The expression of ITGB2 in renal cell carcinoma and its effect on malignant biological behaviors of ACHN cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective：To investigate the expression of ITGB2 (integrinβ2) in renal cell carcinoma (RCC) tissues and cells (ACHN cells) and its effects on the proliferation, migration, invasion and cell cycle of ACHN cells. Methods: The expression level of ITGB2 in RCC tissues and para-cancerous tissues was analyzed by GEPIA database. The tissue samples of 66 RCC patients retained in the biological specimen bank of the Fourth Hospital of Hebei Medical University from 2016 to 2020 were selected for this study. The expression level of ITGB2 in 66 cases of RCC tissues and para-cancerous tissues was detected by immunohistochemical SP and qPCR, and the relationship between ITGB2 and clinical parameters was analyzed. The shRNA with ITGB2 knockdown was constructed and transfected into ACHN cells for functional experiments to detect its effect on the malignant biological behaviors of ACHN cells, and its effect on the cell cycle was detected by flow cytometry. WB was used to detect the effect of ITGB2 knockdown on ITGB2 protein expression in ACHN cells. Results: The relative expression of ITGB2 in RCC tissues was significantly higher than that in para-cancerous tissue (P<0.01), and the expression was related to the clinical stage of RCC (P<0.05). Transfection of shITGB2 into ACHN cells could knock down the gene and protein expression of ITGB2 (all P<0.01). Knockdown of ITGB2 could significantly inhibit the proliferation (P<0.05), migration(P<0.01) and invasion (P<0.05) of ACHN cells but had no significant effect on cell cycle (P>0.05). Conclusion: ITGB2 is highly expressed in RCC tissues and cells and is associated with the clinical stage of RCC. Knockdown of ITGB2 can inhibit the malignant biological behaviors of ACHN cells.

Molecular mechanism of miR-497 in regulating the schizophrenia susceptibility gene EFNB2 / 西安交通大学学报(医学版)

Objective: To verify whether EFNB2 gene is targeted by miRNA-497 using molecular biology methods. Methods: Bioinformatic method predicted that EFNB2 gene is targeted by miRNA-497. PCR methods amplified the fragment in EFNB2 gene 3'-UTR including the putative miRNA-497 binding site. Then the sequences of wide type (WT) and mutant (MUT) were cloned into the pmirGLO luciferase vector, respectively. The DNA sequences of the amplified fragments were identified by restriction enzyme digestion and sequencing, and were consistent with the reference sequence from UCSC. This constructed vector was marked as pmirGLO-EFNB2 vector. Finally, the pmirGLO vector, the pmirGLO-EFNB2 vector, miRNA-497 mimics and negative control (NC) were divided into five groups and transfected into HEK393T cells, and the luciferase activity was tested after 24 h and 48 h by dual luciferase reporter gene assay. Results: The results of DNA sequencing demonstrated that the PCR fragment was successfully cloned into pmirGLO vector. The transfection results showed that the recombinant plasmid of WT and MUT was successfully transfected into HEK293T. The results of dual luciferase activity assay demonstrated that miRNA-497 significantly decreased the reporter gene activity compared with the NC. Conclusion: At the cellular level, the schizophrenia susceptibility gene EFNB2 was verified to be targeted by miRNA-497, which provides a new idea and new clue for subsequent studies on miRNA-497 function in the molecular mechanism of schizophrenia.

Correlation of psoriasis with erectile dysfunction: A meta-analysis / 中华男科学杂志

Objective@#To systematically analyze the correlation between psoriasis and erectile dysfunction (ED).@*METHODS@#We searched the Cochrane Library, EMbase, PubMed, OVID, Medline, VIP, WanFang, China National Knowledge Infrastructure (CNKI), and Chinese Biomedical Literature Database (CBM via SinoMed) for the published literature about the relationship between psoriasis and ED up to June 2016. According to inclusion and exclusion criteria, two researchers respectively extracted the relevant data and made a meta-analysis on the correlation of psoriasis with ED and IIEF-5 scores using the Review Manager 5.3 software.@*RESULTS@#A total of 6 studies were included in this analysis. The analysis with the fixed-effects model revealed a significant correlation between psoriasis and ED (OR = 1.92, 95% CI: 1.53－2.40, P <0.01), and that on 3 of the studies with the random-effects model showed that the IIEF-5 scores were significantly lower in psoriasis patients than in non-psoriasis males (MD = －3.11, 95% CI: －4.85－－1.37, P <0.01).@*CONCLUSIONS@#There is a certain correlation between psoriasis and ED. Psoriasis patients may have a higher incidence of ED though it is to be further confirmed by more higher-quality studies.

Culture of rat corpus cavernosal endothelial cells using modified immunomagnetic beads and cloning / 中华男科学杂志

Objective@#To search for the methods of isolating, purifying and culturing corpus cavernosal endothelial cells (CCECs) from SD rats, observe their growth characteristics, and providing seed cells for the study of erectile dysfunction (ED).@*METHODS@#The corpus cavernosal tissue from the SD rat was digested with 0.1% elastase, followed by purification of CCECs with immunomagnetic beads. After further amplification, monoclonal CCECs were sorted out with the cloning cylinder and their morphological and proliferative characteristics were observed. The von Willebrand factor (VWF) in the CCECs was identified by immunofluorescence staining, the CD31 molecule detected by immumohistochemistry, the purity of the CCECs determined by flow cytometry, and the proliferation of the cells measured with CCK-8 and growth curves.@*RESULTS@#After 7 days of purification and culture, the CCECs were fused into a monolayer under the inverted phase-contrast microscope, arranged like flagstones. The growth curves showed that the CCECs were in latency with a low growth rate at 1－2 days, in the logarithmic growth phase with a rapid rate at 3－4 days, and into the platform phase around the 6th day. VWF was positively expressed in the CCECs with much green fluorescence, and so was CD31 with a large number of brownish particles. The positive rate of the CCECs which were labelled with the VWF purified with magnetic beads combined with cloning cylinders was up to (91.9±3.75)%.@*CONCLUSIONS@#High-purity rat CCECs can be cultured successfully using immunomagnetic beads combined with cloning cylinders, with stable proliferation and passage in the endothelial cell medium.

Updated relationship between sleep and erectile function / 中华男科学杂志

Penile erection (PE) is a physiological phenomenon involving complex mechanisms. PE may occur as reactive erections, psychogenic erections in the conscious state and spontaneous erections during the sleep. Sleep-related PE refers to the erections occurring spontaneously during the sleep with rapid eye movement. Studies have shown a correlation between sleep and PE as well as between sleep disorders and erectile dysfunction but not yet revealed the exact mechanisms. This paper updates the relationship between sleep and erectile function.

Research of helicobacter pylori in gastric cancer development mechanism / 肿瘤研究与临床

Numerous studies showed that Helicobacter pylori (HP) infection and gastric cancer have a correlation, which is a important factor to influence the development of gastric cancer, with scholars' intensive research, it is found that the way of HP acting on gastric cancer is complicated and diverse, this review summarizes the role of HP in mechanism, correlation factor and action way of gastric cancer.

Rapid purification of human albumin with immunomagnetic microsphere technology / 药学学报

<p><b>AIM</b>To develop a rapid and inexpensive method for purification of human albumin, a method of immunomagnetic microspheres (IMMS) based on enzyme-linked immunosorbent assay (ELISA) for the purification of human albumin from human serum.</p><p><b>METHODS</b>Polystyrene magnetic microspheres with carboxyl groups as carriers were prepared, and then the carboxyl groups on the surface of the microspheres were activated by ethylcarbodiimide (EDC). Finally rabbit anti-human serum albumin (HSA) antibodies were covalently bound to it and the complex can specifically capture HSA. After the procedure of capturing HSA, through taking rabbit anti-human albumin protein antibodies as a capture antibody, and goat anti-human albumin protein antibodies as a detection antibody, an ELISA on IMMS was developed, which can determine the recovery yield of HSA from the human serum.</p><p><b>RESULTS</b>The result of the experiment was that the recovery of human albumin with IMMS was (86 +/- 4)%, and IMMS were reused for two other purifying cycles, the results of which were (69.0 +/- 0.6)% and (40.8 +/- 0.8)%, and the purity of the product was about 90%.</p><p><b>CONCLUSION</b>The results above prove that the immunomagnetic purifiying strategy was shown to be efficient and offers an new thought for a large scale production of high-purity HSA.</p>

Anticancer activity of N-(phenylacetyl) doxorubicin combined with folate-targeted PGA / 药学学报

<p><b>AIM</b>To demonstrate the specific killing of folate receptor (FR)-positive tumor cells can be achieved by folate-targeted penicillin-G amidase (PGA) combined with its prodrug substrate N-(phenylacetyl) doxorubicin (DOXP).</p><p><b>METHODS</b>Folic acid was covalently linked to PGA and folate content value was determined by quantitative UV spectrophotometry. The ability of folate conjugated PGA to hydrolyze DOXP was measured by RP-HPLC. Visual demonstration of uptake by FR (+) HeLa and SKOV3 cells was detected by using FITC labeled folate-PGA and a fluorescence microscopy. The cytotoxicity of DOXP towards the cells in the presence or absence of folate-PGA was assayed by using MTT method.</p><p><b>RESULTS</b>The folate-PGA has a specific activity of 29. 8 U x mg(-1) (protein). FR selectivity was confirmed by fluorescence microscopy. The combination of DOXP prodrug with folate-PGA generated higher cytotoxicity towards the FR (+) cells than free doxorubicin. The IC50 was 0.72 micromol x L(-1) for HeLa cells and 0.75 micromol x L(-1) for SKOV3 cells, respectively. Further, the enhanced cytotoxicity reduced greatly with the addition of free folic acid.</p><p><b>CONCLUSION</b>Folate conjugated PGA did not significantly compromise PGA catalytic activity and enabled binding prodrug-activating enzyme PGA to folate receptor expressing cells, and increased the sensitivity of the cells to doxorubicin followed by administration of its prodrug substrate.</p>