Objective To investigate the expression of peripheral blood (PB) CD4+ CD25+ Foxp3+ regulatory T cells (Tregs) in patients with benign end-stage liver disease after liver transplantation and the relationship between levels of PB Tregs and acute rejection. Methods A prospective analysis was performed on 55 consecutive patients who underwent liver transplantation.Fourteen out of 55 cases suffered from acute rejection after liver transplantation were defined as rejection group,while the rest patients were classified into no acute rejection group. PB was obtained from liver transplant patients at different time points longitudinally: pre-transplant, post-transplant within one year and acute rejection. The circulating CD4+ CD25+ Foxp3+ Tregs in PB were measured by flow cytometry. Blood samples were drawn during acute rejection, at the same time, liver biopsies were performed. The circulating CD4+ CD25+ Foxp3+ Tregs were compared between two groups.Results There was no difference between two groups in levels of circulating CD4+ CD25+ Foxp3 + Tregs cells pre-transplant. However, the levels of circulating CD4+ CD25+ Foxp3+ Tregs in rejection group were decreased significantly as compared with no-rejection group (2. 23 % ± 0. 54 % vs. 2. 99 % ±0. 86 %,P＜0.01). The frequency of CD4+ CD25+ Foxp3+ T cells was negatively correlated with rejection activity index (RAI) (r = - 0. 80, P＜0. 01 ). Conclusion Monitoring PB CD4+ CD25+ Foxp3+ Tregs levels may be helpful in evaluating the immune state and act as a more sensitive marker for acute rejection diagnosis in the patients following liver transplantation.

Objective To study the effect of anterolateral calf flap with extra-territorial blood supply in repairing large area of foot skin tissue injury. Methods 40 cases of arge area of foot skin tissue injury were divided into the observation group which were given anterolateral crural flap transplantation and the control group which were given anterolateral thigh flap transplantation. The wound healing condition and walking ability were observed. Results The level of wound healing, Holden walking ability and FAC walking ability were better than control group (P<0. 05). In the observation group, wound healing time was (15. 2 ± 6. 2) d and donor skin crusting time was (9. 4 ± 1. 5) d, which were less than the control group(P<0. 05). Conclusion Anterolateral calf flap with extra-territorial blood supply has positive clinical value as it can improve wound healing and walking function.

Aim To observe the effects of tetrahydro-biopterin ( BH4 ) on nitric oxide ( NO ) production in the kidney of type 2 diabetic nephropathy ( DN) mice, and to find a new target for the treatment of type 2 DN. Methods The 12 week-old db/db mice developed in-to DN phase were divided into 2 groups:DAHP group, subjected to intraperitoneal injection of 150 mg·kg-1 DAHP (n=8);DN group, subjected to intraperitone-al injection of same dose of normal saline containing 5% DMSO ( n = 6 ) . The age-matched db/m mice ( NS group) were subjected to intraperitoneal injection of same dose of normal saline containing 5% DMSO ( n =6 ) . Three groups of mice were treated for 7 days. Then the fasting blood-glucose, serum creatinine, u-rine protein and activity of iNOS were determined by chemical colorimetry. And the iNOS protein in renal cortex was determined by immunohistochemisty and western blot, respectively. BH4 was measured by HPLC method. NO level was determined by Griess method. Results The levels of fasting blood-glucose, serum creatinine, 24h urine volume, 24h urine pro-tein, BH4 , iNOS and NO in DN group were signifi-cantly higher than those in NS group;The levels of ser-um creatinine, urine volume, urine protein, BH4 , iN-OS and NO in DAHP group were significantly lower than those in DN group. Conclusion In the kidney of type 2 DN mice, the increased BH4 contributes to over-production of NO by the increased iNOS expression, and resultes in the increase of urine volume and urine protein.

Objective To probe the indication of biliary tract reconstruction without T-tube in orthotopic liver transplantation.Methods We put forward indications of biliary tract reconstruction without T-tube in orthotopic liver transplantation since January 2004 and there were 102 patients who underwent liver transplantation in our hospital without a T-tube in place after biliary tract reconstruction.The incidence of biliary tract complication was observed in these patients.Results All patients were followed up for more than 6 months.The incidence of biliary tract complication was 4.9 percent(5/102)in this group with 3 patients of intrahepatic difluse bile duct stenosis necessitating liver re-transplantation.The other 2 patients with common hepatic duct nonanastomotic stenosis were healed by ERCP plus stent placement.Conclusions Biliary tract reconstruction without T-tube placement helps to decrease the incidence of biliary tract complications resulting from the T-tube removal.

Objective To study the clinical significance of CD39 on regulatory T (Treg) cells in the peripheral blood (PB) of patients following liver transplantation and to evaluate the relationship between the levels of CD39+ Treg cells in the PB and acute rejection.Methods A prospective study was conducted to compare the CD39+ Treg cells from 76 liver transplant patients with those coming from 20 age-matched healthy individuals.The PB samples were collected within one year at different time points post-transplant.Blood samples and liver biopsies were collected at the time when acute rejection was diagnosed.The percentages of CD39 within the CD4+ CD25+ T cells were measured by using flow cytometry.The liver transplant patients were classified into two groups:the rejection group which consisted of 17 patients who an episode of acute rejection,and the non-rejection group consisted of the remaining 59 patients who had no acute rejection episodes.The percentages of CD39 within the CD4 + CD25 + T cells and the inhibition function of the CD39+ Treg cells were compared between the two liver transplant groups.Results The percentages of CD39 within the CD4+ CD25+ cells were significantly lower in the rejection group during acute rejection as compared to the non-rejection group (P ＜ 0.05).The percentages of CD39 within the CD4 + CD25 + cells were negatively correlated with the Rejection Activity Index (r =-0.86,P ＜ 0.05).The inhibition rate regarding the CD4+ CD25+ CD39+ Treg cells in patients with acute rejection was significantly lower than those without rejection (P ＜ 0.05).Conclusions The percentages of CD39 within the CD4+ CD25+ T cells were significantly lower in the rejection group during acute rejection and were negatively correlated with the RAI.The inhibition rate regarding the CD4+ CD25+ CD39+ Treg cells in patients with acute rejection was significantly lower than those without rejection.

A method for simultaneous determination of 20 Perfluorinated alkyl substances ( PFAS) in animal liver using QuEChERs and HPLC_MS/MS technique was developed. The samples were extracted with 0. 1%hydrochloric acetonitrile and cleaned up with C18 , N_Propylethylendiamine ( PSA ) and graphitized carbon blacks ( GCB ) . The analytes were separated by a reversed phase C18 column and gradiently eluted with a mixed solution of 5 mmol/L ammonium acetate methanol and 5 mmol/L ammonium acetate. The samples were quantified using isotope internal standard and external standard with the matrix matched standard calibration curve method. Good linearity was obtained for all the 20 PFAS at the concentration of 0. 1-10 μg/L with the linear correlation coefficients more than 0. 9995. The limits of detection (LOD) and the limits of quantification ( LOQ) for PFAS were 0. 05-0. 2 μg/kg and 0. 4-0. 5 μg/kg, respectively. The recoveries at three different concentration levels ( 0 . 5 , 2 and 5 μg/kg ) were in the range of 70 . 3% -108 . 1%. The repeatability expressed as relative standard deviations (RSD) was ranged from 2. 1% to 11. 9% (n=6).

Objective To investigate the curative effect of percutaneous transhepatic cholangial drainage(PTCD) on patients with non-anastomotic stricture after liver transplantation when treated with endoscopic retrograde cholangio-pancreatography(ERCP) unsuccessfully.Method The clinical data of 29 patients with non-anastomotic biliary stricture after liver transplantation were retrospectively analyzed,who failed to respond to ERCP and underwent PTCD from January 2005 to December 2007.Result All patients were performed PTCD successfully including cholangiography in 141 cases,drainage tube replacement in 115 cases,and balloon dilation of bile duct stricture in 39cases.The intubation time ranged from 2 months to 65 months.The mean levels (x ± SD) of alanine aminotransferase,aspartate aminotransferase and total bilirubin were 68.0 ± 29.1 U/L,52.6 ± 34.8 U/L,63.2 ± 33.3 μmol/L after treatment in comparison to 178.3 ± 63.3 U/L,144.0 ± 59.1 U/L,154.2 ± 92.0 μmol/L before treatment.Conclusion PTCD,which could improve the symptoms and prolong the survival time of both grafts and patients in spite of inconvenience of intubation,is suggested for patients with non-anastomotic biliary stricture if they are not suitable for liver retransplantation.

ObjectiveTo evaluate the feasibility and efficacy of ex-vivo liver resection combined liver autotransplantation for patients with massive primary liver cancer who underwent complex liver resection.Methods The clinical data of 4 patients suffering from massive primary liver cancer who were admitted to the Beijing Chaoyang Hospital from January 2008 to May 2010 were retrospectively analyzed.Regular liver resection could not be carried out because the first,second and third hepatic hilum of the 4 patients were invaded by the tumors,so ex-vivo liver resection combined liver autotransplantation were performed.ResultsThe operation was successfully carried out for the 4 patients.The operation time,the duration of anhepatic phase and the volume of operative blood loss were 690-840 minutes,250-300 minutes and 400-1400 ml,respectively.Portacaval bypass operation was not performed.After ex-vivo liver resection,the inferior vena eava or hepatic vein and portal vein of the 4patients were repaired,and the allogenous blood vessels were kept to extend the superior vena cava of the remnant liver so as to facilitate the anastomosis of blood vessels and reconstruction of the first hepatic hilum. After operation,the hepatic function of 1 patient was back to normal; 1 patient who stfffered from abdominal hemorrhage received reoperation for hemostasia; 1 patient was found with hepatic dysfunction; 1 patient died of hepatorenal dysfunction at postoperative day 5.Compensatory hypertrophy was observed in the 3 patients who survived at postoperative months 1-2.Of the 3 patients,2 were found with multiple pulmonary metastases at postoperative months 8 and 9,and they died at postoperative mouths 13 and 15.Until April 2012,1 patient survived for 37 months with no tumor recurrence or metastasis. ConclusionsEx-vivo liver resection combined liver autotransplantation provides the technical feasibility for performing complex liver resection for patients. The incomplete compensation of liver function and the short-term recurrence of tumors after operation are still the main issues which hinder the development of this technique.

Objective To explore causes leading to and the timing of liver retransplantation. Methods Among 164 cases of liver transplantation from Jul. 1999 to Dec. 2004, 6 cases underwent retransplantation with an incidence of 3. 65%. Causes included multiple intrahepatic bile duct stricture by ischemic reperfusion injury in 3 cases, hepatic artery stricture and thrombosis, hepatitis B recurrence, outflow obstruction of hepatic veins in one each. Results Clinical symptom improved in 4 cases, and failed to improve in 2 cases. Two cases suffered from intraabdominal bleeding, one biliary leak, one bacterial infection, two mold infection. Two patients died from bacterial and mold infection in four months. Conclusion Ischemic reperfusion injury is main cause resulting in intrahepatic bile duct stricture, liver retransplantation should be performed when the function of graft deteriorates significantly and conservative therapy fails.

Objective To investigate the use of laparoscopic ultrasound to exclude cystic duct obstruction and its related risk factors in laparoscopic cholecystectomy.Methods The data of 28 patients who underwent laparoscopic cholecystectomy in our department for cystic duct obstruction from February 2008 to April 2010 were analyzed.Subtotal resection of gallbladder and exclusion of cysticduct were carried out when the gallbladder triangle anatomy was not clear.An abdominal drain was used.Results All the patients were cured and there was no bleeding,abdominal infection,or jaundice.On univariate analysis,risk factors for cystic duct obstruction were adhesions in Calot triangle,gallbladder atrophy,acute cholecystitis,cystic duct stone incarceration,gallbladder wall thickening and white bile.Adhesion in Calot triangle,acute cholecystitis and white bile were independent risk factors on multivariate analysis.Conclusion Excluding cystic duct obstruction by laparoscopic ultrasound for patients who underwent laparoscopic cholecystectomy for cystic duct obstruction is safe and effective.