Retroviral vectors are the most used gene delivery vehicle in human gene therapy.When the therapeutic gene is packagedfrom producer cell line, one has to determined whether thereplication-competent retrovirus(RCR) is being generated. In this study, two methods, namely the marker rescue assay and RT/PCR, were employed to detect RCR in the cells. While the marker rescue assay can detect RCR with a limit at 6?102CFU/ml, RT/PCR can be used to detect RCR at as low as lCFU/103ml. Combining the specificity of the marker rescue assay and the sensitivity of RT/PCR, both assays together should serve as an adequate test for detecting the generation of RCR in retroviral producer cell lines.

OBJECTIVE To type Staphylococcus epidermidis and analyze its polymorphism by randomly amplified polymorphic DNA(RAPD),and provide the method for analysis of homogeneity of S.epidermidis.METHODS Twenty one collected strains from ophthalmology ward were analyzed using random primer RAPD1 with RAPD.RESULTS On the basis of bands,difference of clinical strains was analyzed.Twenty one strians of S.epidermidis were divided into 5 patterns.Among them type Ⅰ taken 71 percent(15/21).CONCLUSIONS Typing of S.epidermidis by RAPD at molecular level is very rapid,easy and reliable.It can apply to track pathogens of the hospital and study epidemiology of the bacteria.

0. 05). Conclusion: The HCAPl can be expressed in both normal peripheral white blood cells and the leukemic cells, and there is no difference in the protein levels between them.

Aim To study the effects of Shenfu soup onthe ISO -induced myocardial ischemia injury model ofrats and the influence of apelin level changes on myocardial ischemia.Methods The myocardial ischemiainjury model of rats was established by the subcutaneous injection of ISO.For 1 5 days,rats in Shenfugroups were given by gavage 3,6 and 1 2 g · kg -1Shenfu soup and rats in SF injection group were given6.67 g·kg -1 Shenfu injection,rats in sham group andmodel group were given the same volume of distilledwater.Rats were subcutaneously multipoint injectedISO for 5 days on the eleventh administrating day.Theeffects of Shenfu soup on myocardial morphology,serum myocardial enzyme levels,apelin mRNA level inserum and apelin protein level in myocardial tissue,and relationship between myocardial enzyme and apelinexpression were evaluated. Results Shenfu soupcould reduce the degree of myocardial tissue necrosisand compared with model group,CK values and LDHvalues of rats in middleand highdose groups were significantly lower (P <0.05 or P <0.01 ).However,the apelin mRNA level in myocardial tissue and apelinprotein level in serum of the rats in middleand highdose groups compared with model group had increased(P <0.05,P <0.01 ).Meanwhile,serum levels ofapelin and the expression of myocardial enzyme CK andLDH were negatively correlated.Conclusion Shenfusoup has a protective effect on myocardial ischemia injury in rats induced by ISO and the mechanism is involved with the promotion of apelin mRNA and proteinexpression,the inhibition of myocardial enzyme production and the improvement of myocardial ischemia.

Objective To determine the expression profile of serum microRNAs(miRNAs) in early-onset familial Alzheimer's disease (EO-FAD) patients. methods miRNA microarrays were performed to detect the expression profile of serum miRNAs in 2 cases of EO-FAD patients,2 cases of EO-FAD carriers and 2 cases of normal controls.Preliminary bioinformatic analysis was conducted. Result sIt was found that 21 miRNAs were up-regulated and 22 miRNAs were down-regulated in serum of EO-FAD patients,the differences were statistically significant(P<0.05).miR-5704(P=0.0002),miR-4639-3p(P=0.0195),miR-107(P=0.0204) were markedly up-regulated,miR-5572(P=0.0008),miR-204-3p(P=0.0014),miR-542-5p(P=0.0106) and miR-155-5p(P=0.0240) were markedly down-regulated.Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis suggested that the dysregulated miRNAs may be involved in the mechanism of EO-FAD by affecting neurotrophin signaling pathway.Conclusion miR-5704,miR-4639-3p,miR-107,miR-5572,miR-204-3p,miR-542-5p and miR-155-5p may be used as potential biomarkers of EO-FAD,and involved in the mechanism of EO-FAD by affecting neurotrophin signaling pathway.

Objective: This study aimed to investigate the inhibitory effects of Brucea javanica oil oral emulsion (BJOOE) on primary liver cancer induced by diethylnitrosamine (DEN). Methods:Rats were randomly divided into the control group, model group, and BJOOE group. Rats were given free access to water. DEN was administered intragastrically to induce liver cancer in rats. Five weeks later, rats were intragastrically administered with BJOOE for five times per week. The rats were killed after 14 weeks. Abdominal aortic blood samples were collected. The contents of ALT, AST, ALP, γ-GT, and AFP of serum were detected by an automatic biochemical analyzer. The liver index, spleen index, thymus index, and changes in liver cancer nodules of the surface were observed in rats. Changes in the number of liver cancer nodules of the surface were detected by imaging. Results:Compared with the control group, the liver index, spleen index, and number of nodules of the model group significantly increased, whereas the thymus index significantly decreased (P<0.01). The levels of ALT, AFP, AST, ALP, andγ-GT of serum in the model group were significantly higher than those in the control group (P<0.01). Compared with the model group, BJOOE significantly reduced the liver index, spleen index, and number of cancer nodules, but increased the thymus index in the liver of rats with cancer (P<0.01). The levels of ALT, AFP, AST, ALP, andγ-GT of serum in rats with hepatic carcinoma significantly improved (P<0.01 or P<0.05). Conclusion:BJOOE could inhibit primary liver cancer, and the underlying mechanisms are complex.

Objective To explore the early predictive factors of intestinal necrosis in patients with acute superior mesenteric arterial occlusive disease and its significance for the decision of exploratory laparotomy.Methods This retrospective study enrolled 29 patients diagnosed with acute superior mesenteric artery embolism or thrombosis in Peking University People's Hospital between July 1995 and June 2015.Results 12 patients developed intestinal necrosis.Patients with intestinal necrosis had a poorer prognosis than those who did not develop intestinal necrosis (x2 =14.867,P =0.000).In univariate analysis,the early predictive factors for intestinal necrosis were D-Dimer ≥ 600 μg/L (x2 =11.455,P =0.002),INR≥1.2 (x2 =3.948,P =0.047),pH values ＜7.4 (x2 =8.191,P =0.004),BE ＜ -1.0 mmol/L (x2 =8.191,P =0.004),blood lactate ≥ 2.2 mmol/L(x2 =7.535,P =0.006),BUN ≥ 6 mmol/L (x2 =10.076,P =0.002),CK ≥ 80 U/L (x2 =8.191,P =0.004),LDH ≥ 210 U/L (x2 =13.079,P=0.000),AST ≥25 U/L (x2 =10.076,P =0.002),SIRS (x2 =10.076,P =0.002).Multivariate logistic regression analysis found no independent predictive factors of intestinal necrosis in patients with acute superior mesenteric arterial occlusive diseases.Conclusion Intestinal necrosis in acute mesenteric arterial occlusive diseases indicates a poor prognosis.Coagulation abnormalities,liver or kidney dysfunction,metabolic acidosis and SIRS necessitates an immediate exploration.

Objective To analyze the risk factors for pancreatic fistula after pancreaticoduodenectomy.Methods Clinical data of 351 patients who underwent pancreaticoduodenectomy in our hospital from Jan 2001 to Dec 2015 were retrospectively analyzed.Results The overall incidence of pancreatic fistula was 20.2％ (71/351),with grade A in 34 (9.7％) patients,grade B in 25 (7.1％),and grade C in 12 (3.4％).Postoperative hospital stay was significantly prolonged in patients with grade B and C pancreatic fistula.Multivariate analysis using Logistic regression identified three variables as independent factors associated with pancreatic fistula,namely,BMI (OR =2.281),preoperative total bilirubin level (OR =2.180) and soft pancreatic texture (OR =3.653).Preoperative total bilirubin level (OR =2.684),soft pancreatic texture (OR =5.128) and postoperative hemorrhage (OR =9.030) were independent risk factors of grade B and C pancreatic fistula after pancreaticoduodenectomy.Conclusions The incidence of pancreatic fistula after pancreaticoduodenectomy is still high.Pancreatic fistula prolongs the postoperative hospital stay significantly.The incidence of grade B and C pancreatic fistula could be reduced by avoiding postoperative hemorrhage.

Objective To analyze the cause and treatment of the postoperative gastrointestinal hemorrhage after pancreaticoduodenectomy(PD).Methods Clinical data of 213 patients who underwent PD in our hospital from January 1996 to December 2011 and 2 patients who suffered from gastrointestinal hemorrhage after PD transferred to our hospital from other hospitals were retrospectively analyzed.Results The incidence of postoperative gastrointestinal hemorrhage was 8.5％ (18/213),the mortality rate of which was 22.2％ (4/18).Among the twenty patients with postoperative gastrointestinal hemorrhage (including the 2 patients transferred from other hospitals),stress ulcer was the most common reason of gastrointestinal hemorrhage (11/20,55 ％).There were 8 patients suffering from mild hemorrhage who were treated by medications.Seven of 12 patients who suffered from severe hemorrhage underwent reoperation.Univariate analysis showed that duration of operation above 420 min (x2 =3.976,P =0.046) and volume of intraoperative blood loss above 1200 ml (x2 =6.753,P =0.009) were significantly associated with postoperative stress ulcer bleeding.Multivariate logistic regression analysis showed that intraoperative blood loss was the only independent factor associated with postoperative stress ulcer bleeding (OR =5.677,P =0.035).Conclusion Gastrointestinal hemorrhage is one of the common complications after PD.The incidence of stress ulcer bleeding could be reduced by skillful operation and decreasing intraoperative blood loss.Operation should be used properly according to the cause and location of bleeding if hemorrhage could not be stopped by medications.

Objective To investigate the expression of Toll-like receptors(TLRs) in condyloma acuminatum(CA) lesions and their possible roles in the pathogenesis of CA. Methods The expressions of TLR1-10 mRNA level in the lesions of CA and in the cervix scrape cells from the patients with human papillomavirus(HPV) negative chronic cervicitis were detected by real-time quantitative fluorescent PCR. HPV typing was detected by HPV GenoArray test kit. Results Low-risk HPV type 6 and type 11 were the most prevalent types in the forty CA cases with positive rate of 77.5% and 55% respectively. 55% CA patients were found infected with more than two types of HPV. 35% CA patients were concurrently infected with high-risk HPV. The expressions of TLR3, 7, 8 mRNA were higher than other TLRs and the expression of TLR9 mRNA was lower than others in the lesions of CA. No significant differences of the TLR1-10 mRNA levels were found between HPV6 and HPV11 positive CA lesions, so did it between low-risk and high-risk HPV concurrent infected CA lesions. The expressions of TLR1-3, TLR5-8, TLR10 mRNA, especially TLR2, TLR7 and TLR8 in the lesions of CA were significantly higher than that in cervix scrape cells of HPV negative chronic cervicitis. There were no significant differences of TLR4 and TLR9 mRNA levels between the two groups. Conclusion There were higher expressions of some TLRs (3, 7, 8) and lower expression of TLR9 in the lesions of CA. Compared with HPV negative chronic cervicitis, the expressions of TLR1-3, TLR5-8, TLR10 mRNA in the lesions of CA were up-regulated. The expression profile of TLRs in different type of HPV infected CA lesions had no significant differences. Our results suggested that the expression profile of TLRs in CA may be associated with the HPV infection. Whether it was associated with the immune escape mechanism and persistent infection of HPV need further demonstration.