BACKGROUND: Caveolin-1, as the most important functional protein in caveolae, is involoved in a variety of cell biological processes, and is closely related to the occurrence and development of breast cancer. OBJECTIVE: To explore the effect of caveolin-1 on epidermal growth factor (EGF) in fibroblasts after co-cultured with breast cancer cells.METHODS: Caveolin-1 expression in fibroblast lines ESF-1 was interfered with siRNA, and the optimal effect was determined through QRT-PCR and western blot assay. (1) The optimal silencing model of ESF-1-Caveolin-1 SiRNA-N.2 was obtained, which was co-cultured with breast cancer cells BT474 as experimental group, single-cultured ESF-1 and ESF-1-Caveolin-1 SiRNA-N.2 as controls. The expression level of EGF in ESF-1 was detected by QRT-PCR at 24 and 48 hours of culture; the expression level of EGF in the culture medium was detected by ELISA at 48 and 72 hours of culture. (2) ESF-1-Caveolin-1 SiRNA-N.2 (experimental group) and ESF-1 (control group) were respectively co-cultured with BT474, and single-cultured BT474 as blank control group. The proliferation of BT474 was detected by cell counting-kit 8 assay after 24- and 48-hour culture.RESULTS AND CONCLUSION: The mRNA expression level of EGF in the experimental group was significantly higher than that in the other two groups (P < 0.05), and the EGF expression level in the ESF-1-Caveolin-1 SiRNA-N.2 group was significantly higher than that in the ESF-1 group (P < 0.05). The protein expression level of EGF was ranked as follows: experimental group > ESF-1-Caveolin-1 SiRNA-N.2 group > ESF-1 group (P < 0.05). The proliferation of BT474 cells was significantly increased after co-cultured with BT474 cells and ESF-1 siRNA Caveolin-1 cells, especially with BT474 cells (P < 0.05). Our findings suggest that Caveolin-1 siRNA can promote the expression of EGF in fibroblasts, especially co-cultured with breast cancer cells. Furthermore, caveolin-1 siRNA accelerates the proliferation of breast cancer cells after co-cultured with fibroblasts.

This study was aimed to establish the determination method of nineorganic solvent residues of macrop-orous resin. Headspace capillary gas chromatography (GC) was used in the content determination of hexane, benzene, methyl-benzene, o-xylene, m-xylene, p-xylene, styrene, diethyl-benzene and divinyl-benzene of macroporousresin residues in Ginkgo leaf tablets. The results showed that ninemacroporous resin residues in Ginkgo leaf tablets were completely separated. The calibration curves were in good linearity (r=0.9992-0.9998). The average recoveries were 80%-110%. It was concluded that themethod was accurate, quick, and sensitive, which can be used to determine the macroporous resin residues in Ginkgo leaf tablets.

Objective:To investigate the relationship between serum vascular endothelial growth factor (VEGF), peripheral blood microRNA-126 (miR-126) and the number and distribution of cerebral microbleeds (CMBs).Methods:Consecutive patients with non-acute ischemic cerebrovascular disease admitted to the Department of Neurology, the First Affiliated Hospital of Baotou Medical College from June 2019 to June 2020 were enrolled. The clinical data were collected, 3.0 T MRI examination was performed, and susceptibility-weighted imaging was used to detect CMBs. The serum VEGF concentration was detected by enzyme-linked immunosorbent assay, and miR-126 was detected by fluorescence quantitative polymerase chain reaction. Multivariate logistic regression analysis was used to determine the independent influencing factors of CMBs. Multiple linear regression analysis was used to determine the correlation between serum VEGF concentration, miR-126 in peripheral blood and the number of CBMs. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of serum VEGF concentration and relative expression of miR-126 in peripheral blood for CMBs. Results:A total of 193 patients with non-acute ischemic cerebrovascular disease were enrolled, including 110 patients (57.0%) in the non-CMBs group, 20 (10.4%) in the strictly lobar CMBs group and 63 patients (32.6%) in non-strictly lobar CMBs group. The comparison among the three groups showed that age might be a risk factor for strictly lobar CMBs, while higher VEGF, higher cystatin C level, lower relative expression of miR-126 in peripheral blood, hypertension and previous stroke or transient ischemic attack might be the risk factors for non-strictly lobar CMBs. Multivariate logistic regression analysis showed that higher serum VEGF concentration was an independent risk factor for non-strictly lobar CMBs (odds ratio 1.186, 95% confidence interval 1.035-1.358; P=0.014), while the higher relative expression of miR-126 was an independent protective factor for non-strictly lobar CMBs (odds ratio 0.154, 95% confidence interval 0-0.269; P=0.026). Multiple linear regression analysis showed that higher serum VEGF concentration ( r=0.848, P<0.001) and the lower relative expression of miR-126 ( r=-0.043, P=0.035) significantly increased the number of CMBs. ROC curve analysis showed that the area under the curve of serum VEGF for predicting non-strictly lobar CMBs was 0.803 (95% confidence interval 0.741-0.865), the optimal cut-off value was 120.55 ng/L, the sensitivity was 70.7%, and the specificity was 75.5%. Conclusions:In patients with non-acute ischemic cerebrovascular disease, there is a significant correlation between serum VEGF concentration and the relative expression of miR-126 in peripheral blood and the number and distribution of CMBs. Serum VEGF can be used as a biomarker for predicting the presence of non-strictly lobar CMBs.

Aim To study the mimicry peptide of JEV E protein by screening a phage 15-mer peptide library with anti-JEV E protein mAb 2H4. Methods After three rounds biopanning, the enriched positive phage clones were identified by ELISA. 10 positive phage clones were sequenced and compared homologously with JEV E protein. Results The short peptide displayed on screened positive phage could bind specifically to mAb 2H4, and the binding could be inhibited by natural JEV Ag. Amino acid sequences of the 10 positive phage clones were consensus, that is, RQDPQWPYANSTIAR. By homology analysis, two higher homologous sequences STXAR and WXXAXST were found in different regions of JEV E protein. The peptide displayed on positive phage could react specifically with the mouse antiserum against natural JEV Ag . Conclusion This peptide displayed on positive phage may mimic partial antigenicity of JEV E protein.

[ ABSTRACT] AIM:To study the effect of livin gene-modified bone marrow mesenchymal stem cells ( BM-MSCs) transplantation on the cardiac function following acute myocardial infarction in a rat model and the expression of livin , caspase-3, caspase-7 and caspase-9 in the livin gene-modified BM-MSCs.METHODS: The MSCs were obtained by the whole bone marrow culture method , and the apoptosis of the MSCs after infection with adenovirus vector carrying enhanced green fluorescent protein ( EGFP) gene and livin recombinant vector ( rAd-livin) were detected by flow cytometry .The ex-pression of livin, caspase-3, caspase-7 and caspase-9 was detected by Western blot .After permanent left anterior descend-ing artery occlusion , the rats were randomized to receive intramyocardial injection of DMEM without cells ( vehicle group ) , or containing MSCs ( MSCs group ) , MSCs ( EGFP ) ( rAd-control/MSCs group ) or MSCs ( livin ) ( rAd-livin/MSCs group).Left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), the maximum in-creased rate of left ventricular pressure ( -dp/dtmax ) and the maximum decline rate of left ventricular pressure ( +dp/dtmax ) were recorded for evaluating the cardiac functions .RESULTS: The apoptosis of rAd-livin/MSCs was significantly decreased as compared with MSCs and rAd-control/MSCs (P<0.05).Meanwhile, the expression of caspase-3, caspase-7 and caspase-9 was significantly downregulated as compared with the other 2 groups ( P<0.05 ) .The cardiac function in rAd-livin/MSCs group was significantly improved as compared with DMEM group , and those in the other 2 groups got the similar results, but the function in rAd-livin/MSCs group was better improved .Meanwhile, the number of surviving cells in rAd-livin/MSCs group was significantly improved as compared with the other 2 groups .CONCLUSION:The apoptosis of MSCs is decreased after rAd-livin transfection, and the expression of caspase-3, caspase-7 and caspase-9 is also significant-ly downregulated while the expression of livin is significantly upregulated .Transplantation of livin-modified BM-MSCs by lentiviral vector results in better prognosis for treating myocardial infarction by enhancing cell survival .

Since 2014, the United States and Europe has approved all oral, interferon free- regimens that combine with direct-acting antiviral agents. Hence, the sustained virological response rate of patients with chronic HCV genotype 1 infection has improved over 90%, and the treatment modalities has introduced a new era. These drugs, ombitasvir and dasabuvir, received customary authorization of Food and Drug Administration in 2015 and are the first combined direct-acting antiviral agents for treating HCV genotype 1 infection. It has superior application prospects in China because of its high-sustained virological response rate and safety profile. This article reviews the pharmacokinetics, drug interactions, efficacy and safety of this therapeutic regimen.

Objective To examine the expression of human ?-defensin 2 (hBD_ 2 ) recombinant adenovirus expression vector in rat dermal multipotent stem cells (dMSCs) and to observe the antiseptic activity of recombinant hBD_ 2 . Methods The expression of hBD_ 2 in dMSCs was examined by RT-PR, fluorescent immunochemistry and Western blotting, and the concentration of recombinant hBD_ 2 in supernate was measured by ELISA. The antiseptic activity of recombinant hBD_ 2 was assessed by K-B disc agar diffusion test. Results hBD_ 2 could be effectively expressed in dMSCs, and the concentration of recombinant hBD_ 2 in supernate was about 743.6 ng/ml . Recombinant hBD_ 2 in supernate showed antiseptic activity. Conclusion Recombinant adenovirus expression vector of hBD_ 2 could be effectively expressed in dMSCs, and the recombinant hBD_ 2 in supernate showed obvious antiseptic effects toward some standard bacteria lines.

Objective To observe the repair effect of human ?-defensin 2 (hBD 2)-modified rat dermal multipotent stem cells (dMSCs) transplantation on infected wound. Methods Thirty Wistar rats were excised a piece of whole-layer back skin, 3 mm in diameter, then infected the wound with 1?10 8/ml pseudomonas aeruginosa 1 ml, then the rats were injected on the wounded back respectively with dMSCs modified by hBD 2 (n=10), or pure dMSCs (n=10) or none as control (n=10). The repair effect was evaluated by observing the amount of bacteria under the scar, wound healing time and the percentage of remaining wound area. Results The amount of bacteria under the scar in rats that were transplanted with dMSCs modified by hBD 2 was less than that in rats transplanted with dMSCs or controls (P

Objective To analyze hepatitis C virus genotype(HCV GT)1b NS5A resistance-associated variants(RAV)and its related factors,and to provide references for direct-acting antivirals (DAA)agent selection and application.Methods From January 2017 to July 2017,53 hepatitis C patients were selected from the Department of Infectious Diseases of Henan Province People's Hospital. The mutations of L31M and Y93H in NS5A RAV were analyzed in 43 HCV GT1b patients,and their correlations with hepatitis C virus,liver function,platelet and liver fibrosis diagnostic model[APRI, gamma-glutamyl transpeptidase to platelet ratio(GPR),FIb-4]were analyzed.The quantitative data were compared by two independent samples t test,and the qualitative data were compared by chi square test. Results Fifty-three subjects were enrolled,including 43 GT1b(9 males and 34 females)and 10 GT2a(2 males and 8 females).No other genotype was detected.The incidence of NS5A RAV in 43 HCV GT1b patients was 13.9%(6/43),of which L31M and Y93H were 1/43(2.3%)and 5/43(11.6%)with no significant difference(χ2= 1.500,P= 0.219).There were no significant differences in HCV RNA, ALT,AST,albumin,platelets and age between patients with or without mutation(all P> 0.05). Conclusions The incidence of NS5A RAV in HCV GT1b patients is high,but not affected by virus, biochemical factors and liver fibrosis.The detection of NS5A RAV before HCV treatment is helpful for rational selection of DAA,which could reduce the drug resistance.

UNLABELLEDObjective To explore the effects of sperm DNA integrity rate, acrosome integrity rate and acrosome reaction rate on the outcomes of rescue intracytoplasmic sperm injection (ICSI).

METHODSThis retrospective analysis was conducted among 97 infertile couples receiving rescue ICSI due to failure of in vitro fertilization procedures in our Reproductive Medicine Center. Of these 97 women, 41 had clinical pregnancy and 56 did not, and the effects of sperm DNA integrity rate (estimated by DNA fragmentation index, DFI), acrosome integrity rate and acrosome reaction rate on rescue ICSI outcomes were analyzed.

RESULTSNo significant difference was found in paternal age, testosterone value, testicular volume, FSH, female patient' age or the number of eggs retrieved between the two groups (P>0.05), but the infertility years was significantly shorter in the pregnancy group than in the non-pregnancy group (P<0.05). The fertilization rate and cleavage rate were similar between the two groups (P>0.05), but the good embryo rate was significantly higher in the pregnancy group (P<0.05). The sperm DNA integrity or acrosome reaction rate did not differ significantly between the two groups (P>0.05), but the acrosome integrity rate was significantly higher in the pregnancy group (P<0.05). The sperm DNA integrity rate, acrosome integrity or acrosome reaction rate were not correlated with the fertilization rate, cleavage rate or good embryo rate (P>0.05). The pregnancy rate, twin and single fetus rates were 42.3%, 10.3% and 32.0% in this cohort after recue ICSI, respectively.

CONCLUSIONRescue ICSI is an effective treatment after failed in vitro fertilization procedure, and sperm acrosome integrity rate is associated with the outcome of rescue ICSI.

Acrosome ; pathology ; Acrosome Reaction ; DNA Fragmentation ; Female ; Fertilization ; Fertilization in Vitro ; Humans ; Infertility ; Male ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic