Objective To explore the repair method for refractory diabetic wound. Methods A total of 206 patients with refractory diabetic foot ulcers were treated with proper surgical treatments.Results Of all, 106 patients were treated by skin flap (51.5 % ), with one stage wound healing rate of 85.8%; 122 patients were repaired with split-thickness skin graft ( 59.2% ), with survival rate of the graft for 79.5%. Simple toe amputation was made in 34 patients (46 toes). The high level amputation was performed in 56 patients (27.2%). Of all, 132 patients were followed up for 6-18 months, which showed that ulcer recurred in 12 patients (9.1%). Conclusion Timely and effective treatment as well as flap and skin graft repair could reduce high level amputation rate of diabetic foot ulcer and promote the quality of life.

Objective To investigate the flora distribution and drug resistance status in the Affiliated Hospital of Academy of Military Medical Sciences so as to provide experimental data for clinical doctors to use antibiotics more efficiently.Methods The clinical data of pathogenic bacterial infections over nearly one year in our hospital were retro -spectively analyzed .Results There were 3815 strains of pathogenic bacteria isolated from the sample .The percentage of Gram-positive strains was 36.4%while that of Gram-negative bacteria was 63.6%.The most common bacteria were Esche-richia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Staphylococcus epidermidis.In terms of drug tolerance , Enterobacteriaceae remained highily sensitive to carbapenems .The total resistance rate was 2%-5%.The resistance rate of A.baumannii to meropenem and imipenem was 60%.There were still a few pan-drug resistant strains among K.pneumoniae,A.baumannii and P.aeruginosa,but there were no drug resistant strains to vancomycin , tige-cycline and linezolid in Staphylococcus.The resistance rate of Enterococcus faecium to vancomycin was 9%.The bacteria were distributed predominantly in ICU ,Department of Hematology and Department of Oncology .The samples were mainly composed of phlegm specimens .Conclusion The high distribution in the three departments mentioned above is largely re-lated to the diseases being treated .The specimens from the lower respiratory tract show more types of bacteria that are mostly drug-resistant, and the isolating rate of vancomycin resistant Enterococcus and carbapenems resistant K.pneumoniat is com-paratively high .

Many researchers employed mammalian expression system to artificially express cannabinoid receptors, but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports. In present study, we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system. This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide. In addition, the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e., [Symbol: see text]95°C), forming a large molecular weight band when analyzed by immuno-blotting. Only denaturing temperatures [Symbol: see text]75°C yielded a clear band at the predicted molecular weight. Collectively, we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence, and described the requirement for a low sample denaturing temperature in immuno-blot analysis. These findings provide very useful information for efficient mammalian expression and immuno-blotting of membrane receptors.

AIM: To purify human yolk sac mesenchymal ste m cells (hYS-MSC) and investigate its osteogenic and neurogenic differentiation potentials. METHODS: hYS-MSC were separated from yolk sac and purified via p assage culture. The karyotype of hYS-MSCs was analyzed via G-banded characterist ics. Flow cytometric analysis was used to determine the cell cycle and phenotype of hYS-MSC. The AKP expression of hYS-MSC was also tested. Osteogenic different iation of hYS-MSCs was induced by 10 -8mol/L dexamethasone, 10 mmol/L ?-gl ycerophosphate and 50 mg/L vitamin C. Alizarin red S stain was used for identifi cation of mineralization. ?-mecaptoethanol or salviae miltiorrhizae were used t o induce neurogenic differentiation of hYS-MSCs. The expressions of NSE, NF and GFAP were identified by immunohistochemical method. RESULTS: hYS-MSCs could be purified at passages 2 or 3. The cell cycle analysis suggested that hYS-MSCs showed strong proliferational potentials by which the cells kept normal diploid karyotype during the in vitro cultur e. Flow cytometry showed the phenotype of purified hYS-MSCs was uniformly positi ve for CD29, CD44, CD105, and CD166, and negative for reactivity to antigens CD3 4, CD45, or CD86. hYS-MSCs were weakly but clearly positive in AKP. Osteogenic d ifferentiation was appeared after induction of osteogenic differentiation. hYS-M SCs, which were of spindle shape, uniform in size, were induced to pleomorphism osteoblast-like cells which expressed high level of AKP. Aggregates or nodules w ere formed at day 7 and calcium accumulation was detected by alizarin red S stai ning on day 10 or day 14. Neurogenic differentiation of hYS-MSCs was induced by ?-mecaptoethanol or salviae miltiorrhizae. NSE, NF or GFAP positive cells were detected by immunohistochemical staining. CONCLUSIONS: hYS-MSCs have strong proliferation potential and th e normal diploid karyotype is kept during the in vitro culture. The phenoty pe of hYS-MSCs is coincident with adult hMSCs. hYS-MSCs could be induced to dif ferent iate into osteogenic or neurogenic cells.

OBJECTIVETo investigate the effect of semaphorin-3F (SEMA-3F) gene transient transfection on the proliferation of Tca8113 tongue carcinoma cells.

METHODSAfter construction of a full-length SEMA3F expression vector, Tca8113 cells were transient transfected with pEGFP-N1-SEMA3F by Lipofectamine 2000. The expression of SEMA-3F gene was detected by RT-PCR The differences of the expression in the transfected cell groups, empty vector groups and un-transfected cell groups were compared. The survival rates were assayed by methyl thiazolyl tetrazolium (MTT) enzymatic labeling technique. Cell cycle were assayed by flow cytometer (FCM).

RESULTSThe gene was transfected into Tca8113 cells. High expression of SEMA-3F was successfully detected in the transfected cell groups after gene transfection. The cell cycle percentage of G1 stage decreased and S stage increased.

CONCLUSIONSSEMA-3F gene transient transfection may inhibit the proliferation of Tca8113 cells.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Membrane Proteins ; genetics ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; Tongue Neoplasms ; genetics ; metabolism ; pathology ; Transfection

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex

Objective To discuss the application of flipped classroom teaching model in the teaching reform of Science of Tuina Technique based on MOOC; To provide references for promoting teaching quality of Science of Tuina Technique. Methods Flipped classroom teaching model based on MOOC and traditional teaching model were conducted in 2 classes of 2013 in Guangxi University of Chinese Medicine, one experimental groups and one control group. After the end of the course, theoretical knowledge and techniques skill scores of the two classes were compared. Questionnaire was given to students in the experimental class to understand their satisfaction towards flipped classroom based on MOOC. Results The scores of theoretical knowledge, performance demonstration test results, mannequin test and practices mechanics test of experimental group were high than the control group, with statistical significance (P<0.05). The results of questionnaire showed that the satisfaction of students in the experimental group towards flipped classroom based on MOOC was 95. Conclusion Flipped classroom teaching model based on MOOC applied to the teaching of Science of Tuina Technique can improve the students' learning interest and initiative, and then improve academic performance.

Many researchers employed mammalian expression system to artificially express cannabinoid receptors,but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports.In present study,we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system.This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide.In addition,the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95℃),forming a large molecular weight band when analyzed by immuno-blotting.Only denaturing temperatures ≤75℃ yielded a clear band at the predicted molecular weight.Collectively,we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence,and described the requirement for a low sample denaturing temperature in immuno-blot analysis.These findings provide very useful information for efficient mammalian expression and immuno-blotting of mcmbrane receptors.

OBJECTIVETo analyze the experiences on surgical treatment of severe aortic valve stenosis.

METHODSFrom December 1990 to December 2006, 171 patients with severe aortic valve stenosis underwent aortic valve replacement (AVR). There were 135 males and 36 females aged from 10 to 75 years old, with a mean of (45.8 +/- 15.6) years old. The intervals between the first episode of exertion dyspnea and administration to operation were 2 months to 52 years. The pathological lesions of the group were rheumatic aortic valve stenosis in 75 cases, calcified aortic stenosis in 66 cases, bicuspid aortic valve in 26 cases and other congenital aortic valve stenosis in 4 cases. One hundred and twenty-four patients underwent AVR, 7 AVR combined with replacement of the ascending aorta, 5 AVR with coronary artery bypass grafting, 19 AVR with mitral valve plasty (MVP), 8 AVR with plasty of the ascending aorta and 8 AVR with enlargement of the aortic root.

RESULTSThe averaged operation time was (4.4 +/- 0.6) h. Cardiopulmonary bypass (CPB) time was (124.7 +/- 38.5) min and the aorta clamp time was (78.3 +/- 21.7) min. The averaged blood loss during operation was (754.5 +/- 518.4) ml. All the procedures were successfully performed and all patients were weaned off CPB uneventfully. The indication of early complications was 12.3% (21/171), including low cardiac output syndrome in 7 cases, multi-organ failure in 3 cases, endocarditis in 1 case, renal dysfunction in 4 cases, ventricular fibrillation in 1 case, excessive bleeding in 2 cases, III atrial-ventricular block in 2 cases, and mediastinal infection in 1 case. The total mortality was 5.8% (10/171) with the main causes as cardiac failure for 4 cases, arrhythmia for 1 case, multi-organ failure for 4 cases, and infectious endocarditis for 1 case.

CONCLUSIONSSuccessful management of severe aortic valve stenosis requires sophisticated surgical techniques and experienced peri-operative care. Satisfactory results can be achieved if valve replace surgery is performed adequately.

Adolescent ; Adult ; Aged ; Aortic Valve ; surgery ; Aortic Valve Stenosis ; surgery ; Child ; Female ; Heart Valve Prosthesis Implantation ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the etiological role of hMLH1 gene A655 polymorphism in colorectal cancer.

METHODSA case-control study was carried out, including 115 colorectal cancer patients and 135 healthy people as control. Genomic DNA was extracted from peripheral white blood cell from all the subjects. Polymorphism was detected by PCR-based DHPLC analysis and verified by DNA sequencing.

RESULTSThe hMLH1 gene A655G polymorphism was detected in 3.0% of healthy people and 11.3% of colorectal cancer patients (P<0.01), and the difference was significant (P<0.01). The hMLH1 gene A655G polymorphism was detected in 8.2% of tubular adenocarcinoma or tubular-papillary adenocarcinoma and 27.8% of mucinous adenocarcinoma, which was also significant (P<0.05).Meanwhile, hMLH1 gene A655G polymorphism was not associated with age, gender and lymphatic metastasis (all P>0.05).

CONCLUSIONSThe hMLH1 gene A655G polymorphism may play a role in the pathogenesis of colorectal cancer. Determination of the polymorphism may be a potential marker to predict the prognosis of colorectal cancer patients.

Adaptor Proteins, Signal Transducing ; genetics ; Case-Control Studies ; Colorectal Neoplasms ; etiology ; genetics ; DNA Mismatch Repair ; Female ; Humans ; Male ; Middle Aged ; MutL Protein Homolog 1 ; Mutation ; Nuclear Proteins ; genetics ; Polymorphism, Single Nucleotide ; Prognosis ; Sequence Analysis, DNA