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 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 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.

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.

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 explore the effect of Da Vinci robot?assisted laparoscopic radical prostatecto?my on the respiratory function of elderly in Intensive Care Units( ICU) . Methods Thirty?nine elderly patients received Da Vinci robot?assisted laparoscopic radical prostatectomy ( RARP ) from January 2015 to April 2016 and 25 cases received conventional laparoscopic radical prostatectomy from January 2014 to December 2014 ad?mitted into ICU were retrospectively analyzed. Their comorbidities,blood loss and transfusion during surgery,ate?rial blood gas(ABG) analysis and respiratory complications after operation,clinical outcomes between the two groups were compared. Results Compared with conventional laparoscopic radical prostatectomy,RARP group spent more time in surgery((4. 23±1. 44) h vs. (3. 25±1. 31) h,t=2. 783,P<0. 05),more patients need venti?lation(11 vs. 1,χ2=4. 378,P<0. 05) . ABG analysis showed respiratory and metabolic acidosis with lower pH (7. 29±0. 09 vs. 7. 35±0. 05,t=3. 886,P<0. 05),HCO3?((20. 05±2. 50) mmol/L vs. (22. 86±2. 53) mmol/L,t=3. 473,P<0. 05),BE(-5. 11±3. 94 vs.-3. 64±1. 17,t=5. 018,P<0. 05) and higher pCO2(46. 15±8. 31 vs. 40. 25±6. 57,t=2. 475,P<0. 05),Lac((3. 54±1. 99) mmol/L vs. (2. 91±1. 39) mmol/L,t=2. 254,P<0. 05) . Conclusion RARP may cause carbon dioxide retention and respiratory complications on elderly pa?tients. It may reduce postoperative respiratory complications by shortening surgery time,lowering pneumoperitone?um pressure,hyperventilation,recruitment maneuvers and chest physical therapy.

9).Renal pathology revealed 3 patients with class Ⅳ, 1 patient with class Ⅱ. Image results showed diffuse pulmonary infiltration lesions. Bronchial lavage fluid was hemorrhage. Three patients recovered after treatment with corticosteroid and intravenous cyclophosphamide therapy. Conclusions Diffuse alveolar hemorrhage is rare serious complications of lupus nephritis. Aggressive immunosuppressive therapy should be started early for a favorable outcome.

Objective To investigate the performance of dual source CT scanning combined with low-tube-voltage iterative reconstruction (SAFIRE) for demonstrating coronary artery calcium plaques. Methods 70 patients with clinically suspected coronary atherosclerosis and with normal BMI received SAFIRE-3 or filtered back projection (FBP) for reconstruction after undergoing CT scans with tube voltage range of 100 kV. The CT images (average CT values, noise, SNR, and CNR) and subjective scores (number of calcium plaque, calcium plaque adjacent artifacts, and image quality) were compared between the two groups. Results The average CT value of the aortic root did not differ significantly between the two procedures for reconstruction (P > 0.05). Ob jective evaluation of the images (noise, SNR, and CNR) between the two groups was statistically significant (P < 0.05 for all comparisons), so was image subjective rating (number of calcium plaque and rate of calcium plaque edge)(P < 0.05 for all comparisons). SAFIRE-3 reconstruction was markedly superior to FBP reconstruc-tion. Conclusions Low kilovoltage combined with reconstruction of SAFIRE-3 can reduce calcium plaque adja-cent artifacts and enhance quality of images , which is better than FBP in displaying the morphology and adjacent conditions of calcified plaque; therefore it is worth popularizing.