Objective To evaluate the efifcacy and safety of hemostatics of injected gelatin matrix (HIGM) under the guidance of contrast-enhanced ultrasound (CEUS) for treating splenic trauma in canine model. Methods A total of 24 commercial hybrid dogs underwent celiotomy with creation of uniformly blunt splenic trauma lesion of 4.0 cm×4.0 cm×2.5 cm (length, width and depth, respectively) by hemostatic clamp. Subjects were prospectively randomized into two groups. The treatment group was treated with HIGM under the guidance of CEUS and the positive control group received thrombin solution. Conventional ultrasound and CEUS were performed to record the ascites and the splenic lesion areas at 1st, 3rd, 7th, 14th and 21st day. The ifne needle biopsy and splenectomy were performed for histopathologic examination. The weight, free intraperitoneal lfuid and injury site were compared with t test between HIGM and postive group. Results All animals in two groups survived. All dogs stopped hemorrhage after injection of HIGM under CEUS guidance. The area of injury site was (12.91±0.89) cm2, (4.45±0.75) cm2 and (1.38±0.23) cm2 at 1st, 3rd and 7th day and splenic lesions were not found at 14th and 21st day in all dogs (n=12) of HIGM group. The splenic lesion was (16.74±0.91) cm2, (11.26±0.99) cm2, (8.02±0.82) cm2 and (1.58±0.36) cm2 in the postive group at 1st, 3rd, 7th and 14th day and splenic lesions were not found at 21st day in all dogs (n=12). At 7th and 14th day post-injection, lesion areas were statistically significant between two groups (t=27.162, P=0.008;t=15.129, P=0.001). Free intraperitoneal lfuid was (0.91±0.05) cm at 1st day detected by conventional ultrasound and free intraperitoneal fluid was not found at 3rd, 7th, 14th and 21st day in all dogs (n=12) of HIGM group. The free intraperitoneal fluid in thepositive group was (1.96±0.17) cm, (1.30±0.11) cm and (0.81±0.12) cm at 1st, 3rd and 7th day and free intraperitoneal lfuid was not found at 14th and 21st day in all dogs (n=12). At 1st, 3rd and 7th day post-injection, free intraperatitoneal lfuid was statistically significant between two groups (t=20.934, P=0.003; t=41.310, P=0.000; t=22.520, P=0.000). Histopathological examination showed that there was no foreign body and foreign body granuloma and the structure of red pulp was recovered at 7th, 14th and 21st day. Gross anatomy showed that the splenic injury site was recovered completely without complications. Conclusion This study explored the value of HIGM for splenic trauma and provided a preliminary experimental evidence for clinical treatment.

Objective To investigate apoptosis of HepG2 ceils after transfecfion with LIGHT gene and interferon-γ. Methods LIGHT gene and interferon-γ were transfected into HepG2 cells by liposome mediated method. The HepG2 cells were divided into group A (transfected with LIGHT gene or interferon-γ), group B (transfeeted with LIGHT gene and interferon-γ) and group C (non-transfection group). The apoptosis rate of the HepG2 cells and the expression of Bcl-2 and Caspase-8 were detected 12, 24, 48 hours after transfeetion. Results (1) The apoptosis rates of HepG2 cells at hour 12, 24 and 48 after transfeetion were 18.8% ± 3.5%, 25.7%± 2.8% and 36.4% ±3.6% in group A, 23.8% ±2.4%, 31.1% ±2.1% and42.5% ±4.5% in group B, and 8.7% ± 2.1%, 9.3% ± 1.6% and 10.9% ± 1.2% in group C. There was a significant difference in apoptosis rate among the 3 groups (F = 15.69, 53.33, 48.28, P < 0.01). (2) The expression of Bcl-2 in HepG2 cells at hour 12, 24 and 48 after transfection was 16.4% ± 5.0%, 13.4% ± 3.5% and 8.6% ± 2.3% in group A, 14.7%±3.8%, 9.1% ±2.0% and 4.6% ±2.0% in group B, and 25.3% ±6. 3%, 19.8% ±4.4% and 10.1% ±3.8% in group C. There was a significant difference in the expression of Bcl-2 among the 3 groups (F = 6.19, 12.29, 5.81, P <0.05). (3) The expression of Caspase-8 at hour 12, 24 and48 after transfection were 19.3% ±2.4%, 27.2% ±1.9% and 33.7% ±3.0% in group A, 22.7% ±2.2%, 30.9% ±3.1% and 38.2% ±3.2% in group B, and 1.2% ±0.8%, 1.8% ±0.6% and 3.2% ±1.5% in group C. There was a significant difference in the expression of Caspase-8 among the 3 groups (F =71.54, 112. 78, I01.61, P < 0.01). Condusions LIGHT gene can signiticanfly promote cell apoptosis through regulating the expression of Bcl-2 and Caspase-8. Interferon-γ enhanced the effect of LIGHT gene on the apoptosis of HepG2 cells.

Objective To construct an efficient eukaryotic expression recombinant vector of human interleukin-1O(hIL-lO),and observe its expression in rabbit synoviocytes(RSCs).Methods Total RNA was extracted from peripheral blood mononuclear cells(PBMCs)of a patient with drug allergy.Specific Drimers for full-length open reading frames(ORFs)of hIL-10 were designed according to GeneBank(NM 000572).Withtotal RNA as the template,full-length ORFs of hIL-10 were amplified by reverse transcription Dolymerase chain reaction(RT-PCR).RT-PCR products were digested by restrictive endonucleotidase.then inserted into plasmid pcDNA4/HisMaxA.Both restrictive endonucleotidase analysis and DNA sequencing Were carried out for inserts verification.RSCs were transfected with recombinant plasmid expression vector PcDNA4 HisMaxA-hiL10 by liposome-mediated gene transfer methods,then cultured in vitro.The supernatants were collected af-ter transfection for 12 hours,24 hours,48 hours,72 hours,7 days,14 days respectively for IL-10 measure-ment by enzyme linked immunosorbent assay(ELISA).Results Full-length ORFs of hIL-1o(0.54 kb)had been successfully cloned from PBMCs through RT-PCR.The inserts and insert location of pcDNA4 HisMaxA were in a fight way verified by enzyme analysis and DNA sequencing.ELISA results showed that exogenous hIL-10 gene had expressed in the transfected RSCs from 12 hours to 7 days after transfection,and hIL-10level of transfection group significantly higher than that of the control group.Conclusion pcDNA4 HisMaxA-hiL10,the hIL-10 efficient eukaryotic expression vectors,has been suecessfully constructed.

BACKGROUND: The affected liver can be completely removed by liver transplantation,long-term efficacy is superior to liver resection,the 5-year survival rate reaches 70% H1.In addition,liver transplantation can avoid a serious risk for incomplete liver function caused by hepatic resection in the case of liver dysfunction.OBJECTIVE: To retrospectively analyze the treatment effects and importance of orthotoplc liver transplantation for primary hepatic cancer patients.METHODS: A total of 75 patients with primary hepatic cancer treated by orthotopic liver transplantation in Department of Hepatobiliary Surgery,Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA from March 1980 to December 2008 were involved in the analysis for the postoperative survival rates and recurrence of tumors.RESULTS AND CONCLUSION: For all the patients,the total postoperative survival rate in the 1st,2nd and 3rd year was 86.6%,66.7% and 53.3% respectively,the disease free survival rate was 65.2%,53.9%,34.1%.Their mean survival time is 25 months.For the patients in line with Milan standard,the postoperative survival rate in the 1st,2nd and 3rd year was 88.4%,72.5% and57.9% respectively,the disease free survival rate was 77.6%,62.3%,51.8%.Their mean survival time is 39 months.Tumor recurrence occurred within one year in all six patients who were beyond Milan standard.Two patients died in one year after operation,the survival rate at postoperative one year was 66.7% and the remanent four patients all died in the 2nd year after operation.Orthotopic liver transplantation was one of the effective treatments for pdmary hepatic cancer patients.The patients which were measured up to Milan standard would have the best curative effects.

ObjectiveTo study the efficacy of adjuvant percutaneous transhepatic portal vein chemoembolization (PVCE) in the prevention of tumor recurrence after partial hepatectomy for hepatocellular carcinoma.MethodsThe clinical data of 89 patients who received liver resection for hepatocellular carcinoma in our hospital from January 2007 to January 2010 were studied retrospectively.41patients received PVCE (the treatment group) while 48 patients received no PVCE (the control group).Postoperative recurrence and cumulative disease free survivals were compared using the Kaplan-Meier method.ResultsOn follow-up which ranged from 6-42 months,the 1- and 2-year disease free survivals were 76.5％ and 48.0％ in the treatment group,and 53.8％ and 25.8％ respectively in the control group (P＜0.05).The mean disease free survivals were 19.91 (95％ CI,16.09-23.73)and 13.8 months (95 ％ CI,10.95-16.65),respectively.The cumulative disease free survivals in the PVCE group were significantly higher than the control group (P=0.01).Cox multivariate analysis showed that adjuvant PVCE,tumor size,portal vein thrombosis,and postoperative transcatheter arterial chemoembolization (TACE) were independent factors of disease free survival.ConclusionAdjuvant PVCE was effective in preventing postoperative recurrence of hepatocellular carcinoma after partial hepatectomy.

Objective To investigate the impact of blood glucose level on the recurrence of liver cancer after laparoscopic surgery. Methods The clinical data of 98 patients with primary hepatocellular carcinoma from January 2012 to January 2015 were retrospectively analyzed. All patients were treated by laparoscopic radical resection of hepatocellular carcinoma. Patients were divided into elevated blood glucose group (n = 23) and control group (n = 75) according to whether the fasting blood glucose was ≥6.1 mmol/L. The recurrence of liver cancer in 1 year and 2 years after operation was compared. The factors influencing the recurrence of liver cancer were analyzed by univariate and multivariate analysis. Results The recurrence rates were 47.82% and 21.33% respectively in the patients with elevated blood glucose and the control group. The recurrence rates were 73.91% and 36.00%respectively in the 2-year postoperative patients with blood glucose and 1 year and 2 years. The recurrence rate was higher than that of the control group, the difference was statistically significant (P < 0.05). Logistic multivariate analysis showed that fasting blood glucose was high, Child-Pugh grade B, intraoperative blood transfusion, lymphatic invasion, high clinical pathology stage, postoperative alpha-fetoprotein (AFP) high, no postoperative adjuvant therapy (P < 0.05). Conclusion The recurrence rate of patients with elevated liver cancer after laparoscopic surgery is high, and fasting blood glucose is high, Child-Pugh grade is B grade, blood transfusion is high, there is lymphatic invasion, high clinical pathology stage after AFP high, no postoperative adjuvant therapy for its postoperative recurrence of risk factors, should strengthen the monitoring of high-risk patients, reduce postoperative recurrence rate.

Objective To explore the influence of child head injury under different impact angles by applying the finite element model of six-year-old child pedestrian as specified in the European New Car Assessment Programme (Euro NCAP). Methods Based on the finite element model of 6-year-old pedestrian with detailed anatomical structure as specified by the Euro NCAP (TB024), four groups of simulation experiments were set up to explore the mechanism of head injury in children under different impact angles. The initial position for head mass center was on the longitudinal center line of the car. The initial speed of the car was 40 km/h. The car contacted with the model from the direction of the right (0°), the front (90°), the left (180°) and the back (270°). The kinematics differences and head impact responses were compared, and injuries of the facial bone and skull were analyzed. Results Through the analysis of head contact force, acceleration of head mass center, resultant velocity of head mass center with the vehicle, head injury criterion (HIC15), facial bone fracture and skull stress distribution, it was found that the risk of head fracture and brain contusion under back impact and front impact was higher than that under side impact. The risk of head fracture and brain contusion was highest under back impact, while the lowest under side impact. Conclusions Child pedestrian head injury was the largest under back impact. The results have important application values for the assessment and development of car-pedestrian collision protection device.

Objective: our study was to investigate the effect of hepaCAM gene on cell cycle of bladder cancer cell line T24 and BIU-87. Method: Adenovirus vector with hepaCAM was infected into T24 and BIU-87 cells. The expression of hepaCAM mRNA and protein were measured byRT-QPCR and Western-blot. Immunofluorescence detected the cellular localization of hepaCAM. The distribution of cell cycle was analyzed by FCM. The protein expression of cyclinD1 was measured by Westein-blot. Result: The expression of hepaCAM mRNA and protein was increased in two cell lines infected with pAdH5-hepaCAM. The G0/G1 cycle was increased in infected cells, and the protein expression of cyclinD1 was decreased in same cells. Immunofluorescence revealed that hepaCAM protein localized on cytoplasm in well-spread cells, otherwise it localized on the junction of each cell. Conclusion: T24 and BIU-87 cells that infected pAdH5- hepaCAM was blocked in G0/G1 cycle, and it can decreased the protein expression of cyclinD1.

Four salts of ticagrelor, ticagrelor-3,5-dinitrobenzoic acid, ticagrelor-pyrazinamide, ticagrelor-D-proline and ticagrelor-L-proline were prepared by solvent suspension and liquid-assisted grinding to improve the solubility of ticagrelor. The compounds were characterized by powder X-ray diffraction, Fourier transform infrared spectroscopy, differential scanning calorimetry, nuclear magnetic resonance spectroscopy, elemental analysis, and the intermolecular salt-bonding forces were analyzed. The equilibrium solubility of salts and pure drug in hydrochloride buffer pH 1.2 and phosphate buffer pH 6.8 were measured by high-performance liquid chromatography. Ticagrelor was salted with 3,5-dinitrobenzoic acid, pyrazinamide, D-proline, L-proline all in a stoichiometric ratio of 1∶1; with the exception of ticagrelor-D-proline, the solubility of the other three salts provided significantly improved solubility in hydrochloride buffer pH 1.2, and the equilibrium solubility of ticagrelor-3,5-dinitrobenzoic acid was increased by approximately 1.7 folds as compared to pure drug. Salt-forming technology is convenient and can improve the solubility of ticagrelor.

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