AIM: To evaluate the therapeutic value of endoscopic metal stent implantation for malignant biliary obstruction. METHODS: Clinical data of 212 patients with malignant biliary obstruction undergoing endoscopic biliary drainage were retrospectively analyzed. Based on the clinical and imaging data,malignant biliary obstruction was identified and according to drainage effect of endoscopic nasobiliary drainage and endoscopic retrograde biliary drainage (ERBD),endoscopic metal biliary endoprothesis was conducted and compared with palliative endoscopic drainage and ERBD. RESULTS: The success rate of one-off operation was 99.5%. For jaundice release,68.4% of the cases were satisfactory,27.5% were released,and 4.1% had no effect. The effective rate of jaundice release (satisfactory + release) in obstruction at low site was 96.6% and that at high site was 82.4%. Average duration of obstruction release was 289 days and average survival was 310 days. The complication rate was 6.7% with a mortality rate of 1.2%. CONCLUSION: Based on rigorous indication selection,EMBE is as a safe and effective therapeutic means to remove malignant biliary obstruction. EMBE may replace palliative biliary surgery for patients with mid-late stage biliary tumors.

Objective To study the effects of survivin antisense RNA on SGC7901 cell’s apoptosis and chemosensitivity to taxotere, and to investigate its effect on the expression of multi-drug resistance gene-1 (MDR-1). Methods Survivin antisense eukaryotic vector anti-pcDNA3-svv was transfected into SGC7901 cell lines by lipofectamine and positive clones were screened out then. Survivin protein and MDR-1 mRNA were measured by western blot and RT-PCR, respectively. Apoptosis that was induced by anti-pcDNA3-svv was observed by electronic microscope, and the sensitivity of SGC7901 cell to taxotere was examined by MTT. Results The expressions of survivin protein and MDR-1 mRNA in transfected SGC7901 cells both decreased more significantly than that of non-transfected cells (P

BACKGROUND: During xenogenic liver transplantation, major histocompatibility antigen can induce immunological rejection, and immunosuppressant can cause adverse effect on organism. Recently, treatment prior to transplantation induces immune tolerance, which is perspective for organ transplantation.OBJECTIVE: To investigate the correlation between thymus induction and immunological rejection during liver transplantation. METHODS: A total of 40 male SD rats of clean grade were selected as donors. Moreover, 30 male Wistar rats of clean grade and 10 male SD rats of clean grade were selected as recipients. The donor rats were divided into allogeneic gene transplantation, allotransplantation, cyclosporine, and thymus induction groups, with 10 rats in each group. The modified Kamada and improved two-cuff technique was used to establish a stable rat orthotopic liver transplantation model. The cyclosporine group was given cyclosporine (50 mg/kg) for 5 successive days. Thymus induction group was injected with major histocompatibility antigens (50 pL) for 5 successive days. Other groups were not given any interventions. Survival time of rats was recorded in each group. Pathological observation and mixed lymphocyte cultured were performed at days 3, 7, 14, 21, and 28 after transplantation. RESULTS AND CONCLUSION: Survival time was longer in the thymus induced group compared with other groups (> 60 days), damaged level was mild, local immunological rejection was reduced, and lymphocytes were decreased. The effect after liver transplantation was similar to allogeneic gene transplantation but superior to cyclosporine intervention (P < 0.05). This suggested that thymus induction relieved immunological rejection following liver transplantation.

BACKGROUND: Most patients who underwent liver transplantation would suffer acute rejection or transplanted liver failure resulted by chronic rejection, therefore, inducing specific immune tolerance via varied pathways is the ideal method to solve this problem. OBJECTIVE: To treat rat transplanted liver by injecting transforming growth factor β1 (TGF-β_1) plasmid, and to analyze the relationship between TGF-β1 and allograft rejection from gene level. METHODS: A total of 30 male, Wistar rats were served as allogenic liver donors, and 10 male, SD rats served as syngeneic donors Totally 40 male SD rats were served as liver recipients, and divided into 4 groups by order number table: ailogenic transplantation, syngeneic transplantation, ciclosporin, and ciclosporin plus TGF--β_1 groups. In each group, rat orthotopic liver transplantation model was established by modified Kamada and improved two-cuff technique. After modeling, rats were received cyclosporine 1-5 days in the cyclosporine group, or intraperitoneal injected ciclosporin for 1-5 days, combined with TGF-β_1 plasmid 0-2 days in the cyclosporine plus TGF-β_1 group. No intervention was performed in the other groups. The survival time of rats were recorded, and the pathological changes was detected at days 3, 7, 14, 21, and 28 after transplantation, then the mixed lymphocyte culture was performed. RESULTS AND CONCLUSION: The survival time of rats in syngeneic transplantation group and cyclosporine plus TGF-1,β_1 group was more than 60 days, which was obviously greater than that of allogenic transplantation and cyclosporine groups (P< 0.05). The histopathologic slide showed that there was moderate and severe acute rejection, with evident intrahepatic inflammatory cell infiltration in the allogenic transplantation and cyclosporine groups. Few rejections were observed in the syngeneic transplantatior group, which was close to the normal lever tissues. Mixed lymphocyte culture of the cyclosporine plus TGF-β_1 group was superior to the syngeneic transplantation group or cyclosporine group (P < 0.05). The results demonstrated that cyclosporine combined with local injection of TGF-β_1 plasmid can relieve post-transplant immune rejection.

BACKGROUND: A small number of mesenchymal stem cells (MSCs) present in bone marrow, which would gradually drop with age. OBJECTIVE: To verify the effect of adherent method on culture of bone marrow-derived MSCs. METHODS: Under anaesthesia, bone marrow cells were obtained from femur and tibia of rats, cultured by DMEM containing calf serum, placed in an incubator containing 5% CO_2 at 37 ℃. The culture medium was renewed after 24 hours, and remained periodical medium change with once per week. The weakly adherent cells were passaged. The cell morphology, growth curve, and the expression of cell-surface markers were identified by flow cytometry and immunocytochemical staining. RESULTS AND CONCLUSION: After 24 hours of culture, the cells could adhere to the walls with fusiform or triangle shapes, proliferated faster after 2-3 days, and presented whirlpool-like or clustering. The cells reached a logarithmic growth phase after 2 days, and into the late stationary phase after 12 days, which covered the bottle after 15 days. The cultured cells were positive to CD90 and CD54. The results verified that bone marrow-derived MSCs can be isolated by adherent method. This method is easy operation, and can maintain cell activity preferably.

BACKGROUND:Tumor necrosis factor-α(TNF-α), a main cytokine inducing chondrocyte apoptosis of osteoarthritis, plays a regulatory role in the process of osteoarthritis. OBJECTIVE:To compare the rat models of chondrocyte apoptosis induced by different mass concentrations of TNF-α, thus providing theoretical basis for further study on the regulation of drugs on chondrocyte apoptosis. METHODS:Chondrocytes were isolated from the knee cartilage of 4-week-old Sprague-Dawley rats of clean grade by mechanical l col agenase digestion and were then incubated with different mass concentrations of TNF-αto induce apoptosis. The morphology of chondrocytes was observed under inverted phase contrast microscope, cel s were identified using immunohistochemical staining of type II col agen, as wel as the cel viability and apoptosis were detected by MTT and DAPI, respectively. RESULTS AND CONCLUSION:(1) In vitro, the cytoplasm of chondrocytes was stained brown-yel ow by using immunohistochemical staining of type II col agen. (2) At 48 hours, the apoptosis rate of chondrocytes induced by 10, 20 and 30μg/L TNF-αwas significantly higher than that of the 0μg/L TNF-α(P<0.01), and the apoptosis rate of chondrocytes induced by 40μg/L TNF-αwas significantly higher than that of the 10μg/L TNF-α(P<0.01). (3) The viability of chondrocytes induced by 10, 20 and 40μg/L TNF-αwas significantly lower than that of the 0μg/L TNF-α(P<0.01). In detail, the viability of chondrocytes induced by 20μg/L TNF-αwas lower than that of the 10μg/L TNF-α(P<0.05);the viability of chondrocytes induced by 40μg/L TNF-αwas significantly lower than that of the 10 and 20μg/L TNF-α(P<0.01, P<0.05). (4) These results suggest that 20μg/L TNF-αcan successful y induce the chondrocyte apoptosis model.

Objective To explore the characteristic and clinical meaning of myocardial bridge in patients with multi-slice spiral CT(MSCT) coronary angiography.Methods 875 patients with suspected or diagnosed coronary artery disease were studied with MSCT coronary angiography,579 cases were male and 296 cases were female,ranged from 30~87 years old in age with average of 60 years old.The heart was scanned with retrospectively ECG-gated,reconstruction from phase for all original tomographic source images at 30%~40% R-R phase interval according to heart rate ≥75 bpm and at 40%~50% R-R phase interval according to heart rate ≤75 bpm.The images of maximum intensity projection,multiplanar reconstruction and vulume reconstruction were gained to show left and right main coronary artery and their main branchs on mulliple angle.Results The myocardial bridges were detected in 89 cases,the positive rate was 10.2%,36 cases(40.5%)had different grade stenosis(≤50%).The myocardial bridges located at middle segment of anterior descending artery(79.8%),the thickness of myocardial bridge was 0.06~0.55 cm,the atherosclerosis plaques were found in 20 cases,the vascular stenosis was approximate to 50%.39 cases with simple myocardial bridges had angina,21 cases hadn’t angina.23 cases with myocardial bridges accompanied by coronary artery atherosclerosis had angina.Conclusion MSCT coronary angiography may clearly detect the myocardial bridge,which can provide more worthy information for clinical diagnosis and treatment.

ObjectiveTo evaluate the fourth-generation ultrasonic lithotripsy system for treating the urinary tract stones.Methods243 cases of urinary tract stones who received treatment of fourth-generation EMS ultrasonic lithotripsy system were analyzed retrospectively.ResultsImmediate phase I lithotrotrisy was performed successfully in 227 cases and the successful rate of phase I was 93.3％ (227/243).Delayed phase Ⅱ lithotripsy was performed for 16 patients.Complications happened in 2 cases,one case occurred hydropneumothorax during operation,required indwelling thoracic drainage tube processing,the other had severe bleeding which conservative treatment was ineffective,cured by the intervention of super-selective renal artery embolization.Conclusion Fourth generation of ultrasonic lithotripsy for treating stones on different anatomical locations of urinary system was safe,practical and efficient,worthy of clinical application.

BACKGROUND:Bone marrow stromal stem cel s have multilineage differentiation capacity and can differentiate into transparent chondrocytes under certain conditions, which can provide new thoughts for treatment of osteoarthritis. OBJECTIVE:To investigate the effects of Tougu Xiaotong Granule containing serum on the cartilage differentiation of bone marrow stromal stem cel s. METHODS:Bone marrow stromal stem cel s from Sprague-Dawley rat limbs were cultured in vitro, and those cel s at passage 3 were used in the study. Cel s were divided into six groups:saline serum group, Tougu Xiaotong Granule water extract group, Tougu Xiaotong Granule alcohol extract group, chondroinductive group, Tougu Xiaotong Granule water extract and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group. The Sox9, col agen Ⅱ, and col agen X mRNA and protein expression levels were tested by real-time polymerase chain reaction and western blot analysis. RESULTS AND CONCLUSION:After cel s were intervened with drug-containing serum for 14 days, the Sox9, col agen Ⅱ, and col agen X mRNA and protein expression in Tougu Xiaotong Granule water extract group, Tougu Xiaotong Granule alcohol extract group, chondroinductive group, Tougu Xiaotong Granule water extract and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group were significantly higher than that in saline serum group (P<0.05, P<0.01). In addition, the chondroinductive group, Tougu Xiaotong Granule water extract serum and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group showed significantly higher expression levels than Tougu Xiaotong Granule water extract serum group, Tougu Xiaotong Granule alcohol extract group (P<0.01). Sox9 expression in Tougu Xiaotong Granule water extract serum and chondroinductive group were significantly higher than that in the chondroinductive group. Experimental findings indicate that, Tougu Xiaotong Granule containing serum can accelerate bone marrow stromal stem cel s differentiate into cartilage cel s by up-regulation of Sox9 expression.

BACKGROUND:Previous studies have found that electroacupuncture can delay articular cartilage degeneration mediated by JAK-STAT signaling pathway through upregulating the expression level of transforming growth factor β1 as well as mRNA expression levels of STAT3, Smad3 and LepR. In the meanwhile, electroacupuncture can inhibit the mRNA expression of p38 and Fas mRNA mediated by MAPK signaling pathways, further inhibiting the apoptosis of chondrocytes. OBJECTIVE: To explore the effect of electroacupuncture on the degeneration of articular cartilage in rats with knee osteoarthritis based on Ras-Raf-MEK1/2-ERK1/2 signaling pathway. METHODS:120 male healthy Sprague-Dawley rats aged 2 months olds were selected and randomly divided into normal, model, 15-minite electroacupuncture and 30-minute electroacupuncture groups (n=30 per group). The rats in the latter three groups received the intra-articular injection of 4% papain bilaterally, and the remaining rats received no intervention. At 2 weeks after modeling, the latter two groups were respectively given 15- and 30-minute electroacupuncture, five times weekly for consecutive 12 weeks. The morphology of the cartilage was observed by hematoxylin-eosin staining, the expression level of interleukin-1β in the synovium was detected by ELISA assay, and the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were detected by western blot analysis. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that: in the model group, the cartilage surface was rough, the cartilage layer became thinner, and the cartilage structure was damaged with incomplete tidal line; in the 15- and 30-minute electroacupuncture groups, the cartilage structure was complete with clear layers and complete tidal line. ELISA showed that the expression level of interleukin-1β in the model group was significantly higher than that in the normal group (P< 0.01), and the level in the 15- and 30-minute electroacupuncture groups was significantly lower than that in the model group (P < 0.05). Western blot assay found that compared with the normal group, the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were increased in the model group. However, all above protein levels except ERK1/2 in the 15- and 30-minute electroacupuncture groups were significantly lower than those in the model group (P < 0.01,P < 0.05). To conclude, electroacupuncture inhibits the degeneration of articular cartilage in osteoarthritisvia Ras-Raf-MEK1/2-ERK1/2 signaling pathway and downregulating the expression level of interleukin-1β.