Objective To explore the effect of the recombinant plasmid pcDNA3.1/C-sis on fulminant hepatic failure (FHF) in rats.Methods 48 h after recombinant plasmid pcDNA3.1/C-sis being imported into rat liver by using the method of fluid mechanics,FHF in rats was induced by endotoxin (LPS)+D-galactosamine (D-GaIN).With fluorescence quantitative PCR and Western blotting,C-sis expression was tested.The apoptosis of rat liver was detected by using HE staining and measuring Caspase-3 activity.The expression changes of Bcl-2 and Bax were examined through using Western blotting.The mortality rate of rats was calculated during 24 h observation period.Resnlts Compared with the normal control group and FHF+ empty plasmid group,C-sis mRNA and protein expression levels were increased significantly in the FHF+C-sis plasmid group,there were statistically significant differences (P＜0.01).Compared with the normal control group,the apoptotic hepatocytes were increased in the FHF + Ringer's solution injection group and FHF+ empty plasmid group;compared with FHF+ empty plasmid group,the apoptotic hepatocytes in the FHF+C-sis plasmid group were decreased.Compared with the normal control group,Caspase-3 expression level was increased in the FHF+ Ringer's solution injection group (P＜0.01);compared with the FHF+ empty plasmid group,Caspase-3 expression level in the FHF+C-sis plasmid group was decreased (P＜0.05).Compared with the normal control group,Bcl-2 expression level was decreased significantly (P＜0.01),and Bax expression level was increased significantly (P＜0.01) in the FHF+Ringer's solution injection group;compared with the FHF+ empty plasmid group,the Bcl-2 expression level was increased (P＜0.05),and Bax expression was decreased (P＜0.05)in the FHF+C-sis plasmid group.During the 24 h observation period,all rats in the normal control group were alive;the mortality rates of the FHF+ Ringer's solution injection group and FHF+ empty plasmid group were 70.0％ and 80.0％ respectively,while that of the FHF+C-sis plasmid group was only 20.0％.Conclusion C-sis gene could inhibit FHF in rats induced by LPS+D-GalN.

Objective:Expression of protein TSLP and selection of full human anti-TSLP single chain Fv ( scFv).Methods:The cDNA of TSLP was amplified.The amplified target gene and the expression vector pET 101/D-TOPO were ligated , and then transformed into E.coli BL21.The protein was induced to expression by IPTG and purified and identified.The biotinylated TSLP protein was used as antigen to select of human TSLP scFv from a constructed human scFv library by phage display .Results: The size of amplified cDNA of TSLP was about 423 bp,and that of expressed protein was about 26 kD.Dot blot and Western blot results showed that the expressed protein was correct.The constructed human scFv library was enriched for three rounds using biotinylated TSLP as antigen by phage display.ELISA results showed that 35% scFvs had binding activity with TSLP.The scFvs with good binding activity were selected and identified by Western blot and sequencing.Conclusion: The full human scFvs against for TSLP were selected suc-cessfully.

Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.

Objective:To single amino acid mutation of the full human scFvs against TSLP to enhance its affinity.Methods: The specific scFvs against TSLP was screened in our previous study and here the three-dimensional structures of TSLP and anti-TSLP scFvs were simulated by Discovery Studio system,then the molecular docking was made.The amino acids of binding epitope were randomly mutated and the mutated amino acids were selected which could remarkably improve the affinity of scFvs.The primers were designed based on the sequence of mutation amino acids and the scFv sequences were mutated by the overlapping extension PCR.The DNA of mutated scFvs was ligated with the expression vector pLZ16 and transformed into E.coli DH5αF′.Then the scFvs were expressed and the scFvs with improved affinity were selected by ELISA and BIAcore.Results: The five scFvs with single amino acid mutation were screened out by DS system,which could elevate the affinity of scFvs.The mutated anti-TSLP-scFvs were amplified by PCR,which size was about 1 000 bp.The mutated scFvs with correct sequence were expressed,and the mutated scFvs with improved affinity were detected by ELISA and BIAcore.The affinity of selected mutated scFv (M4) has been about 10 times higher than the scFv nonmutation.Conclusion: The affinity of anti-TSLP-scFv has been improved successfully.

Objective To investigate the clinical value of high frequency ultrasonography with E-flow imaging in the evaluation of fingertip's microcirculation changes in patients with systemic sclerosis(SSc).Methods Twenty-four SSc patients and 29 healthy subjects were involved.High frequency ultrasonography with E-flow imaging was used to observe the configuration and distribution of digital arteries in the last segment of left and right middle finger.Peak systolic velocity (PSV),end diastolic velocity (EDV),mean velocity(MV),vascular resistance index (RI) and pulsatility index(PI) of digital palmar propria arteries,nail bed arteries and finger ventral arteries were measured.Results In control group,rich blood supply was revealed within the fingertips.Digital palmar propria arteries,nail bed arteries and finger ventral arteries and their small branches were displayed clearly and continuously by E-flow imaging.While in SSc patients,the definition and continuity of fingertip's small vascular flow images were not as good as that in the control group,with the distribution of blood flow markedly reduced.compared with the control group,PSV,EDV and MV of digital palmar propria arteries,nail bed arteries,finger ventral arteries were decreased in SSc group(P＜0.01),but both RI and PI were increased(P＜0.05).There were no statistically significant differences between left and right fingertip's arteries index in normal control group (P＞0.05).But PSV,EDV and MV of left digital palmar propria arteries in SSc group were higher than that of the right(P＜0.05),whose differences bear statistic significance.Conclusions High frequency ultrasonography with E-flow imaging is sensitive and reliable to reflect fingertip's microcirculation changes and provide a new method to assess microvascular changes in SSc patients.

Caliciviridae Infections ; epidemiology ; China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Male ; Norovirus ; Schools

Objective To investigate the feasibility and clinical effect of immediate ureterovesical reimplantation after ureteral rupture during laparoscopic hysterectomy. Methods From August 2010 to December 2015, 5 cases of ureteral rupture during the operation of laparoscopic hysterectomy were treated with immediate ureterovesical reimplantation under laparoscopy. Results All operations were successfully performed without transversion to open surgery. No patients with urinary leakage occurred. The mean follow-up were 21 months (range 3-60 months) . No cases with ureteral stricture were observed. Slight urine reflux was found in two patients, of whom obvious hydronephrosis and renal damage were not found. Conclusion Immediate ureterovesical reimplantation under laparoscopy is a feasible, safe and minimal invasive method for treatment of ureteral ruputure during laparoscopic hysterectomy.

Objective To evaluate the feasibility and safety of a delivery pathway for the performance of percutaneous left atrial appendage (LAA) occlusion in experimental canine models. Methods Transseptal puncture was performed via femoral vein approach under fluoroscopic and angiographic guidance in 12 experimental dogs. A pigtail catheter was advanced into the left atrium (LA), which was followed by LA angiography. The diameters of the neck of LAA were measured on LAA angiogram obtained in appropriate projection. After the delivery sheath was advanced along the wire into LA, a pigtail catheter was inserted into the ostium of the LAA and the sheath was then advanced over the pigtail into the LAA. LAA angiography was then performed through the delivery sheath to confirm the position of the delivery sheath. One hour after the procedure both electrocardiography (ECG) and transthoracic echocardiography (TTE) were carried out in five dogs to check the results, immediately after which the five dogs were sacrificed to macroscopically observe the damages of the puncture site of inter-atrial septum as well as inside the LA and LAA. One hour and 2 weeks after the procedure TTE was conducted in the remaining 7 dogs and these dogs were followed up for one month. Results One dog died of pericardial tamponade during the operation. In 8 dogs the LAA was clearly displayed in the projection position of right anterior oblique (RAO) 30°/cranial (CRA) 20°,while in 3 dogs the LAA was well visualized in the projection position of RAO 30° , and in one dog in the projection position of RAO 30°/caudal (CAU) 20°. The diameter of LAA neck was (13.6 ± 5.2) mm. The delivery sheath was safely advanced into the LAA along the pigtail catheter in all dogs, and no air embolism, thrombus or pericardial tamponade occurred. Hematoma at puncture point of groin occurred in 2 dogs, which was absorbed through pressure dressing. Macroscopic examination of the heart performed immediately after the operation showed that no bloody pericardial effusion was found, and mild hematoma at posterior wall of LA was seen in one dog and mild damage of the upper-margin intima of LAA was noted in 2 dogs. The mean fluoroscopy time was (10.1 ± 2.5) minutes and the mean operation time was (58 ± 12) minutes. TEE showed no pericardial effusion 2 weeks after the procedure. During the follow-up period of one month no sudden death, stroke or infection occurred. Conclusion This method of placing the delivery sheath into the LAA is clinically safe and effective, and it can reliably establish a pathway to advance the LAA occluder into LAA.

Objective To study the relationship between serum UA level and early outcome in acute ischemic stroke (AIS) patients. Methods Four hundred and twenty-one AIS patients admitted to the First Affiliated Hospital of Xi'an Jiaotong University from March 2015 to March 2016 were divided into good outcome group (n = 232) and poor outcome group (n = 189) according to their modified Rankin scale (mRS) score. Their demographic data, risk factors for vascular disease, laboratory testing parameters, imaging and clinical data and NIHSS score were recorded and compared. The relationship between serum UA level and early poor outcome in AIS patients was analyzed by unconditioned logistic regression analysis. Results The incidence of AF and cerebral infarction in the territory of anterior cerebral artey and middle cerebral artery, SBP, serum TC,LDL and urea levels,NIHSS and mRS score, and mortality were significantly higher and the hospital stay time was significantly longer while the serum UA level and GCS score were significantly lower in poor outcome group than in good outcome group (P＜0.05,P＜0.01). Unconditioned logistic regression analysis showed that SBP,NIHSS score and serum UA level were the major risk factors for the early poor outcome in AIS patients (OR = l.017,95％CI:1.003-1.031,P = 0.018;OR = 1.274,95％CI:1.178-1.378,P=0.000;OR=0.993,95％CI:0.989-0.996, P = 0.000). Conclusion The low serum UA level is related with the early poor outcome in AIS patients.

OBJECTIVESTo investigate a survey about acceptance of central slaughtering of live poultry in residents of Guangzhou.

METHODSWe conducted a telephone survey by sampling residents with fixed-line telephone and with normal hearing, whose age is more than 15 years, by Mitofsky-Waksberg two-stage method during Jan 6(th) to 8(th), 2014. 358 residents finished the telephone questionnaire by 12 320 health hot line. We investigated the acceptance rate of city-wide central slaughtering permanently. We compared the difference between the respondents and the 2010 Guangzhou census data by Cohen's effect sizes (w) and weighted by population age and sex. We used χ(2) test to compare the acceptance rate of central slaughtering in residents with different characteristic. We used multiple logistic regression analysis to analyze the factors.

RESULTSThe difference in gender and age was small between respondents and the 2010 Guangzhou census data (w value was 0.13, 0.28, respectively), but that in education and marital status was large (w value was 0.52, 0.31, respectively). 49.0% (95% CI: 43.7%-54.3%) accept city-wide central slaughtering permanently. The acceptance rate of city-wide central slaughtering permanently in those who bought fresh, chilled and frozen poultry in their family in previous year was 54.3% (133/245), 60.0% (57/95) and 59.8% (49/82), respectively. It was more than those who didn't buy fresh, chilled and frozen poultry (38.1% (43/113), 44.9% (118/263) and 45.7% (126/276); χ(2) values were 8.15, 6.40 and 5.03; P values were 0.004, 0.011 and 0.025, respectively). The acceptance rate of city-wide central slaughtering permanently in those who deem fresh poultry taste better than live poultry was 64.9% (24/38). It more than those who deem not (47.0%, 151/320) (χ(2) = 4.22, 6.02, P = 0.040, 0.014, respectively). The acceptance rate of city-wide central slaughtering permanently in the male (OR = 2.68, 95% CI: 1.64-4.37) and those who deem getting sick due to buying live birds from LPM (OR = 1.72, 95% CI: 1.05-2.82), who can accept only fresh poultry carcass supply (OR = 2.39, 95% CI: 1.33-4.30), Who bought live poultry in their family in previous year (OR = 0.29, 95% CI: 0.11-0.74), who will decrease the consumption after ban on live poultry sale (OR = 0.50, 95% CI: 0.30-0.83) was 58.6% (109/186), 59.0% (92/156), 60.7% (139/230), 44.9% (132/295), 36.6% (68/186), respectively.

CONCLUSIONIn the early stage of avian influenza A(H7N9) epidemic in Guangzhou, the rate of acceptance of central slaughtering permanently in residents was not so high. Who deem getting sick due to buying live birds from LPM, who could accept only fresh poultry carcass supply and the male more accept city-wide central slaughtering permanently.

Animals ; Attitude to Health ; Birds ; Epidemics ; Humans ; Influenza A Virus, H7N9 Subtype ; Influenza in Birds ; Influenza, Human ; Male ; Meat-Packing Industry ; Poultry ; Surveys and Questionnaires