group ⑤ was divided into 5 nmol, 10 nmol and 15 nmol subgroups according to the dose of p38 MAPK AS-ODNs (n=10).The dose of p38 MAPK S-ODNs was 15 nmol.All the rats were sacrificed 6 h and 2 d after the sham operation or the last time of global brain ischemia reperfusion.Western blot and immunohistochemistry analysis were used for detecting the expression of p-p38 MAPK and GLT-1 protein.Results CIP moderately up-regulated the expression of p-p38 MAPK and significantly up-regulated the expression of GLT-1 protein, inhibited the excessively up-regulation of p-p38 MAPK and the down-regulation of GLT-1 induced by ischemic insult.p38 MAPK AS-ODNs significantly inhibited the up-regulation of p-p38 MAPK and GLT-1 protein in a dose-dependent manner during the induction of brain ischemic tolerance by CIP.Conclusion p38 MAPK AS-ODNs inhibit the up-regulation of GLT-1 during the induction of brain ischemic tolerance induced by CIP.

Objective To observe the mode of RET proto-oncogene mutation in a pedigree with multiple endocrine neoplasia type 2A (MEN2A).Methods Six members from a MEN2A family,including the proband,were enrolled.Genomic DNAs of these members were extracted from peripheral blood lymphocytes for polymerase chain reaction(PCR),PCR products of 21 exons of the RET proto-oncogene were purified and a direct gene sequence analysis was performed.DNA sequencing was performed on the related exon of the other family members after verifying the mutation site.Results The female proband sufferd from pheochromocytoma and medullary thyroid carcinoma since the age of 45,two missense mutations of TGC(Cys) to TCC(Ser) at codon 634 and CTG(Leu) to TTT(Phe) at codon 633 in exon 11 of the RET proto-oncogene were detected in the proband,while the other members remain unchanged.Conclusions Analysis of the RET proto-oncogene identifies a united mutation of TGC (Cys) to TCC (Ser) at codon 634 and CTG(Leu) to TTT(Phe) at codon 633 in the proband.The former is a proven mutation related to MEN2A,while the latter has never been reported before.

OBJECTIVE:To establish a method for the simultaneous determination of paeoniflorin and paeonol in Dirda pill, and provide reference for improving its quality control standard. METHODS:HPLC was performed on the column of ZORBAX C18 with mobile phase of acetonitrile-0.1% phosphoric acid (gradient elutio) at a flow rate of 0.8 ml/min,detection wavelength was 230 nm(paeoniflorin),274 nm(paeonol),columne temperature was 35 ℃,injection volume was 20 μl. RESULTS:The linear range was 0.514 8-1.544 5μg for paeoniflorin(r=0.999 2)and 0.643 2-1.960 4μg for paeonol(r=0.999 8);the limits of quantifi-cation were 0.135 6,0.126 4 μg,limits of detection were 0.067 8,0.063 2 μg;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 95.78%-97.27%(RSD=0.62%,n=6) and 97.38%-98.38%(RSD=0.42%,n=6). CONCLUSIONS:The method is simple with good reprosucibility and high sensibility,and can be used for the simultaneous deter-mination of paeoniflorin and paeonol in Dirda pill.

Gallbladder carcinoma is the most common malignancy in biliary tract.Early dissemination,rapid and silent progression and delayed diagnose could portend a dismal prognosis.The 5-year survival rate is less than 10％.Clarifying the etiological feature can improve the early diagnosis.Comprehending surgical indications of benign lesions can provide the methods for rational prophylactic resections.Standardized treatment of accident gallbladder carcinoma could improve the patients' prognosis.Efficient serologic markers could be used for early diagnosis and screening.

Patients with liver cirrhosis have different clinical manifestations and prognoses, and it is necessary to accurately predict the clinical endpoints of liver cirrhosis. Liver pathology can directly display the change in liver structure and thus plays an important role in predicting clinical endpoints. This article summarizes the application of histological staging systems and parameters in predicting clinical endpoints and describes the significance of histological features after etiological treatment in predicting clinical prognosis.

Objective To investigate the mechanisms of hypoxia inducible factor-2 alpha (HIF-2a) regulating human umbilical vein endothelial cells (HUVECs) under hypoxic conditions.Methods The experimental study was adopted.(1) HUVECs in logarithmic growth phase were taken:HUVECs without any disposals as control group,HUVECs with shRNA transfection control as shRNA control group,HUVECs with HIF-2α shRNA transfection as HIF-2α shRNA group and HUVECs with HIF-2α shRNA transfection then added rhAng-2 as HIF-2α ± rh-Ang-2 group.(2) Western blot testing:the expressions of Ang-2 and HIF-2α proteins in HUVECs were cultured under hypoxia conditions at 0,2,4,8,12,16,20 hours,and the levels of which were detected in the control group,shRNA control group and HIF-2α shRNA group.(3) Enzyme-linked immunosorbent assay(ELISA):the level of Ang-2 protein in supernatant of HUVECs was detected in the control group,shRNA control group and HIF-2α shRNA group.(4)The amounts of endothelial cell tubes in HUVECs among the 4 groups were detected by tube formation experimental testing.(5) Transwell method was performed to detect the amounts of cells migration in HUVECs and hepatoma cells SMMC-7721 migration intervened by supernatant of HUVECs among the 4 groups.Measurement data with normal distribution were presented as x ± s,repeated measurement data were analyzed by the repeated measures ANOVA,comparison among groups and pairwise comparison were conducted respectively by the one-way ANOVA and Dunnett's test.Results (1) Western blot test:the expression levels of Ang-2 and HIF-2α proteins in HUVECs under hypoxia conditions at 0,2,4,8,12,16,20 hours were 0.110 ±0.011,0.120 ±0.020,0.210 ±0.070,0.410 ±0.100,0.520 ± 0.090,0.790±0.130 1.010 ±0.220 and 0.180 ±0.090,0.410 ±0.070,0.470 ±0.110,0.470 ±0.070,0.580 ± 0.120,0.690 ± 0.140,0.920 ± 0.130,respectively,and which were increased after culturing under hypoxia conditions and had an ascending tendency as the hypoxia time extended,with statistically significant differences (F =403.550,3 265.587,P ＜ 0.05).The expression levels of Ang-2 and HIF-2α proteins in the control group,shRNA control group and HIF-2α shRNA group were 1.030 ±0.180,1.070 ±0.120,0.210 ± 0.070,and 0.940 ± 0.110,0.930 ± 0.190,0.170 ± 0.021,respectively,showing statistically significant differences (F =290.242,26.688,P ＜ 0.05).(2) The results of ELISA:the expression levels of Ang-2 in the control group,shRNA control group and HIF-2α shRNA group were (433.2 ±9.7)ng/L,(438.3 ± 2.6)ng/L,(114.6 ± 4.2) ng/L,with a statistically significant difference (F =2 642.180,P ＜ 0.05).(3) The results of tube formation experiments:the number of endothelial cell tubes in the control group,shRNA control group,HIF-2α shRNA group and HIF-2α ± rh-Ang-2 group were 48.3 ± 2.5,47.4 ± 3.1,19.7 ± 1.5 and 38.3 ± 2.1,respectively,with a statistically significant difference (F =148.196,P ＜ 0.05).(4) The results of Transwell method:① the number of HUVECs migration in the control group,shRNA control group,HIF-2α shRNA group and HIF-2α + rh-Ang-2 group were 140.3-± 3.5,142.7 ± 2.1,42.7 ± 3.1 and 78.1 ± 4.2,respectively,showing a statistically significant differences (F =212.205,P ＜ 0.05).②The results of Transwell method:the number of SMMC-7721 cells migration after intervening using four different supernatant in the control group,shRNA control group,HIF-2α shRNA group and HIF-2α ± rh-Ang-2 group were 106.7 ± 5.5,102.7 ± 6.6,63.0 ± 3.3 and 96.7 ± 2.1,respectively,showing a statistically significant difference (F =55.122,P ＜ 0.05).Conclusion HIF-2a could not only affect HUVECs formation but also promote SMMC-7721 cells migration via regulating Ang-2 expression.

Objective To summarize and analyze the dynamic change of HBsAg levels in patients with chronic Hepatitis B (CHB) after receiving nucleos(t)ide analogues (NAs) as antiviral treatment.Methods Patients who were performed quantitative Hepatitis B surface antigen(qHBsAg) from July 30, 2012 to December 30,2016 in Peking Union Medical College Hospital were retrospectively enrolled.qHBsAg, HBV DNA, HBeAg were collected and analyzed at baseline and at 192-week follow-up every 24 weeks.qHBsAg and HBeAg were assessed with chemiluminesent microparticle immuno assay(CMIA).HBV DNA was assessed with PCR and COBAS Amplicor.Results 60 patients were included.Patients in HBeAg-positive group had higher HBV DNA than that in HBeAg-negative group (P<0.05)at baseline and the two groups both were under detection limit after 48 weeks.BaselineqHBsAg in HBeAg positive-group and negative-group were (3.43±0.73) log10 IU/mL, (3.08±0.47) log10 IU/mL respectively.qHBsAg in HBeAg-positive group was higher than that in HBeAg negative-group on all follow-ups(P<0.05) except 48weeks.However on 168 weeks and 192 weeks, difference between the two groups was statistically significant(P<0.05).In HBeAg-positive group,quantitative HBeAg dropped significantly during antiviral treatment.Conclusions HBV replication can be suppressed in the process of long-term NAs treatment in CHB patients.However qHBsAg decline is not so obvious, which indicates that HBsAg cleavence is difficult,and long-term NAs therapy is still necessary.

Objective To investigate the effects of miR?23a?3p on proliferation, migration and apoptosis on human acute myeloid leukemia ( AML) cells by targeting SMC1A. Methods Microarray analysis was used to screen differentially expressed microRNAs and mRNAs in human AML cells. Real?time fluorescence quantitative PCR (RT?qRCR) was used to detect the expressions of miR?23a?3p and SMCA in human AML cell line U937. TargetScan database was used to analyze the correlation between miR?23a?3p and SMC1A. Double luciferase reporter gene was used to detect the interaction between miR?23a?3p and SMC1A. The effect of miR?23a?3p expression on the proliferation of U937 cells was detected by clonal assay. The migration, apoptosis, cell cycle and caspase?3 activity of U937 cells regulated by miR?23a?3p were detected by cell scratch assay and flow cytometry, respectively. Western blot was used to detect the expressions of Bax and Bcl?2 in U937 cells. Results Compared with human normal monocyte SC group (1.00), the expression of miR?23a?3p in U937 cells was up?regulated (2.56±0.78) ( P＜0.01), while the expression of SMC1A was down?regulated (0.48±0.56, P＜0.01). miR?23a?3p specifically bond to SMC1A 3′UTR and regulated the expression activity of SMC1A. Overexpression of miR?23a?3p promoted the proliferation and migration of U937 cells and inhibited the apoptosis of U937 cells, while up?regulation of SMC1A inhibited the proliferation and migration of U937 cells and promoted the apoptosis of U937 cells. The percentages of G0／G1 phase, G2／M phase and S phase cells in the negative control group were ( 37.48 ± 0.21)%, (16.78±0.18)% and (45.74±0.15)% respectively, and those in the miR?23a?3p mimics group were (19.96±0.11)%, (41.69±0.24)% and (38.24±0.34)%, respectively. The difference was statistically significant (all P＜0.05). The proportions of G0／G1 phase, G2／M phase and S phase cells in the group of miR?23a?3p mimics+pcDNA3.1?SMC1A were (36.88± 0.21)%, ( 30.44± 0.33)% and ( 32.88± 0.16)%, respectively, without significant difference when compared with those of the miR?23a?3p mimics group ( P＞0.05). The relative expression levels of Bax and Bcl?2 protein in the negative control group were 0.55±0.45 and 0.31±0.54, respectively. Overexpression of miR?23a?3p inhibited the expression of Bax protein in U937 cells (0.23± 0.13, P＜0.001), promoted the expression of Bcl?2 protein ( 0.50 ± 0.23, P＜0.01), while SMC1A increased the expression of Bax protein in U937 cells (0.40± 0.11, P＜0.01), and inhibited the expression of Bcl?2 protein (0.37± 0.15). In the negative control group, caspase?3 activity was (25.82± 0.89)%.Overexpression of miR?23a?3p inhibited caspase?3 activity in U937 cells (3.64±0.56)%, P＜0.01, while up?regulation of SMC1A promoted caspase?3 activity in U937 cells ( 15.29 ± 0.85)%, P＜0.01. Conclusion miR?23a?3p can inhibit the proliferation and migration and promote apoptosis of human AML cells by targeting SMC1A.

Objective To investigate the effects of miR?23a?3p on proliferation, migration and apoptosis on human acute myeloid leukemia ( AML) cells by targeting SMC1A. Methods Microarray analysis was used to screen differentially expressed microRNAs and mRNAs in human AML cells. Real?time fluorescence quantitative PCR (RT?qRCR) was used to detect the expressions of miR?23a?3p and SMCA in human AML cell line U937. TargetScan database was used to analyze the correlation between miR?23a?3p and SMC1A. Double luciferase reporter gene was used to detect the interaction between miR?23a?3p and SMC1A. The effect of miR?23a?3p expression on the proliferation of U937 cells was detected by clonal assay. The migration, apoptosis, cell cycle and caspase?3 activity of U937 cells regulated by miR?23a?3p were detected by cell scratch assay and flow cytometry, respectively. Western blot was used to detect the expressions of Bax and Bcl?2 in U937 cells. Results Compared with human normal monocyte SC group (1.00), the expression of miR?23a?3p in U937 cells was up?regulated (2.56±0.78) ( P＜0.01), while the expression of SMC1A was down?regulated (0.48±0.56, P＜0.01). miR?23a?3p specifically bond to SMC1A 3′UTR and regulated the expression activity of SMC1A. Overexpression of miR?23a?3p promoted the proliferation and migration of U937 cells and inhibited the apoptosis of U937 cells, while up?regulation of SMC1A inhibited the proliferation and migration of U937 cells and promoted the apoptosis of U937 cells. The percentages of G0／G1 phase, G2／M phase and S phase cells in the negative control group were ( 37.48 ± 0.21)%, (16.78±0.18)% and (45.74±0.15)% respectively, and those in the miR?23a?3p mimics group were (19.96±0.11)%, (41.69±0.24)% and (38.24±0.34)%, respectively. The difference was statistically significant (all P＜0.05). The proportions of G0／G1 phase, G2／M phase and S phase cells in the group of miR?23a?3p mimics+pcDNA3.1?SMC1A were (36.88± 0.21)%, ( 30.44± 0.33)% and ( 32.88± 0.16)%, respectively, without significant difference when compared with those of the miR?23a?3p mimics group ( P＞0.05). The relative expression levels of Bax and Bcl?2 protein in the negative control group were 0.55±0.45 and 0.31±0.54, respectively. Overexpression of miR?23a?3p inhibited the expression of Bax protein in U937 cells (0.23± 0.13, P＜0.001), promoted the expression of Bcl?2 protein ( 0.50 ± 0.23, P＜0.01), while SMC1A increased the expression of Bax protein in U937 cells (0.40± 0.11, P＜0.01), and inhibited the expression of Bcl?2 protein (0.37± 0.15). In the negative control group, caspase?3 activity was (25.82± 0.89)%.Overexpression of miR?23a?3p inhibited caspase?3 activity in U937 cells (3.64±0.56)%, P＜0.01, while up?regulation of SMC1A promoted caspase?3 activity in U937 cells ( 15.29 ± 0.85)%, P＜0.01. Conclusion miR?23a?3p can inhibit the proliferation and migration and promote apoptosis of human AML cells by targeting SMC1A.

Liver biopsy has been regarded as the gold standard for the assessment of liver fibrosis regression. In 2017, Liver Research Center, Beijing Friendship Hospital, proposed a new classification called PIR classification for the evaluation of liver fibrosis regression in patients with chronic hepatitis B after antiviral therapy, which was also called “Beijing classification”. This classification is new breakthrough based on conventional staging and grading systems, quantitative assessment methods for liver fibrosis, and the concept of liver biopsy. This article discusses the prospects and shortcomings of PIR classification.