Aging ; psychology ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Wistar

Objective To establish human choriocarcinoma JeG-3 cell line resistant to floxuridine (FUDR)and describe the characteristics of this FUDR-resistant subline.The thymidylate synthase (TS) expression level in FUDR-resistant subline was also discussed.Methods The FUDR-resistant sub-line JeG-3/FUDRA was established by intermitted exposure to grads increased FUDR.Reversed microscope was used to observe the morphological changes in FUDR-resistant sub-line.Population doubling time was calculated and compared based on the growth curve of these two cell lines,cell cycles and chromosomal ploidy were assayed with flow cytometry methods.The chemo-luminescence assay was used to detect the hormone secretion by two kinds of cell lines.The resistant index (RI) was measured by cell counting kit-8 (CCK-8)assay.Quantitative RT-PCR was used to detect the mRNA expression level of TS and we also detected the TS mRNA expression level in different doses exposed subline.Results The RI of JeG-3/FUDRA was 31.62.Compared with the JeG-3 cell,the FUDR-resistant cell line had gross changes in morphological,cell growth,cell cycles and chromosomal numbers.The ability of human chorionic gonadotrop(hCG) and progesterone secretion was lower in JeG-3/FUDRA subline.The trend of TS mRNA expression was:while exposed to low concentration of FUDR,the TS mRNA expression level was downregulated,then followed the increasing dose of the drug,the expression level of TS mRNA ascended gradually.When the terminal concentration was reached,the expression level of TS mRNA in JeG-3/FUDRA subline was higher than that of JeG-3 cell line (P＜0.05).Conclusions We established the FUDR-resistant subline of JeG-3 successfully.The TS mRNA expression level is stage-related to the different concentration and different phase in FUDR exposure.Our data suggested that TS mRNA expression level may not be used as a biomarker to predict the chemosensitivity in FUDR-based chemotherapy.

It is important to pay more attention to students’ basic skills training and their comprehensive abilities cultivating in microbiology experiment teaching. Explorations and reforms in enforcing the students’ base and cultivating their abilities were carried out in order to improve teaching quality and train specialized talents with high quality.

Posterior capsule opacification (PCO) is the most common complication after cataract surgery.How to prevent and treat PCO is an urgent problem we need to solve at present.Non-coding RNA(ncRNA) is a kind of RNA, which can not encode proteins.Studies have shown that non-coding RNA is closely related to the occurrence and development of human diseases.This paper has collected the progress of research on different kinds of ncRNA in PCO and may raise new ideas and methods on the prevention and treatment of PCO.

Objective To investigate the analgesic effect of salmon calcitonin(sCT)and its effect on expression of calcitonin receptor(CT-R)in periaqueductal gray(PAG). Methods Rat models of neuropathic pain were prepared by chronic constriction injury(CCI). Thermal withdrawal latency(TWL)and mechanical nociceptive threshold(MNT)were measured using hot plate test and yon Frey monofilaments test. The distribution of CT-R in PAG was detected by immunohistochemical method. CT-R protein was quantitatively determined by western blotting. Fourty male SD rats were randomized into 5 groups: normal group, sham-CCI group, CCI group, CCI plussubcutaneous sCT group, and CCI plus microinjection of sCT into PAG group. Results TWL, MNT, andexpression of CT-R in PAG showed no difference between normal group and sham-CCI group(P＞0. 05). TWL and MNT in CCI group were significantly lower than those in normal group(P＜0.05), and expression of CT-R in CCI group was significantly higher than that in normal group(P＜0.05). TWL, MNT and expression of CT-R in CCI rats increased significantly after sCT therapy(P＜0. 05), and the effect was more marked in PAG injection group than subcutaneous injection group(P＜0.05). Conclusions sCT raises the pain threshold and increase the expression of CT-R in PAG of CCI rats, while PAG injection showed more marked effect than subcutaneous injection.

In this study, we explored the mechanism of Huganning tablet (HGNP) in the treatment of nonalcoholic fatty liver disease (NAFLD) based on network pharmacology and computer-aided drug design. Firstly, the potential ingredients and targets of HGNP were identified from TCMSP database, Swiss Target Prediction database, Chinese pharmacopoeia (2015) and literatures, and then the targets of HGNP intersected with NAFLD disease targets that obtained in GeneCards database to acquired potential targets. The bioconductor bioinformatics package of R software was used for gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. The network of “potential ingredient-key target-pathway” was formed in Cytoscape software to study the interactions between potential ingredients of HGNP, key targets, pathways and NAFLD. Based on the results of network pharmacology, the molecular docking analysis of the key targets and potential active ingredients in HGNP tablets with top degree in the network was conducted using Discovery Studio 2020 software, followed by molecular dynamics simulations, binding free energy calculation, drug-likeness properties analysis and ADMET (absorption, distribution, metabolism, excretion and toxicity) properties prediction. In vitro, HepG2 cells were used to establish steatosis model, and the effects of five key compounds on hepatocyte steatosis were analyzed by oil red O staining and triglyceride (TG) content determination. The results showed that 141 ingredients and 151 potential targets were obtained. A total of 2 526 items and 151 pathways were identified by GO and KEGG enrichment analysis. The molecular docking suggested that five components, isorhamnetin, salvianolic acid B, emodin, resveratrol and rhein, exhibited strong binding ability with key targets [retinoic acid receptor RXR-alpha (RXRA), tumor necrosis factor (TNF), glycogen synthase kinase-3 beta (GSK3B), serine/threonine-protein kinase 1 (AKT1)]. It was further verified that isorhamnetin and salvianolic acid B bind to key targets with good structural stability and binding affinity based on molecular dynamics simulations and binding free energy calculations. The drug-likeness properties, pharmacokinetic properties and toxicity of five key compounds were more comprehensively analyzed through drug-likeness properties analysis and ADMET properties prediction. In vitro, all five compounds, isorhamnetin, salvianolic acid B, emodin, resveratrol, and rhein, improved hepatocyte steatosis of HepG2 cells, confirming the reliability of the present study. In conclusion, based on network pharmacology, computer-aided drug design and in vitro validation, this study investigated the mechanism of HGNP for the treatment of NAFLD at multiple levels and provided a basis for its clinical application.

Objective To investigate the effect of antisense oligonucleofide(PCNA ASON)and vascuiar endothelial growth factor (VEGF) gene traneduction on restenosis after balloon angiopasty.Memods chinese rabbits were randomly divided into control group(I),PCNA ASON(II),VEGF only (Ⅲ),PCNA ASON+VEGF(Ⅳ)groups.Each group included 7 rabbits.Restenosis wasevaluated by pathology immunohistochemistry and Western-blotting for the expression of PCNA,and the depth and area oftunica media and tunicca intima were measured. Results All rabbits experienced restenosis on different severities,especially in control group.Lesions were less severe in PCNA ASON and VEGF groups than in controls.The proliferation of smooth muscle and intima significantly ameliorated in PCNA ASON and VEGF combination group that in PCNA ASON or VEGF groups(P<0.01).But the difference between the PCNA ASON and VEGF group was not significant(P>0.7). Conclusion PCNA ASON and VEGF gene transductionn are effective in preventing restenosis after balloon angioplasty caused vessel injury in rabbits.

Cell Membrane ; metabolism ; Gastrointestinal Stromal Tumors ; diagnosis ; metabolism ; Humans ; Immunohistochemistry ; Proto-Oncogene Proteins c-kit ; analysis

BACKGROUND:Uterine arterial embolization is a minimal y invasive technique and widely used for the treatment of several obstetrics and genecology diseases. However, reports about the influence of uterine arterial embolization on the pathophysiological microvessel density and angiogenesis changes in the endometrium have been very limited.
OBJECTIVE:To investigate the influence of uterine artery embolization on endometrial microvessel density and neovascularization.
METHODS:Sixty female guinea pigs were randomly divided into control group (n=15) and uterine artery embolization group (n=45). In the uterine artery embolization group, trisacryl gelatin microspheres (Embosphere) was used to perform uterine artery embolization and animals were further divided into three subgroups including E1, E2, and E3 (n=15 in each subgroup). Uterine specimens were col ected at 7-15 days, 16-30 days, and 31-45 days after uterine artery embolization respectively in the three subgroups.
RESULTS AND CONCLUSION:Microspheres were visible in the lumen of primary branch of uterine artery, subserosal arteriole and intramuscular arteriole. Statistical analysis demonstrated that CD34-positive microvessel density of the endometrial basal layer significantly decreased after uterine artery embolization, and reverted to the normal level (P<0.05). Increase of neovascularization with CD105-postive microvessel density presented after uterine artery embolization, and then returned to the normal (P<0.05). These findings indicate that uterine artery embolization may lead to a temporal decrease of microvessel density in the endometrium which wil recover over time as a result of the increase of CD105-postive neovascularization.