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.

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.

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 .

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.

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.

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.

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

OBJECTIVETo investigate the association of the single-nucleotide polymorphism (SNP) IVS10+12 G>A in hMSH2 gene with colorectal cancer in a Chinese population of Jiangsu province.

METHODSA case-control study to investigate whether this SNP affects the risk of developing colorectal cancer was conducted. Subjects included 108 colorectal cancer patients and 180 healthy individuals. Peripheral white blood cell DNA was obtained from all subjects. The hMSH2 gene IVS10+12 G>A was genotyped using a PCR-based DHPLC, the existence of IVS10+12 G>A was verified by DNA sequencing.

RESULTSThe allele frequency of the IVS10+12 G>A in the hMSH2 gene in the healthy individuals was 51.7%. There was significant difference in the frequency of the IVS10+12 G>A between patients and healthy controls (P<0.05), and between familial patients and healthy controls (P<0.05). There was also significant difference of the frequency of the IVS10+12 G>A between patients younger than 50 years, and patients with high consumption of fried food and pickled vegetable and healthy controls respectively (P<0.05).

CONCLUSIONThis SNP may be associated with colorectal cancers in Chinese. Further investigation with larger sample size is needed.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; China ; Colorectal Neoplasms ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; MutS Homolog 2 Protein ; genetics ; Pedigree ; Point Mutation ; Polymorphism, Single Nucleotide ; Young Adult

OBJECTIVETo study the changes in pathogenic causes and the prognosis of aortic valve replacement (AVR).

METHODSThe clinical data of 1026 patients undergoing AVR from December 1980 to December 2006 were analyzed retrospectively. The mortality, morbidity, changes in pathogenic causes and risk factors were analyzed.

RESULTSThe postoperative mortality and complication morbidity were 4.3% and 10.6% respectively within 30 days followed operation. Main causes of operative death were heart failure, multi organ failure and endocarditis. The major risk factors for operative death were left ventricle ejection fraction less than 0.4, endocarditis, valve regurgitation and emergency operation before AVR. Late mortality was 0.54% patient-year (3.4%), most of whom died of heart failure, endocarditis and arrhythmias. Patients underwent reoperation 0.22% patient-year (1.4%), with the causes of endocarditis and perivalvular fistula.

CONCLUSIONSMorbidity of rheumatic damage in aortic valve has decreased, while valve degeneration has increased gradually in the recent years. Avoiding prosthesis-patient mismatch, good postoperatively guide and prevention of endocarditis can improve the prognosis of AVR.

Adolescent ; Adult ; Aged ; Aortic Valve ; surgery ; Female ; Follow-Up Studies ; Heart Valve Diseases ; surgery ; Heart Valve Prosthesis Implantation ; methods ; mortality ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; mortality ; Prognosis ; Retrospective Studies ; Risk Factors ; Treatment Outcome