Objective:To study e ects of Saikosaponin a(SSa) on tumour necrosis factor-alpha(TNF-?) release and its receptor expression in cultured hippocampal astrocytes induced by pentylenetetrazol(PTZ).Methods:The in vitro cultured primary hippocampal astrocytes were randomly divided into control group(group A),PTZ-induced group(group B)PTZ10mmol/L+SSa groups(group C and group D,the SSa concentrations were 1.25mg/L and 0.625mg/L respectively).The extracellular uid TNF-? level and the expression of tumour necrosis factor receptor type 1(TNFR1) in hippocampal astrocytes were respectively detected by ELISA and Western-blot after PTZ-induced 2 hours.Results:the TNF-? level and TNFR1 expression of group B were signi cantly higher than that of group A,group C and group D(P

To set up a new rapid method for the rapid determination of influenza virus H5N1 and H7N9 basing on the Multi-Analyte Suspension Array (MASA) technology. Sequence analysis and design of degenerate primers and specific probes were set in the comparison and analysis of H5, N1, H7 and N9 genes. In combination with MASA technology, these primers and probes were used for the determination of samples of H5N1 and H7N9 and other subtypes ( H1N1, PH1N1, H5N2, H3N2 and H9N2). We developed a rapid determination method. This method had high specificity and sensitivity that could detect H5N1 and H7N9 at one time, and could detect samples that containing 10 copies of H5N1 and H7N9. This determination method could be used for rapid determination of influenza virus H5N1 and H7N9 at one time.
Humans ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza A Virus, H7N9 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; virology ; Oligonucleotide Array Sequence Analysis ; methods

We aimed to develop a real-time polymerase chain reaction (PCR) detection method for the Reston subtype of the Ebola virus. The NP gene of the Reston subtype of the Ebola virus was selected as the detection object. Sequences of different subtypes of Ebola viruses were aligned using Clustal W software. The most unique and conserved regions of the Reston subtype of the Ebola virus were recruited as candidate sequences for specific primers. Primer Express and Primer Premier 5. 0 software were used to filter the optimal pair of primers for detection. Real-time PCR was carried out using optimized parameters and positive DNA prepared by serial (tenfold) dilution of a recombinant plasmid and by plotting a standard curve. In addition, the reproducibility, accuracy, and specificity of the assay were tested. Results showed that the sensitivity of detection of the Reston subtype of the Ebola virus by real-time PCR could reached 10(2) copies/microL. The linear relationship (R2) reached 0.997, the slope of the standard curve was -0.3101, and amplification efficiency was 110.145%. A sharp and narrow melting peak appeared at 79.94 degrees C for all standards in different dilutions. In conclusion, a fast and sensitive real-time PCR detection system for the Reston subtype of the Ebola virus was developed. This system could be used as a supplementary diagnostic and monitoring approach for basic and clinical studies on the Reston subtype of the Ebola virus. The detection system does not require expensive technology or specialist operators.
DNA Primers ; genetics ; Ebolavirus ; classification ; genetics ; isolation & purification ; Hemorrhagic Fever, Ebola ; virology ; Humans ; Real-Time Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To investigate the effect of recombinant intestinal trefoil factor(rITF)on the intestinal mucosa of acute necrotizing pancreatitis(ANP)rats and explore the mechanism.Methods 60 male SD rats were randomly divided into control group(n=20),ANP group(n=20),rITF group(n=20).ANP was induced by retrograde injection of 5%sodium taurocholate(100μl/100 g)into biliopancreatic duct.Rats in rITF group were injected with rITF(0.5 mg/100 g)before and after ANP induction.The rats in ANP group and control group received same amount of normal sailne.Each group were sacrificed 12 h or 24 h later.respectively.Blood sample was taken to determine the serum level of amylase.Terminal ileum was resected to observe the pathologic changes;immunohistochemistry method was applied to detect the activity of NF-κB;the expression of TNF-α mRNA,ICAM-1 mRNA in intestinal mucosa was measured by RT-PCR.Results Intestinal injury score of ANP group was higher than that ofcontrol group(P＜0.05),intestinal injury score of ANP 12 h group was higher than that of rITF 12 h group(P＜0.05),but there was no significant difference between ANP 24 h group and rITF 24 h group.The number of positive NF-κB cells was 26±4,55±8,49±4,respectively;the relatively expression of TNF-α mRNA in terminal ileum was 0.050±0.005,1.040±0.031 and 0.792±0.0256,respectively;the relatively expression of ICAM-1 mBNA was 0.045±0.010,0.795±0.037 and 0.400±0.031,respectively,in control group,ANP group and rITF 12 h group.The corresponding values in the ANP group were significantly increased compared with those values in the control group (P＜0.05 or P＜0.01).Conclusions rITF may protect the intestinal mucosa.the mechanism may include inhibit NF-κB activation,down-regulate TiNF-α mBNA,ICAM-1 mRNA expression.

Objective To study the risk factors of severe acute pancreatitis(SAP) complicated with MODS at the early stage.Methods One hundred and seven SAP patients served as A group,who admitted from Janurary 1995 to December 1999.One hundred and thirty patients served as B group who admilted from Janurary 2000 to 2005.Age,sex,Ranson' score,APACHEⅡ ,CT score,biliary tract obstruction,hypoxia,lung infection,shock,abdomen compartment syndrome,hyperlipemia,pleural effusion,and mortality were analyzed.Results Twenty-five in the A group and 28 in B group two were complicated with MODS at the early stage.There existed difference in Ranson's score,APACHEⅡ,CT score between patients with and without MODS(P

OBJECTIVE:To prepare Geinstein (GEN) solid dispersion,and improve the dissolution rate of GEN in vitro. METHODS:Using PVP K30,PEG6000,and PEG4000 as carriers,GEN solid dispersion was prepared by solvent melting meth-od,and its dissolution in vitro was investigated. The structure of the solid dispersion was characterized by FTIR and DSC. RE-SULTS:GEN solid dispersion prepared with PEG4000 as carrier was better than those with other carriers in dissolution,and drug-carrier ratio (1:5) was the best. The results of DSC and FTIR showed that GEN in solid dispersion took amorphous form. CONCLUSIONS:GEN solid dispersion is prepared successfully and significantly improve the dissolution of GEN in vitro.

Objective:To evaluate the correlation between the prognosis of patients with hilar cholangiocarcinoma and the degree of bile duct dilatation in MRCP .Methods:The clinical data of 89 patients with hilar cholangiocarcinoma undergoing radical operation at Tianjin Nankai Hospital from Jan 2009 to Dec 2013 were analyzed retrospectively.Results:Tumor size ( P=0.024), Bismuth-Corlette classification ( P=0.048) and tumor stage ( P=0.013) were related factors of biliary dilatation. Tumor differentiation ( P=0.002), R 0 resection ( P=0.002) and biliary dilatation ( P<0.001) were independent predictors of disease-free survival (DFS). Conclusion:The imaging evaluation of the degree of biliary dilatation has a certain predictive value for the prognosis of patients with hilar cholangio-carcinoma.

Objective To analyze the clinical features and the effect of therapy on cutaneous emphysema of chest wall and/or pneumomediastinum complicated in severe acute triphosgene poisoning patients.Methods Among 81 triphosgene poisoning patients,5 complicated with cutaneousemphysema of chest wall and/or pneumomediastinum were analyzed in respect of the clinical data including age,gender,arterial blood gas (ABG),modes of mechanical ventilation support and so on.Results Five patients consisting of 3 males and 2 females,aged (23.20 ± 5.17) years,were complicated with cutaneous emphysema of chest wall and/or pneumomediastium with a prevalence rate of 0.06％.Of them,4 were alleviated completely and 1 died of acute respiratory distress syndrome (ARDS).There was no significant difference in arterial blood gas analysis (ABG) between patients with cutaneousemphysema and/or pneumomediastinum and patients without ( P ＞ 0.05 ).Conclusions Triphosgene-induced acute lung injury treated with mechanical ventilation support with high PEEP is highly suggested as high risk factor for the formation of cutaneous emphysema of chest wall and/or pneumomediastinum in severe acute Triphosgene poisoning patients.It is very important to set the PEEP level of mechanical ventilation support as low as possible for avoidance of alveolar rupture.

Objective To investigate the effects of advanced oxidation protein products (AOPP) in type 2 diabetic ocular muscles palsy (DOMP). Methods 58 DOMP patients and 50 type 2 diabetes patients were included in the research. Hemoglobin A1c (HbA1c), blood glucose (FPG), fasting insulin (FINS) and triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL) were measured and recorded. Homeostasis model assessment was performed to evaluate the status of insulin resistance (HOMA-IR), basal insulin secretion (HOMA-β) and the insulin sensitivity index (ISI). Serum AOPP was measured by enzyme linked immunosorbent assay. Unconditional logistic regression model was used to evaluate the influencing factors of DOMP. Results The DOMP group showed higher levels of plasma AOPP, TG, LDL, FPG, FINS, HbA1c and HOMA-IR, but lower levels of HDL, HOMA-β and ISI than those of the T2DM group. Unconditional logistic regression analysis revealed that AOPP was an independent risk factors for DOMP (OR =3.01, P = 0.002). Conclusion AOPP may be involved in the pathogenesis of DOMP. AOPP could be a useful indicator for monitoring the development of DOMP and for evaluating its severity.

Objective To investigate the protective effect of Gabexate mesilate(GM)on D-galactosamine- lipopolysaccharide-indneed acute liver failure in rats.Methods The model of acute liver failure in rats was produced by injection of D-galactosamine(D-GalN)and lipopolysaccharide(LPS).The alanine aminotransferase(ALT),aspartate aminotransferase(AST)in serum and malondiadehyde(MDA)content,superoxide dismutase(SOD)and glutathione peroxidase(GSH-PX)activities in liver homogenate were assayed by spectrophotometry.The levels of turnout necrosis factor-?(TNF-?),interleukin-?(IL-?)and interleukin-6(IL-6)were determined by ELISA method.Hepatic pathological examination was observed.Results 25 mg?kg~(-1),50mg?kg~(1),100 mg?kg~(-1) of GM significantly decreased the serum transaminase activities,the infiltration of inflammatory cells,and MDA content,hut didn't reduce SOD and GSH- PX activities in liver homogenate.GM significantly reduced TNF-?,IL-1?and IL-6 levels in serum.Conclusions GM showed significant protective effects on acute liver failure in rats.