An analytical method for determination of tetramethylpyrazine (TMP) in ligusticum chuanxiong hort by RP-HPLC coupled with countercurrent extraction used for the preconcentration of trace TMP was established. The column was packed with Zorbax ODS (250 mm×4 mm，5 μm). The mobile phase consisted of methanol water (1 % acetic acid)(35∶65)，the flow rate was 0.8 ml/min and detection was performed at 300 nm. The calibration curves showed good linearity over the range of 0.41 μg/ml to 4.1 μg/ml，γ＝0.9993，the recoveries were 92.3 %. The method was simple，fast，sensitive and accurate and could be applied to the analysis of ligusticum chuanxiong hort. The results showed that the content of TMP in ligusticum chuanxionghort were different with different sources.

Objective To improve the technique of pancreatoduodenectomy in order to facilitate the(management) of complications and direct observation on follow up.Methods Pancreatoduodenectomy and Child′s method of digestive tract reconstruction was performed in 42 paltents.A blind loop of jejunum 5 to 6 cm in length was constructed beyond the pancreatojejular anastomosis and it was fixed to the subcutaneous(tissue) of the adjacent abdominal wall.Results Thirty-two cases recovered uneventfully,and 10 cases had complications induding pancreatic and biliary leakage and hemorrhage.These complications were successfully treated under direct vision by choledochoscope passed into the blind jejunal loop.This method of observation was used for long-time follow up in 22 cases,and revealed ercurrent tumor(n=5),bile duct stricture(n=4) and bile duct ascariasis(n=1).Conclusions This operative method did not cause new(complications) and it can be combined with the traditional operation.Postoperatively,direct observation and management of leakage of pancreatojejular anastomosis and biliointestinal anastomosis and hemorrhage can be accomplished,and the anastomoses and pancreatic stump can be directly observed at followup.

Objective] To find a rapid method for diagnosing Acanthamoeba keratitis and identifing Acanthamoeba . [Methods] 10% potassium hydroxide(KOH) wet mount preparations, Acanthamoeba culture, inverted phase contrast microscopy,and pathological examination using H.E. staining and PAS staining. [Results]Using corneal scrapings and corneal materials obtained from surgery,7 cases and 5 cases of Acanthamoeba keratitis were diagnosed by 10% KOH wet mount preparations. 6 strains of Acanthamoeba were isolated in corneal materials of 6 cases by protozoa culture method. The cysts, trophozoites and pseudopods on the trophozoites of Acanthamoeba were directly observed under the inverted phase contrast microscope. The cysts and trophozoites of Acanthamoeba were seen by H.E. staining and PAS staining with 20 h. [Conclusion] Acanthamoeba keratitis could be rapidly diagnosed by 10% KOH wet mount preparations and inverted phase contrast microscopy. Acanthamoeba organisms could be directly observed and identified under inverted phase contrast microscope.

Objective To discuss the value of choledochoscopy via subcutaneous jejunal blind loop in the diagnosis and treatment of severe complications after pancreatoduodenectomy. Methods A fiber choledochoscope (FCC) was introduced through a subcutaneous jejunal blind loop. Under direct vision, the pancreatojejunal or choledochojejunal anastomotic leakages were coated with biological glue, or the anastomotic bleedings were stopped with hemostatic drugs. Results Five cases of intestinal leakage, 3 cases of biliary leakage and 2 cases of anastomotic bleeding were all cured. Follow-up for 1～3 years in the 10 cases found no recurrence of tumors. Choledochojejunal anastomotic stricture occurred in 1 case 1 year after the surgery and was cured by dilatation under fiber choledochoscope. In the remaining 9 cases, the pancreatic juice and bile fluid were excreted normally. Conclusions Application of FCC through the subcutaneous jejunal blind loop can be employed in the diagnosis and treatment of complications after pancreatoduodenectomy and in the long-term follow-up of pancreatojejunal or choledochojejunal anastomotic stoma under direct vision.

Objective To evaluate the effectiveness of scanning with labeled autologous leukocytes for the detection of abdominal inflammation in equivocal patients. Methods From July 1998 to April 2000 sixteen patients with equivocal abdominal inflammation were recruited into this study. ResultsThere were 10 cases with positive scanning. The diagnosis of intraabdominal infection was consequently confirned by laparotomy and or good response to antibiotic therapy.The 6 cases with negative result were eventualy proven to have no intraabdominal infection. The average radioactivity of imaging and paired non imaging district was determined ( P

Objective To investigate the effects and mechanisms of the CXCR3-inhibitor genistein on the acute rejection reaction of pancreas transplantation in a rat model. Methods Three groups of rats underwent pancreas transplantation, group one: the syngeneic transplantation group; group two: heterogous transplantation group without genistein-treatment; and group three: genistein-treatment (10 mg/kg,i. p.×7d) transplantation group. The grafts of three groups were harvested at day 7 after operation for histopathology and immunohistochemistry analysis of CXCR3, the serum levels of IFN-γ, IL-2 were examined by ELISA. Results There was no acute rejection reaction of pancreas transplantation in group one and no expression of CXCR3 ,the serum levels of IFN-γ,IL-2 were (2.76±0.66) pg/ml and (11.83±1.86) pg/ml. In group two, there was significant acute rejection reaction and strong expression of CXCR3, the serum levels of IFN-γ, IL-2 were (32.64±2.52) pg/ml and (29.59±1.71 ) pg/ml, which were significantly higher than those in group one (P<0.05). Compared with group two, the damages of graft tissue in genistein-treatment group alleviated and the lymphocytes infiltration decreased, the expression of CXCR3 was weakly positive, the serum levels of IFN-γ, IL-2 were (15.94±1.95) pg/ml and (22.62±1.76) pg/ml, which were significantly lower than those in group two(P<0.05), but was higher than those in group one (P<0.05). Conclusions Genistein could mitigate the acute rejection of pancreas transplantation effectively through inhibiting CXCR3 expression.

Objective To analyse our clinical experience in pediatric living donor liver transplantation (PLDLT).Methods The clinical data of 45 patients who underwent PLDLT in our hospital from April 2005 to April 2014 were retrospectively studied and their preoperative,intraoperative and postoperative data were analyzed.Results All donors recovered well.The graft to recipient weight ratio (GRWR) ranged from 1.0％ ～ 6.4％ (2.5％ ± 1.2％).Size reduction of graft were performed in 2 patients.An interposition venous conduit from the confluence of the native right and left portal vein (PV) to the graft PV was carried out in 1 patient,venous grafts for revascularization of the tributaries of the middle hepatic vein from segment Ⅴ and Ⅷ were used in 3 patients,and a venous patch for revascularization of the left hepatic vein was used in 2 patients.Hepatic artery re-reconstruction was performed in 3 patients after hypoperfusion was detected on intraoperative Doppler ultrasound.The postoperative complications included acute rejection (n =2),vascular complications (n =7),biliary complications (n =11),and infectious complications (n =27).The 1-,2-and 5-year survival rates were all 84.4％.Seven of 45 recipients died within one year post transplantation,with 3 patients who died of vascular complications,and 4 patients who died of infection.The differences in age [(50.8 ± 49.8) months vs (12.6 ± 14.9) months],body weight [(16.2 ± 10.5) kg vs (7.3 ± 1.7) kg],serum total bilirubin [(177.0 ± 126.5) μmol/L vs (301.9 ± 110.6)μmol/L],Pediatric end-stage liver disease (PELD) score (16.1 ± 12.1 vs 26.2 ± 11.3) and GRWR (2.2％ ± 0.8％ vs 4.2％ ± 1.6％) between the survival and the dead groups were significant (P ＜ 0.05).Conclusions PLDLT is an effective method to treat children with end-stage liver disease.Using a multidisciplinary approach in the preoperative management,excellent surgical techniques,and proper postoperative management are extremely helpful to improve postoperative survival rate.

Objective To investigate the characteristics of infection and risk factors after pediatric living donor liver transplantation (PLDLT).Method Form April 2005 to April 2014 the clinical data of 45 cases of PLDLT in General Hospital of Chinese People's Armed Police Forces were retrospectively investigated,and the difference between the patients after PLDLT with infection and those without infection was analyzed.Result Eighty-four infections occurred in 27 (60.0％) of 45 patients,including 25 cases of bacterial infections,14 cases of viral infections,and 3 cases of candida albicans infections.Most infections occurring within 3 months after PLDLT have been found to be caused by bacteria,viruses and fungi.The trough level of tacrolimus (Tac) was in target therapeutic window in 16/20 infected patients and more than 10 ng/mL in 4/20 infected patients within 3 months after PLDLT,and there were 12/16 infected patients with the trough level of Tac of more than 10 ng/mL 3 months post-PLDLT,with the difference being significant (P＜0.05).Multivariate analysis revealed that post-transplant infection was significantly related with the factors as weight＜10 kg,age ＜12 months,biliary intestinal anastomosis,pediatric end-stage liver disease (PELD) score,ChildPugh score,total bilirubin,blood loss per kg body weight and graft to recipient weight ratio (GRWR).Logistic regression analysis suggested that PELD score was independent risk factor of infection after PLDLT.Conclusion The infection after PLDLT has their special characteristics.The proper irnmunosuppressive protocol and control of above risk factors are helpful to decrease the incidence of infection after PLDLT.

Objective To assess the effects of the risk factors of grafts from donors after cardiac death (DCD) on the prognosis of liver transplantation (LT).Methods In this retrospectively study,215 cases of LT using DCD donor grafts were performed at our institution from September 2013 to January 2017.Due to the loss to follow-up in 4 cases,211 cases were enrolled in the study.The following DCD donor data were collected:gender,age,primary disease,ABO blood type,body mass index (BMI),medical history (fatty liver,hypertension),ICU hospitalization time,mechanical ventilation time,warm ischemia time,cold ischemia time,and indexes of routine laboratory test before donation.Statistical analyses using the Kaplan-Meier method,log-rank test,multivariate step-wise Cox regression were performed.Results Of the 211 donors,univariate analysis showed that the overall 6-month,1-,and 3-year survival rate after DCD LT was 88％,84％,and 82％,respectively.Univariate analysis showed that donor serum sodium level ＜136 mmol/L (P =0.018) and cold ischemia time ＞9 h (P =0.013) were all significant risk factors affecting overall survival after DCD LT.Additionally,donor BMI ＞30 kg/m2 (P =0.011) and donor age ＞60 years (P =0.025) were significantly associated with postoperative complications.Multivariate analysis showed that donor serum sodium level (P=0.025) was an independent risk factor of survival after DCD LT.Conclusion To select suitable DCD liver allografts and control risk factors of donor can help to improve outcomes of recipients.

Objective CD4 +IL-17 +cells are a group of newly discovered effector CD4 +T cells, which may play a key role in the pathogenesis of cancer.This study aims to investigate the expres-sion of CD4 +IL-17 +cells in pancreatic cancer and its correlation with the clinicopathological characteristics and prognosis of the dis-ease as well as the clinical significance of the cells in the microenvironment of pancreatic cancer. Methods We collected tumor tis-sue and tumor-adjacent normal tissue samples from 51 pancreatic cancer patients.We determined the expressions of CD34 and vascular endothelial growth factor ( VEGF) and measured the proportion of IL-17 +cells in the cancer tissue using immunohistochemistry and the fluorescence activated cell sorter, respectively, followed by analysis of their correlation with tumor angiogenesis, clinicopathological pa-rameters, and survival time of the patients. Results The percentage of CD4 +IL-17 +cells in tumor tissue was positively correlated with microvessel density (r =0.534, P<0.05) and the expression of VEGF in the tumor tissue (r=0.356, P<0.05).IL-17 +cells were expressed more highly in the tumorous than in the tumor-adjacent normal tissue (P<0.05), and the expression level was correla-ted with the stage of tumor, node, and metastasis (TNM) and lymph node metastasis (P<0.05), but not with the patients′gender or age, tumor size, tumor location, histological grade, or local invasion (P>0.05).Fifty (98.0%) of the patients were successfully followed up for 2-67 months, which revealed a median survival time of 16.6 ±4.8 months, significantly longer in those with a higher expression of intratumoral IL-17 +cells (P<0.05).Univariate analysis showed an association of the survival rate with the tumor size, TNM stage, lymph node metastasis, and level of intratumoral IL-17 +cells, while multivariate analysis showed the TNM stage to be an independent prognostic factor for the survival of the pancreatic cancer patients. Conclusion The expression of CD4 +IL-17 +cells in the tumor tissue is positively correlated with tumor angiogenesis, while that of IL-17 +cells with the clinicopathological parameters and survival time of the patients and therefore may serve as an important immune indicator for the prognosis of pancreatic cancer.