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.

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 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 observe the effects of Chinese herbal compounds (alkaloid of Coptidis Rhizoma 40%, total saponin of panax notoginseng 35%, and polysaccharide of Ophiopogonis Radix 25%) on levels of blood glucose, fructosamine, and insulin of diabetic rats, and explore mechanism of action.Methods Ten rats were randomly chosen as blank control group, and other rats were used to establish diabetic rat models by STZ intraperitoneal injection. Successfully molding rats were randomly divided into Chinese herbal compounds high, medium, low dose groups, metformin group, and model group, 10 rats in each group. Each rat received gavage for continuous 4 weeks. Then rat blood was collected by cutting head and taking caudal vein. FBG was detected by glucose oxidase method;insulin level was detected by radio-immunity double antibody method;fructosamine content was detected by fructosamine method.Results Compared with the blank control group, model group the levels of FBG, froctosamine content increased, while fasting insulin level were lower after 4 weeks'medication. Compared with the model group, the levels of FBG, froctosamine content in Chinese herbal compounds high and medium dose groups and metformin group were lower, while fasting insulin level increased after 4 weeks'medication (P<0.05).Conclusion TCM herbal compounds can increased the basic insulin level by decreasing the contents of FBG and fructosamine.

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 investigate the clinicopathological features of tubular aggregate myopathy.Methods Eight patients as experimental group were diagnosed with tubular aggregate myopathy in Department of Neurology,People' s Liberation Army General Hospital,between March 2000 and March 2013.The data were retrospectively analyzed.Enzyme histochemical techniques and electron microscopy were taken to observe the muscle structures.Results The detection rate of tubular aggregate was 0.374％ (8/2 137).All of the 8 patients with tubular aggregate myopathy were male.Five patients presented with episodes of muscle weakness,while 3 patients presented chronic progressive muscle weakness.The main clinical features of all patients were muscle weakness.The creatine kinase level was mildly elevated in 4 patients,while it was normal in the other 4 patients.Electromyogram showed myogenic damage in 5 patients and normal in 3 patients.All of the 8 patients denied family history.By light microscopy,hematoxylin-eosin staining showed that tubular aggregates were multiple basophilic subsarcolemmal substance.Tubular aggregate stained red with modified Gomori trichrome stain,reacted intensely for nicotinamide adenine dinucleotidetetrazolium reductase,and remained unstained on oil red O,periodic acid Schiff,sudan black B,acid phosphatas,adenosine triphosphatase.By electron microscopy,tubular aggregates were densely packed tubules predominantly in the subsarcolemmal region.Conclusion Enzyme histochemical staining and electron microscopy show special features of tubular aggregate myopathy,are the critical techniques for the diagnosis of the disease.

Objective To describe the changes of thrombosed venous sinus on MRI after administration of contrast material and evaluate the sensitivity and specificity of thread-like enhancement around sinus to diagnose thrombosis in the corresponding sinus.Methods Patients with cerebral venous sinus thrombosis (CVST) admitted to our department from January 2005 to December 2014 and undergone a MRI scan with administration of contrast material were included in this study.The enhancement features of venous sinus were studied in the plane parallel to the interested sinus.The features of enhancement were classified as peripheral thread-like enhancement, partial enhancement and complete enhancement.The proportion of these three type of enhancement in thrombosed sinuses and normal sinuses were described and compared.The sensitivity and specificity of peripheral thread-like enhancement to diagnose thrombosis in corresponding sinus were calculated.The proportion of each type of enhancement was also described and compared in acute (≤ 7 d), subacute (8-30 d) and chronic (≥ 31 d) stage after onset of symptoms.Results Peripheral thread-like enhancement, partial enhancement and complete enhancement were all found in both thrombosed and normal sinuses.There was a significant difference of enhancement features between normal and thrombosed sinus in superior sagittal sinus (100％ (30/30) vs 60％ (27/45), x2 =13.789, P =0.001), left trans verse sinus, and right sigrnoid sinus.The sensitivity and specificity of peripheral thread-like enhancement to diagnose thrombosis in the corresponding sinus were 10.5％-44.4％ and 53.3％-76.7％ respectively.There was no significant difference of contrast features at different stage after onset.Conclusion The value of peripheral thread-like enhancement to diagnose CVST is limited because of low sensitivity and specificity.

There are lack of specific treatment for diabetic peripheral neuropathy (DPN)right now.However, TCM intervention highlights the distinct advantage for the disease.Modified Huangqi-Guizhiwuwu Tang is one of the important recipes for that.But there are no uniform standards curative effect evaluation for it, and some clinical trials do not have some new evaluation standards.We summary the clinical research literatures in recent years and analyze the existing problems, therefore, provide new recommendations as views for medical researchers in the design of clinical trials.

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.