Objective To investigate the effect of nicotine on coagulation abnormalities in endotoxemic rats.Methods Ninety-six male SD rats weighing 200-250 g were randomly divided into 4 groups (n=24 each): group normal saline (group NS);group LPS;group nicotine(group NIC)and group α-bungarotoxin (α7 nicotinic acetylcholine receptor antagonist, group α-BGT) . Endotoxemia was induced by LPS 10 mg/kg injected via femoral vein in LPS, NIC and α-BGT groups. In group NIC nicotine 400 μg/kg was injected intraperitoneally at 30 min before LPS injection. In group α-BGT α-BGT 1 μg/kg was injected intraperitoneally at 15 min before intraperitoneal nicotine. Prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(Fib),antithrombin (AT),von Willebrand factor(vWF),plasminogen activator inhibitor-1(PAI-1),D-dimer,platelet count and TNF-α were measured before (baseline) and 2, 4 and 6 h after LPS injection.Results PT and APTT were significantly prolonged and plasma Fib and AT concentrations and platelet count were significantly decreased, while plasma PAI-1, D-dimer, vWF and TNF-α concentrations were significantly increased after LPS administration in group LPS as compared with group NS. Nitotine pretreatment significantly attenuated the LPS-induced changes in group NIC.The effect of nicotine was counteracted by α-BGT. Conclusion Nicotine can attenuate coagulation abnormalities induced by LPS by acting on α7 nicotinic acetylcholine receptor.

BACKGROUND:Studies have shown that tibial plateau posterolateral fractures can be treated by posterolateral knee approach, however, there is no clear anatomical research to deeply evaluate and analyze this approach. OBJECTIVE:To evaluate the efficacy and safety of posterolateral knee inverted“L”shaped pathway in invloving tibial plateau posterolateral fractures. METHODS:Eight fresh frozen adult corpses, 16 knees side, were al dissected using posterolateral inverted“L”shaped pathway. During the dissection, the exposure range was observed and important parameters of anatomical structure were measured. RESULTS AND CONCLUSION:The pathway may be ful y exposed to the posterolateral aspect of tibial plateau and posterior cruciate ligament tibial insertions. The operations completed by out team did not create any obvious interference to superior tibiofibular joint, fibular head and posterolateral corner structure. The exposed mean length of common peroneal nerve in incision was 56.48 mm, with a mean angle of 14.7° tilt towards the axis of the fibula. The mean distance between the neck of the fibular and fibular head tip was 31.26 mm, an average of 42.18 mm to the joint line. The mean distance between the opening of the interosseous membrane and the articular surface was 48.78 mm. The divergence of the fibular artery from the posterior tibial artery was an average of 76.46 mm from articular surface. These results confirm that posterolateral inverted“L”shaped pathway met the requirements of anatomical reduction and buttress fixation for posterolateral tibial plateau fracture. Exposure of the common peroneal nerve can be minimized or even avoided by modifying the skin incision. Because the popliteal artery branches anterior tibial artery passed through interosseous membrane hole and peroneal artery and then separated from the posterior tibial artery, pathways dissection to distal deep area should be carried out careful y. Placement of a posterior buttressing plate carries a high vascular risk if the plate is implanted beneath these vessels.

Objective To investigate the rule of the cerebral tissues aquaporin-4(AQP-4) expression in acute and chronic hepatic failure mice.To study the molecular biologic mechanism of the diffusion weighted imaging(DWI).Methods Sixty five male Sprague-Dawley rats were divided into 3 groups randomly,including acute(n=25),chronic hepatic failure(n=25)and control group(n=15). Thioacetamide(TAA)intraperitoneal injection produces the acute and chronic hepatic failure models.All rats in groups were examined with MR DWI.We Observed the distribution of abnormal signal on DWI.The DWI single values of top and lateral cortex of parietal lobe,peripheral region of lateral ventricle in the highest hyperintensity section of brain were measured.Blood ammonia values were examined.The pathologic and immuno-histochemistry and RT-PCR examination for brain specimen were performed.All date were analyzed with statistical methods.Results The mean values of blood ammonia were significantly different (P0.05).Conclusions Increase of the blood ammonia was the main cause for the brain energy metabolic abnormality and AQP-4 mRNA and protein expression.The hyperammonemia was the key factor in the occurrence and development of the hepatic brain edema.The abnormal findings in DWI signal could reflect the range and degree of the brain edema and AQP-4 protein expression.

Objective The purpose of this study was to develop a high-throughput micro-plate radiobinding assay (RBA) of glutamic acid decarboxylase antibody (GAD-Ab) and to evaluate its clinical application. Methods 35labeled GAD65 antigen was incubated with sera for 24 h on a 96-well plate, and then transferred to the Millipore plate coated with protein A, which was washed with 4℃ PBS buffer, and then counted by a liquid scintillation counter. The GAD-Ab results were expressed by WHO standard unit (U/ml). A total of 224 healthy controls, 162 patients with type 1 diabetes mellitus(T1DM) and 210 patients with newly diagnosed type 2 diabetes (T2DM) were recruited. A total of 119 TI DM and healthy cases with gradually changing GAD-Ab levels were selected to compare the consistency of micro-plate RBA with conventional radioligand assay (RLA). Blood samples were obtained from the peripheral vein and finger tip in 32 healthy controls, 35 T1DM and 24 T2DM patients, and tested with micro-plate RBA and then compared with the conventional RLA to investigate the reliability of finger tip sampling. Linear correlation,student's t-test, variance analysis and receiver operating characteristic (ROC) curve were performed using SPSS 11.5. Results (1) The optimized conditions of micro-plate RBA included 2 μl serum incubated with3 ×104 counts/min 35S-GAD for 24 h under slow vibration, antigen-antibody compounds washed 10 times by 4℃ PBS buffer, and radioactivity counted with Optiphase Supermix scintillation liquid. (2)The intra-batch CV of the micro-plate RBA was 3.8%- 10.2%, and the inter-batch CV was 5.6%- 11.9%. The linearity analysis showed a good correlation when the GAD-Ab in serum samples ranged from 40.3 to 664 U/ml and the detection limit of measurement was 3.6 U/ml. The results from Diabetes Autoantibody Standardization Program (DASP) 2005 showed that the sensitivity and specificity for GAD-Ab were 78% (39 positive among 50 new-onset T1DM) and 98% (2 positive among 100 healthy controls). The results of GAD-Ab obtained with micro-plate RBA and RLA were closely correlated (r=0.915,P<0.001) with a high concordance level of 97.5% and a Kappa value of 0.95. (3)TI DM and T2DM patients showed higher positive rates for GAD-Ab than the healthy controls(46.9% and 5.2% vs 0.89% ,X2=123.5 and 10. 1 ,P <0.001 and <0.01, respectively). (4)The consistency of GAD-Ab measurement with RBA using finger tip blood and RLA measurement using venous blood was 96.7% (r =0.946,P <0.001, Kappa value: 0.905). Conclusions The micro-plate RBA of GAD-Ab has high sensitivity, specificity and reproducibility, and can be measured with finger tip blood sampling. It might be a better alternative for clinical practice.

Objective To investigate the changing characteristics of myocardial movement in patients with dilated cardiomyopathy by three?di?mensional speckle tracking echocardiography(3D?STE). Methods The peak systolic global longitudinal train(GLS),global radial strain(GRS), global circumferential stain(GCS)and global area strain(GAS)of left ventricle were measured by 3D?STE technology in 69 patients with dilated cardiomyopathy. According to the left ventricular ejection fraction(LVEF),all patients were divided into group A(35%≤LVEF<50%)and group B (LVEF<35%). The differences of measurements were compared between two groups. The correlation between global myocardial strain in all direc?tions and left ventricular ejection fraction was analyzed. Results The GLS,GRS,GCS and GAS were significantly higher in group A than those in group B(P<0.01). The GLS,GRS,GCS and GAS were correlated with LVEF in group A(r=-0.871,0.610,-0.423,-0.797;P<0.05).The GCS,GRS and GAS were correlated with LVEF in group B(r=-0.517,0.368,-0.438;P<0.05). There was no significant correlation between GLS and LVEF in group B. Conclusion 3D?STE technology can be applied to evaluate the change of the myocardial movement. GLS is a promis?ing marker of the prognosis in patients with DCM.

Objective:To explore the clinical features of recurrence after choledocholithotomy and to analyze the risk factors.Methods:The clinical data of 730 patients with choledocholithiasis who were treated in our hospital from January 2005 to July 2016 were analyzed retrospectively,550 cases who were received choledocholithotomy were defined as laparotomy group,30 cases with laparoscopic common bile duct exploration (LCBDE) were defined as the LCBDE group,and 150 cases with endoscopic sphincterotomy (EST) were defined as EST group.The recurrence rate of the three groups were compared.The patients of three groups were divided into recurrence group (n=227) and non recurrence group (n=503) according to the recurrent situation,then the clinical features and risk factors of recurrent patients were analyzed by univariate and multivariate Logistic regression analysis.Results:The recurrence rate of EST group was 38.67％,which was significantly higher than that of LCBDE group with 26.67％ and the laparotomy group with 29.27％,and there was statistical difference (P＜0.05).The results of univariate analysis showed that there were statistically significant differences in age,history of HBV infection,jaundice,abnormal total bilirubin,peripapillary diverticulum,biliary infection,biliary stricture,papillary stenosis,sphincter of Oddis dysfunction,history of biliary surgery,cholecystectomy,bile duct diameter ≥ 15 mm,bile duct angle ≤120°,operation type,stone quantity ≥ 2 grains,stone diameter ≥ 10 mm,with or without gallstones (P＜0.05).The results of Logistic multivariate regression analysis showed that age,having peripapillary diverticulum,having history of biliary surgery,bile duct diameter ≥ 15 mm,stone quantity ≥ 2grains and EST operation type were the independent risk factors of the recurrence after choledocholithotomy (P＜0.05).Conclusion:There are many risk factors of recurrence after choledocholithotomy,and operation method should be based on the size and the number of the stones,and the constitution of patients.Preventive measures should be strengthened to control the recurrence after choledocholithotomy.

Objective To study the structure of class 1 integrons in 90 strains of Pseudomonas aeruginosa isolated during two periods of 1992-1996 and 2003-2005,and to get information about the structure changing of class 1 integrons by comparing their structures in two different periods.Methods Routine PCR and long PCR were performed to amplify the class 1 integrons and the gene cassettes they carried, followed with sequencing and blast via GenBank.Results Thirteen out of 41 strians ioslated during the period of 1992-1996 were positive on class 1 intergrons.Long PCR showed that the class 1 integron was 1868 bp in length and contained 2 resistance genes averagely.Six types of resistance genes of qacEA1 (n=6), sull (n=14),aadA1 (n=2),aadB (n=1),PSE-1 (n=2) and tetA (n=1) were found in these integrons, which consisted of 5 patterns of resistance cassette arrangements.Nineteen strains were proved to carry class 1 integrons in 49 isolates from 2003-2005.The mean DNA sequence length of them was 3383 bp with 3.6 resistant genes in averagely,10 types of resistance genes,qacEA1 (n=18),sull (n=25),aadA1 (n=6), aadB (n=7),aacA4 (n=2),PSE-1 (n=3),VEB-1 (n=4),OXA10 (n=1),cm1 A (n=1) and tetA (n =2),were identified in these integrons,which were composed of 9 patterns of resistance cassette arrangements.Conclusion In terms of produce length and resistance cassettes carried in the integrons, greater complexity is found in the structure of class 1 integrons in strains isolated during 2003-2005 than those during 1992-1996.

Objective To study hemodynamics of cerebral blood flow in newborns with asphyxia.Method Bilateral cerebral electrical admittance plethysmogram(BCEAP) was used to explore characteristics of cerebral blood flow in 20 healthy newborns and 20 newborns with asphyxia respectively from first to fourth day after birth.Results Both the ratio of Hs to b-S(Hs/ b-S) and index of admittance differential loop(ADL) Ⅰ+Ⅱ decreased significantly in newborns with asphyxia compared to normal control from first to fourth day after birth(P

OBJECTIVETo explore the methods of isolation, cultivation, purification, identification of the fetal mice testis Leydig cell and to observe its biological characteristics in vitro.

METHODSLeydig cells were isolated by 0.03% collagenase (type I) from fetal mice testis and cultured in DMEM/F12 medium. The identity and purity of Leydig cell were assessed by 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD). Cell viability was measured by trypan blue. Testosterone level in the medium of cultured Leydig cells was measured in various culture phases and cell density by radioimmunoassay.

RESULTSThe purity of Leydig cell was (45.10 +/- 1.66)% before culture, and (81.17 +/- 2. 32)% 72 h after culture. The level of testosterone secreted by Leydig cells could be detected in the medium and its level was associated with the density and time of cultured Leydig cells. The secretion capacity of testosterone by single Leydig cell decreased gradually during the culturing period.

CONCLUSIONThe fetal Leydig cells isolated from fetal mice testis have high purity. It can be cultured and kept the secretion ability of testosterone for a few days in vitro. This system can provide a valuable model for further study on the cellular function of the Leydig cells of fetal mice.

Animals ; Cell Separation ; Cells, Cultured ; Leydig Cells ; cytology ; physiology ; Male ; Mice ; Mice, Inbred Strains ; Testis ; embryology ; Testosterone ; secretion

Objective To investigate the clinical application of a new thermosensitive embolic agent in interventional arterial embolization treatment, and to evaluate its effectiveness and safety in treating inoperable advanced hepatocellular carcinoma (HCC). Methods Prospective, open and single center clinical study of a new thermosensitive embolic agent was conducted. A total of 10 patients with clinically confirmed inoperable advanced HCC were treated with percutaneous arterial embolization using thermosensitive embolic agent; a maximum of 4 therapeutic cycles were performed until the disease progressed. Objective effect was evaluated at (30 ±7) days after the treatment according to mRECIST standard; the progression free survival (PFS) time and the survival period (OS) were recorded. The adverse reactions and adverse events were determined by NCI-CTC 3.0 version standard. Results Ten patients (23 target lesions in total) were enrolled in this study. After the treatment, complete remission (CR) was obtained in one patient, partial remission (PR) in 3 patients, stable disease (SD) in 5 patients, and progress disease (PD) in one patient. The objective response rate (CR+PR) was 40%, and the disease control rate (CR+PR+SD) was 90%. The PFS of hepatic lesions was 18.4 weeks (95%CI:14.15-22.65), the cumulated median survival period was 46.2 weeks (95%CI:38.18-54.22). The most common adverse reactions included pain, fever, hepatic function damage, small amount of ascites, diarrhea, etc. Conclusion The new thermosensitive embolic agent is not sticky to vessel and is visible under X-ray; it has reliable embolization effect, and the therapeutic results can be easily evaluated. This embolic agent is not perfect, and it has some limitations in clinical use. The adverse reactions are mild, which can be easily tolerated by the patients. It is worth trying to carry out large sample and randomized controlled studies in order to open up more areas for arterial chemoembolization therapy of tumors.