Objective To explore the risk factors and prevention measures for anastomotic leakage after rectal radical resection.Methods The clinical data of 404 patients with rectal radical resection were analyzed retrospectively and the risk factors for anastomotic leakage were analyzed.Results Thirty-one patients (7.67％,31/404) were subjected to anastomotic leakage.The mean leakage time was 6.5 (3-14) d postoperatively.The muhivariate Logistic regression analysis showed that preoperative hemoglobin (OR =3.023,95％ CI:1.101-8.303,P=0.031 8),tumor size (OR =2.543,95％ CI:1.075-6.018,P=0.033 7) and tumor distance from anal verge (OR =3.160,95％ CI:1.387-7.199,P=0.006 2) were the risk factors for anastomotic leakage.Conclusions Preoperative hemoglobin,tumor size and tumor distance from anal verge are significant factors for anastomotic leakage.Therefore correction of anemia,improvement of surgical technique and suitable use of preventive diversion stoma ane all benefit for prevention of anastomotic leakage after rectal radical resection.

Objective To observe the curative effect of using artificial liver bilirubin specific adsorption for treatment of patients with hyperbilirubinemia and its effect on nursing.Methods A prospective study was conducted, 146 patients with hyperbilirubinemia admitted to Mianyang Central Hospital from January 2015 to December 2016 were enrolled, and they were divided into an observation group (77 cases) and a control group (69 cases) according to random number table method. The observation group was treated by medical treatment and the artificial specific liver bilirubin adsorption, while the control group only treated by medical therapy. The changes of levels of liver function indexes alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), direct bilirubin (DBil) before and after treatment and clinical curative effect were observed in the two groups.Results Before treatment, there were no statistical significant differences in levels of the ALT, AST, TBil, DBil between the two groups (allP > 0.05), after treatment, the above indexes were significantly decreased compared to those before treatment, and the degrees of decrease in observation group were more obvious than those in control group [ALT (U/L): 341±42 vs. 455±37, AST (U/L): 120±35 vs. 197±37, TBil (μmol/L): 185.4±20.6 vs. 302.6±30.6, DBil (μmol/L): 42.6±10.8 vs. 87.5±11.6, allP < 0.05]. The total effective rate in observation group was obviously higher than that of control group [62.3% (48/77) vs. 40.6% (28/69),P < 0.05].Conclusions Based on liver protection, symptomatic and supportive medical treatment, using artificial liver bilirubin specific adsorption for treatment of patients with hyperbilirubinemia is safe and effective, and in addition, close observation and careful nursing is beneficial to the reduction of incidence of complications and elevation of therapeutic efficiency.

The supercondensed DNA, a special kind of topological structure of plasmid DNA, was firstly found in E. coli SD108(topA+ gyrB225). Now, this structure is also found in E. coli DM800(topA- gyrB225). The result indicates that the formation of supercondensed DNA is related with decrease of the activity of gyrase in vivo. Topoisomerase Ⅳ was proved to relax the supercondensed DNA completely in vitro, which suggested that the supercondensed DNA and the supercoiled DNA could transform to each other in cells. The supercondensed DNA samples were analyzed by atomic force microscopy and compared to supercoiled DNA. The results showed that the length of supercondensed DNA decreased about 30% and the width and height of double-strand increased about 60%, which indicates that the structure of double-strand of supercondensed DNA is much more similar to A-DNA than B-DNA. The results also showed that chloroquine intercalation did not change the supercoiling level of supercondensed DNA, but made it knot and compact.

Objective To establish the reference intervals of neuron specific enolase (NSE) in tumor markers in cerebrospinal fluid .Methods According to National Committee for Clinical Laboratory Standards document C 28‐A2 ,120 samples were collected to establish reference intervals .Then ,the established intervals were evaluated according to China National Accreditation Service for Conformity Assessment document CL38 :2012 .Results The biological reference interval of NSE in cerebrospinal fluid was 0-3 .14 ng/mL .There was no significant correlation between cerebrospinal fluid NSE level and age ,sex (P> 0 .05) .Conclusion Method used in this study could be ensured reliable ,accurate ,scientific and practical ,desirable for clinical requirement and with great poten‐tial for clinical application .

OBJECTIVETo investigate the genotypes , allele frequencies and dynamic distribution on resistance associated esterase genes of Culex pipiens complex in Hangzhou.

METHODSThe PCR-restriction fragment length polymorphism (PCR-RFLP) assay was applied to type the resistance associated esterase genes, and dynamic surveillance on frequencies of the resistance associated esterase gene of natural population of Culex pipiens complex in Hangzhou during 2003-2005, and phenotype of the resistance associated esterase genes were detected by esterase starch gel electrophoresis technique.

RESULTSThe PCR-RFLP assay of esterase allele genes for three consecutive years disclosed four esterase genotypes, namely, the world-wide highly active homozygous Est beta 1(1) (50%-54%), homozygous Est beta 2 (29%-34%), heterozygous Est beta 1(1)/beta 2 (5%-10%) and Est beta N (3.13%) of a new homozygous genotype. The research of the resistance associated esterase genes phenotype in natural population of Culex pipiens complex in Hangzhou in 2005 with esterase starch gel electrophoresis technique revealed four major types, namely, Est beta 1(1) (61%), Est alpha 2/beta 2 (12%), Est alpha 8/beta 8 (7%) and sensitive phenotype (29%).

CONCLUSIONThere should be various resistance associated esterase genotypes in natural population of Culex pipiens complex in Hangzhou. During the period of 2003-2005, Est beta 1(1) was the major type; Est alpha 2/beta 2 was the second. Est beta N was a new esterase genotype detected in 2005 only with a mere percentage of 3.13%. As for its resistance to the new insecticide, a follow-up study should be needed. The molecular typing of the amplified esterase gene should be consistent with the resistance associated esterase genes phenotype.

Alleles ; Animals ; China ; Culex ; genetics ; physiology ; Esterases ; analysis ; genetics ; Gene Frequency ; Genotype ; Insecticide Resistance ; genetics ; Phenotype

OBJECTIVETo prepare the armored RNA containing M gene of influenza H3N2.

METHODSThe vector pAR-1 was constructed from expression vector pET30b in which the bacteriophage MS2 DNA fragment, containing the genes for maturase and coat protein and the pac site, was inserted. The M gene fragment of influenza A was inserted into the HindIII site downstream of the pac site on the pAR-1, which formed a new recombinant plasmid pAR-2. After the prokaryotic expression was carried out, armored RNA AR-2 containing M gene was obtained. AR-2 was purified, and then was quantified by real time RT-PCR. Moreover, the stability of AR-2 was checked.

RESULTSAR-2 was expressed successfully. AR-2 remained stable under various storage environments. Approximately 8.9 x 10(11) copies of AR-2 particles can be purified from one milliliter of culture.

CONCLUSIONIt showed that AR-2 was stable and RNase-resistant, which, as a virus surrogate, would be used as RT-PCR standards, controls and training or proficiency samples.

Influenza A Virus, H3N2 Subtype ; genetics ; Plasmids ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; standards ; Viral Matrix Proteins ; genetics

OBJECTIVETo investigate the heterozygous genotype and molecular characteristics of Organophosphorus resistance associated with heterozygous Estbeta2 of esterase B2 gene from natural population of Culex pipiens complex.

METHODSGenomic DNA was extracted from natural populations of Culex pipiens complex in Hangzhou. The PCR-restriction fragment length polymorphism (PCR-RFLP) assay was applied to type the resistance associated esterase gene. Estbeta2 of esterase B2 gene was identified by PCR-RFLP, and the genotyping for heterozygous Estbeta2 was carried out after restriction enzyme digesting by Bfm I endonuclease.

RESULTSThe DNA was isolated from 207 Culex pipiens respectively, while 156 PCR samples showed positive and the positive rate was 75.36% (156/207). The PCR-RFLP assay of esterase B2 gene revealed that the Estbeta2 was accounted about 28.20% (44/156) in 156 positive samples. There were two genotypes identified, namely homozygous Estbeta2 (90.90%, 30/33) and heterozygous Estbeta2 (9%, 3/33), heterozygous Estbeta2 was in existence of a hybrid form as which combined with Estbeta2 and a subtype (Estbeta2/Estbeta2(1)).

CONCLUSIONHeterozygous Estbeta2 of Organophosphorus resistance associated with esterase genotype was determined in natural population of Culex pipiens, and a genotyping method was established.

Animals ; Culex ; enzymology ; genetics ; Genes, Insect ; Genotype ; Heterozygote ; Insecticide Resistance ; genetics ; Insecticides ; pharmacology ; Organophosphorus Compounds ; pharmacology ; Phenotype ; Serine Endopeptidases ; genetics

Purpose: Neoadjuvant chemotherapy is strongly recommended for advanced gastric cancer due to good local control and a high rate of R0 dissection with this strategy. Minimally invasive techniques such as laparoscopy-assisted or total laparoscopic approaches is becoming more and more acceptable in the treatment for gastric cancer. However, the safety and efficiency of total laparoscopic D2 gastrectomy (TLG) for advanced gastric cancer after neoadjuvant chemotherapy have not been well evaluated. Methods: A retrospective study in a single center from 2014 to 2016 was conducted. A total of 65 locally advanced gastric cancers were treated by laparoscopy-assisted gastrectomy (LAG) or TLG. Parameters which include operation time, blood loss, complications, hospital stay, 3-year overall survival, and 3-year disease-free survival were used for comparison. Results: The time of operation in the TLG group was shorter than in the LAG group (P = 0.013), blood loss was less (P = 0.002) and time to first flatus was shorter (P = 0.039) in the TLG group than that in the LLG group. Intraoperative and postoperative complications were comparable in both groups. No significant difference was found in 3-year overall and disease-free survival. Conclusion For patients with locally advanced gastric cancer after neoadjuvant chemotherapy, laparoscopic D2 gastrectomy can be considered as a safe and efficient alternative. A further multicenter prospective randomized controlled study is needed to elucidate the applicability of this technique for advanced gastric cancer.

OBJECTIVETo study the infection status and pathogenic features of human metapneumovirus (hMPV) among children with acute respiratory tract infection in Hangzhou.

METHODSA total of 372 children less than 14 years old with acute respiratory tract infections were recruited as subjects from the pediatric clinic or intensive care unit (ICU) of 3 hospitals in Hangzhou during November 2009 to January 2010, and November 2010 to January 2011. A total of 372 specimens were collected, including 351 respiratory swab, 9 nasopharyngeal aspirate material, 8 endotracheal aspirate material and 4 sputum. The total nucleic acid was then extracted from the specimens, and the nucleoprotein (N) gene of hMPV was amplified by RT-PCR, whose positive products were sequenced and analyzed. Africa green monkey kidney cells (Vero-E6) were applied to culture hMPV among the positive samples; meanwhile fluorescence quantitative RT-PCR was adopted to test other respiratory virus infection.

RESULTSOut of 372 patients, 42 (11.2%) were positive for N gene of hMPV. The positive rate of hMPV among boys was 11.5% (26/226), and correspondingly 10.9% (16/146) among girls. The difference showed no statistical significance (χ(2) = 0.026, P > 0.05). The youngest patient was only 2 month-old and the eldest patient was 14 years old. The median of the patients' age was 24 months. Fifteen positive samples amplified by RT-PCR were sequenced, and all turned out to be subtype B1; whose similarity to GD165 found in Guangdong was 98.1% - 99.5% and similarity to BJ1897 in Beijing was 87.8% - 89.2%. The co-infection rate between hMPV and other respiratory virus was 45.2% (19/42); most of which was between hMPV and respiratory syncytial virus, whose rate at 26.1% (11/42).

CONCLUSIONhMPV was the single genotype relevant with the acute respiratory tract infection disease among children in Hangzhou district; however, the co-infection with other respiratory virus did exist.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Genotype ; Humans ; Infant ; Male ; Metapneumovirus ; genetics ; isolation & purification ; Paramyxoviridae Infections ; epidemiology ; virology ; Respiratory Tract Infections ; epidemiology ; virology

OBJECTIVETo analyze the prognostic factors for chronic lymphocytic leukemia (CLL) with typical and atypical immunophenotype. The parameters analyzed included sex, age, Binet stages, absolute lymphocyte count (ALC), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein, CD38 expression and cytogenetic aberrations.

METHODSAccording to the clinical guideline and scoring system for CLL in Britain, among 77 patients, 61 patients with score 5 called typical immunophenotype CLL, 16 with score 4 or 3 were atypical immunophenotype CLL. Multiparameter flow cytometry was employed for immunophenotypic analysis in 77 CLL patients for CD5, CD19, CD23, FMC7, sIg, CD20, CD79b expression and ZAP-70 protein and CD38. IgVH mutation status was detected by multiplex RT-PCR and sequencing of the purified PCR amplification products. Fluorescence in situ hybridization (FISH) and a panel of probes were used to detect cytogenetic aberrations.

RESULTSThere was no significant difference between the two groups in sex, age, ZAP-70 and IgVH mutation status (P=0.398, P=0.189, P=0.268 and P=0.131, respectively). The incidence of ALC> or =50 x 10(9)/L, Binet B + C, CD38> or =30% in atypical CLL patients (43.8%, 87.5% and 43.8%, respectively) were higher than that in typical group (16.4%, 36.1% and 16.4%, respectively) (P=0.026, P<0.01 and P=0.026, respectively). The proportion of typical patients (26.8%) with a 13q14 deletion as sole abnormality was higher than that of atypical patients (7.6%), and that with deletion of 11q22 or 17p13 was lower than that of atypical patients (12.2% vs 46.2%) (P=0.022).

CONCLUSIONThere were obvious differences between the typical immunophenotype CLL and atypical CLL in ALC, Binet stages, CD38 expression level and cytogenetic aberrations.

ADP-ribosyl Cyclase 1 ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Immunophenotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; immunology ; metabolism ; Male ; Middle Aged ; Mutation ; Neoplasm Staging ; Prognosis ; ZAP-70 Protein-Tyrosine Kinase ; metabolism