ObjectiveTo set up the porcine combined liver/small bowel orthotopic allotransplantation model.MethodThe donors including liver, about 2 to 3?m proximal jejunum, duodenum and a sma ll part of pancreas head were transplanted to the hybridized long-white recipie nt pigs after the resection of their livers and the majority of small bowels.ResultWithout using immunosuppressive drugs, the median survival time of the animals w as 92?h with the longest survival time being 196?h, and there was about 75?% of the animals lived for more than 24?h after operation.ConclusionThe successful establishment of the combined liver/small bowel transplantation m odel will do great favor to our future clinical practice and further study of th e combined liver/small bowel transplantation.

0. 05), respectively. Conclusion: Repeated +Gz stress may disorder the ratio of ET and NO. This imbalance may be one of the causes which lead to pathology changes in masseter musscles.

Objective To study the impact of preprocedure intravenous urography (IVU) on the extracorporeal shock wave lithotripsy(ESWL) for proximal ureteral stones.Methods One hundred patients with solitary radiopaque proximal ureteral stones on plain radiographs and no severe hydronephrosis on ultrasonographic examination were allocated randomly to two treatment groups.IVU group (n=50) had IVU before the start of ESWL,whereas patients in control group (n=50) underwent ESWL without IVU.Postop- erative success,the stone-free rates and complications were evaluated in both groups. Results Seven patients in IVU group were excluded from the study. The success rate [95.3%(41/43) in IVU group vs 94.0% (47/50) in control group],stone-free rate [83.7% (36/43)vs 86.0% (43/50)] and complication rate[27.9% (12/43 ) vs 26.0% (13/50)]were similar in two groups (P＞0.05).Conclusions It is not necessary to obtain an IVU for patients who have solitary radiopaque proximal ureteral calculi on plain radiographs with no severe hydronephrosis on uhrasonographie examination before scheduling them for ESWL,thus minimizing the cost,avoiding exposure to contrast medium,and reducing radiation exposure.

Objective To isolate and indentify fetal hepatic progenitor/stem cells, and study the feasibility and effectiveness of their transplantation on acute liver injury in nude mice. Methods The primitive cells isolated from 13.5dpc pregnant mouse fetal hver by way of enzyme digesting were cultured in vitro and liver specific markers as AFP, CK19, Albumin, c-Met, were identified by immunofluorescence and RT-PCR on colonies. 4 × 105 cells were transplanted into nude mice with carbon tetrachloride induced acute liver injury. Hapatic functions were measured pre- and -post transplantation on days 1,2,4, 7. Meanwhile, hepatic pathology was studied. Results Compared to control group the hepatic functions gradually recovered in transplant group, on days 1,2,4 after fetal hepatic stem cell transplantation. The hepatic pathology significantly improved in stem cells transplantation group. Conclution Fetal hepatic progenitor/stem cell were successfully yielded by enzyme digest. Stem cells transplantation improved the hepatic function and pathology in acute hapatic injury model of nude mice.

Objective To investigate the prevention and mechanism of Extracorporeal shock wave lithotripsy(ESW) induced renal Injury by pre-treating kidneys with low-energy shock waves(LESW).Methods Forty healthy female domestic rabbits were surgically managed to the mono-nephron models and random divided into 4 groups consisting of ten each: Control,LESW,ESW and ESWL plus LESW pretreated groups.LESW group received 100 LESW,ESW group received 1500 standard ESW,and same dose on ESW group except 100 LESW pretreatment in ESW plus LESW pretreated group.The rabbit kidney tissues were obtained 24 hours after ESW.Activity of superoxide dismutase(SOD) and malondialdehyde(MDA) levels in the renal tissue,and the level of N-acetyl-β-D-glucosaminidase(NAG) in urinary were measured.Renal cell apoptosis was detected by TdT-mediated dUTP Nick End Labelling(TUNEL).Results The MDA,the urinary level of NAG and rate of apoptosis in the LESW groups were reduced(P<0.01),and the activity of SOD increased significantly(P<0.05) as compared with ESW group,and these changes in LESW group had no statistics difference compared with the control group(P>0.05).Conclusions LESW pretreatment protocol substantially limits the renal injury that often caused by ESW.LESW may suppress oxidative stress and antagonize the process of renal cellular apoptosis.

Objective To investigate the cytotoxic effects and mechanism of PNP-CD chimeric gene vector originated from PNP/MeP-dR system on HCC cells. Methods The fusion suicide gene PNP-CD obtained by site directed mutagenesis technique was subcloned into pcDNA3.0 to construct a eukaryotic expression vector containing a chimeric gene, pcDNA3.0/ PNP-CD. After being identified by recombinant enzyme, PCR and subsequent sequencing, it was transfected into HepG2 cells by liposome-mediation method. The G418-resistant cellular clone with stable transfection of pcDNA3.0/PNP-CD, HepG2/PNP-CD was established by selection. The expression of PNP-CD gene was also verified by RT-PCR and Western blotting. The curve of cellular growth was assayed by Trypan blue exclusion. The cellular sensitivity of HepG2/PNP-CD to its specific prodrugs and its bystander effects were also assayed by MTT method. Results The chimeric gene, PNP-CD, was inserted into pcDNA3.0 correctly, and the stable expression of pcDNA3.0/PNP-CD in HepG2 cells was confirmed.This cellular clone was highly sensitive to its corresponding prodrugs. It was indicated that its bystander effects with the synergetic treatment of its specific prodrugs were substantially higher than those caused by the same vector with the administration of only a single prodrug, MeP-dR. Conclusion The bi-functional fusion suicide gene vector, pcDNA3.0/PNP-CD, yields powerful cytotoxic effects on HCC cells in the presence of the synergetic treatment of its specific prodrugs, which would be a high-performance therapeutic vector in gene therapy for liver cancer.

Objective With ordinary acupuncture as the control, to observe the feasibility and effectiveness of electroacupuncture at Jiaji (EX-B2) points in treating primary trigeminal neuralgia.Method Forty patients with primary trigeminal neuralgia were randomized into a group of electroacupuncture at Jiaji points and an ordinary acupuncture group, 20 cases in each group. In the electroacupuncture group, Jiaji of C2and T1on the affected side were selected, while the points were selected by following the Acupuncture-moxibustion Therapeuticsin the ordinary acupuncture group. Carbamazepine (CBZ) was taken as the basic treatment for the two groups. A treatment course (3 weeks) was observed, and follow-up study was conducted every 4 weeks for a total of 12 weeks. The dose of CBZ, Visual Analogue Scale (VAS) and Short-form McGill Pain Questionnaire (SF-MPQ) scores at each time point were recorded. The changes of the index in the two groups at each time point were compared.ResultAt the end of the treatment (the third week), the dose of CBZ, VAS and SF-MPQ scores all declined in the two groups, and the between-group differences were statistically insignificant (P>0.05). The follow-up study in the 7th week showed that the data of the observed indexes all decreased in the two groups, while the electroacupuncture group presented a more significant efficacy, and the between-group differences were statistically significant (P<0.05). The follow-up studies in the 11th and 15th weeks showed that the data of the observed indexes continued to decline in the electroacupuncture group but began to increase in the ordinary acupuncture group, and the between-group differences were statistically significant (P<0.001).Conclusion Electroacupuncture at Jiaji points and ordinary acupuncture both can reduce the dose of CBZ for patients with primary trigeminal neuralgia, improve pain and other discomforts as well as the negative emotions, but electroacupuncture at Jiaji can produce a more significant long-term efficacy compared to the ordinary acupuncture.

ObjectiveTo isolate and culture human adipose derived mesenchynal stem cells (hADSCs) and study the potential of hADSCs to differentiate into hepatocyte-like cells. MethodsTo ensure the removal of contaminating hematopoietic cells, hADSCs were selected based on plastic adherence. Cell surface antigen was confirmed by flow cytometry; ultramicrostructure was detected by transmission electron microscopy; adipogenic and osteogenic differentiation of hADSCs was analyzed by oil Red O staining and yon Kossa staining. hADSCs were exposed to differentiation medium containing EGF,FGF,OSM, HGF,TSA in vitro. 10% CCl4 (100 μ1/20 g body weight )was injected into immune-deficient BALB/c-nu mice and hADSCs (5 × 105 cells) were simultaneously administrated from the tail vein. Blood samples were collected and concentration of aminotransferase and direct bilirubin were detected. Administration without hADSCs was used as a control. One month later, we sacrificed the mice and liver sections were examined by histochemical immunofluorescence with human ALB specific antibodies. ResultshADSCs exhibited fibroblast-like morphology, and expressed CD73, CD90,CD105, and were lacking of CD34 and CD45. Adipogenic and osteogenic differentiation showed that hADSCs have the capacity of multidifferentiation. The differentiated cells showed hepatocyte-like cell morphologies and hepatocyte-specific markers including albumin (alb) and α-fetoprotein (AFP). The bioactivity assays revealed that these hepatocyte-like cells could uptake low-density lipoprotein (LDL).Histochemical immunofluorescence showed that hADSCs were incorporated into injured livers. Some human alb-positive cells were found in liver sections after implantation of undifferentiated hADSCs. Transaminase activity in the experimental group was lower than in the control group. Conclution hADSCs can differentiated into functional hepatocyte-like cells and can relieve CCl4 induced BALB/c-nu acute liver injury.

Objective Accumulating reports have suggested that hepatic stellate cells (HSCs) exhibit immunosuppressive ability and may be responsible for the occurrence and development of hepatocellular carcinoma (HCC).The mechanisms through which HSCs affect T-cell-induced adaptive immune responses and the relationship with the regulatory T cells (Treg cells) were studied.Methods We isolated HSCs from wildtype mice to demonstrate the influence of HSCs on T-cell proliferation and explored their effect on Treg cells through mixed leukocyte reactions (MLRs) in vitro.Results We found that activated HSCs could induce T-cell hyporesponsiveness in adaptive immune response by inhibiting the proliferation of T cells andincreasing the quantity of Treg cells.Conclusion Activated HSCs may lead to hypoergia of T cells in adaptive immune reaction and up-regulate the expression of Treg cells,thus facilitating immunotolarance.

Objective To determine immune modulatory activity of activated hepatic stellate cells( HSCs) in hepatocellular carcinoma and immune response in tumor microenvironment. Methods Cell proliferation was measured by BrdU incorporation with a microtiter plate reader at 450 nm. The effect of HSCs on T cell proliferation was measured by MLR. Mouse hepatic cancer cell line H22 were implanted on the backs of BALB/c mice to establish the subcutaneous transplanted tumor model. Then the mice were sacrificed after 20 days for anatomical and size determination. Furthermore, Paraffin-embedded tissue was removed by serial tissue sectioning and immunohistochemically examined for expression of T lymphocyte subsets. T lymphocyte subsets in splenocytes were detected by FCM. Apoptotic mononuclear cells were evaluated by FITC-labeled Tunel assay. Results We determined that HSCsCM promoted hepatocellular carcinoma(HCC) cell line proliferation and HSCs inhibit T cell proliferation by MLR in vitro. We also examined normal immune mice to assess the immunosuppression of HSCs in the development of HCC. In the co-transplantation with HSCs group, T cells and their subtypes decreased obviously in the tumors and the spleen. The results showed that co-transplanted HSCs can induce more PD-L1 expression and more mononuclear cell apoptosis in tumor tissue. Conclusion Our results demonstrated that HSCs promote HCC progression partially because of their immune regulatory activity. Our data supply new information to support an integral role for HSCs in promoting HCC progression and suggest that HSCs may serve as a therapeutic target for HCC.