Objective To explore the diagnosis value of serum cystatin C (Cys C),neutrophil gelatinase-associated lipocalin (NGAL) and urinary interleukin-18 (IL-18) levels in critically cases from pediatric Intensive Care Units (ICU).Methods 114 cases from June 2015 to January 2017 in Xidian Group Hospital were enrolled in the study.According to whether cases secondary to AKI,they were divided into AKI group (64 cases) and non-AKI group (50 cases).At same time 50 healthy cases were selected as the control group.The Serum creatinine (Scr),urinary nitrogen (BUN),Cys C,NGAL and urinary IL-18 levels were measured in each subject.The relationship between the indexes of AKI group was analyzed by Pearson analysis.The value of serum Cys-C,NGAL and urinary IL-18 in the diagnosis of AKI was analyzed by receiver operating characteristic (ROC) curve.Results Serum Cys-C,NGAL and urinary IL-18 levels of the AKI group were higher than that of the non-AKI group and the control group,the difference had statistical significance (t=3.137～28.642,t1 =7.653～33.672,all P＜0.05).The levels of serum Cys-C and NGAL were positively correlated with Scr levels (Beta=0.273,0.514,all P＜0.05).The urinary IL-18 levels were negatively correlated with serum albumin (Alb) (Beta =-0.342,P＜0.05).The area under the ROC carve (AUC) of serum Cys-C,NGAL and urinary IL-18 was 0.872 (95％ CI:0.831～0.936),0.823 (95％ CI:0.641～0.925) and 0.714 (95％CI:0.598～0.833).Conclusion The levels of serum Cys-C and NGAL were elevated earlier than Scr,and it can be used as reliable indicators of early diagnosis of early cases AKI in ICU.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Female ; Genes, Viral ; Genotype ; Hepacivirus ; genetics ; Hepatitis C, Chronic ; epidemiology ; Humans ; Male ; Middle Aged ; Young Adult

Objective:To explore the mechanism of EV71 antagonizing IFN signaling pathway.Methods: RD cells were infected or un-infected with EV71.Then the cells were treated with or without IFN-β.The four groups (the control group,the EV71 group,the IFN-β group,the EV71+IFN-β group) were detected by molecular biology techniques.The expression of interferon stimulated genes (ISGs) were detected by Real-time PCR,while the protein levels of STAT1 and IRF9 were examined by Western blot assay.By preparing the cytosolic and nuclear fractions,the translocation of p-STAT1 was monitored through Western blot assay.Results:Compared with the IFN-β group,the mRNA level of OAS1,MX1 and ISG54 in the EV71+IFN-β group was down regulated by 47%, 50% and 48%,respectively,indicating that EV71 inhibited the expression of ISGs.The results also showed that EV71 did not effect the protein level and phosphorylation of STAT1.Moreover,we found that p-STAT1 was translocated into neuclear in IFN-β group,while p-STAT1 was located in the cytoplasm in the EV71+IFN-β group.And the expression of IRF9 was boviously down regulated in EV71+IFN-β group compared with that in IFN-β group,suggesting that EV71 blocked the expression of IRF9 induced by IFN-β.Conclusion:EV71 inhibited the IFN signaling pathway by downregulating the expression of IRF9 induced by IFN-β.

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

BACKGROUNDAlthough it is recognized that bronchial smooth muscle cells (BSMCs) play a key role in airway remodeling during chronic asthma, it is not well understood how BSMCs exert their inflammatory functions. The extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway is an important signaling pathway in chronic asthma, but its influence on secretion by BSMCs has not been well-studied. We investigated the impact of ERK1/2 signaling pathway on secretion by BSMCs in a rat model of chronic asthma in this study.

METHODSTo create a rat model of chronic asthma, Wistar rats underwent ovalbumim (OVA) injection and eight weeks of inhalation. BSMCs were isolated and cultured in vitro. Epidermal growth factor, PD98059 and ERK1/2 antisense oligonucleotide were used to explore the role of ERK1/2 signaling pathway. The expression of P-ERK1/2 (phospho-ERK1/2) in BSMCs was analyzed by Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Secretion of BSMCs was detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSPhospho-ERK1/2 expression was increased in BSMCs of chronic asthmatic rats compared with the controls. PD98059 inhibited expression of phospho-ERK1/2 protein, while treatment with an antisense oligonucleotide inhibited the expression of P-ERK1/2 mRNA and protein. BSMCs obtained from the chronic asthma group secreted significantly greater quantities of growth factors (transforming growth factor (TGF)-beta(1), vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF)), cytokines (regulated upon activation, normal T cell-expressed and secreted (RANTES) and eotaxin), and extracellular matrix (fibronectin and collagen I) compared with normal controls. Epidermal growth factor stimulated secretion in both groups, but the response of the chronic asthma group was more intense. Both PD98059 and antisense oligonucleotide suppressed secretion by BSMCs in chronic ashmatic rats. Antisense oligonucleotide reduced the level of RANTES nearly to that of normal controls, while PD98059 could not.

CONCLUSIONThese results suggest that ERK1/2 signaling pathway may play an important role in the augmented secretion of BSMCs in chronic asthmatic rats, and ERK1/2 antisense oligonucleotide effectively inhibits the process.

Animals ; Asthma ; metabolism ; Bronchi ; secretion ; Chemokine CCL5 ; secretion ; Chronic Disease ; Disease Models, Animal ; MAP Kinase Signaling System ; physiology ; Male ; Mitogen-Activated Protein Kinase 1 ; physiology ; Mitogen-Activated Protein Kinase 3 ; physiology ; Myocytes, Smooth Muscle ; secretion ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; secretion ; Vascular Endothelial Growth Factor A ; secretion

To investigate the regulatory effect of extracellular signal-regulated kinase (ERK) signaling pathway on airway smooth muscle cell (ASMC) proliferation in chronic asthmatic rats, the rat model of chronic asthma was established, and ERK agonist epidermal growth factor (EGF) and inhibitor PD98059 were used in the cell culture. ASMC proliferation was examined by flow cytometry analysis, methyl thiazolyl tetrazolium (MTT) colorimetric assay, [(3)H]-thymidine (TdR) incorporation and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. The expressions of ERK mRNA, ERK protein, phosphorylated ERK1/2 (p-ERK1/2) protein were observed by RT-PCR and Western blot. The results showed that in chronic asthmatic group, compared with that in the control group, the percentage of cells at G(0)/G(1) phase was significantly decreased and the percentage of cells at S+G(2)/M phase was significantly increased. Absorbance (A(490)), DNA synthesis and the expression of PCNA protein in ASMCs in chronic asthmatic group were significantly increased. The expressions of ERK mRNA, ERK1/2 protein, p-ERK1/2 protein and the activation ratio of ERK in ASMCs in chronic asthmatic group were significantly increased compared with those in the control group. After treatment with PD98059, the percentage of cells at S+G(2)/M phase, A(490), DNA synthesis and the expression of PCNA protein in ASMCs in chronic asthmatic group were significantly decreased; the expressions of ERK mRNA, ERK1/2 protein, p-ERK1/2 protein and the activation ratio of ERK in ASMCs in chronic asthmatic group were significantly decreased compared with those in the control group. After treatment with EGF, the percentage of cells at S+G(2)/M phase, A(490), DNA synthesis and the expression of PCNA protein in ASMCs in chronic asthmatic group were significantly increased compared with those before treatment; and PD98059 markedly inhibited the effect of EGF. These results suggest that the endogenous proliferation activity of ASMCs in chronic asthmatic rats significantly increases compared with that in the control rats, and ERK1/2 participates in this process. The ERK signaling pathway might play an important role in regulating ASMC proliferation, leading to asthmatic airway remodeling.
Animals ; Asthma ; enzymology ; pathology ; Bronchi ; pathology ; Cell Proliferation ; Cells, Cultured ; Chronic Disease ; Colorimetry ; Enzyme Activation ; Extracellular Signal-Regulated MAP Kinases ; analysis ; genetics ; physiology ; Flow Cytometry ; Myocytes, Smooth Muscle ; pathology ; Rats

This work was designed to explore the role of extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway in migration of bronchial smooth muscle cells (BSMCs) of chronic asthmatic rats. To make chronic asthma model, Wistar rats underwent ovabumin (OVA) injection and eight-week inhalation. BSMCs were cultured in vitro. The expression of ERK1/2 in BSMCs was analyzed by immunocytochemistry, Western blot and RT-PCR. Migration of BSMCs was detected by both plate test and Boyden cell test. Results showed: (1) With Western blot technique, the ratio of p-ERK1/2 to total ERK1/2 in chronic asthmatic group was obviously higher than that in the control group (0.55 +/- 0.05 vs 0.48 +/- 0.04, n=10, P<0.01). (2) With RT-PCR, the relative A values of ERK1 and ERK2 mRNA in airways of chronic asthmatic rats were 1.83 +/- 0.24 and 1.07 +/- 0.11, respectively, which were significantly increased compared with that in the control group (0.58 +/- 0.14 and 0.51 +/- 0.12, n=10, P<0.01). (3) In plate test, the migration of BSMCs of chronic asthmatic rats was 2.9 times of that in the control group and reached 5.0 times by epidermal growth factor (EGF) stimulation, but decreased to 1.7 times by 30 mumol/L PD98059. (4) In Boyden cell test, the migration of BSMCs of chronic asthmatic rats was 1.9 times of that in the control group, and reached 3.1 times by EGF stimulation, but decreased to 1.45 times by 30 mumol/L PD98059. Our results indicate that the migration ability of BSMCs of chronic asthmatic rats increases, and ERK1/2 signaling pathway may play an important role in this process.
Animals ; Asthma ; chemically induced ; physiopathology ; Bronchi ; pathology ; Cell Movement ; physiology ; Cells, Cultured ; Male ; Mitogen-Activated Protein Kinase 1 ; metabolism ; physiology ; Mitogen-Activated Protein Kinase 3 ; metabolism ; physiology ; Myocytes, Smooth Muscle ; pathology ; Ovalbumin ; Rats ; Rats, Wistar ; Signal Transduction ; physiology

Objective To study the acceptability of pre-exposure prophylaxis (PrEP) to prevent the transmission of HIV among men who have sex with men (MSM) in Guangxi, China.Methods Snow-balling methods were used to recruit 650 MSM in Guangxi. Questionnaires and interview were administrated to these 650 men, using a self-designed questionnaire and face to face interviews to collect information on HIV-related risk behaviors, knowledge and acceptability of PrEP.effective, safe and free of charge', 597 (91.9%) of the 650 MSM claimed that they would accept it,who refused to use it, most of them said that were afraid of the side-effect and doubted on the effectiveness of PrEP. Data from logistic regression analysis showed that those who had found partners through friends (OR=6.21, P=0.020) and those who would advise his friend to use PrEP (OR=39.32, P=0.000) were more likely to accept PrEP. Those who thought they could protect themselves from HIV infection (OR=0.32, P=0.010) or not having sex with the ones who refused to use a condom (OR=0.34, P=0.010) were less likely to accept PrEP. Conclusion Effectiveness, safety and cost seemed to be the main influential factors related to the acceptability of PrEP. Peer education might improve the acceptability of PrEP.

Objective: To identify the etiology and source of infection in a diarrhea outbreak in Yunnan in May 2017 and to provide the evidence for formulating prevention and control measures. Methods: Epidemiological investigation was carried out on the epidemic situation of diarrhea in the village of Lvchun County in Yunnan Province, the field sampling, laboratory testing and data analysis were also performed. Results: Among the 44 patients, 11 of the 13 samples were positive for rotavirus nucleic acid in group A, and the positive rate was 84.62%. The survey showed that the water supply pipe was damaged and polluted by human and livestock manure and domestic sewage. The trend of the damaged water pipe was basically the same as the case distribution, and the rainfall was significantly related to the number of the disease. Conclusions This event was an outbreak of diarrhea caused by group A rotavirus. The direct pollution of drinking water caused by rainfall may be the risk factor of this outbreak. The health management of rural drinking water should be strengthened and the health knowledge and education of preventing intestinal infectious diseases should be promoted.