Objective To summarize the clinical experience of laparoscopic appendectomy(LA).MethodsA retrospective analysis was conducted on the clinical data of 683 patients with appendicitis treated by LA,including 258 cases of chronic appendicitis,425 cases of acute appendicitis(87 cases of complicated appendicitis).ResultsThe procedure was completed in all the patients without conversion to open surgery.The operation time was 30 min in average(15-150 min).The average hospital stay was 4d (2-15d).Postoperative complications included 26 cases of incision infection,3 cases of abdominal abscess,1 case of appendical mesenterium hemorrhage,1 case of small intestine fistula and 1 case of appendical stump fistula.The patients were followed up for 2 to 12 months with a mean of 6 months,during which no intestinal adhesive obstruction occurred.Conclusion LA is safe and effective for appendicitis,with advantages of minimal trauma,lighter pain,quicker recovery,shoter hospital stay,less complications and so on.For complicated appendicitis,the abundant surgical experience and proficient endoscopic technique are indispensable to reduce the occurrence of complications effectively.

Objective:To build the model of the gene FKBP38 (FK506 binding protein 38) conditional knock out in liver.Methods:Transgenic mouse whose FKBP38 gene was flanked with loxP was constructed by embryo microinjection.The FKBP38 gene was deleted by breeding mice harboring two loxP sites in FKBP38 (FKBP38fl/fl) with the mice bearing the expression ofCre recombinase mice driven by an album promoter.Afterward,the genotype of FKBP38 conditional knockout mice was analyzed.Results:①Relative hepatic FKBP38 mRNA levels showed significant difference between FKBP38 conditional knockout mice (FKBP38-/-) and wild type(P＜ 0.001).②Relative hepatic FKBP38 protein expression levels of FKBP38 conditional knockout mice (FKBP38-/-) were significantly different with wild type(P＜0.001).③Relative phosphorylation of hepatic p70 S6K and 4E-BP-1 protein of FKBP38 conditional knockout mice (FKBP38-/-) showed no significant difference,with slight decrease in phosphorylation of 4E-BP-1,compared with wild type.④No significant difference in expression of hepatic Bcl-2 between FKBP38-/-and wild type.Conclusions:The mouse model of the gene FKBP38 (FK506 binding protein 38) conditional knock out in liver is successfully built.

Constitution of TCM is based on the study of individual constitution and physical health,and it has also drawn attention from the medical profession in the world.This commentary has reviewed about the development of constitution of TCM over the past few years,summed up the past 30 years,especially analysis the new research fruits during 2005-2006,expounded the important status of Chinese medical constitution in the development of Chinese medicine and the significance in socio-economic sustainable development,and put forward the direction of development and research for Chinese medical constitution.

Gap junction has been considered allowing direct transfer of small metabolites between cells to maintain homeostasis in multicellular organisms. Deficient gap junction involved in genesis and development of many central neurological disorder, and may be a potential target for the treatment of them.

Phospholipase D(PLD) is ubiquitous in bacteria,fungi,and mammal.In pathogenic microorganisms,PLD can be pathogenic determinant and play a role in spore generation.In mammalian cells,PLD functions in several signal transduction pathways,such as membrane transportation,mitosis regulation,and actin cytoskeleton regulation.In the process of pathogens invasion host cells,both of the pathogen and host cells’ PLD will be activated and a series of cascade reaction will be generated.During this process,pathogen’s PLD can regulate the polymerization and reorganization of its own actin filaments and induce the polymerization or reorganization of the host cell actin filaments near the foci,thus to promote the phagocytosis of the pathogen by host cell.Investigating the role of PLD activation in the infection will be significance for further understanding the molecular mechanism of pathogen-host cell interaction.

Objective To construct p66shc gene interfering lentivirus vectors recombination and transfect it to 293T cells ,RNA interfering was carried out to induce p66shc gene silence ,so as to provide basis for further study of the p66shc function .Methods Screening of three RNA targets which were named after p66shc‐shc1 ,p66shc‐shc2 ,p66shc‐shc3 ,cloned into the pLenR‐GPH vec‐tor ,which contained green fluorescent protein(GFP) and transformed into DH5αcells .The positive clone were picked out for right sequencing and transfected to 293T cells with pRsv‐REV ,pMDlg‐pRRE ,pMD2G .The expression of GFP in inverted fluorescence microscope confirmed the virus packaging success .Fluorescence quantitative PCR and Western blot technology were used to investi‐gate the expression of p66shc at the molecular and protein levels ,p66shc‐shc1 target of effective silencing p66shc gene was selected to prepare for subsequent tests .Results The shRNA lentivirus vector was constructed which could express p66shc and was trans‐fected into 293T cells successfully .Fluorescence quantitative PCR and Western blot technology were used to investigate p66shc gene silence by RNA interference .Conclusion The lentivirus RNAi vector of targeted expression p66shc could induce p66shc gene si‐lence at the molecular and protein levels after transfected into 293T cells by RNA interference .

To establish an HPLC method for determination of dactylorhin A and militarine in Cremastrae Pseudobulbus/Pleiones Pseudobulbus. The analysis was achieved on an Alltech Prevail C18 column (4. 6 mm x 250 mm, 5 microm) using a mobile phase of acetonitrile (A), water (B) gradient elution in a total run time of 35 min (0 min, 20:80; 30 min, 55:45; 35 min, 55:45) and a diode array detector was set at 224 nm. The flow rate was 0.8 mL x min(-1). The assay displayed good linearity over the concentration range of 0.257-9.95 microg (r = 0.999 8), and 0.128-10.27 microg (r = 0.999 9), respectively. The average recoveries (n = 9) were 94.70% and 102.8% for dactylorhin A and militarine, respectively. The method is accurate, quick, simple and reproducibility. It can be used for the quality control of Pleione bulbocodioides and Pleione yunnanensis.
Chromatography, High Pressure Liquid ; Glucosides ; analysis ; Orchidaceae ; chemistry ; Succinates ; analysis

Objective To confirm whether the open-ended coaxial line is effective in detection of the differences in dielectric properties between colorectal cancer tissues and surrounding normal tissues and evaluation of the depth of tumor invasion. Methods The open-ended coaxial line system at frequencies ranging from 50 to 500 MHz in 98 freshly excised colorectal cancerous specimens obtained from the operating theatre of Zhujiang Hospital, was used to detect both the relative permittivity and conductivity on the serosal surface of the carcinoma nidus, the mucosa of the carcinoma nidus, and the mucosa of the surgical resection margin. Pathological examinations were conducted on each specimen after surgery. Results The values for relative permittivity and conductivity of the colorectal cancerous mucosa were significantly higher than those of the normal mucosa (P < 0.01). For the tumor which had invaded or penetrated the serosa (stage ≥ T3), the dielectric properties of both the cancerous serosa and mucosa were higher than the one restricted to muscularis propria or less intestine wall (stage < T3) over the measured frequency range, and there existed statistical differences at the common frequencies of 213 MHz and 426 MHz. Conclusion The open-ended coaxial line system may result in fast and effective diagnostic differentiation between cancerous and normal colorectal tissues as well as reasonable assessment of the tumor infiltration depth.

Objective Investigate the prognosis of patients undergoing liver transplantation (LT) for end-stage autoirnmune liver disease (ALD).Method The clinical data of 45 patients with endstage ALD undergoing LT from April 2001 to March 2015 in the first affiliated hospital of Zhejiang University were analyzed retrospectively.The postoperative cumulative survival rate of the recipients was calculated,and the causes of death were analyzed.The postoperative rejections,new onset viral hepatitis and ALD recurrence were also analyzed.Result In 45 ALD recipients,33 cases survived and the postoperative 5-year cumulative survival rate was 78.8％.Causes for 12 dead cases were mnultiple organ failure,liver graft failure,respiratory complications,hemorrhage and hepatic artery embolization.In 45 ALD recipients,6 cases suffered rejection after operation with the incidence bing 13.3％.One case suffered new onset hepatitis B infection 8 years after opcration.One recipient suffered primary disease (primary biliary cirrhosis) recurrence 2 years after operation,and 1 recipient with primary disease (primary sclerosing cholangitis) developed into overlap syndrome.They all survived for a long term after active treatments.Conclusion Most LT recipients with endstage ALD can obtain a long term survival.Attentions should be paid to the immunosuppressive regimens in early period after LT,prevention of infection,rejection and postoperative new onset viral hepatitis,and timely diagnosis of primary disease recurrence.

Objective To examine the effects and mechanisms of siRNA targeting aquaporin 4 (AQP 4) in combination with hyperbaric oxygen therapy(HBO) on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage.Methods Rats were randomly divided into four groups,the control group,the hyperbaric oxygen group,the AQP-4 siRNA group and the combination therapy group (24 rats).Thrombin Ⅶ was injected into the caudate nucleus to establish the hemorrhage model.Construction of siRNA targeting aquaporin 4 was conducted.The mRNA expression of AQP-4 was detected by RT-PCR at day 3.Changes in brain moisture and blood-brain barrier perme ability were measured by a wet/dry weight method and Evans blue fluorometry.The nerve cell apoptosis rate was analyzed by Annexin V andTdT-mediated dUTP-biotin nick end labeling (TUNEL).The expression of proteins including AQP-4,MMP-2,MMP-9,Bcl-2 and caspase-3 was detected by Western Blotting.All the animals were given a score for their nerve function at day 3.Results AQP-4 siRNA treatment obtained better effects than HBO in decreasing the brain edema leveland silencing AQP-4 mRNA(P＜0.05)while,the combination therapy group achieved the best results(P＜ 0.05).Compared with the control group,the percentage of apoptotic cells decreased in all the three treatment groups,with the most marked decrease observed in the combination treatment group(4.24± 0.04)％(F=13.76,P=0.001).The expression of AQP-4,MMP-2,MMP-9 and caspase-3 was lower (P＜0.05) and the expression of Bcl-2 was higher(P＜0.01)in the combination treatment group than in the other three groups.Compared with the control group,all the other three groups received better scores on nerve function defect evaluation at day 3 after hemorrhage(P＜0.05),with the combination treatment group again achieving the most favorable score (4.7 ± 1.1) (F=7.21,P =0.013).Conclusions Targeted siRNA interference combined with hyperbaric oxygen can effectively reduce cerebral edema after cerebral hemorrhage,inhibit neuronal apoptosis and promote neuron function recovery.The underlying mechanisms may be related to down-regulation of AQP-4,MMP 2,MMP-9 and caspase-3 expression and up-regulation of Bcl-2 expression.