Background Keratoconus is a bilateral,noninflammatory,gradually progressive corneal disorder characterized by progressive thinning and steepening of the central cornea.It is significant to investigate keratoconusrelated pathogenic gene for elaborating the pathogenesis and establishing early diagnosis standard and taking clinical measurement.Objective The aim of the study was to explore the relationship of visual system homeobox gene (VSX1) polymorphism and the risk of sporadic keratoconus.Methods This study was approved by Ethic Commission of First Hospital of Xi' an.Written informed consent was obtained from each subject prior to enrollment.A case-controlled study was conducted.One hundred and one Han nationality patients with sporadic keratoconus were included in this study.These keratoconus patients were clinically diagnosed by slit lamp examination and corneal tomography.Single nucleolide polymorphism (SNP) of VSX1 gene was assayed and classified using the MassARRAY SNP technique.Demography and relevant risk factors were collected from each subject by questionnaire.Eighty healthy volunteers served as controls.Chi-square test and Binary logistic regression were used to evaluate the difference in the distribution of allele frequency and genotype frequency and to analyze the association with keratoconus risks.Results SNP of two genes was found in the Chinese Han population (rs743018 (c.843+140 C＞T) and rs6138482(R217H C＞T)).There were no significant differences in the genotype frequency and allele frequency of the SNP of two genes in the keratoconus group in comparison with the normal control group (P＞0.05).After adjustment by age and sex,SNP of two genes was not significantly associated with the risk of keratoconus (regression model:rs743018 (C＞T) adjusted:P=0.35,OR=0.72,95％ CI:0.37-1.43 ;rs6138482 (C＞T) adjusted:P =0.48,OR=0.76,95％ CI:0.35-1.64).Conclusions Gene polymorphisms of rs743018(c.843+140 C＞T) and rs6138482(R217H C＞T) in the Chinese Han population is not associated with the risk of keratoconus.Due to the racial difference in genotype and allele frequency,the role of the VSX1 gene in the pathogenesis of keratoconus still remains controversial,and further study needs to be developed.

The incidence of septic acute kidney injury (AKI) is increasing, it has become a major threat to human health because of its acute onset, poor prognosis, and high hospital costs. The most common cause of AKI in critical-care units is sepsis. Septic AKI is a complex and multi-factorial process; its pathogenesis is not fully understood. In sepsis, the destruction of mucosal barriers, intestinal flora disorders, intestinal ischemia/reperfusion injury, use of antibiotics, and lack of intestinal nutrients lead to an inflammatory reactions that in turn affects the metabolism and immunity of the host. Such changes further influence the occurrence and development of AKI. New technology is enabling various detection methods for intestinal flora. Clinical application of these methods in septic renal injury is expected to clarify the relationship among pathogenesis, disease progression mechanism, and intestinal flora.

Objective To study the effect of advanced glycation end products (AGEs) on the proliferation, cell cycle and apoptosis of SH-SY5Y cells, and the mRNA expression of AD related protiens. Methods Bovine serum albumin-AGEs (BSA-AGEs) were prepared by incubation of BSA with glucose in vitro. SH-SY5Y cells were incubated with different concentrations of BSA-AGEs. Cell proliferation was measured by MTT assay; cell cycle and apoptosis were determined by flow cytometry; mRNA levels of APLP1, IL-6 and HMGB-1 were determined by RT-PCR. Results After incubation with BSA-AGEs for 48 h, SH-SYSY cell proliferation was significantly inhibited and cell apoptosis was induced; the cells were found to be arrested at G1/G0; these effects were dose-dependent. RT-PCR results showed that mRNA levels of IL-6, APLP1 and HMGB-1 were significantly up-regulated. Conclusion BSA-AGEs inhibit the cell proliferation and induce the apoptosis of SH-SY5Y cells, and stimulate the expression of some pro-inflammatory cytokines, suggesting a possible important role of AGEs in pathogenesis and development of AD.

OBJECTIVETo find a new method of treating hepatocellular carcinoma with melittin by way of using the melittin gene.

METHODSThe recombinant adenoviruses carrying the melittin gene and alpha-fetoprotein (AFP) promoter (Ad-rAFP-Mel) were constructed through a bacterial homologous recombinant system. The efficiency of the adenovirus mediated gene transfer and the inhibition effect of Ad-rAFP-Mel on the proliferation of hepatocarcinoma cells were determined by X-gal staining and MTT assay respectively. The tumorigenicity of hepatocarcinoma cells transfected by Ad-rAFP-Mel and the antitumor effect of Ad-rAFP-Mel on the transplanted tumors in nude mice were detected in vivo.

RESULTSThe mRNA of the melittin gene was transcripted in HepG2 hepatocellular carcinoma cells transducted by Ad-rAFP-Mel. The efficiency of adenovirus mediated gene transfered to BEL-7402 hepatocarcinoma cells was 100% when the multiplicities of infection (MOI) of Ad-rAFP-Mel was 10 in vitro and was high in vivo as well. The inhibitive rates of Ad-rAFP-Mel and Ad-rAFP for BEL7402 cells were 66.2%+/-2.7% and 2.9%+/-2.3% (t = 30.83) by MTT assay. The inhibitive rates of Ad-CMV-Mel for BEL7402, SMMC7721 and L02 cells were 58.9%+/-9.6%, 65.9%+/-3.8%, 31.7%+/-1.2%, respectively, and those of the Ad-rAFP-Mel were 6.2%+/-2.7%, 16.1%+/-6.6%, 7.5%+/-3.3%, respectively (t = 1.27; t = 11.31, and t = 12.12, vs. Ad-CMV-Mel group in same cells). The tumorigenicity rates of hepatocarcinoma cells transfected by Ad-rAFP-Mel were decreased. A significant antineoplastic effect was detectd on transplanted tumor in nude mice by intratumoral injection of Ad-rAFP-Mel.

CONCLUSIONAd-rAFP-Mel can inhibit specifically the proliferation of AFP-producing human hepatocarcinoma cells in vitro and in vivo. It suggests that animal toxin gene can be used as an interesting antitumor gene.

Adenoviridae ; genetics ; Animals ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Liver Neoplasms, Experimental ; pathology ; Male ; Melitten ; biosynthesis ; genetics ; pharmacology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transcription, Genetic ; drug effects ; alpha-Fetoproteins ; biosynthesis ; genetics

OBJECTIVETo observe the induced apoptosis of recombinant adenovirus carrying melittin gene (Ad-rAFP-Mel) for hepatocellular carcinoma cell line (BEL-7402).

METHODSThe morphological observe, DNA electrophoresis, TUNEL and Flow cytometry assay were used to study the apoptosis of BEL-7042 cell line transfected by Ad-rAFP-Mel.

RESULTSThe morphological changes and apoptosis of BEL-7402 transfected by Ad-rAFP-Mel were confirmed with microscopy and DNA electrophoresis, TUNEL, Flow cytometry assay. The DNA ladder could be demonstrated on DNA electrophoresis in Ad-rAFP-Mel group. The apoptosis rates of BEL-7402 cells in Ad-rAFP-Mel, Ad-rAFP, and control groups were (21.5+/-2.4)%, (10.5+/-4.4)% and (3.0+/-1.4)% respectively by TUNEL assay (F = 38.0, P < 0.05) and were (7.3+/-0.5)%, (3.9+/-0.1)% and (0.8+/-0.1)% respectively by flow cytometry assay (F = 415.1, P < 0.05).

CONCLUSIONIt seems that melittin inducing apoptosis might be one of the antitumor mechanisms.

Adenoviridae ; genetics ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Gene Expression ; drug effects ; Gene Silencing ; drug effects ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Liver Neoplasms ; pathology ; Melitten ; biosynthesis ; genetics ; pharmacology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transcription, Genetic ; drug effects ; Transfection

Breast Neoplasms ; pathology ; Cell Culture Techniques ; methods ; Cells, Cultured ; Female ; Humans ; Immunohistochemistry ; methods ; Quality Control

Objective:To construct HIV-1 pseudovirus containing enhanced green fluorescent protein ( EGFP ) gene. To understand the interaction between the virus and the cells. Methods: HIV-1 pseudovirus containing EGFP gene was constructed by lentiviral packaging systems, and its EGFP gene was amplified using RT-PCR. The level of genomic integration and transcription of HIV-1 pseudovirus containing EGFP gene were detected on iDCs infected with HIV-1 pseudovirus. At the same time, research on expression of the EGFP gene in iDCs infected with HIV-1 pseudovirus was performed. Results:The EGFP gene of HIV-1 pseudovirus was detected through RT-PCR. The EGFP gene was identified in iDCs infected with HIV-1 pseudovirus through PCR and RT-PCR. The EGFP was observed in iDCs infected with HIV-1 pseudovirus under fluorescence microscopy. Conclusion: HIV-1 pseudovirus containing EGFP gene has been successfully produced. The HIV-1 pseudovirus that we constructed can infect iDCs,then its RNA can integrate into the genome of iDCs in the way of reverse transcription,and the EGFP gene could express in the iDCs after infected with HIV-1 pseudovirus.

BACKGROUND: Previous studies on whether or not levosimendan improved the prognosis of patients with sepsis and septic shock have been inconsistent. We aimed to provide an updated analysis of the therapeutic value of levosimendan in adult patients with sepsis and septic shock, in order to provide evidence-based medical evidence for its use. METHODS: PubMed, Embase, Cochrane Library, Wanfang Data, and CNKI were searched until August 2018 without language restriction. Randomized controlled studies of levosimendan with either inotropic drugs or placebo for the treatment of sepsis or septic shock were enrolled. The primary outcome was mortality, and cardiac index and serum lactate levels were the secondary outcomes. RESULTS: A total of 20 randomized controlled studies were included in this meta-analysis, including 1467 patients, with 738 patients in the experimental group (levosimendan group) and 729 patients in the control group (other inotropic drugs or placebo). There were no significant differences in mortality between the levosimendan and control groups (fixed-effect relative risk [RR] = 0.90, 95% confidence interval [CI] [0.79, 1.03], P = 0.13). Levosimendan increased the cardiac index (VMD [weighted mean difference] = 0.51, 95% CI [0.06, 0.95], P = 0.02); and serum lactate levels were lower (VMD = -1.04, 95% CI [-1.47, -0.60], P < 0.00001). CONCLUSIONS Based on current clinical evidence, levosimendan does not reduce mortality in adult critically ill patients with sepsis and septic shock. Physicians should use levosimendan with caution in patients with sepsis and septic shock.

Objective To optimize extraction technology for Sihuang Shaoshang ointment by orthogonal test,and establish the HPLC fingerprint of Sihuang Shaoshang ointment to provide references for the quality evaluation of the preparation.Methods Taking Sihuang Shaoshang ointment as the model drug,the fingerprints were established by HPLC method,the common peak areas were analyzed by principal component analysis,and the total factor scores were used as the evaluation index.The technological parameters such as extraction times,extraction time and liquid-solid ratio were optimized by orthogonal test,and the optimal extraction process parameters were screened out.Results The common mode of fingerprint was set up with 12 common peaks and the two principle components with the accumulative contribution rate of 84.751% extracted by principal component analysis were screened out to calculate the comprehensive scores of 9 samples.The optimal extraction process was as follows:10-fold amount of water,extracting 3 times,and extracting 2 h for each time.Conclusion The established method is simple and effective,which can provide a reference for the extraction process optimization of Sihuang Shaoshang ointment.

Aim To screen and study the effects of Tao Hong Si Wu decoction(THSWD)-mediated treat-ment on circular RNA(circRNA)expression profiles in rats with middle cerebral artery occlusion(MCAO),and investigate the possible roles and molecular mecha-nisms of THSWD.Methods Next-generation RNA sequencing was conducted to identify circRNA expres-sion profiles in MCAO rats after treatment with THSWD and compared with the MCAO model group and control group.Bioinformatics analysis was performed to predict the potential target microRNAs and mRNAs.Gene On-tology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses for the potential target mRNAs were applied to explore the potential roles of differentially expressed circRNAs.RT-qPCR was performed to verify circRNAs with significant differences in expression.Results We identified 87 significantly differentially expressed circRNAs between the MCAO group versus the control group,and 86 sig-nificantly differentially expressed circRNAs between the MCAO group versus the THSWD group.respective-ly.Among them,17 circRNAs induced by the MCAO model were reversed via treatment with THSWD.To demonstrate the roles of mRNAs targeted by DECs,the GO and KEGG databases were used.Further analysis revealed that five circRNAs may play important roles in the development of MCAO.Conclusions The com-prehensive expression profile of circRNAs in rats with middle cerebral artery occlusion after THSWD treat-ment is determined for the first time,suggesting that the therapeutic effect of THSWD on MCAO may be a-chieved by regulating the expression of circRNAs.