Pancreatic fistula is a common complication after pancreaticoduodenectomy(PD),and other complications occurred secondary to it such as hemorrhage and severe abdominal infection are also the main cause of death after PD.Proper selection of anastomotic methods is essential to the success of PD.The most common anastomotic methods can be divided into pancreaticojejunostomy and pancreatogastrostomy,but the effects of these methods have always been controversial consistently.In this paper,the effects of pancreaticojejunostomy and pancreatogastrostomy in the prevention of postoperative pancreatic fistula were synthetically analyzed based on recent research results.

Objective To establish the crystal violet plaque assay for detection of virus titer of recombined Tiantan vaccinia AIDS vaccine,and provide more stable method of virus titration for rTV AIDS Vaccine.Methods Optimized the concentration of Vero cells,the time and temperature of virus adsorption,and the time of determination for CPE,then established the crystal violet plaque assay for virus titer of rTV.Counting and analysis the plaques by BioSpot Reader,then analyzed the relativity of plaques counted with BioSpot Reader and manual; Several lots rTV AIDS Vaccine and Tiantan vaccinia were titrated by the method of plaque formation-hemadsorption assay,neutral red and crystal violet plaque assay,then analyzed the relativity of the results of three methods ; meanwhile,the virus titer of samples were determine repeatedly by the crystal violet plaque assay,then calculated the coefficient of variation( CV),and verified the precision of the method; SPSS17.0 was used in statistical analysis of the experimental results.Results When the concentration of Vero cells was 5.0×105-9.O×105 cells/ml,virus been adsorbesd 2 h at 37℃,then cultivated 72 h after adding the culture medium containing methyl cellulose.Plaques counted by BioSpot Reader was highly related with counted by manual (r =0.985),so BioSpot Reader counting can objectively reflect the virus plaques with various size,and reduce the error by manual counting; compared the virus titration for different lots of rTV AIDS vaccine and Tiantan vaccinia with three methods,the crystal violet plaque assay was highly related with plaque formation-hemadsorption assay (r =0.997,P＜0.01 ) and neutral red plaque assay(r=0.980,P＜0.01 ).Conclusion Crystal violet plaque assay was established for virus titration of rTV AIDS Vaccine.

Prior to implantation, the blastocyst has to hatch out of its zona pellucida to invade the endometrium .In mammals including humans , failure of blastocyst hatching leads to infertility .Blastocyst hatching is believed to be regulated by a variety of autocrine and paracrine molecules such as proteases , cyclooxygenase-2, p38 mitogen-activated protein ki-nase, activin A and Wnt signal pathway .This article reviews the mechanisms of the key molecular regulators involved in mammalian blastocyst hatching and hatching-assisting methods , which can help clarify the mechanism of blastocyst hatching and the treatment of infertility due to failure in blastocyst hatching .

A novel high performance liquid chromatographic method was developed for the determination of 4-O-methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation.Plasma samples were treated by one-fold volume of methanol and acetonitrile to remove the interference proteins.A reverse phase column of SHIMPACK VP-ODS (150 mm× 4.6 mm,5.0 μm) was used to separate 4-O-methylhonokiol in the plasma samples.The detection limit of 4-O-methylhonokiol was 0.2 μg/L and the linear range was 0.012- 1.536 mg/L.The good extraction recoveries were obtained for the spiked samples (84.7％,89.3％ and 87.7％ for low,middle and high concentrations of added standards,respectively).The relative standard deviation of intra-day and inter-day precisions was in the range from 0.6％ to 13.5％.The pharmacokinetic study of 4-O-methylhonokiol was made and the results from the plasmaconcentration curve of 4-O-methylhonokiol showed a two-apartment open model.This work developed a sensitive,stable and rapid HPLC method for the determination of 4-O-methylhonokiol and the developed method has been successfully applied to a pharmacokinetic study of 4-O-methylhonokiol.

Objective To analyze clinical effects of gastric tube operation for treatment of esophageal cancer in elderly patients and evaluate its clinical efficacy and safety.Methods A total of 171 patients aged 60-72 years with esophageal cancer in our hospital were selected.They were randomly divided into treatment group and control group.Treatment group was treated with gastric tube operation,and control group was traditionally treated with complete replacement of esophagus with stomach.The quality of life,patient satisfaction and safety of operation were evaluated after 3 weeks,6 months and 12 months after operation,respectively.Results The operation for esophageal cancer in 171 patients were successful.At 3 weeks after the operation,the score of life quality in treatment group and control group were both low [(67.3±9.6) vs.(65.3±8.4)],and there were no significant differences between the two groups (P＞0.05).At 6 months and 12 months after operation,the scores of life quality were (89.2±8.3) and (90.3±9.6) in treatment group,and (66.5± 10.4) and (60.5 ± 11.2) in contol group,respectively.There were statistical differences between the two groups after 6 months and 12 months of operation (P＜0.05).The complication rate in treatment group (6.9％) was much lower than that in control group (30.6％),and there were significant differences between the two groups (x2 =18.43,P＜0.01).The patient satisfaction was in no differences between the two groups at 3 weeks after operation (P＞ 0.05),while there were statistically significant differences at 6 and 12 weeks after the operation (P＜0.05).Conclusions Gastric tube operation in treatment of elderly patients with esophageal cancer,can effectively improve the life quality,and prevent the occurrence of postoperative complications,which is worthy of clinical application.

A novel high performance liquid chromatographic method was developed for the determination of 4-O- methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation. Plasma samples were treated by one-fold volume of methanol and acetonitrile to remove the interference proteins. A reverse phase column of SHIM- PACK VP-ODS （150 mm ×4.6 mm, 5.0 μm） was used to separate 4-O-methylhonokiol in the plasma samples. The detection limit of 4-O-methylhonokiol was 0.2 μg/L and the linear range was 0. 012 - 1. 536 mg/L. The good extraction recoveries were obtained for the spiked samples （84.7%, 89.3% and 87.7% for low, middle and high concentrations of added standards, respectively）. The relative standard deviation of intra-day and inter-day precisions was in the range from 0.6% to 13.5%. The pharmacokinetic study of 4-O-methylhonokiol was made and the results from the plasmaconcentration curve of 4-0-methylhonokiol showed a two-apartment open model. This work developed a sensitive, stable and rapid HPLC method for the determination of 4-O-methylhonokiol and the developed method has been successfully applied to a pharmacokinetic study of 4-O-methylhonokiol.

Objective To investigate the predictive value of platelet parameters on distant metastasis of different clinical stages of non‐small cell lung cancer (NSCLC) .Methods A retrospective analysis was performed on the inpatients with lung cancer in our hospital from April 2013 to January 2014 .The parameters of platelet count (PLT ) ,mean platelet volume (MPV) ,platelet distribu‐tion width (PDW) ,platelet ‐large cell ratio(P‐LCR) ,platelet hematocrit (PCT ) ,etc .were detected respectively .Results The re‐sults showed that PLT ,MPV ,P‐LCR and PCT in the lung cancer group were higher than those in the benign lung tumor group and the healthy control group ,the differences were statistically significant (P< 0 .01) ,while PDW had no statistically significant differ‐ence among these groups(P> 0 .05) .With the TNM stage increase ,the increased platelet proportion in the stage Ⅲ and Ⅳ was sig‐nificantly higher than that in the stage Ⅰ and Ⅱ (32 .35% ,42 .85% vs .19 .35% ,22 .85% ,P< 0 .01) .The platelet five parameters in the stage Ⅳ had statistical difference compared with those in the stage Ⅰ and Ⅱ (P< 0 .05) ;but PDW and P‐LCR in the stage Ⅲhad no statistical difference compared with those in the stage Ⅰ and Ⅱ (P> 0 .05) ;PLT ,MPV and PCT in the stage Ⅳ had statisti‐cal difference compared with those in the stage Ⅲ (P< 0 .05) .Conclusion The patients with lung cancer often stimulate the bone marrow megakaryocytes to produce platelets by a particular mechanism .As the staging increase ,the proportion of increased platelet is more obvious .Continuously and dynamically observing platelet parameters is conducive to judge the development and prognosis of the disease .

Objective The purpose of this study was to determine the relationship between microecological balance and the syndrome type of acute upper respiratory infection.Methods The nature and quantity of pharyngeal flora were assayed in 31 cases of wind-cold acute upper respiratory infection and 36 cases of wind-heat acute upper respiratory infection,with 30 healthy cases served in control group.Results There was a significant rise of concentration of pharynx flora in acute upper respiratory infection than that in control group,whereas there was a significant decrease of diversity of pharynx flora in acute upper respiratory infection than that in control.Conclusion Imbalance of pharyngeal micro-ecology is one of the major factors leading to acute upper respiratory infection,manifested as insufficiency of genuine Qi failure in guarding.

BACKGROUND: LIM Mineralization protein 1 (LMP-1), an intracellular non-secretory protein, plays roles in bone calcification. Presently, it is found that LMP-1 can promote an increase in bone morphogenetic protein 2 and transforming growth factor ? 1. This indicates that LMP-1 may recruit a mass of ossified factors to participate in the differentiation of osteoblasts. OBJECTIVE: To construct human LMP-1 gene adenovirus recombinant with AdEasy adenovirus vector system, and to detect LMP-1 expression in infected rabbit bone marrow mesenchymal stem cells (BMSCs). DESIGN, TIME AND SETTING: An opening experiment was performed at the Central Laboratory of South West Hospital, Third Military Medical University of Chinese PLA from March 2006 to February 2007. MATERIALS: Three New Zealand rabbits were used to isolate bone marrow mesenchymal stem cells. Plasmid pIRES2-EGFP-LMP-1 carrying human LMP-1 gene was kept in Department of Orthopedics, Second Hospital Affiliated to Chongqing Medical University. AdEasy was presented by Dr. Tong-Chuan He from USA. Human embryo kidney 293 cells were gifted by Wang from Department of Clinical Laboratory of Chongqing Medical University. METHODS: LMP-1 gene with a sequence encoding His-tag was amplified by using pIRES2-EGFP-LMP-1 plasmid as a template for polymerase chain reaction (PCR) with a specially designed downstream primer. The target gene was cloned to the pMD18-T vector for sequencing. Once verified, the gene was cut out by double endonucleases, connected to the shuttle vector pAdTrack-CMV. The newly constructed vector was linearized by PmeⅠ following efficient homologous recombination with the backbone vector pAdEasy-1 in BJ5183. The correct recombinant pAd-LMP-1 was linearized with Pac I and transfected to HEK293 cell by means of mediated Lipofectamine. The titer of virus was measured after amplification and purification. The mRNA and protein expression of LMP-1 was detected in BMSCs, which were infected with Ad-LMP-1 at the most appropriate MOI, were detected by reverse transcription (RT)-PCR and Western blot, respectively. MAIN OUTCOME MEASURES: Plasmid pAd-LMP-1 identification, its titre and efficiency of infection. mRNA and protein expression of LMP-1 were detected by RT-PCR and Western blot, respectively. RESULTS: The recombinant plasmid pMD18-T-LMP-1 carrying LMP-1 gene with His-tag was successfully constructed. After packaging and amplification of the recombinant adenovirus, the 3.5?109 efu/ml titer of Ad-LMP-1 was obtained by CsCl gradient purification. The optimal efficiency infection was 50%-70%, which was get after Ad-LMP-1 infected BMSCs for 3 days at the most appropriate MOI 150. The mRNA and protein expression of LMP-1 in infected BMSCs had been proved. CONCLUSION: The recombinant adenovirus containing human LMP-1 gene with His-tag is successfully constructed. The BMSCs infected with recombinant adenovirus Ad-LMP-1 can effectively express LMP-1.

Virtual slides were applied in experiment teaching of pathology on trial in clinic medicine college of Sichuan university.The resuhs from the Survey showed that students and teachers preferred virtual slides in learning microscopic lesions.Virtual Slides can help us save time,promote quality of observation,carry out discussion based teaching and manage teaching documents.It can be used in network teaching after marking the lesions.