The expression cDNA library of A. tabescens was constructed by SMART technique, which useλTriplEx2 as a vector. The titer and the percentage of the constructed library were about 1.0 × 106pfu/mland 98.3% respectively, and the titer and the capacity of the amplified library were about 3.1 × 108pfu/mland 4.2 × 1010. The library was used to provide expressed sequence tags (ESTs). 147 Expressed SequenceTaqs (ESTs) were gained from 176 clones, which were selected randomly and sequenced at the 5'end. Thesequences were submitted to the EMBL database. Blasting the sequences in the GenBank, 43 of them werefound that they have significant similarity with data in GenBank. EST AJ620046 was has significantsimilarity with the arabinosidase of Bacteroides thetaiotaomicron. Using SMART-RACE a full-length cDNA ofAJ620046 was successfully obtained. In order to initially characterize the biochemical properties ofAJ620046, the ORF of AJ620046 named AF was cloned and expressed in Pichia Pastoris yeast.Recombinant pHIL-S1-AF constructed by inserting AF into pHIL-S1 was transformed into Pichia PastorisGS115. Preliminary experiments indicated that AJ620046 was expressed as a 32 kDa protein in recombinantyeast.

The three-dimensional visualization model of human body duct is based on virtual anatomical structure reconstruction with duct angiography,which realizes virtual model transferred from two-dimensional,planar and static images into three-dimensional,stereoscopic and dynamic ones repectively.In recent years,the multi-duct segmentation and division of the same specimen (or organ) is the focus of attention shared by surgeons and clinical anatomists.On the basis of 4.22 g/cm3 body bone density,this study has screened out metal oxide contract agent with different density for infusion and modeling,as well as compared and analyzed the effects of three-dimensional image of CT virtual bronchoscopy (CTVB),three-dimensional image of CT maximum intensity projection and three-dimensional model.This experiment result showed synchronously infusing multi-duct of same specimen (or organ) with contrast agent in different densities could reconstruct three-dimensional models of all ducts once only and adjust threshold to develop single or multiple ducts.It was easier to segment and observe the duct structure,anastomosis,directions and crossing in different parts,which was beyond comparison with three-dimensional image of CTVB.Although the existing three-dimensional duct reconstruction techniques still cannot be applied in living bodies temporarily,this study focused on a creative design of ducts segmentation in different density,which proposed a new experimental idea for developing multi-duct three-dimensional model in living body in the future.It will play a significant role in disease diagnosis and individual design in surgical treatment program.Therefore,this study observes the three-dimensional status of human duct with the application of contrast agent fillers in different density,combined with three-dimensional reconstruction technology.It provides an innovative idea and method for constructing three-dimensional model of digital multi-duct specimen,and the ultimate goal is to develop the digitized virtual human and precise medical treatment better and faster.

Objective To explore the tumor cell apoptosis induced by the effect in vitro of eukaryotic ex pression of full-length TRAIL cDNA and its extracellular region. Methods The eukaryotic expression vectors for both forms of the cDNA acquired from the fetal heart cDNA library were constructed. After gene transfecting, the stable expression cell lines were obtained by G418 screening. Results The supernatants from tansfectants could induce apoptosis of different tumor cell lines in vitro, and an enhanced effect was observed by adding TFAR19 (TF-1 cell apoptosis-related protein 19), a novel apoptosis gene product discovered in our laboratory. Conclusion Eukaryotic expression products of TRAIL can induce apoptosis of the tumor cells, and TFAR19 could enhance the effect on apoptosis of tumor cells.

The study aimed to explore the changes of the immunocyte quantities and cytotoxicity in bone marrow, and how many hematopoietic progenitor cells retained after the bone marrow cells were activated by different combinations of various cytokines. Bone marrow cells were divided into four groups and were cultured in vitro: (1) Control: no cytokines were added. (2) IL-2 group: bone marrow cells were activated by IL-2. (3) CD3-AK group: bone marrow cells were activated by IL-2 and CD3-McAb. (4) CIK group: IFN-gamma, IL-1, IL-2 and CD3-McAb were added. CFU-GM assay and CD3 phenotype detection were performed before and after activating culture in all groups. The changes of cell quantities during culture and cytotoxicity of cultured cells were tested. CD3 positive cells markedly increased in both CD3-AK and CIK groups. The cell numbers and cytotoxicity of CD3-AK and CIK groups were higher than those of control or IL-2 group obviously after culture (P < 0.05). CFU-GM were decreased in all groups after culture and there had no significant difference among four groups. The combination of IL-2 and CD3-McAb not only stimulates the proliferation of marrow immunocytes and increases their cytotoxicity but retains enough hematopoietic progenitor cells as well. This combination of cytokines can be used to purge autologous bone marrow in vitro.

Adult ; Hepatic Encephalopathy ; therapy ; Hepatitis ; complications ; therapy ; Humans ; Liver Failure, Acute ; therapy ; Liver, Artificial ; Plasma Exchange

To study immunophenotype and cytotoxicity of the immunocytes in bone marrow and peripheral blood after activation by combined cytokines, mononuclear cells (MNC) of bone marrow and peripheral blood were activated by IFN-gamma, IL-1, IL-2 and McAb-CD3 in vitro. The cell amount and morphology during culture were observed. Cytochemical staining and immunophenotype analysis were done before and after culture in two groups of MNC. Cytotoxicity was tested by MTT method. The results showed that the cell number of two groups increased obviously in culture (P < 0.05), while the peripheral blood mononuclear cells increased more markedly (P < 0.05). The cytochemical staining showed POX decrease, but PAS increase in two groups. The positive ratios of CD3(+), CD56(+) and CD38(+) cells in two groups increased obviously after culture (P < 0.05), but there was no significant difference between those two groups. CD3(+) CD56(+) cells increased obviously in peripheral blood mononuclear cells activated by cytokines (P < 0.05), but CD3(+) CD56(+) cells did not increase in bone marrow mononuclear cells. There was no significant difference between two groups' cytotoxicity. It was concluded that IFN-gamma, IL-1, IL-2 and McAb-C D3 increased cell number and cytotoxicity of both bone marrow and peripheral blood mononuclear cells that can be used in cell immunotherapy.
ADP-ribosyl Cyclase ; immunology ; ADP-ribosyl Cyclase 1 ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; immunology ; Bone Marrow Cells ; cytology ; drug effects ; immunology ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; Cell Count ; Cell Division ; drug effects ; Coculture Techniques ; Cytokines ; pharmacology ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; HL-60 Cells ; Humans ; Immunophenotyping ; Interferon-gamma ; pharmacology ; Interleukin-1 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Membrane Glycoproteins ; Time Factors

OBJECTIVETo investigate the clinical effects and prevent the complications of posterior and anterior decompression and internal fixation in the revision of cervical anterior internal fixation failure.

METHODSFrom 2008 January to 2011 December, 17 patients with cervical anterior internal fixation failure were treated with posterior and anterior decompression and internal fixation. There were 12 males and 5 females, aged from 26 to 68 years old with an average of 44.1 years. The lower screw loosening was found in 6 cases, the upper screw loosening in 5 cases, titanium mesh caving in 3 cases, the upper screw breakage in 2 cases, the lower screw breakage in 1 case. Informations of bone fusion were observed by X-ray, CT, MRI. Clinical effects were evaluated by modified JOA score.

RESULTSAll the revision operations were successfully completed. One case with poor blood coagulation function before operation resulted in postoperative hematoma and occurred neurological symptoms; after hematoma removal and fresh frozen plasma infusion later, neurological symptoms of the patient disappeared. All patients were followed up from 6 to 38 months with an average of (22.4±10.0) months. Postoperative at 2 weeks, 3 months, and final follow-up, JOA score had obviously improved and respectively was 13.1±1.6, 13.4±1.6, 14.2±1.5. All internal fixation locations were good after revision,and obtained bone fusion at 10 months after operation, with an average fusion time of 6 months.

CONCLUSIONThe combined posterior and anterior decompression and internal fixation in the revision of cervical anterior internal fixation failure is safe, can achieve thoroughly decompression, maintain the cervical curvature, reconstruct the three column stability, and it may be used for the patients of cervical anterior fixation failure.

Adult ; Aged ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged

AIM To investigate the improving effect of Suoquan Capsules (Linderae Radix,Alpiniae oxyphyllae Fructus and Dioscoreae Rhizoma) on mice with diabetic cystopathy and its mechanism of action.METHODS Sixty mice were randomly assigned into normal group (n =8) and model group (n =52);the diabetic models of the latter were induced by high-fat feeding combined with streptozotocin (STZ) injection,then modeled mice (n =32) were randomly divided into model,Mecobalamin Tablets,low-and high-dose Suoquan Capsules groups.The influences of Suoquan Capsules on fasting blood glucose (FBG),glycated serum protein (GSP) level and general conditions were observed.The bladder leak point pressure (BLPP) was determined.Histopathological staining was performed on urinary bladder.And the expressions of substance P and NK1 receptor were detected by double immunofluorescent staining.RESULTS There were no significant differences in FBG,GSP,body weight,food intake and water consumption among various groups.The high-dose Suoquan Capsules significantly decreased urine volume of mice.Compared with the model group,the treatment with Suoquan Capsules markedly increased BLPP,the expressions of substance P and NK1 receptor were significantly increased,and the histopathology of bladder in mice was obviously improved.CONCLUSION Suoquan Capsules improves the diabetic cystopathy in mice,and its mechanism maybe related to the up-regulation of substance P and NK1 receptor expressions in bladder tissue.

Objective To observe the clinical efficacy of Sanjiao Tiaoqi Acupuncture in the treatment of post-stroke respiratory dysfunction.Methods Seventy-two patients with post-stroke respiratory dysfunction were randomly divided into observation group and control group,36 cases in each group.The control group was given routine treatment,and the observation group was treated with Sanjiao Tiaoqi Acupuncture on the basis of the control group,both groups were treated for 14 consecutive days.After 2 weeks of treatment,the clinical efficacy of the two groups was evaluated,and the changes of white blood cell count(WBC),C-reactive protein(CRP)and clinical pulmonary infection score(CPIS)were observed before and after treatment.The changes of diaphragmatic activity were compared before and after treatment between the two groups.Results(1)After treatment,the WBC and CRP levels of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the WBC and CRP levels,with statistically significant differences(P＜0.05).(2)After treatment,the CPIS scores of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving CPIS scores,with a statistically significant difference(P＜0.05).(3)After treatment,the diaphragm mobility of patients in the two groups was significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving diaphragm mobility,and the difference was statistically significant(P＜0.05).(4)The total effective rate was 91.67％(33/36)in the observation group and 75.00％(27/36)in the control group.The efficacy of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion Sanjiao Tiaoqi Acupuncture for post-stroke respiratory dysfunction can significantly promote the absorption of inflammatory factors in patients and improve diaphragm mobility,with remarkable clinical efficacy.

BACKGROUNDThe purpose of this article was to clarify the optimal management concerning transjugular intrahepatic portosystemic shunts (TIPSs) and surgical shunting in treating portal hypertension.

METHODSAll databases, including CBM, CNKI, WFPD, Medline, EMBASE, PubMed and Cochrane up to February 2014, were searched for randomized controlled trials (RCTs) comparing TIPS with surgical shunting. Four RCTs, which were extracted by two independent investigators and were evaluated in postoperative complications, mortality, 2- and 5-year survival, hospital stay, operating time and hospitalization charges.

RESULTSThe morbidity in variceal rehemorrhage was significantly higher in TIPS than in surgical shunts (odds ratio [OR] = 7.45, 95% confidence interval[CI]: (3.93-14.15), P < 0.00001), the same outcomes were seen in shunt stenosis (OR = 20.01, 95% CI: (6.67-59.99), P < 0.000001) and in hepatic encephalopathy (OR = 2.50, 95% CI: (1.63-3.84), P < 0.0001). Significantly better 2-year survival (OR = 0.66; 95% CI: (0.44-0.98), P = 0.04) and 5-year survival (OR = 0.44; 95% CI: (0.30-0.66), P < 0.00001) were seen in patients undergoing surgical shunting compared with TIPS.

CONCLUSIONSCompared with TIPS, postoperative complications and survival after surgical shunting were superior for patients with portal hypertension. Application of surgical shunting was recommended for patients rather than TIPS.

Gastrointestinal Hemorrhage ; Humans ; Hypertension, Portal ; surgery ; Portasystemic Shunt, Surgical ; adverse effects ; Portasystemic Shunt, Transjugular Intrahepatic ; adverse effects