Objective To explore the reliable method for primary cultivation and identification of satellite cells of mice skeletal muscles in vitro.Methods Cell suspension was obtained from posterior limb skeletal muscles of newborn mice by mixed enzymatic digestion,and the muscle satellite cells were separated by Percoll-density gradient centrifugation and purified with differential adherence method.Cells growth was observed under an inverted microscope and the growth curve was drawn.Satellite cells were identified with monoclonal antibody against desmin and ?-actin.Results The primary satellite cells were round in shape,and began on adherence one day after cultivation,and the cells distributed uniformly,most of the cells were roundish.After cultured for 48 hours,the cells began on proliferation,and then became bigger and started to division,bubble cell arrangements with 2-3 or more cells were observed.At the third or fourth day of cultivation,the cells were in exponential phase of growth,and the round cells predominated,many clusters of cell clones were found.The satellite cells formed monolayer sheet(50%-70%) 7-10 days after cultivation,the fusiform cells predominated,but there were still some round cells in this phase.The cell proliferation decreased after cultured for 10-12 days.Immunocytochemical staining showed that the obtained cells expressed myogenic marker desmin and skeletal muscle-specific protein ?-actin.Conclusions High purity muscle satellite cells could be obtained by mixed enzymatic digestion,Percoll-density gradient centrifugation and differential adherence method.Immunocytochemical staining of desmin and ?-actin is the effective method to identify muscle satellite cells.

To study the dynamic process of myocardial uptake of thiopentai in the isolated rabbit hearts. Method: Thiopental at doses of 500?mol, 1500?mol and 500?mol was given sequentially to the perfused rabbit heart in a total time of 15 min. The outflow concentration of thiopental was measured with high performance liquid chromatography and the left ventricular +dp/dtmax served as a effective parameter. Resuh: The disposition and elimination of thiopental can be best described hy a two-compartment open model. It can disposed into myocardium rapidly (T_(1/2)?=0.5?0.1 min), but elimination was relatively slow (T_(1/2)?=25.3?10.1 min). The transfer rate was slower from peripheral to central compartment than from central to peripheral compartment. The tbeoritical maximum depressant effect of thiopental on + dp/dt (Emax) was 19.0 4-11.2 kPa.s~(-1) corresponding to 1/10 E_0. Conclusion: The myocardial uptake of thiopental can be fitted to a two-compartment open model with rapid disposition and relative slow elimination process.

Objective To analyze and summarize the clinical , pathological features of 16 patients with centronuclear myopathy.Methods All of the 16 patients performed clinical examination and sporadic patients and a proband with family history ( n=6 ) performed serum creatine kinase , electromyography and open muscle biopsies , and the specimens were used for a standard series of histological and histochemical stainings.The clinical and pathological features of these patients were analyzed.Results The proportion of centronuclear myopathy in suspected myopathy cases was 0.127%(6/4 724) in our department.The onset time ranged from infancy to adulthood.The common initial symptoms included mild to moderate weakness of lower limbs and bilateral ptosis ( n =4 ).The other symptoms were facial weakness ( n =1 ) and ophthalmoplegia (n=1).There were 12 patients performing distal weakness exceeding proximal weakness . One family with autosomal dominant inheritance of 11 patients had a later age of onset than the sporadic ones and manifested bilateral ptosis , bilateral lower limbs weakness , especially in distal muscle.Muscle biopsies showed pronouncedly increased amount of fibers with centrally placed nuclei with predominant type Ⅰfibers and a clear perinuclear halo surrounding the centrally placed nuclei and an appearance of spoke of a wheel in some fibers.Conclusions This series of centronuclear myopathy patients manifest clinical heterogeneity.Muscle biopsies show features of centralized nuclei pronounced increase , type Ⅰfibers predominance , etc.These can provide evidences for the diagnosis of the disease.

Objective To examine the content of the 69 iPhone applications(Apps) for smoking cessation that were distributed through the online iTunes store and explore Apps′level of adherence to the U .S .Public Health Service′s 2008 Clinical Practice Guideline for Treating Tobacco Use and Dependence (called guidelines) for the purpose of developing high quality Apps ,as of De-cember 29 ,2012 .Methods All Apps was categorized into 5 types based on National Tobacco Cessation Collaborative .In order to get the Apps′level of adherence to guidelines ,the content of every application was compared with guidelines respectively .Results Apps identified for smoking cessation were found to have low levels of adherence to key guidelines .Conclusion It is necessary that current and future Apps should be developed around evidence-based practices and guidelines to get reliable Apps for smoking cessa-tion .

The medical law science interdisciplinary talents play an crucial role in solving medical service dispute and alleviating contradictory. Based on the questionnaire survey and the interview to the court and law offices as well as the analysis of the society's demand for the knowledge structure and ability of medicine law professionals, we proposed the construction of the training mode for medicine law professionals with the ability to adapt to the social demand.

Lactate and succinate were produced by Corynebacterium acetoacidophilum from glucose under oxygen deprivation conditions. To construct knockout mutant, lactate dehydrogenase gene (ldh) of C. acetoacidophilum was deleted by double-crossover chromosome replacement with sacB gene. Comparing with the wild strain ATCC13870, ldhA-deficent mutant produced no lactate with glucose consumption rate decreased by 29.3%, while succinate and acetate concentrations were increased by 45.6% and 182%, respectively. Moreover, the NADH/NAD+ rate was less than 1 (about 0.7), and the activities of phosphoenolpyruvate carboxylase and acetate kinase of the ldhA-deficent mutant were enhanced by 84% and 12 times, respectively. Our studies show that succinicate and acetate production pathways are strengthened by blocking lactate synthesis. It also suggests that improving NADH supply and eliminating acetate generation are alternative strategies to get high succinate-producer.
Corynebacterium glutamicum ; genetics ; metabolism ; Glucose ; Industrial Microbiology ; L-Lactate Dehydrogenase ; genetics ; metabolism ; Lactic Acid ; metabolism ; Oxygen ; metabolism ; Phosphoenolpyruvate Carboxylase ; Succinic Acid ; metabolism

Objective To analyze the surgical and pathologic features of isolated ascending aortitis without evidence of rheumatologic or autoimmune diseases by comparing that of Takayasu's arteritis.Methods Consecutive 965 excised ascending aortas were reviewed and 40 cases with aortitis were selected from the pathological archives of past 20 years in Fuwai Hospital.The clinical history,laboratory and angiographic data,surgical findings,steroid therapy and followup results of these 40 cases were reviewed.Pathological parameters included the thickness of aortic wall and its various layers,inflammation activity,multinuclear giant cells,inflammatory necrosis,laminar necrosis and atheroselerosis.The t test and Chi square analysis were used to compare the means and the rates between the two groups.Results Twenty-five of 40 cases were diagnosed with isolated aortitis and its prevalencei was 2.6%(25/965),while 8 cases with Takayasu's arteritis and 7 cases with other vasculitis were confirmed.The age of isolated ascending aortitis was older than that of Takayasu's arteritis[(46±12)y vs(34±14)y,t=2.30,P>0.05] while the male/female ratio was similar in both groups(1.1 vs 1.0,t=0.01,P>0.05).Their main clinical manifestation was aortic aneurysm with a bigger aortic diameter in isolated aortitis than that in Takayasu's arteritis [(59±18)vs(46±12)mm,P>0.05].The asymptomatic cages mostly were found in isolated ascending aortitis (28%vs 0,x2=2.98,P>0.05).The erythrocyte sedimentary rate was normal in isolated aortitis but increased in Takayasu's arteritis[(15±17)mm/1 hvs[27±25)mm/1 h,U=48.50,P<0.05 ].Inflammatory edema,thickening and calcification were noted in more than half cases of both groups.but slighter thickening of aortic wall was foand in isolated ascending aortitis in contrast to the remarkable thickening in Takayasu artefifis[(2506±493)vs (3300±430)μm,t=-3.00,P<0.01].Giant cells,inflammatory necrosis,laminar necrosis and accompanied atherosclerosis were more common in isolated ascending aortitis.Aortic regurgitation was common but inflammatory invasion wege found in few aortic valves. Steroid was prescribed in 5 cases with isolated ascending aortitis and none of them was reported with complications while aortic perivalvular leakage occurred in 3 of 20 cases without steroids therapy. In the Takayasu's arteritis cases, new aneurysm of abdominal aorta was found in 1 of 3 cases with steroid therapy. Meanwhile, perivalvular leakage and subclavian artery stenosis occurred in 2 of 5 cases without steroid. Conclusion Isolated aortitis is more common than Takayasu's arteritis in ascending aortic diseases and there is some definite difference in clinical and pathological aspects between these two diseases. Since the effect of steroid therapy is uncertain and in order to avoid overtreatment, we suggest that steroid should not be given to cases with isolated ascending aortitis after surgery and clinical and angiographic follow-up should be emphasized.

Objective To investigate the drug resistance and risk factors of hospital-acquired pneumonia (HAP) induced by imipenem-resistant Acinetobacter baumannii.Methods Clinical data on 114 patients with Acinetobacter baumannii-related HAPs admitted in Wujiang First People' s Hospital in Suzhou during January 2013 and December 2014 were retrospectively analyzed.According to the results of drug sensitivity test,patients were divided into imipenem-resistant group and non imipenem-resistant group.Drug resistance to 20 commonly used antibiotics was observed in two groups,and multivariate Logistic regression analysis was performed to identify the risk factors of imipenem-resistant Acinetobacter baumannii infection.Results Among 114 strains ofAcinetobacter baumannii,66 strains (57.89％) were imipenem-resistant and 48 strains (42.11％) were non-imipenem-resistant.The resistance rates to β-lactams,quinolones and aminoglycosides were significantly higher in imipenem-resistant group than those in non-imipenem-resistant group (P ＜ 0.01),and no tigecycline-resistant strain was found in both groups.Univariate analysis showed that acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score ≥ 15,plasma level of albumin ≤ 25 g/L,intensive care unit (ICU) stay,indwelling gastric tube,deep venous catheterization,establishment of artificial airway,mechanical ventilation time ≥ 7 d,use of broad-spectrum antibiotics ≥ 14 d and combined use of antibiotics were risk factors of imipenem-resistant Acinetobacter baumannii related HAP (x2 =13.06,6.86,25.40,15.09,17.87,21.46,17.94,6.91 and 10.10,P ＜0.01).Multivariate Logistic regression analysis revealed that establishment of artificial airway [OR =72.014,95％ confidetial interval (CI):19.566-265.061,P ＜ 0.01],and use of broad-spectrum antibiotics ≥ 14 d (OR =3.892,95％ CI:1.092-13.879,P ＜ 0.05) were independent risk factors of imipenem-resistant Acinetobacter baumannii related HAP.Conclusion Imipenem-resistant Acinetobacter baumannii strains are highly resistant to most antibiotics.Strict control of invasive procedures and long-term combined use of antibiotics may reduce the occurrence of imipenem-resistant Acinetobacter baumannii related HAPs.

Objective To explore the validity of simulative employment training course on enhancing the core competitiveness of university students.Method We investigate the students with simulative employment trainin, graduates and recruiters through interview, symposium and questionnaire survey. Results The comprehensive quality and occupational capacity of students that completed simulative employment training had been improved. In the simulative employment training course,98%students thought their interview skills had been promoted, 90%recruiters were satisfied with the students,95%graduates who had worked consider that the simulative employment training were helpful. Conclusions Simulative employment training is effective for enhancing the core competitiveness of university students.

Objective To investigate the expression of B-cell activating factor ( BAFF ) and its specific receptor BAFF-R in patients with B-cell non-Hodgkin′s lymphoma ( B-NHL) and to analyze the cor-relations between BAFF and the development of B-NHL.Methods RTQ-PCR and Western blot assay were used to measure the expression of BAFF and its specific receptor BAFF-R in patients with B-NHL.Fluores-cence immunocytochemical staining was used to determine the localization of BAFF and BAFF-R in Raji cells, a B-NHL cell line.The expression of BAFF in tumor tissues from patients with B-NHL of different his-tologic subtypes was measured by immunohistochemistry.WST proliferation and TUNEL assays were used to evaluate the effects of BAFF and BAFF-R on the proliferation, survival rate and apoptosis of Raji cells.Lin-ear correlations between the concentrations of lactate dehydrogenase ( LDH) and the expression of BAFF and BAFF at mRNA and protein levels in patients with B-NHL were analyzed.Results BAFF and its specific receptor BAFF-R were expressed in Raji cells and played an important role in the survival and proliferation of B-NHL cell line.The expression of BAFF in tumor cells from patients with B-NHL varied with the different histologic subtypes of B-NHL.Patients with small B-cell malignant lymphoma, large B-cell lymphoma ( LBCL) , mucosa-associated lymphoid tissue lymphoma ( MALT lymphoma) and follicular lymphoma showed higher levels of BAFF, while those with mantle cell lymphoma showed lower levels of BAFF.Compared with the healthy subjects, patients with B-NHL showed significantly increased expression of BAFF at mRNA and protein levels.The levels of LDH were closely related to the expression of BAFF at mRNA and protein lev-els.Conclusion BAFF and its specific receptor BAFF-R might play an important role in the growth and survival of malignant B cells.