BACKGROUND:Hepatocyte transplantation can improve biochemical parameters and survival of animals with acute liver failure.However,cell sources.immunological rejection,distribution and histomorphological alternation of transplanted hepatocytes are important issues for its wide application in clinic.OBJECTIVE:To observe early-stage histomorphological and ultrastructural changes of SV40LT antigen gene transfected hepatocytes after transplanted into rat spleen.DESIGN,TIME AND SETTING:Randomized controlled animal trial was performed at Laboratory of National Hepatobiliary and Enteric Surgery Research Center.Ministry of Health from March to December 2001.MATERIALS:Sixty male Wistar rats were selected for hepatocyte transplantation.METHODS:Sixty rats were randomly divided into 4 groups(n=15).Primary cell group and primary cell plus cyclosporine A group were intrasplenically transplanted with primarily cultured hepatocytes;SV40LT antigen gene group and SV40LT antigen gene plus cyclosporine A group were intrasplenic transplanted with SV40LT antigen gene transfected hepatocytes.Twenty-four hours before and 14 days after transplantation,the rats in primary cell group and SV40LT antigen gene group were injected with 0.5 mL normal saline through tail vein daily,while the other groups were injected with cyclosporine A (10mg/kg per day).MAIN OUTCOME MEASURES:The spleen of one rat was harvested every day postoperatively for light microscopic and electron microscopic examinations to observe survival rate,histomorphological and ultrastructural features of transplanted hepatocytes for 14 days.RESULTS:Compared with primary cell group and SV40LT antigen gene group,the survival rate of transplanted hepatocytes in the other groups was significantly increased(P<0.05),but the histomorphological and ultrastructural changes were minor.There were no significant differences in the survival rate between SV40LT zntigen gene transfected hepatocytes and primarily cultured hepatocytes during the first week after transplantation(P>0.05),while the survival rate in SV40LT antigen gene transfected hepatocytes was significantly higher than primarily cultured hepatocytes 8-14 days postoperatively(P<0.01).CONCLUSION:Under treatment of cyclosporine A,the SV40LT antigen gene transfected hepatocytes can maintain a stable ultrastructure and a long survival rate.

BACKGROUND: Hepatocyte transplantation can improve biochemical parameters and survival of animals with acute liver failure. However, cell sources, immunological rejection, distribution and histomorphological alternation of transplanted hepatocytes are important issues for its wide application in clinic. OBJECTIVE: To observe early-stage histomorphological and ultrastructural changes of SV40LT antigen gene transfected hepatocytes after transplanted into rat spleen. DESIGN, TIME AND SETTING: Randomized controlled animal trial was performed at Laboratory of National Hepatobiliary and Enteric Surgery Research Center, Ministry of Health from March to December 2001. MATERIALS: Sixty male Wistar rats were selected for hepatocyte transplantation. METHODS: Sixty rats were randomly divided into 4 groups (n=15). Primary cell group and primary cell plus cyclosporine A group were intrasplenically transplanted with primarily cultured hepatocytes; SV40LT antigen gene group and SV40LT antigen gene plus cyclosporine A group were intrasplenic transplanted with SV40LT antigen gene transfected hepatocytes. Twenty-four hours before and 14 days after transplantation, the rats in primary cell group and SV40LTor antigen gene group were injected with 0.5 mL normal saline through tail vein daily, while the other groups were injected with cyclosporine A (10 mg/kg per day). MAIN OUTCOME MEASURES: The spleen of one rat was harvested every day postoperatively for light microscopic and electron microscopic examinations to observe survival rate, histomorphological and ultrastructural features of transplanted hepatocytes for 14 days. RESULTS: Compared with primary cell group and SV40LT antigen gene group, the survival rate of transplanted hepatocytes in the other groups was significantly increased (P 0.05), while the survival rate in SV40LT antigen gene transfected hepatocytes was significantly higher than primarily cultured hepatocytes 8-14 days postoperatively (P

Objective: To construct a recombinant adenovirus(rAdCDES) which is capable of both direct and indirect treatment to mammary cancer and enhancement to the antitumor effect of radiation. Methods: A method of homologous recombination in bacteria was used to construct prAdCDglyES. The recombination adenovirus was transfected to 293 cells by liposome, in which rAdCDES was packaged and generated. The growth curve and MTT methods was used to detect the growth inhibition effect of rAdCDES on MCF-7; rAdCDES was directly injected into established MA737 tumors-bearing mice for observing difference in tumor size and survival days of mice and enhancement of the antitumor effect of radiation. Results: The inhibiting rate of rAdCDES on MCF-7 cell was (83. 1?8. 1)% and had significant difference compared with control was (19.2 ? 7.8)% (P

Objective To summarize the experience of diagnosis and treatment of solid-pseudopapillary tumor of pancreas (SPTP).Methods The clinical data of 15 patients with SPTP who were treated in our hospital from March,2003 to March,2006 were analyzed retrospectively.Results The 15 cases were all women, and the average age was 29.4 years.The chief manifestations were abdominal mass, abdominal pain or other abdominal discomfort. None of the 15 cases had history of pancreatitis or abdominal trauma, and no long history of drinking or smoking. Six cases were negative for CEA,CA50,CA199,CA125 and other tumor markers.Solid and solid-cystic masses in pancreas or solid and solid-cystic tumors in retroperitoneum were found both by B-mode ultrasonography and CT examinations. Preoperative fasting blood sugar was within normal limits. The tumor in 8 cases was located in the pancreatic head, in 6 cases was in the body and tail of pancreas, and in 1 case was in the neck of pancreas. The diameter of the tumors was 2.5-10 cm. No metastasis was found in the abdominal cavity or liver. Local excision was performed in 6 cases, distal pancreatectomy was performed in 5 cases, including 2 cases combined with splenectomy, and pancreaticoduodenectomy was performed in 3 cases, segmental pancreatectomy was performed in one patient with tumor in the neck of pancreas. The 15 cases showed typical pathologic manifestation of SPTP by microscopy. At followed up for 16-52 months, no evidence of recurrence or metastasis in these cases was found.Conclusions Solid-pseudopapillary tumor of pancreas primarily affects young women, and it may be located in any part of pancreas. Surgical resection is recommended as the treatment of choice, and the prognosis is good.

Objective To explore the efficacy difference of biliary stent combined with 125I seed intracavity irradiation and palliative biliary drainage in the treatment of obstructive jaundice caused by pancreatic head cancer. Methods The clinic date of 95 patients with pancreatic head cancer who underwent palliative surgery from June 2010 to June 2015 were analyzed retrospectively. 46 of the cases were treated with percutaneous biliary metal stent implantation combined with 125I seed intracavity irradiation (stent group), and the other 49 cases were treated with palliative biliary enterostomy (surgery group). The levels of jaundice, liver function, tumor size, survival time and hospitalization cost were compared between the patients before and after treatment. Paired t-test was applied for statistical analysis. Results Both the two treatment coulel effectively relieve jaundice and improve liver function. The mean TBIL of surgery group and stent group was (29.4±9.6) and (21.0±8.0)μmol/L after three months treanment, (40.3±11.0) and (24.4±9.6)μmol/L after six months treatment, respectively. Tumor maximum diameter of the two groups were (37.9 ± 4.5) mm and (33.5 ± 6.0)mm after three months treatment, (45.9 ± 5.0) mm and (37.0 ± 6.5) mm after six months, respectively. Each time point between them had statistical significance (t=-4.235,-5.476,-3.654,-6.702, P<0.05). The mean survival time was (12.83 ± 0.80) months in the stent group and (9.06±0.49) months in the surgery group, the difference was statistically significant (t=-3.49, P<0.01). The mean hospitalization cost of the stent group was (38 453.0 ± 7 282.0) yuan and the surgery group was (43 057.0 ± 7 667.4) yuan, the difference was statistically significant (t= 2.759, P<0.05). Conclusion For inoperable pancreatic head cancer patients, compared with palliative biliary enteric drainage, percutaneous biliary stent placement combined with 125I seed intracavity irradiation can effectively relieve biliary obstruction, improve liver function, inhibit tumor growth and prolong survival time.

Objective To investigate the effects of captopril on intubation response and the mechanism. Methods Thirty ASA Ⅰ - Ⅱ patients aged 25-60 yr, weighing 45-70 kg, scheduled for elective surgery under general anesthesia with tracheal intubation and mechanical ventilation were randomly divided into 2 groups: captopril group ( n = 15) and control group ( n = 13) . Premedication consisted of intramuscular phenobarbital 0.2 g and atropine 0.5 mg. In captopril group, captopril 0.3-0.35 mg?kg-1 was injected intravenously 10 min before induction while in control group normal saline 10 ml was given instead. Anesthesia was induced with droperidol 0.05 mg?kg-1 , fentanyl 1 ?g?kg-1 , thiopentone 5-6 mg?kg-1 . Intubation was facilitated with succinylcholine 1.5 mg ? kg-1 . Laryngoscopy and tracheal intubation was successfully performed within 30 seconds. Mechanical ventilation was started and enflurane inhalation was begun and maintained at 2.1 % , SBP, DBF, HR, EGG and SpO2 were continuously monitored. Blood samples were taken from peripheral vein before induction (T0 ) , at intubation (T1 ), 1-1.5 min (T2) and 5 min (T3) after intubation for determination of plasma concentrations of angiotensin- Ⅰ (A 1 ), angiotensin- Ⅱ (A Ⅱ ), aldosterone (ALD), noradrenaline (NE), adrenaline (E), atrial natriuretic polypeptide (ANP) and thromboxane B2(TXB2) .Results (1) In captopril group SBP, DBF, HR, and heart rate-systolic BP product (RPP) remained unchanged at intubation, while in control group the parameters were significantly increased at T1 or T2 as compared with the baseline values ( P

Objective To investigate the clinical value of improved percutaneous tramhepatic cholangiographic drainage(PTCD)and percutaneous placement of biliary metallic stents on the treatment of malignant obstructive jaundice.Methods Thirty three patients with malignant biliary obstruction were treated by the improved PTCD guided by ultrasound followed by pereutaneous placement of biliary tract metallic stcnts 1～3 weeks later.Results Improved PTCD WaS succeeded in all the patients,30 patients(90.9%)accepted percutaneous placement of biliary metallic stents successfully,3 patients who failed in stents placement accepted PTCD again for palliative management of iallndice.Thofle who failed in stent placement were found to have bleeding from the PTCD tube,but recovered several days later.Jaundice alleviated in all the patients.The patients were followed up for 1～29 months,medium follow-up period is 14.8 months.Six cases were found lever and jaundice,among that,two ageepted improved PTCD again,one recovered after antibioic treatment,the remaining 3 patients failed in further treatment due to the metastasis of the tumor.Condusion Pereutaneous placement of biliary metallic stents after improved PTCD is a safe and effeetive method to relieve malignant biliary obstruction,it can improve life quality,prolong survival time.This method could be a good choice for treating the patients with mnlignant biliary obstruction without operation.

Objective:To investigate the effect and mechanism of Wogonin on CD3AK cell proliferation and cytotoxicity to SMMC-7721.Methods:CD3AK cells were cultured from peripheral blood mononuclear cells ( PBMC) in vitro by a variety of cytokines for 7 d,and treated with different concentrations of Wogonin for 48 h.CD3AK cells proliferation was measured by CCK-8 assay.SMMC-7721 cell growth was detected by MTT.The expression of perforin (PFP),granzyme B (GrB) and CD107a on CD3AK cells were measured by flow cytometry ( FCM).The cytotoxicity to SMMC-7721 cells was detected by LDH release assay.The expression of ERK1/2 on CD3AK cells was detected by Western blot.The mobility of SMMC-7721 cells was detected with transwell chambers.The merge of SMMC-7721 cells were measured with Wound healing assay.Results:Wogonin could significantly promote CD3AK cells proliferation, especially at 3.2 mg/L (23%higher than that of control group,P<0.05).The highest cytotoxicity to SMMC-7721 was also at the con-centration 3.2 mg/L (60.4%).The expression of PFP,GrB,CD107a were significantly higher than that of control group( P<0.05).The expression of ERK1/2 was obviously improved,especially at 12.5-0.8mg/L.After treated with Wogonin 50,12.5,3.2,0.8,0.2 mg/L for 48 h,the lowest transwell cell was at 12.5 mg/L and lowest merge rate was at 3.2 mg/L.Conclusion:Wogonin could promote CD3AK cell proliferation and enhance the cytotoxicity to SMMC-7721.Wogonin could also inhibit SMMC-7721 cell growth,migration and cell merge.The mechanism may be related to activated ERK1/2 and increase the expression of PFP,GrB,CD107a.

Objective To evaluate the clinical application of the treatment on malignant obstructive jaundice by combination of percutaneous biliary stent placement and 125 I particles intraluminal implanation.Methods As a prospective study , 32 patients with malignant obstructive jaundice who either have no opportunity for radical operations or unwilling to be surgically treated were enrolled to be treated with percutaneous biliary stent placement and 125 I particles intraluminal implanation.Biochemical routine tests , blood routine tests , tumor markers , coagulation function , color ultrasound , CT and magnetic resonance cholangiopancreatography were conducted prior to the operation to obtain general information of the clinical status of the patients and the tumor and the site of obstruction.Percutaneous transhepatic cholangial drainage was performed first under the B -type ultrasound system.After one week , biliary stents were placed under DSA.According to the stent expansion presented by real-time DSA imaging , 125 I particles were implanted simultaneously or afterwards.Routine biochemical tests and cholangiopancreatography under DSA were conducted in one week , one month and three months after the implantation.Variance analysis was performed with repeated measurements to compare the difference of liver function indexes pre -and post-operation.Meanwhile, 125I particle displacement, falling off and stent patency were observed.After three months, the tumor size was measured by CT.Student t-test was used to compare the tumor sizes of pre-and post-operation.Results The symptoms of jaundice resolved and the general physical conditions improved in 32 patients substantially.The total bilirubin level decreased from preoperative level of (302 ±169)μmol/l to the level of (108 ±50)μmol/l at one week following the surgery , and the indirect bilirubin level decreased from preoperative level of ( 250 ±160 )μmol/l to the level of ( 85 ±43 )μmol/l at one week following the surgery(F=76.32,58.23,P<0.01).The maximal diameters of the tumors decreased from preoperative size of (3.78 ±1.14)cm to the size of (3.14 ±1.28)cm at three months following the surgery, and the minimal diameters of the tumors decreased from preoperative size of ( 2.80 ±0.88 ) cm to the size of ( 1.93 ± 1.00)cm at three months following the surgery, and the differences were statistically significant (t=5.11, 6.73,P<0.05).By using Kaplan-Meier survival curve, the average survival periods were ( 9.9 ± 0.7) months.Conclusions Percutaneous biliary stent placement and 125 I particles intraluminal implanation have definite short-term effects in prolonging survival time , stent patency time and improving the living standard of the patients.The technique is safe and simple.It only needs small incision , has no absolute contraindications and can be applied repeatedly .

AIM:To investigate the regulatory effects of phosphatylinositol 3-kinase/protein kinase B (PI3K/PKB) signaling pathway on the expression of osteopontin ( OPN) in transforming growth factor-β1 ( TGF-β1 )-induced hu-man hepatic stellate cells .METHODS:Human hepatic stellate cell line LX-2 was cultured in DMEM and stimulated by TGF-β1 at the final concentration of 2.5, 5, 10 and 20 μg/L for 24 h or at final concentration of 10 μg/L for 12 h, 24 h and 48 h.LX-2 cells were pretreated with wortmannin , a specific inhibitor of PI3K/PKB signaling pathway , at final con-centration of 0.1 μmol/L for 1 h, followed by incubation with TGF-β1 at final concentration of 10μg/L for 24 h.The cells were collected.The expression of OPN was detected by real-time PCR and Western blotting .RESULTS: In LX-2 cells, the expression of OPN was apparently elevated when incubated with TGF-β1 .With the increase in TGF-β1 concentration or the extension of incubation hours , the expression of OPN was increased gradually in a dose-and time-dependent manner with certain limits.LX-2 cells pretreated with wortmannin and incubated with TGF-β1 had a significant decrease in the OPN expression as compared with control group (P<0.01).CONCLUSION:The expression of OPN in TGF-β1-induced LX-2 cells is regulated by the PI3K/PKB signaling pathway.