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 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

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

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.

Objective To improve the success rate of puncture among senile patients with difficulties in venipuncture by applying the near-infrared visualization puncture technology.Methods This study selected 160 elderly patients with difficulties in peripheral venipuncture of department of General Surgery and Geriatric Medicine Department by random sampling to carry out peripheral venipuncture at Xuanwu Hospital of Capital Medical University from June 2015 to December 2016. Those patients were divided into control group and observation group with the random number table, 80 patients in each group. Patients in control group received peripheral venipuncture by nurses' personal experience along with assessing vessel by routine visual method and touch method without any accessory equipment. Patients in observation group received peripheral venipuncture by the location of infrared vessel imager. The infrared vessel imager was used to evaluate the vessel shape and depth to understand vascular branches so as to locate target vessel. And then, this study compared the success rate, time and site of punctures between two groups.Results The success rate of first puncture of observation group was 91.25% significantly higher than that of control group (χ2=43.495,P<0.01). The puncture time [(3.7±1.8) min]of observation group was less than that of control group with a significant difference (t=4.629, P<0.01). In the aspect of site of punctures, there was 72.5% of patients with punctures in observation group in forearm vein higher than that in control group (χ2=46.920,P<0.01).Conclusions The near-infrared visualization puncture technology can improve the success rate of puncture in elderly patients with difficulties in peripheral venipuncture, short the puncture time and help to achieve forearm venipuncture with the first choice. This technology can also relieve the pain of elderly patients, save consumptive material cost and reduce nurses' pressure on puncture. It is worth to be used widely in clinical application.

Objective To investigate the relationship of procollagen?lysine 2?oxoglutarate 5?dioxygenase 2 ( PLOD2 ) expression and the clinical characteristics of osteosarcoma, and explore the potential mechanism of tumour metastasis promoted by PLOD2. Methods The expression of PLOD2 in osteosarcoma tissues and paired adjacent tissues were detected by immunohistochemistry and qRT?PCR. Correlation of PLOD2 expression in osteosarcoma with the clinical pathologic features was analyzed by Chi square test and Kaplan?Meier analysis.Fibrillar collagen formation and collagen deposition in the tumor tissues were detected by picrosirius red staining. We transfected U?2OS cells with LV?vector, LV?over／PLOD2, sh?NC and sh?PLOD2. The expression of PLOD2 was detected by qRT?PCR. The impact of POLD2 on U?2OS cell invasion was determined by wound?healing assay and Transwell migration assay. The expressions of PLOD2／FAK／JAK2?STAT3 signal pathway related proteins were detected by western blotting. Results The high expression level of PLOD2 in osteosarcoma tissues was 72.5%, significantly higher than 0% in paired adjacent noncancerous tissues ( P＜0.01), the expression of PLOD2 was positively correlated with lymph node metastasis, pulmonary metastasis and poor outcome (P＜0.01). The same results were also observed in qRT?PCR assay. The median survival time of patients with high expression of PLOD2 protein was 13 months, significantly shorter than 32 months of patients with low expression of PLOD2 ( P＜0.05). The result of picrosirius red staining showed that the percentage of collagen fiber deposition in the osteosarcoma tissue with high level of PLOD2 was (74.43+9.63)%, significantly higher than ( 9.67± 1.28)% in tissue with low expression of PLOD2 (P＜0.001).The result of wound?healing and Transwell migration assay showed that over?expression of PLOD2 markedly promoted the invasion, however, knockdown of PLOD2 suppressed the invasion of U?2OS cells ( both P＜0.01).The result of western blotting showed that over?expression of PLOD2 significantly increased the expression levels of p?FAK, p?JAK2, p?STAT3, but knockdown PLOD2 decreased the levels of p?FAK, p?JAK2, p?STAT3 in U?2OS cells. Conclusions Up?regulation of PLOD2 in osteosarcoma is correlated with lymphatic and distant metastasis. PLOD2 promotes invasion and metastasis of osteosarcoma might through FAK／JAK2?STAT3 signal pathway.