Objective To analyze the therapeutic effect of percutaneous transhepatic cholangial drainage (PTCD) in the treatment of bile duct obstruction in patients with malignant hilar bile duct carcinoma,and to discuss the clinical application and practical value of PTCD.Methods A total of 55 patients with malignant biliary obstruction were divided into the PTCD group (30 cases who recieved percutaneous transhepatic cholangial drainage) and the control group (25 cases who recieved endoscopic stent implantation).Observed the preoperative and postoperative biochemical indexes of PTCD group,including serum total bilirubin (TB),serum direct bilirubin (DB),serum alanine aminotransferase (ALT) and serum glutamic acid amino turn shift of aspartate aminotransferase(AST) and serum alkaline phosphatase(AKP).Compared the effect rate and postoperative survival time of the two groups through postoperative follow-up.Results The TB,DB,ALT,AST and APK of PTCD group one week after operation changed obviously compared with the relative index before opreation with statistically significant differences (P<0.05), which indicated a significant improvement of biochemical indicators.The treatment efficiency of the PTCD group and the control group were 83.3% and 64.0% respectively, and survival time of the two groups were(7.5±2.6)months and(4.8±2.8)months respectively.Results of the PTCD group was significantly better than that of the control group,and the differences were statistically significant(P<0.05).Conclusion All the patients with PTCD get better biochemical indicators and longer postoperative survival time,and the interventional therapy PTCD can be used as an effective clinical treatment method for bile duct obstruction with malignant hilar bile duct carcinoma.

Objective To study the clinical effect of total parathyroidectomy on the patient with secondary hyperparathyroidism related to chronic renal failure. Methods The clinical data of 12 cases of total parathyroidectomy were retrospectively analyzed. All changes between preoperation and postoperation were compared, that included the clinical presentations, serum calcium and phosphate, plasma alkaline phosphatase (AKP), parathyroid hormone (iPTH), blood haematocrit (HCT), blood-lipid(TG) and complications. Results The clinical symptoms and signs were markedly improved in all cases. A postoperative decrease in the laboratory indexes of serum calcium and phosphate, AKP, iPTH, HCT was also observed(P<0.05). But the difference of TG did not reach statistical significance(P>0.05). Hypocalcaemia occurred in all patierts in different degrees. Plasma iPTH maintained at high level in 1 case and recurrence happened in 1 case after operation. Conclusions Total parathyroidectomy is an effective treatment for severe uremic secondary hyperparathyroidism and can improve the patient's life quality.

Objective To detect the protective effect of sufentanil preconditioning on hepatic ischemia-reperfusion injury in rats and the role of c-Jun N-terminal kinase signal pathway in the mech-enism.Methods One hundred and sixty-two SD rats(in either gender,weighing 250-300 g)were ran-domly divided into seven groups:Sham-operated group (group S,n = 30 ),ischemia-reperfusion group (group IR,n =30),sufentanil preconditioning group (group SF1:1 μg/kg,n =30;group SF5:5 μg/kg,n =30;group SF10:10 μg/kg,n =30),SP600125 group (group SP,n =30),and dimethyl sulphoxide control group (group DMSO,n =6),different doses of sufentanil was administered 30 min before hepatic ischemia in group SF1,SF5 and SF10.Blood and liver samples were collected from each group at 0(T1 ),1 (T2 ),2 (T3 ),4 (T4 ),and 6 (T5 )hours after reperfusion.Serum alanine amin-otransferase (ALT)and aspartate aminotransferase (AST)were measured by an automatic biochemi-cal analyzer.Malondialdehyde (MDA)and superoxide dismutase (SOD)in liver tissue was measured. Liver sample was stained with HE to observe the hepatic pathological changes.Immunohistochemical method was used to determine the expression of JNK and western blotting was used to detect the ex-pression of P-JNK.Results Compared with group S,levels of AST,ALT increased significantly in group IR,SF1,SF5,SF10 at T1-T5 and in group SP,DMSO at T3 (P <0.05 ).Compared with group IR,levels of AST,ALT decreased significantly in group SF1,SF5,SF10 at T1-T5 and in group SP at T3 (P <0.05).Compared with group S,levels of MDA,SOD increased significantly in group IR,SF1, SF5,SF10 at T1-T5 and in group SP,DMSO at T3 (P < 0.05 ).Compared with group IR,levels of MDA,SOD decreased significantly in group SF1,SF5,SF10 at T1-T5 and in group SP at T3 (P <0.05).Compared with group SF1 and SF5,levels of MDA,SOD decreased significantly in SF10 at T4 . Compared with T1 ,the expression of p-JNK in group IR increased significantly at T3 (P < 0.05 ). Compared with group S,the expression of p-JNK in groups IR,SF1,SF5,SF10,SP,DMSO increased significantly at T3 (P < 0.05 ).Compared with group IR,the expression of p-JNK in groups SF1, SF5,SF10,SP decreased significantly and that in groups SF5,SF10 were less than that in group SF1 (P <0.05 ).The expression of p-JNK in group SF10 was less than that in group SF5 (P < 0.05 ). Conclusion Sufentanil preconditioning can reduce the hepatic ischemia-reperfusion injury and the dos-age of 10 μg/kg was the most effective.The protective mechanisms may inhibit JNK pathway and re-duce the expression of JNK.

Objective To compare the number of lymph node dissected by intraoperative lymphatic mapping guided D2 gastrectomy and that by standard D2 gastrectomy plus lymphadenectomy in patients of advanced gastric cancer. Methods In this study 20 advanced gastric cancer cases received intraoperative peritumor injection of carbon nanoparticles suspension ( group 1 ) and D2 lymphadenectomy was guided by the black-stained lymph nodes. 21 cases undergoing standard D2 lymphadenectomy served as controls (group 2). The number of lymph nodes removed and the condition of lymphatic metastasis in two groups, blackstained lymph nodes in group 1, and postoperative complications were compared. Results The average lymph nodes dissected in group 1 (35. 1 ± 13.4) were higher than in control group (26.2 ±7.8). The differences were statistically significant (t =2. 126, P =0. 034). The number of removed N2 and N3 lymph nodes in group 1 were more than that in control group. The total black-stained ration of lymph nodes was 52. 7% in group 1. The positive rate of lymph nodes was higher in black-stained lymph nodes (27.6%) than in unstained lymph nodes ( 10. 8% ) in group 1 and in control group ( 16. 9% ). The differences were also statistically significant ( x2 = 6. 034, P = 0. 016; x2 = 5. 142, P = 0. 023 ). Postoperative afferent loop obstruction developed in one case in group 1. Conclusions Lymphatic mapping guided D2 radical gastrectomy plus lymphadenectomy increases the number of lymph nodes dessected and improves the efficiency of positive lymph nodes excision for patients of advanced gastric cancer.

Objective To study hepatic blood flow exclusion for the resection of liver tumors involving hepatic hilar region. Methods The clinical data of 16 cases of liver tumors involving hepatic hilar region from January 2005 to March 2008 were retrospectively analyzed. Liver tumors were resected by the technique combining hepatic portal control ( Pringle's maneuver) and normothermie total hepatic vascular exclusion (NHVE). The relation of liver tumors to major vessels, episodes and durations of hepatic blood flow exclusion, intraoperative blood loss and blood transfusion, postoperative complications were analyzed. Results The technique combining Pringle's maneuver and NHVE was used in 16 cases. The mean episodes and durations of Pringle's maneuver were (3.8±1.6) min and (46.6±28.8) min, respectively. The mean episodes and durations of NHVE were (1.6±0.4) min and (23.5±8.2) min, respectively. The mean amount of intraoperativ blood loss was (1250±320) ml, blood transfusion (860±245) ml. Major hepatic vessel injuried were repaired intraoperatively including inferior vena cave in 4 cases, main hepatic veins in 2 cases and portal veins in 2 cases. The serum alanine transaminase(ALT) and bilirubin raised in different degrees after operation, and recovered gradually to normal level. There was no postoperative mortality and serious postoperative complications. Conclusions Alternative use of hepatic blood flow exclusion combining Pringle maneuver and NHVE reduces the time of total hepatic vascular exclusion, improves safety for resection of liver tumors involving hepatic hilar region.

Objective: To investigate the effects of adipose-derived stem cell released exosomes (ADSC-Exos) on wound healing in diabetic mice. Methods: The ADSCs were isolated from the adipose tissue donated by the patients and cultured by enzymatic digestion. The supernatant of the 3rd generation ADSCs was used to extract Exos (ADSC-Exos). The morphology of ADSC-Exos was observed by transmission electron microscopy. The membrane-labeled proteins (Alix and CD63) were detected by Western blot, and the particle size distribution was detected by nanoparticle tracking analyzer. The fibroblasts were isolated from the skin tissue donated by the patients and cultured by enzymatic digestion. The 5th generation fibroblasts were cultured with PKH26-labeled ADSC-Exos, and observed by confocal fluorescence microscopy. The effects of ADSC-Exos on proliferation and migration of fibroblasts were observed with cell counting kit 8 (CCK-8) and scratch method. Twenty-four 8-week-old Balb/c male mice were used to prepare a diabetic model. A full-thickness skin defect of 8 mm in diameter was prepared on the back. And 0.2 mL of ADSC-Exos and PBS were injected into the dermis of the experimental group ( n=12) and the control group ( n=12), respectively. On the 1st, 4th, 7th, 11th, 16th, and 21st days, the wound healing was observed and the wound healing rate was calculated. On the 7th, 14th, and 21st days, the histology (HE and Masson) and CD31 immunohistochemical staining were performed to observe the wound structure, collagen fibers, and neovascularization. Results: ADSC-Exos were the membranous vesicles with clear edges and uniform size; the particle size was 40-200 nm with an average of 102.1 nm; the membrane-labeled proteins (Alix and CD63) were positive. The composite culture observation showed that ADSC-Exos could enter the fibroblasts and promote the proliferation and migration of fibroblasts. Animal experiments showed that the wound healing of the experimental group was significantly faster than that of the control group, and the wound healing rate was significantly different at each time point ( P<0.05). Compared with the control group, the wound healing of the experimental group was better. There were more microvessels in the early healing stage, and more deposited collagen fibers in the late healing stage. There were significant differences in the length of wound on the 7th, 14th, and 21st days, the number of microvessels on the 7th and 14th days, and the rate of deposited collagen fibers on the 14th and 21st days between the two groups ( P<0.05). Conclusion: ADSC-Exos can promote the wound healing in diabetic mice by promoting angiogenesis and proliferation and migration of fibroblasts and collagen synthesis.

Objective: To summarize the research progress of mesenchymal stem cells derived exosomes (MSCs-EXOs) in wound repair in recent years. Methods: The literature about the role of MSCs-EXOs in wound repair at home and abroad was extensively consulted. The mechanism of MSCs-EXOs in wound repair and its clinical application prospects were summarized and analyzed. Results: MSCs-EXOs can inhibit early inflammatory reaction, promote angiogenesis, proliferation, and migration of epithelial cells, regulate collagen synthesis, and inhibit scar proliferation in the later stage of wound healing. Compared with MSCs, MSCs-EXOs have many advantages, such as high stability, easy storage, non-tumorigenicity, no proliferation, easy quantitative use, and so on. It has broad clinical application prospects. Conclusion: MSCs-EXOs can promote wound repair and hopefully develop into a clinical product to promote the repair of acute or chronic wounds.

Objective To investigate the suppression of mrp1 and MRP1 induced by small interfering RNA and the restoration of sensitivity to chemotherapeutic drugs in the multidrug-resistant hepatocellular carcinoma cell lines HepG2/mrp1. Methods mrp1-targeted small interfering RNA duplexes were designed and composed and introduced into multidrug-resistant hepatocellular carcinoma cell lines HepG2/mrp1. The suppression of mrp1 mRNA and its gene product MRP1 was examined by RT-PCR and flow cytometry (FCM), respectively. MTT assay was performed to measure the reverse effect of small interfering RNA based on the results of ICs0. Results The overexpression of mrp1 mRNA and MRP1 was effectively suppressed by small interfering RNAs. The level of mrp1 mRNA in the transfected HepG2/mrp1 cells was reduced to (86.36±2.76)% and MRP1 to (89.38±3.76)%compared with those of the controls. The resistance to ADR was reversed five-fold, which indicated the restoration of sensitivity to drugs. Conclusion Small interfering RNA can inhibit mrp1 expression effectively and reverse the multidrug resistance mediated by MRP1.

Objective To evaluate the effects of isoflurane postconditioning on angiogenesis during cerebral ischemia-reperfusion ( I∕R) in rats and the role of Shh signaling pathway. Methods Thirty-two clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 220-280 g, were divided into 4 groups ( n=8 each) by a random number table method:sham operation group ( Sham group) , I∕R group, isoflurane postconditioning group ( ISO group) , and isoflurane postconditioning plus Shh signaling pathway specific inhibitor cyclopamine group ( ISO+CYC group) . Cerebral ischemia was produced by inserting a 3-0 nylon thread with a rounded tip into the internal jugular vein. The nylon thread was threaded cranially until resistance was met. Occlusion was maintained for 1. 5 h followed by 24 h reperfusion. Neurological deficit was scored at 24 h of reperfusion. Rats were then sacrificed, and brains were removed for determination of cerebral infarct volume ( by TTC) and expression of glioma-associated oncogene homolog 1 ( Gli1) , vascu-lar endothelial growth factor ( VEGF) and transmembrane phosphoglycoprotein protein ( CD34) in cerebral cortex (by Western blot) and for examination of the pathological changes (by Nissl staining). Results Compared with Sham group, the neurological deficit score and cerebral infarct volume were significantly in-creased, and the expression of Gli1, VEGF and CD34 in the cerebral cortex was up-regulated in I∕R and ISO groups ( P <0. 05) . Compared with I∕R group, the neurological deficit score and cerebral infarct vol-ume were significantly decreased, and the expression of Gli1, VEGF and CD34 in the cerebral cortex was up-regulated ( P<0. 05) , and the pathological changes of brain tissues were significantly attenuated in ISO group, and no significant change was found in the parameters mentioned above in ISO + CYC group ( P>0. 05) . Compared with ISO group, the neurological deficit score and cerebral infarct volume were signifi-cantly increased, and the expression of Gli1, VEGF and CD34 in the cerebral cortex was down-regulated in ISO+CYC group ( P<0. 05) . Conclusion The mechanism by which isoflurane post-conditioning attenuates cerebral I∕R injury is related to activating Shh signaling pathway and promoting angiogenesis in rats.

Objective:This study aims to explore the potential effects of adipose-derived stem cell-extracellular vesicles(ASC-EVs) on TGFβ-Smad signaling pathway during myofibroblast trans-differentiation in vitro. Methods:ASCs were isolated from liposuction and flow cytometry was used to detect the surface protein markers. ASC-EVs were isolated from the supernatant of the third to fifth generation ASCs, and the microscopic morphology was observed by transmission electron microscope. The particle size distribution was detected by nano-particle tracking analyzer NanoSight and the membrane surface marker proteins CD63, Alix and TSG101 were detected by flow cytometry. The uptake of EVs by dermal fibroblasts co-cultured with PKH67 fluorescence labeled ASC-EVs was observed by confocal microscope. Dermal fibroblasts were continuously induced by TGFβ1 for five days, and ASC-EVs at the dose of 50 and 100 μg/ml were added. The expression of α-SMA and Smad-2/3/4 were detected by immunofluorescence staining, RT-PCR and Western Blot.Results:The results of flow cytometry showed that the surface markers CD73, CD49d, CD90 and CD105 of the third generation ASCs, were positive, and CD34 and CD45 were negative. Under transmission electron microscope, ASC-EVs was a round membranous vesicle with clear edge and surrounded by bilayer phospholipid membrane. The particle size of more than 95% of the ASC-EVs was distributed between 30 nm to 261 nm, with an average of (166.0±86.1)nm. The specific marker proteins of extracellular vesicle, CD63, Alix and TSG101 were highly expressed. Under confocal microscope, ASC-EVs with green fluorescence were uptake by dermal fibroblasts and distributed in the cytoplasm, and part of ASC-EVs was distributed around the nucleus.TGF-β1 induced a significant increase in the expression of α-SMA in dermal fibroblasts, and the addition of 50 or 100 μg/ml ASC-EVs reduced the expression of α-SMA genes and proteins, but did not show a dose-dependent manner. The gene and protein expression changes of Smad-2/3/4 were consistent with α-SMA. In addition, ASC-EVs could significantly reduced the content of type Ⅰ collagen in the supernatant, but had no significant effect on the secretion of type Ⅲ collagen.Conclusions:The mechanism of ASC-EVs inhibiting scar hyperplasia may be closely related to the suppression of TGF-β-Smad signaling pathway in dermal fibroblasts, inhibition of myofibroblast trans-differentiation and reduction of type Ⅰ collagen secretion.