ObjectiveEvidence-based medicine was used to make an individualized treatment plan for rare solitary splenic metastasis of an ovarian carcinoma in a female patient. MethodsAfter clinical problems were discovered, evidence was collected from the Cochrane Library, PubMed, EMBASE, ScienceDirect, CNKI and Wanfang databases according to our predefined search strategy. The search was done until October, 2009. The validity of the evidence was assessed. The evidence, combined with the doctor experience and the patient value, was applied to the patient. ResultsA total of 15 English literature and 11 Chinese literature were identified. A rational treatment plan was made upon a critical evaluation of the data. There were 5 review articles in PubMed, EMBASE and ScienceDirect databases and no RCT in the Cochrane Library. And there was one retrospective cohort study inCNKI and Wanfang databases. The others were all case reports. After 20 month follow-up, the treatment protocol was proven correct. Conclusion The patient has rare disease, and the evidence intension for the evidence-based medicine was low. It can not present the efficacy of the evidence-based medicine treatment sufficiently, though it could provide some suggestion for rare disease which was lack of large RCT for evidence-based Methods .

Objective To evaluate the radical effect and prognosis of laparoscopic resection for colorectal cancer in China.Methods Articles of non.randomized comparative studies(NRCs)of laparoscopic resection and open Burgery for colorectal cancer which were published before October 2007 were retrieved,and correlated indexeswere extraeted for meta.analysis.Results The mean quality score of the 14 articles selected was 18.92±1.27.The basic characteristics of patients in laparoscopic resection group were similar to those in open surgery group.Compared with open surgery group,the incised length of the intestine in the laparoscopic resection group was shorter by 0.66 cm.and the distance between distal margin of resection and tumor was farther by 0.26 cm.The 2-year survival rate of patients in laparoscopic group Was 1.67 times higher than that of open surgery group.There was no significant difference upon follow-up rate,tumor diameter,number of resected lymph nodes,local recurrence rate and distal metastasis rate between the 2 groups.Conclusions The results of meta-analysis show that laparoscopic resection has the sanle radical effect as open surgery for colorectal cancer.but the 2-year survival rate of patients treated by laparoscopic resection is comparatively higher.

Objective To investigate the effects of growth-discordant twin pregnancies on neonatal thyroid stimulating hormone (TSH) level and congenital hypothyroidism (CH).Methods A total of 3 444 live-birth twin neonates born between January 1,2012 and December 30,2014 in Ningbo City were enrolled.Blood samples via heel puncture were collected and tested.Incidence of CH in singleton and twin neonates was compared.Deviation of birth weight larger than 25％ in twin neonates was set as the criteria for discordant growth.TSH and 17 α-hydroxylase levels in CH twins and normal twins,with or without discordant growth,were compared.Chi-square and non-parametric statistics were performed for data analysis.Results The incidence of CH in twin neonates was 0.56％ (19/3 444),higher than that in singleton neonates [0.09％ (203/225 712),x2=76.225,P＜0.01].Among nineteen CH twins,CH occurred in both twins in eight cases (four twins) and in one of the twins in eleven cases.The gestational age at birth in the eight CH twins were less than 37 weeks,with four males and four females;five were low birth weight infants;one twin were dichorionic,and three twins were monochorionic.In the eleven cases of CH occurring in one of the twins,the gestational age was less than 37 weeks in nine cases,eight were low birth weight infants,six were male and five female;seven were monochorionic and four were dichoronic twins.Five cases of temporary hypothyroidism were all low birth weight infants among the growth-discordant twins.CH cases in growth-discordant group had lower birth weight than their normal twins [M(P25-P75),2 100 (1 800-2 600) vs 2 770 (2 530-2 960) g,Z=4.369],and a higher TSH level [15.4 (11.8-18.5) vs 6.4 (4.8-7.9) mU/L,Z=6.339] (both P＜0.05).In normal twins with or without discordant growth,the neonates with a lower birth weight had a higher TSH level [3.6(2.5-4.7) vs 2.4(1.8-2.9) mU/L,Z=0.962] in weight consistent group,compared with 6.0(4.4-7.8) vs 3.4(1.9-4.1) mU/L in weight inconsistent group (Z=4.369),both P＜0.05.Conclusions In the growth-discordant twins,neonates with a lower birth weight have a higher TSH level and a higher risk of temporary hypothyroidism.

Objective To establish a double-antibody sandwich ELISA for the rapid detection of shiga toxin typeⅡ ( StxⅡ) in shiga toxin-producing Escherichia coli ( STEC) infection. Methods A pool of murine hybridomas was used to screen out the optimal antibody pair for the establishment of double-anti-body sandwich ELISA. The established ELISA system was used to detect StxⅡin the culture supernatants of 16 clinical strains of STEC. Specificity and sensitivity of the established ELISA system were also evaluated. Results Two antibodies, S2D8 and S2C6, were successfully screened out, based on which the double-anti-body sandwich ELISA was set up. StxⅡand its variants rather than StxⅠwas detected in the culture super-natants of STEC with a lowest detection limit of 4 ng/ml. Its performance was consistent with that of commer-cial colloidal gold test kit, indicating the characteristics of good specificity and sensitivity. Conclusion The S2D8/S2C6-based ELISA laid a foundation for researches which designates the shiga toxin as a potential can-didate on the diagnosis and therapy of STEC infection.

Objective The feasibility of predicting the B-cell epitopes of human Neutrophil Gelatinase-Associated Lipocalin (NGAL) was discussed by applicating bioinformatics technology.Linear epitope molecules that have diagnostic value were screened and these recombinant linear multi-epitope peptides were constructed,and expressed.The immunogenicity of the recombinant linear multi-epitope peptides were also identified.Methods NGAL amino acid sequence was got from GenBank in the Department of Clinical Laboratory of the Second Affiliated Hospital of Nanjing Medical University in July 2015,the Predicted,ABCpred,BepiPred,BcePred,and Lasergene softwares were used to predict the linear B cell epitope prediction.The predict epitopes were constructed and prokaryotic expressed,and then the single epitope antigens which could reacted with commercially available polyclonal NGAL antibody were screened out by Western blot.Finally,the multi-epitope peptide was constructed,expressed,and identified through immunoreactions.Results Eight possible epitopes were obtained after prediction.pET32a-N1-N8 prokaryotic expression vector were used to express the predict epitopes.After purification and Western blot analysis,three of the epitopes have strong antigenicity,and then a soluble fusion protein was expressed and obtained from the multi-epitope prokaryotic expression vector pET22b-Ngal_MEP1.The fusion protein was successfully purified by Ni2 + affinity column.Western blot analysis showed that the fusion protein had a strong antigenicity.Conclusions The constructed multi-epitope linear NGAL antigen peptides can obtain high soluble expression in prokaryotic expression system,and have a strong immunoreactivity,which can be used in subsequent antibody preparation.

Objective To investigate the feasibility and technique in laparoscopic cholecystectomy and exploration of common bile duct (CBD) in patients who have previous subtotal gastrectomy.Methods From January 2001 to October 2010,22 patients who had a history of subtotal gastrectomy received laparoscopic cholecystectomy and CBD exploration in our hospital.In addition,these patients received cholangioscopic lithotomy,electrohydraulic lithotripsy,T-tube drainage,or primary suturing of the CBD wound.Results The procedures were completed successfully in 18 patients.In 4 patients,conversion to laparotomy was necessary because of multiple stones in a stenosed CBD (n=1),severely congested CBD wall with edema (n=1),CBD carcinoma associated with impacted stones (n=1) or Mirzzi syndrome (n =1).T tube drainage was employed in 16 patients,and primary suturing in 2 patients at the end of the operation.There was no hemorrhage,biliary leakage or abdominal infection.At a mean follow-up of 13 (range 3 to 60) months there was no residual stone,biliary stenosis or cholangitis.In 3 patients Stone recurred,and another 3 patients were lost to follow-up.Conclusions A history of subtotal gastrectomy is not a contradiction to laparoscopic cholecystectomy and CBD exploration.With choledocoscopy,surgery can be safe and feasible even for patients with previous gastectomy.

Objective This study explored the expression of polyclonal protein SUZ12 in patients with intrahepatic cholangiocarcinoma(ICC),and its role in predicting the survival and treatment of ICC patients.Methods The expression of SUZ12 and p16INK4a was detected by immunohistochemical assay in 207 liver tissue samples including ICC patients,BilIN-1,-2,-3 and non-tumor-like cholangiocarcinoma.The expression of these proteins was assessed to be related to the pathological characteristics of the ICC patients receiving chemotherapy and the outcome of survival as well as the subsequent chemotherapy response.Results The expression level of SUZ12 was gradually increased from non-neoplastic bile duct tissue to BilIN-1,-2,-3 and ICC.The expression of p16INK4a protein was expressed in non-neoplastic-like cholangiocarcinoma,but it decreased gradually in BilIN-1,-2,-3 and ICC tissues.SUZ12 expression was associated with undifferentiated ICC,lymph node metastasis and advanced cancer.Kaplan-Meier curve analysis showed that ICC patients with high expression of SUZ12 had a significant reduction in overall survival and disease-free survival in comparison with ICC patients with the low expression of SUZ12.SUZ12 expression was significantly associated with overall survival of patients receiving adjuvant gemcitabine-based chemotherapy(AGC).Conclusion SUZ12 expression is able to predict the overall survival and disease-free survival of ICC patients with adjuvant gemcitabine-based chemotherapy.

The dynamic changes of immunoreactive dynorphin1-13 (ir-Dyn1-13) contents in 18 burned patients were studied by radioimmunoassay. It was found that ir-Dyn began to increase gradually at the onset of burn and remained at a relatively high levels by the time of preliminary wound healing. Plasma ir-Dyn increased gradually after bum and decreased abruptly before death in one severely burned patient who died of respiratory failure at the third week. The results suggest that Dyn may have a protective effect on the burned patients.

Objective To study the academic level of randomized control trials(RCTs)published from 2004 to 2009 in Chinese Journal of Hepatobiliary Surgery.Methods Published work about RCTs in 72 issues of Chinese Journal of Hepatobiliary Surgery was searched by finding relevant articles published during 2004-2009.Correlated indexes including literature received time, literature publishing time, the total number of citations, number of citations in Chinese, number of citations in English, research funding situations, the number of authors, article affiliations in all RCTs, together with single(or multiple)center study situation, positive conclusions, the starting time, the end time,and number of patients recruited in clinical RCTs were extracted for aggregate analysis.Results During the past six years, a total of 135 articles of RCT were published in Chinese Journal of Hepatobiliary Surgery, accounting for 8.7% in all clinical studies.The average number of RCTs in each issue was 1.88.The average time for publishing time delay was 450 days.The total number of citations in RCTs was 1252 and the average number of RCTs for citations was 9.3.53 RCT studies were financed, accounting for 39% of the total number of research papers.Beijing Friendship Hospital attached to the Capital University of Medical Sciences had published 8 papers of RCT and it is the institution of publishing the largest number of such papers in the journal.The number of multi-center study for RCTs was one and all RCT studies had significant conclusions.The average research duraton in RCTs was 45 months.The total number of patients in 33 clinical RCTs was 2705 and the average number of patients recruited in each RCT was 82.Conclusion Chinese Journal of Hepatobiliary Surgery put emphasis on the strongest evidence and the most valuable RCT researches that have great significance in the field of domestic hepatobiliary surgery for clinical treatment and experimental research.

OBJECTIVETo investigate the effect of antibacterial peptide LL-37 on the integrity of Acinetobacter baumannii biofilm.

METHODSA model of Acinetobacter baumannii biofilm in vitro was constructed by plates and crystal violet staining method, and the minimal inhibitory concentration of LL-37 was determined by broth dilution. The biofilm morphology was observed by scanning electron microscopy and biofilm formation was analyzed by the crystal violet staining of the adherent biofilm in the presence of antibacterial peptide LL-37.

RESULTSIn the Acinetobacter baumannii biofilm model, the minimal inhibitory concentration of LL-37 was 64 µg/ml; LL-37 caused structural damage of the biofilm at a low concentration of 2.5 µg/ml. The biofilm was decreased gradually as the concentration of LL-37 increased.

CONCLUSIONLL-37 even at a concentration far below its minimal inhibitory concentration can cause structural damage of Acinetobacter baumannii biofilm in vitro.