Objective:To investigate the effect of microRNA-146a (miR-146a) on apoptosis of human retinal microvascular endothelial cells (HRMECs) induced by high glucose and its possible molecular mechanism.Methods:HRMECs were cultured in vitro with 5.5 mmol/L D-glucose in the normal control group and 25 mmol/L D-glucose in the high glucose group for 48 hours, respectively.Normally cultured HRMECs were transfected by miR-146a mimics in the high glucose+ miR-146a mimics group or corresponding mimics control in the high glucose+ mimics control group by lipofection and cultured with 25 mmol/L D-glucose for 48 hours, respectively.Real-time fluorescent quantitative polymerase chain reaction (PCR) was performed to detect the expression level of miR-146a.MTT assay and flow cytometry were used to detect the activity and apoptosis of HRMECs, and Western blot was employed to detect the expression levels of apoptosis-associated protein B-cell lymphoma factor-2 (Bcl-2), Bcl-2 related X protein (Bax) and nuclear factor-κB (NF-κB) signaling-related proteins NF-κB p65 and p-NF-κB p65. Results:The relative expression levels of miR-146a were 1.00±0.10, 0.22±0.02, 0.21±0.02 and 0.88±0.09, and the cell viability was (100.00±10.06)%, (68.41±6.67)%, (67.91±6.74)% and (90.46±8.97)%, and the apoptosis rates were (3.11±1.02)%, (27.28±3.56)%, (27.44±4.03)% and (7.29±2.11)% in the normal control group, high glucose group, high glucose+ mimics control group and high glucose+ miR-146a mimics group, respectively.The relative expression levels of miR-146a and the cell viability were significantly lower, and the cell apoptosis rate was significantly higher in the high glucose group than those in the normal control group, with statistical significant differences (all at P<0.05). The relative expression levels of miR-146a and the cell viability were significantly higher, and the cell apoptosis rate was significantly lower in the high glucose+ miR-146a mimics group than those in the high glucose group and the high glucose+ mimics control group, and the differences were statistically significant (all at P<0.05). The relative expression levels of Bax and p-NF-κB p65 protein were significantly higher, the relative expression level of Bcl-2 protein was significantly lower in the high glucose group than those in the normal control group, showing statistically significant differences (all at P<0.05). The relative expression levels of Bax and p-NF-κB p65 protein were significantly lower, and the relative expression level of Bcl-2 protein was significantly higher in the high glucose+ miR-146a mimics group than those in the high glucose group and the high glucose+ mimics control group, and the differences were statistically significant (all at P>0.05). There was no significant difference in the relative expression of NF-κB p65 protein among the groups ( F=0.106, P=0.955). Conclusions:Overexpression of miR-146a may inhibit the apoptosis of HRMECs induced by high glucose, and its mechanism may be related to the inhibition of NF-κB signaling pathway activation.

Objective To investigate the feasibility of VTQ value of spleen in noninvasive assessment of staging of liver fibrosis in patients with hepatitis B.Methods VTQ values of spleen were measured in 56 patients with hepatitis B by using ARFI technique and liver biopsy was performed.The patients were then grouped according to the stages of liver fibrosis stage.The results were analyzed by the intergroup comparison,Pearson Correlation analysis,and receiver operating characteristic curve (ROC curve).Results There was positive association of the VTQ value with the stages of liver fibrosis,whose correlation coefficient was 0.83 (P ＜ 0.05);There was no significant difference between the spleen VTQ value and the liver fibrosis S1 and SO (P ＞ 0.05),whereas there were significant differences among other groups (P ＜ 0.05).The area under the ROC curve was 0.95,the cut-off value was 2.79m/s,and the specificity and sensitivity were 81.8％ and 91.7％ respectively.Conclusions The VTQ value measured by ARFI technique has a better value in noninvasive diagnosis of stages of hepatitis B liver fibrosis.

In order to search for new antiplatelet agents with higher potency, a series of tetramethylpyrazine ( TMP) /chalcone hybrids ( 2-26) were synthesized and evaluated based on the principle of bioisostere and hybrid-ization. They exerted inhibitory activity against adenosine diphosphate ( ADP )-induced and arachidonic acid ( AA)-induced platelet aggregation to varied extent. Among them, compound 8 was the most potent with IC50 of 0. 14 mmol/L on ADP-induced platelet aggregation ( 9. 1 folds of TMP and 10. 5 folds of chalcone ) and 0. 09 mmol/L on AA-induced platelet aggregation ( 8. 8 folds of TMP and 10. 0 folds of chalcone) , which was superior to clinically used anti-platelet drug aspirin ( ASP, IC50 =0. 15 mmol/L) .

Objective To provide guide for prevention and cure of peritonitis in peritoneal dialysis(PD) by comparing the causative organisms and clinical outcome of PD related peritonitis in younger and elderly patients in our center.Methods All patients who developed PD related peritonitis between January 2006 and December 2013 in Wuhan NO.1 hospital were included.According to their age,episodes were divided into younger patients group (＜ 65 years) and elderly patients group (≥65 years).The microbiology and clinical outcome of PD related peritonitis were compared,and the related risk factors of the treatment failure were analyzed.Results Three hundred and sixty-six episodes of peritonitis occurred in 258 patients during the study period.The overall rate of peritonitis was 1 episode in 76.8 patient-months.Elderly patients had higher incidence of peritonitis (1 episode every 56.4 months vs 1 episode every 88.7 months,P=0.001),higher incidence of fungus infection (9.6％ vs 3.9％,P=0.026) and higher mortality (46.2％ vs 14.0％,P=0.001) than that in younger patients.Cox regression analysis showed that longer duration of PD treatment and fungal peritonitis were both risk factors of the treatment failure.Conclusion Elderly patients had higher incidence of peritonitis,higher incidence of fungus infection and higher PD-related mortality than younger patients.

Aim: To explore the in situ intestinal absorption in rats of ZLR-8, an insoluble NO-donor drug, and to compare the intestinal absorption enhancement by spray-dried emulsion. Methods: Intestine of rats was cannulat-ed for in situ perfusion. UV and HPLC methods were used to monitor phenolsulfonphthalein and ZLR-8, respec-tively. The effects on ZLR-8 absorption of the intestinal segments, the concentration of ZLR-8 and the pH of the circulating perfusate were studied. The absorption of ZLR-8 suspension was compared to that of the spray-dried emulsion. Results: 1-h in situ intestinal perfusion of the spray-dried emulsion allowed the estimation of the absor-tion percentage to be (23. 54 ± 1. 40) %, (15. 95 ± 0. 09) %, (12. 30 ± 0. 74) %, (3. 98 ± 0. 12) %, respec-tively; the absorption rate constants in duodenum, colon, jejunum and ileum to be (0.248 6 ±0.046 0) h~(-1), (0. 143 7 ±0. 036 0) h~(-1), (0. 069 2 ±0. 001 3) h~(-1), (0. 020 8 ±0. 000 4) h~(-1), respectively. Significant differ-ences in absorption characteristics were found among intestinal segments. In the range of 3. 4-9. 4, pH of the per-fuate had significant influence on the absorption of ZLR-8, and better absorption appeared at pH of 5. 4 to 7. 4. It was found that the absorption rate constant was unaffected by ZLR-8 concentration. However, the absorption amount was proportional to ZLR-8 concentration. Compared to the ZLR-8 suspension, the in situ intestinal absorption of ZLR-8 in rats given the spray-dried emulsion increased significantly. Conclusion: It was only found that ZLR-8 administered in suspension has minor absorption in rat duodenum while no apparent absorption occurred in other segemnts. ZLR-8 in spray-dried emulsion was fairly absorbed in the rat intestinal segments. Passive diffusion was invloved in the absorption of ZLR-8. Spray-dried emulsion significantly enhanced the intestinal absorption of ZLR-8 in rats.

Background and purpose:Drug resistance is a major cause of failure in lung cancer chemotherapy. This study aimed to investigate the effect of YAP on doxorubicin resistance in lung cancer and its underlying mechanism. Methods:Doxorubicin resistant lung cancer cell clones were established from parental sensitive cancer PC9 cell line via in vitroinduction, and the expression of YAP was analyzed. YAP was down-regulated via shRNA to different levels. MTS assay was employed to determine cell proliferation and drug sensitivity. Flow cytometry was used to determine cell cycle distribution, apoptosis and uptake of Rh-123. Western blot and quantitative real-time polymerase chain reaction (QRT-PCR) assay were used to determine the expression of ABCB1, ABCC1, p53, Runx2, ITGB2 and ErbB4. The phosphory-lation of serine/threonine kinase (AKT) was determined as well.Results:Doxorubicin resistant PC9/Adr cell clone was obtained with over-expressed YAP. Expression of YAP in PC9/Adr cells was down-regulated to different levels via shR-NA. After YAP silencing, cell proliferation was reduced, while sensitivity to doxorubicin was increased. The cell cycle was significantly halted by G0/G1 phase. Doxorubicin induced-apoptotic rate and cellular uptake of Rh-123 were increased,with positive correlation to YAP silencing level. Western blot and QRT-PCR results showed that after YAP silencing, ABCB1, ABCC1, Runx2, ITGB2, and ErbB4 proteins were down-regulated, while the expression of p53 was up-regulated. Phosphorylation of AKT was inhibited as well.Conclusion:Over-expression of YAP is involved in doxorubicin resistance in PC9/Adr cell line. Silencing of YAP could restore doxorubicin sensitivity. The mechanism involves regulation of drug resistance-related genes and promotion of apoptosis.

This study was performed to prepare immobilized β-glucosidase and snailase, then optimize and compare the process conditions for conversion of icariin. Immobilized β-glucosidase and snailase were prepared using crosslink-embedding method. The best conditions of the preparation process were optimized by single factor analysis and the properties of immobilized β-glucosidase and snailase were investigated. The reaction conditions including temperature, pH, substrate ratio, substrate concentration, reaction time and reusing times of the conversion of icariin using immobilized β-glucosidase or snailase were optimized. Immobilized β-glucosidase and snailase exhibited better heat stabilities and could remain about 60% activity after storage at 4 degrees C for 4 weeks. The optimized conditions for the conversion of icariin were as follows, the temperature of 50 degrees C, pH of 5.0, enzyme and substrate ratio of 1 : 1, substrate concentration of 0.1 mg x mL(-1), reaction time of 6 h for β-glucosidase and 2 h for snailase, respectively. In 5 experiments, the average conversion ratio of immobilized β-glucosidase and snailase was 70.76% and 74.97%. The results suggest an effect of promoted stabilities, prolonged lifetimes in both β-glucosidase and snailase after immobilization. The immobilized β-glucosidase and snailase exhibited a higher conversion rate and reusability compared to the free β-glucosidase and snailase. Moreover, the conversion rate of immobilized snailase was higher than that of immobilized β-glucosidase. The process of icariin conversion using immobilized β-glucosidase and snailase was moderate and feasible, which suggests that immobilized enzymes may hold a promise for industrial usage.

Objective To analyze the relationship between biochemical level and severity levels and clinical,duration of disease in patients with Parkinson Disease (PD).Methods 69 patients with PD and 69 healthy persons of similar sex and age were selected in the research.Serum uric acid and lipids levels were examined and compared.Results The serum uric acid,triglycerides,total cholesterol and low-density lipoprotein cholesterol (LDL-C) were (322.48 ± 66.18) μmol/L,(1.22 ± 0.86) mmol/L,(4.70 ± 0.92) mmol/L and (3.00 ± 0.85) mmol/L in control group,and (384.23 ± 88.28) μmol/L、(1.64 ± 0.94) mmol/L、(5.37 ± 1.31) mmol/L、(3.53 ± 1.03) mmol/L in control group.The differences are significant (t =-4.68,P =0.000;t =-2.74,P =0.007;t =-2.74,P =0.007;t =-3.49,P =0.001;t =-3.27,P =0.001).Serum UA concentration and high-density lipoprotein cholesterol (HDL-C),LDL-C were lower in patients with Parkinson's disease in duration of disease more than 3 years than those in duration of disease less than 3 years (t =3.373,P =0.001;t =2.440,P =0.017).The serum UA levels of any stages of PD patients were lower than the control group (P ＜ 0.05) according to Hoehn-Yahr staging.All lipid levels in early and middle stage PD disease patients were lower than those in control group (P ＜ 0.05).Serum UA,total cholesterol and HDL-C in female PD patients were (305.69 ± 54.25) μmol/L,(4.99 ± 0.95) mmol/L,(1.25 ± 0.27) mmol/L,and (339.76 ± 73.40) μmol/L,(4.41 ± 0.81) mmol/L,(1.06 ± 0.19) mmol/L in male patients.The difference is significant (t =2.198,P =0.031;t =-2.721,P =0.008;t =-3.266,P =0.002).Multivariate logistic regression models assessed lower uric acid concentrations is the risk of PD (OR =1.01,95％ CI 1.004 ～ 1.015,P =0.001).Conclusion Biochemical level changed differently in Parkinson disease and uric acid reduction could be a risk factor for PD.

Objective:To evaluate the heart hemodynamics in fetuses with premature ductus arteriosus constriction or closure using fetal heart quantification (FHQ).Methods:The clinical data of 50 singleton fetuses with ductus arteriosus constriction ( n=35) or ductus arteriosus closure ( n=15) who underwent echocardiography in Department of Ultrasound, Beijing Anzhen Hospital were retrospectively analyzed, from May 2013 to January 2020. Fifty healthy singleton fetuses were randomly selected as the control group. The ductus arteriosus diameter (DA), pulsatility index (PI), diameter of the left atrium(LA) and right atrium(RA), diameter of the left ventricle (LV) and right ventricle (RV), tricuspid regurgitation/right atrium area ratio (TR/RA Ratio), pressure gradient of tricuspid regurgitation (PG of TR), and heart/chest ratio were measured using conventional fetal echocardiography; the correlations among the parameters were analyzed. Speckle-tracking analysis was used to analysis and compute the LV and RV global spherical index (GSI), fractional area change (FAC) and global strain (GS), the LV ejection fraction(EF) and stroke volume (SV). These variables and their correlations were compared and analyzed. Results:Compared with the control group, the GS and FAC of the LV and RV in the ductus arteriosus constriction or closure groups were lower ( P<0.05) while the LV-SV was higher ( P<0.05). The FAC, GS, and EF values of the LV were higher in the premature ductus arteriosus closure group than in the ductus arteriosus constriction group ( P<0.05), while the RV-FAC was lower ( P<0.05), the RV-GS and LV-SV showed no significant changes ( P>0.05). Correlation analyse showed that the PI was positively correlated with DA( r=0.364, P<0.05); the PG of TR was negatively correlated with DA( r=-0.414, P<0.05); the TR/RA Ratio was negatively linearly correlated with PI( r=-0.388, P<0.05), and positively correlated with RV/LV Ratio ( r=0.369, P<0.05); the other parameters were not significantly correlated with the DA or PI ( P>0.05). Conclusions:Fetal heart hemodynamics in the premature ductus arteriosus constriction or closure groups change significantly, FHQ can provide valuable information for the evaluation of the fetal heart with ductus arteriosus constriction or closure.

AIM: To study the effect of Jiunaoning Injection on apoptosis and the changes in cytosolic calcium induced by hypoxia/hypoglycemia in SH-SY5Y human neuroblastoma cells. METHODS: The apoptosis and the changes in cytosolic calcium were induced by hypoxia and hypoglycemia .Apoptosis was measured via flow cytometry(FCM), and cytosolic calcium was determined by Fluo-3 fluorometry. RESULTS: The hypoxia/hypoglycemia induced apoptosis and increased the cytosolic calcium in SH-SY5Y cells. Jiunaoning Injection inhibited the apoptosis of the SH-SY5Y cells and decreased the concentration of cytosolic free calcium. CONCLUSION: Hypoxia/hypoglycemia induced apoptosis and increase in cytosolic free calcium in SH-SY5Y cells, which were significantly inhibited by Jiunaoning Injection.