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.

Objective To evaluated the role of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in the diagnosis of thoracic tuberculosis.Methods The study was retrospective,from September 2009 to September 2012,38 patients who underwent EBUS-TBNA were finally diagnosed of thoracic tuberculosis,with enlarged hilar or mediastinal Iymph nodes on chest enhanced computed tomography(≥ 1.0 cm).Patients in whom EBUS TBNA was nondiagnostic subsequently underwent surgical biopsy.All the patients had a minimum of 6 months clinical and radiologic follow-up.Results EBUS-TBNA was performed on a total of 88 lymph node stations in 38 patients.Of the enlarged lymph nodes,60(68.18％) were located in the mediastinal region and the remaining 28 (31.82 ％) around the hilum or interlobar area.Of the 38 patients,EBUS-TBNA achieved definitive diagnosis in 34 patients(89.47％).EBUS was well tolerated by all of the patients with no complications.Conclusion EBUS-TBNA is a safe procedure with a high yield for the diagnoses of thoracic tuberculosis.

Objective To compare open reduction and cannulated screws combined with ascending branch of lateral femoral circumflex artery iliac bone flap with closed reduction and cannulated screw fixation for the treatment of displaced femoral neck fracture in young adults.Methods From January 1992 to January 2007,totally 204 cases of displaced femoral neck fracture were treated.Group A:one hundred and three cases were treated with open reduction and 2 cannulated screw fixation,covered by the vascular pedicled iliac bone flap with ascending branch of lateral femoral circumflex artery.Garden classification:type Ⅲ.68 cases,type Ⅳ:35 cases.Group B:one hundred and one cases,treated with closed reduction and three cannulated screw fixation.Garden classification:type Ⅲ in 62 cases and type Ⅳ in 39 cases.Results Two groups were followed up for 5-15 years.The healing time of group A was (73 ±21.2) days,including 4 cases of nonunion fracture (3.9％) and 5 cases of femoral head avascular necrosis occurred (4.9％) ; The healing time of group B was (89 ± 12.5) days,including 12 cases of nonunion fracture (11.9％) and 18 cases of femoral head avascular necrosis occurred (17.8％).The healing time,the rate of nonunion of fracture and the rate of femoral head necrosis were significantly different of these 2 groups.Conclusion The treatment of open reduction and 2 cannulated screw fixation,covered by the vascular pedicled iliac bone flap with ascending branch in young adults can effectively prevent nonunion of fractures and osteonecrosis of the femoral head.

Objective To measure the parameters of anterior bow of the femoral medullary cavity by means of 3D spiral CT reconstruction and to provide the anatomical basis for the design of the femoral component prosthesis.Methods 105 healthy Chinese left femurs were scanned in standard neutral position by 3D spiral CT.Parameters of the anterior bow of the femoral medullary were measured by medical imagine software.Results The parameter values of the male femur were statistically bigger than those of the female,including the length of the femoral medullary and the distance from the point of maximum curvature to plane above lesser trochanter (P＜0.001).No significant differences between male and female in the position index of the point of maximum curvature were observed (P＞0.05).The relative curvature of the female femoral anterior arch was statistically bigger than that of the male (P＜0.001).The femoral curvature subtense and the length of the femoral medullary showed no significant correlation (r=0.106,P=0.281).Conclusion There are certain anatomic differences in the anterior bow of the femoral medullary between male and female.Appropriate length of femoral component should be considered in total hip replacement operation.And it is necessary to design the femoral components fitting the features of the femurs according gender difference.

OBJECTIVE:To establish the optimal technical condition for the extraction of volatile oil from the volatile herbs of Huoxiang zhengqi prescription with supercritical CO2 fluid(SFE-CO2).METHODS:The technical condition for the extraction of volatile oil from the volatile herbs of Huoxiang zhengqi prescription were optimized using L9(34)orthogonal design table with extraction pressure,the extraction temperature,the pressure and temperature of separator Ⅰ as factors and with the yield of the volatile oil as index.RESULTS:A highest extraction yield of volatile oil from the volatile herbs of Huoxiang zhengqi prescription was achieved(2.057%)under an optimal technical condition as follows:with extraction pressure at 25 MPa,the extraction temperature at 30 ℃,the pressure for separator Ⅰ at 9 MPa and the temperature of separator Ⅰ at 45 ℃.CONCLUSION:Extraction of volatile oil from the volatile herbs of Huoxiang zhengqi prescription by SFE-CO2 is characterized by high extraction yield,stable,pure operation,safe,and by which the biological activity of the volatile oil can be maintained,thus it is a potential extraction and separation method.

OBJECTIVE: To analyze the differences of sleep respiratory parameters recorded by PSG and synchronous blood pressure measured by ambulatory blood pressure monitor between obstructive sleep apnea (OSA) patients with hypertension (HT) and simple OSA and simple HT; To determine the characteristics of sleep respiratory parameters and blood pressure changes in patient with OSA accompanied HT. METHOD: We chose the patients who were diagnosed simple HT (n=45) and simple OSA (n=50) and OSA with HT (n=56), Compared the sleep respiratory parameters and blood pressure changes between the three groups. Meanwhile the correlations about the sleep respiratory parameters and synchronization blood pressure were analyzed. RESULT: Compared with simple HT and simple OSA, OSA with HT has higher apnea hyponea Index (AHI) (P<0. 001), oxygen desaturation index (ODI), awake index (AI), wake after sleep onset (WASO) and the proportion of non-rapid eyemovement sleepl (N1) in total sleep time(TST), has lower mean arterial oxygen saturation (MSaO2), lowest arterial saturation oxygen (LSaO2), the proportion of slow wave sleep (SWS) and rapid eyemovement sleep (REM) in TST (P<0. 05). There were positive correlations between the systolic/diastolic blood pressure (SBP/ DBP) and AHI, ODI, AI, WASO and N1/TST (P<0. 05). Compared with simple OSA, the mean day systolic blood pressure (dMSP), mean night systolic blood pressure (nMSP), mean day diastolic blood pressure (dMDP), mean night diastolic blood pressure (nMDP) and mean night diastolic blood pressure (nMDP) were significantly decre- sed, meanwhile the difference between the average systolic/diastolic blood pressure day and night were significantly increased after continuous positive airway pressure (CPAP) treatment. OSA with HT has higher There were negative correlations between the SBP/DBP and MSaO2, LSaO2 (P<0. 05). Blood pressure mainly changed in the later sleep stage when the REM phase was increased. Blood pressure changes were characteristic of increasing DBP and decreasing SaO2. CONCLUSION There are significant differences between simple OSA and OSA with HT in the sleep respiratory parameters, which are closely related with changes of blood pressure in sleep stage; AHI is the high risk factor of the OSA with HT. PSG is a effective factor in estimating the OSA accompanied HT course of development and prognosis.
Blood Pressure ; Blood Pressure Monitoring, Ambulatory ; Continuous Positive Airway Pressure ; Humans ; Hypertension ; complications ; Polysomnography ; Sleep ; Sleep Apnea, Obstructive ; complications ; diagnosis ; Sleep Stages

In order to compensate for the deficiencies of current medical education, improve the comprehensive quality of medical students, the First Clinical College, Tongji Medical College of Huazhong University of Science and Technology took the lead in carrying out the health management practice of rela-tives and friends, and made a beneficial exploration of humanistic education for medical students. Relatives and friends were regarded as the specific practice objects, and by telephone and network at school, face-to-face communication during the vacation, medical students established, managed and track the health records of relatives and friends. The students could practice while learning, which made medical students, in im-proving the knowledge structure, enhance their emotional communication with friends and relatives, improve their doctor-patient communication ability, establish the concept of humanistic care, and cultivate their occupation moral respect for life and their dedication to medicine and the sense of responsibility and mission to the society. It provides a new strategy for medical humanities education.

Objectives To explore the associations between drug efflux pump gene expression and phenotypic drug resistance as well as gene mutation patterns related to drug resistance of Mycobacterium tuberculosis.Methods Forty-five Mycobacterium tuberculosis isolates resistant to one or more of drugs including isoniazid, rifampicin, streptomycin and ethambutol, and 26 isolates all sensitive to the above four drugs from Tianjin Tuberculosis Control Institute in 2007 were involved in this study. Direct sequencing was applied to detect the mutations in the corresponding resistance genes(isoniazid:katG, inhA, oxyR-ahpC, ndh, rifampicin:rpoB, streptomycin:rpsL, rrs, and ethambutol:embB, embC and embA). After RNA extration and reverse transcription, real-time PCR was conducted to assess the expressions of putative drug efflux pump genes Rv1410c, Rv2136c, Rv0783c and Rv2136c, and Students' t test and ANOVA analysis were used to analyze the expression differences in Mycobacterium tuberculosis with different phenotypic drug resistance and drug resistance related gene mutation patterns.Results Compared to pan-sensitive isolates[(5.67±3.29)×10-5], Rv1410c showed higher expression in streptomycin[(8.48±6.33)×10-5, t'=2.18, P<0.05], isoniazid[(8.43±6.38)×10-5, t'=2.20, P<0.05], rifampicin[(9.59±7.27)×10-5, t'=2.29, P<0.05], multi-drug[(10.37±7.86)×10-5, t'=2.34, P<0.05] resistant isolates, and in isoniazid + streptomycin resistant isolates[(9.39±6.81)×10-5, t'=2.43, P<0.05];Rv2136c showed higher expression in isoniazid resistant[(3.51±2.43)×10-5, t'=2.03, P<0.05], multidrug-resistant isolates[(4.21±2.94)×10-5, t'=2.22, P<0.05] and resistant to isoniazid+streptomycin[(3.81±2.46)×10-5, t'=2.28, P<0.05] isolates . The expression of Rv0783c in rifampicin resistant isolates with rpoB 531 mutations [(5.41±3.03)×10-6] was higher than those with wild type of rpoB 531[(2.29±1.62)×10-6, t=2.81, P<0.05].Conclusions The expression of Rv1410c and Rv2136c are associated with mutiple-drug resistance of Mycobacterium tuberculosis.The expression of Rv0783c in rifampicin resistant isolates is associated with mutation in rpoB 531.

Objective To investigate the effect of hepatitis B virus (HBV) on the immune function of natural killer (NK) cells.Methods Healthy human peripheral blood-derived NK cells were cultured alone,or co-cultured with plasmacytoid dendritic cell (pDC) (NK∶ pDC=5∶1) for 48 h with or without HepG 2.2.15-derived HBV.Cell activation was assessed by flow cytometry.The specific enzyme linked immunosorbent assay (ELISA) and intracellular cytokine staining (ICS) were used to determine the cytokine production,the cytotoxic effect of NK cells on the carboxyfluorescein diacetate succinimidyl ester (CFSE)-labelled K562 target cells as well as the granzyme and perforin levels in NK cells.Paired results were analysed using Wilcoxon signed rank test.Results HBV did not affect interleukin (IL)-12/IL-18 and IL-18/interferon α (IFNα)-induced interferon γ (IFNγ) production by NK cells when NK cells were cultured alone (both P＞0.05).However,HBV significantly inhibited pDC-induced IFNγ production by NK cells (146.1 pg/mL),while CpG enhanced pDC-induced IFNγ production by NK cells significantly (1135.4 pg/mL,P=0.0005).HBV did not affect pDC-induced NK cell activation and cytotoxicity to K562 target cells as well as intracellular granzyme and perforin levels.ConclusionHBV does not activate but inhibits pDC-induced NK cell function,which may contribute to the persistence of HBV infection.

BACKGROUND:Seed cells and scaffold are two key factors for cartilage defects after osteonecrosis of femoral head using tissue-engineered method. OBJECTIVE:To explore the feasibility of Bio-gide col agen membrane combined with dog bone marrow mesenchymal stem cells into chondrocytes. METHODS:Bone marrow mesenchymal stem cells were isolated from beagle dogs by whole bone marrow blood centrifugation method and adherence screening method in vitro and cultured. Morphological changes in cells were observed, and identification was done using cellsurface antigens. Bone marrow mesenchymal stem cells of passage 3 were induced by chondrocyte induction medium to differentiate into chondrocytes (experimental group). cells cultured in normal medium were considered as control group. 3-(4, 5-Dimethylthiazol-2-yl) 2, 5-diphenyl tetrazolium bromide assay was used to measure growth curve of chondrocytes. cells underwent typeⅡcol agen immunohistochemistry and toluidine blue staining. The coculture of bone marrow mesenchymal stem cells at passage 3 and Bio-gide col agen membrane were observed under an inverted phase contrast microscope and scanning electron microscope. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells with high purity and high viability were obtained by whole bone marrow blood centrifugation method and adherence screening method. cells grew wel and had strong amplified ability, and successful y differentiated into chondrocytes. Numerous bone marrow mesenchymal stem cells adhered on the Bio-gide col agen membrane, showing a tendency of multi-layer growth. cells and Bio-gide col agen membrane seem to blend into an integrant part. Cel processes appeared and connected each other and gradual y wrapped the Bio-gide col agen membrane, with the presence of obvious cel matrix secretion. These results suggested that bone marrow mesenchymal stem cells can grow and differentiate into chondrocytes on the Bio-gide col agen membrane.