Objeetive To analyze the causes and to explore prevention and management of bile leakage after laparoscopic common bile duct exploration with choledochoscopy followed by primary suturing of choledochal incision.Methods The clinical data of 52 patients with bile leakage after laparoscopic common bile duct exploration choledochoscopy and primary suturing of choledochal incision carried out for choledocholithiasis between June 2011 to June 2013 were retrospectively studied.Results All the 52 patients successfully underwent the laparoscopic surgery and left hospital.The operation time was (101 ± 26) minutes (range 55～ 145 minutes).The intraoperative blood loss was (36±28) ml (range 10～ 100 ml).All the patients were ambulatory after the first postoperative day.The recovery time of postoperative gastrointestinal function was (49.8 ± 12.5) hours (range 37 ～ 74 h).The total hospitalization time was (10.8 ± 2.5) days (range 7 ～ 15 days).The average hospitalization days after surgery was (5.7 ± 1.7) days.The average hospitalization cost was (24 827 ± 3 776) yuan.There were two patients who developed intraoperative bile leakage which was treated with further suturing.Five patients developed postoperative bile leakage and they were cured after unobstructed drainage for 5 days through conservative treatment.After a follow-up of 1 ～ 2years,there was no recurrent lithiasis.The stone clearance rate was 100％.There was no bile duct stricture or other complications.Conclusion In expert hands and with proper selection of patients,laparoscopic common bile duct exploration,choledochoscopy and primary suturing of choledochal incision were safe,effective and feasible for choledocholithiasis.

Objective To investigate the predominant genotypes of β-lactamase in Klebsiella pneu-moniae isolates and the mechanism of antibiotic and histidine kinase inhibitor in affecting β-lactamase gene expression .Methods Tube dilution method and E-test were used to detect the susceptibility of K.pneumon-iae isolates to β-lactam antibiotics .The major genotypes of β-lactamase in β-lactam antibiotic-resistant iso-lates were identified by PCR and sequencing method .Disk diffusion test was performed to analyze the activity ofβ-lactamase .The effects of cefotaxime and histidine kinase inhibitor closantel on the expression of β-lac-tamase gene were evaluated by real-time fluorescent quantitative RT-PCR.Results All of the 118 β-lactam antibiotic-resistant K.pneumoniae isolates expressed β-lactamase, 90.7%(107/118) of which were KPC-2, TEM-1, CTX-M-14, SHV-11 and/or OXA-1 gene positive.The positive rates of TEM-1 (71.0%) and SHV-11 (64.5%) were significantly higher than that of the other three genotypes ( P<0.05).68.2%(73/107) of the isolates possessed two or more than two β-lactamase genotypes, and 30.8%(33/107) iso-lates were identified as TEM-1+SHV-11 genotype.Except for KPC-2 mRNA, the levels of TEM-1, SHV-11, CTX-M-14 and OXA-1 mRNA were significantly up-regulated by cefotaxime at MIC/4 (P<0.05), but were inhibited by 100μmol/L closantel (P<0.05).Conclusion TEM-1 and SHV-11 are the majorβ-lactamase genotypes carried by K.pneumoniae strains isolated from Zhejiang area , and TEM-1 plus SHV-11 ( TEM-1+SHV-11) is the predominant carrying mode of the β-lactamase genotypes .Sublethal dose of cefotaxime can enhance the expression of β-lactamase genes in K.pneumoniae, while the histidine kinase inhibitor , closan-tel, can block the increased expression of β-lactamase genes induced by cefotaxime .

OBJECTIVETo explore the effect of microneedle combined with Lauromacrogol on skin capillary network.

METHODS24 male Leghone (1.5-2.0 kg in weight) were randomly divided into three groups as group A (microneedle combined with Lauromacrogol), B (microneedle combined with physiological saline) , and C(control). The cockscombs were treated. The specimens were taken on the 7th, 14th, 21th , and 28th day postoperatively. HE staining, immunohistochemical staining and special staining were performed for study of the number of capillary and collagen I/III , as well as elastic fibers.

RESULTSThe color of cockscombs in group A became lightening after treatment. The number of capillary decreased as showing by HE staining. The collagen I and III in group B was significantly different from that in group A and C (P < 0.05). Special staining showed proliferation of elastic fibers in group B.

CONCLUSIONSIt indicates that microneedle combined with Lauromacrogol could effectively reduce the capillary in cockscomb without any tissue fibrosis. Microneedle can stimulate the proliferation of elastic fiber, so as to improve the skin ageing process.

Animals ; Capillaries ; anatomy & histology ; Chickens ; Comb and Wattles ; blood supply ; drug effects ; Male ; Needles ; Polyethylene Glycols ; pharmacology ; Punctures ; instrumentation ; methods ; Random Allocation ; Skin Aging

Objective To investigate the mechanism of linezolid resistance in methicillin-resistant Staphylococcus epidermidis ( MRSE) strains isolated in Hangzhou area. Methods Twenty-three linezolid-re-sistant Staphylococcus epidermidis ( LRSE) strains were isolated from the patients with septicaemia, urinary tract infection or infectious pleuritis in several hospitals in Hangzhou area during May, 2013 to April, 2015. The minimal inhibition concentrations ( MICs) of thirteen different antibiotics against the LRSE strains were detected by using E-test. Pulsed-field gel electrophoresis ( PFGE) and cluster analysis were performed for homology analysis. Correlations between linezolid resistance in the LRSE strains and G2576T mutation at theⅤ functional region of 23S rRNA gene or cfr gene were determined by PCR and sequencing analysis. Re-sults The MICs of linezolid to 15 LRSE strains were higher than 256 μg/ml while to the other 8 LRSE strains were 6 or 8 μg/ml. All of the 23 LRSE strains were resistant to both oxacillin and cefoxitin. Among the LRSE strains with high linezolid MICs (MIC>256 μg/ml), LRSE1 and LRSE2, LRSE3-LRSE6 and LRSE9-LRSE12 respectively belonged to the same clone line and came from the same hospital. All of the 23 LRSE strains carried the cfr gene. Moreover, the G2576T mutation at theⅤfunctional region of 23S rRNA gene was detected in the 15 LRSE strains with high linezolid MICs ( MIC>256 μg/ml ) , but not in the 8 strains with lower linezolid MICs ( MIC=6 or 8 μg/ml) . Conclusion There are significant differences in linezolid resistance among LRSE strains isolated in Hangzhou area. The LRSE strains are methicillin-resist-ant coagulase negative Staphylococcus ( MRCNS) strains and widely distributed. The linezolid resistance in LRSE strains is related to the G2576T mutation at theⅤfunctional region of 23S rRNA gene and cfr gene.

ObjectiveTo explore the effect of the Breviscapine (Bre) on rabbit's cardiac muscles after ischemic preconditioning (IP).MethodsThe myocardial ischemic reperfusion model was made with 32 New Zealand white rabbits by silk thread passed around the left circumflex coronary artery and the apex. Model animals were randomly divided into four groups: myocardial ischemia-reperfusion (I/R) group, Bre+I/R group, IP group and Bre+IP group. The changes of the endothelin (ET), nitrous oxide (NO) and the enzymes of the cardiac muscle were measured, and the areas of myocardium infarction were analyzed.ResultsBre and IP could decrease the content of ET, the enzymes of the cardiac muscle and myocardial infarction area; increase the content of the NO. Bre+IP could strengthen the role of protecting the ischemic myocardial cells.ConclusionThe Bre can protect the ischemic cardiac muscle. The Bre+IP can strengthen the protective effect of the IP.

Objective To investigate the polymorphism of human cytomegalovirus（HCMV）UL146 gene in clinical strains,and to evaluate its clinical diagnostic and therapeutic value of gene.Methods The UL146 gene of clinical strains was examined by quantitative polymerase chain reaction（Q-PCR）or general polymerase chain reaction（PCR）.Positive samples of PCR amplification were sequenced and analyzed.Results High variability of UL146 gene was found among 28 HCMV strains.According to phylogenetic analysis,all sequences of UL146 in clinical strains could be divided into three types and four subtypes.Chemokine ELRCXC region was highly conserved in all sequences.Conclusion HCMV-UL146 genes showed a high degree of polymorphism,and its encoded chemokine ELRCXC region was highly con-served.The relationship between HCMV-UL146 gene′s polymorphism and different clinical symptoms of HCMV infection was unclear.

Objective To systematically investigate the molecular epidemiological profiles of human papillomavirus (HPV) in patients with condyloma acuminata(CA). Methods Two hundred and one samples of HPV DNA isolated from CA were PCR amplified by the PGMY09/11 primer system. The PCR products were simultaneously hybridized to 37 specific HPV probes immobilized on a nylon strip and then genotyped. All DNA templates were further PCR amplified using HPV 6 and 11 type specific primers for verification. Results All samples were HPV DNA positive consisting of totally 31 genotypes, the types of which were type 11(53.7%, 108/201), 6(43.8%, 88/201), 16(6.5%, 13/201), 52(6.0%, 12/201), 33(5.5%, 11/201), cp6108 (5.5%, 11/201) and 42 (5.0%, 10/201). The samples infected with a single and mixed types of HPV accounted for 60.2% (121/201) and 39.8% (80/201) respectively. Consistent results were found with the detection of HPV6 and 11 between hybridization assay and type-specific PCR. Conclusions At least 31 HPV genotypes are associated with CA. HPV 11 predominates while 68, 40, 54, 67, 73, 82, 35, 64 and 83 are rare in CA. Type cp6108 is detected in CA for the first time with a high prevalence. HPV26, 69, 70, 71,72 and IS39 might be not associated with CA. CA infected with a single and mixed HPV types accounts for 60.2% and 39.8%, respectively.

Adenoma ; genetics ; metabolism ; pathology ; surgery ; Adult ; Carcinoma, Papillary ; genetics ; metabolism ; pathology ; Diagnosis, Differential ; Exons ; Female ; Follow-Up Studies ; Humans ; Ki-67 Antigen ; metabolism ; Male ; Mutation ; Nuclear Proteins ; metabolism ; Proto-Oncogene Proteins B-raf ; genetics ; Thyroglobulin ; metabolism ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Thyroid Nuclear Factor 1 ; Transcription Factors ; metabolism

Objective Docetaxel is commonly used in chemotherapy for patients with mucoepidermoid carcinoma (MEC) in the salivary gland after operation,but it cannot kill all residual tumor cells and prevent recurrence.Cancer stem cells (CSCs) may be responsible for tumor recurrence and drug resistance.Based on the theory of CSCs,the authors investigated the effects of docetaxel on cancer stem cell-like cells in the MEC cell line MC3.Methods MC3 cells in the logarithmic growth phase were divided into a negative control and a docetaxel (40 ng/mL) group.The colony formation rate of the MC3 cells was calculated with the soft agar cloning technique,the growth of the cells detected by MTT assay,the protein and mRNA expressions of CD44 and Oct4 determined by immunocytochemistry and flow cytometry,and tumorigenicity observed by nude mouse tumorigenicity assay.Results Compared with the negative control group,the MC3 cells treated with docetaxel exhibited significant increases in the colony formation rate ([9.14±0.75] vs [33.47±1.30]％,P＜0.05),the protein expressions of CD44 (14.47±0.15 vs 99.50±0.30,P＜0.05) and Oct4 (1.37±0.06 vs 14.60±0.36,P＜0.05),and the mRNA expressions of CD44 (0.207±0.009 vs 0.651±0.015,P＜0.05) and Oct4 (0.223±0.008 vs 2.228±0.005,P＜0.05).At 2 months after injection of 1×103,1× 104,and 1×105 MC3 cells,tumor formation was observed in 0,1,and 3 of the nude mice,respectively,but not in the negative control group.Conclusion MC3 cells surviving docetaxel treatment have the properties of stem cells,and docetaxel can enrich cancer stem cells in the MC3 cells,which plays a key role in tumor recurrence.