Objective To construct a cDNA substractive library of dermal papilla cell (DPC) in patchy and normal area of alopecia areata with technique called suppression substractive hybridization. Methods Total mRNA was isolated form DPC of patchy and normal area. Moreover, double strand cDNAs were synthesized in turn. cDNA from patch and normal area then were restricted and divided into two groups and ligated to the specific adaptor 1 and adaptor 2 respectively. After cDNAs from patchy and normal area hybridized with each other twice and underwent two times of nested PCR, then PCR products were ligated with arms of T/A plasmid vectors to set up the substractive library. Results cDNA substractive library of DPC in patchy and normal area of alopecia areata was set up successfully with highly subtractive efficiency. The amplified library contained 120 positive clones, in which 90 positive subclones contained 100 500bp inserts. Conclusion The cDNA substractive library of DPC in patchy and normal area of alopecia areata is a highly efficient one and lays a solid foundation for screening and cloning new and specific genes from patchy and normal DPC of anagen hair follicle in DPC of alopecia areata.

We applied bilingual teaching in internal medicine for students in our school from 2003 in order to cultivate medical students of high quality and to improve their ability in using professional foreign language.We analyzed our experiences from the aspects of teaching object,selection of teachers and teaching methods and summarized some problems during the process.We got the conclusions that only through strengthening bilingual teaching materials compiling,training young teachers and adopting diversified bilingual teaching means,can we achieve better bilingual teaching effect.

Objective To investigate the clinical application of statins for secondary prevention and its efficacy in patients with ischemic stroke.Methods A total of 300 patients with ischemic stroke were divided into statins group (n=91) and control group (n=209) according to whether statins was received or not.Serum lipid parameters,liver and kidney function,carotid artery intimamedia thickness (IMT) and plaque area were compared between the two groups.Results In 209 patients not taking statins,68 cases (32.5％) were not advised by clinic doctor; 55 cases (27.8％)were reluctant to take statins in fear of adverse reactions; 77 cases (36.8％) discontinued stains when serum lipids were normal; 3 cases (1.4％) developed skin allergic reaction and the skin rash that disappeared after stains discontinuance; 3 cases (1.4％) had elevated liver enzyme levels and the enzyme levels were declined to normal after stains discontinuance.The levels of total cholesterol (TC),triglyceride (TG) and low density lipoprotein cholesterol (LDL-C) were significantly lower in statin group than in control group [(4.93±0.45) mmol/L vs.(5.87±0.38) mmol/L,(1.68±0.61)mmol/L vs.(2.01±0.58) mmol/L,(2.49±0.57) mmol/L vs.(3.76±0.44) mmol/L,t=18.60,4.46,20.94,P=0.000,0.007,0.000,respectively].High density lipoprotein (HDL-C) level was significantly higher in stain group than in control group [(1.36±0.31) mmol/L vs.(1.03±0.25)mmol/L,t=9.75,P=0.001].There were no significant differences in liver and kidney function parameters such as alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase,serum total bilirubin,direct bilirubin,indirect bilirubin,glutamyl transferase,blood urea nitrogen and serum creatinine between the two groups (all P＞0.05).The carotid IMT and carotid atherosclerotic plaque area were significantly lower in stain group than in control group [(1.62 ± 0.23) mm vs.(1.74±0.31) mm,(0.57±0.12) cm2 vs.(0.65±0.18) cm2,t=3.32,3.88,P=0.008,0.002,respectively].Conclusions Most patients with ischemic stroke are still not taking statin for secondary prevention.Statins can improve the blood lipid levels and carotid arteriosclerosis status,and no obvious adverse reactions are observed.

OBJECTIVETo investigate the clinical efficacy of Panlongqi tablet (Chinese characters) combined with lumbar facet joint release for lumbar spinal stenosis of type Fengshi Bizu (Chinese characters).

METHODSSince February 2012 to February 2013, 120 patients with lumbar spinal stenosis of Fengshi Bizu (Chinese characters) syndrome were retrospectively studied. According to different treatment methods, 120 patients with lumbar spinal stenosis were divided into Panlongqi tablet (Chinese characters)group and control groups, respectively. In Panlongqi tablet (Chinese characters)group, 60 patients were treated by Panlongqi tablet (Chinese characters) combined with lumbar facet joints release solution including 26 males and 34 females with an average age of (60.40±3.36) years old ranging from 46 to 65 ; the course of the disease was 2 to 15 years (averaged 7.6 years). In control group the other 60 patients were treated with lumbar facet joint release including 24 males and 36 females with an average age of (61.20±2.47) years old ranging from 48 to 63; the course was 3 to 14 years (averaged 6.9 years). The clinical effect of patients were evaluated by JOA and ODI score before treatment, at 4 weeks and 3 months after treatment.

RESULTSAll patients were followed up for 4 to 7 months (means 5.6 months). After 3 months,7 cases in control group recurrenced symptoms,only 1 case in Panlongqi tablet (Chinese characters) group recurrenced. At 4 weeks and 3 months of follow-up, ODI score and JOA score of Panlongqi tablet group were much better than those of the control group.

CONCLUSIONFor lumbar spinal stenosis of type Fengshi Bizu (Chinese characters),which were treated with lumbar facet joint release with Panlongqi tablet(Chinese characters), supplemented by back muscle exercise, in relieving waist and low back pain symptoms and improving functional status of lower lumbar spine, can obtain satisfactory clinical outcome, is a good method of conservative treatment for such diseases.

Aged ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Exercise Therapy ; Female ; Humans ; Lumbosacral Region ; physiopathology ; Male ; Middle Aged ; Punctures ; Retrospective Studies ; Spinal Stenosis ; drug therapy ; physiopathology ; therapy

Objective The purpose of this study was to edit and psychometrically equate a set of Mandarin bisyllabic word lists. Methods 964 bisyllabjc words were recorded by male talker of standard Mandarin,352 words were selected to compose 10 lists. Percentage of correct word recognition was measured for each word at four intensity levels using 20 normal hearing subjects. The order of the presentation of the lists was randomized for each subject. U-sing Statistica7.0 Performance-Intensity function for each word was plotted,slopes and thresholds of them were calculated. 242 words were chosen after that, The thresholds and slopes of these words were in Guassian distribution. These words were ligitally adjusted and included in six Mandarin bisyllabic word lists of 40 words each. Two of them were for practice, four of them for test. 36 subjects with normal hearing served in our equivalent test. The orders of the presentation of the lists were randomized far each subject and results were expressed as thresholds. Results Two-factor ANO-VA was used to compare the thresholds of the 4 lists, F=1.978,P = 0.209. Conclusion These lists were considered to be primarily equivalent with each other.

Objective:To construct the yeast expressive vector of rmFⅦ, in which mFⅦ was mutated to inhibit coagulation without affecting the affinity for TF, and express it in Pichia pastoris. Methods:The full length cDNA encoding mFⅦ was amplified from a mouse liver by RT-PCR method, site-direct mutated and restriction enzyme digested as design. Cloning into pPIC9K, electroporation of Gs115, in vivo screen of multiple inserts by G418 resistance, BMGY/BMMY are used for induction and expression of rmFⅦ in pichia pastoris. These proteins were also screened for functional activity. Results: Three different rmFⅦ-pPIC9K yeast expression vectors and it's aim protein were obtained,two kinds of proteins were found to be functional active as design. Conclusion:rmFⅦ protein can be expressed in pichia pastoris and it might facilitate the development of tumor-target molecule, and novel anti-agiogenesis drug study.

The feasibility of bone marrow stromal cells autologous transplantation for rabbit model of dilated cardiomyopathy induced by adriamycin was studied. Twenty rabbits received 2 mg/kg of adriamycin intravenously once a week for 8 weeks (total dose, 16 mg/kg) to induce the cardiomyopathy model with the monitoring of cardiac function by transthoracic echocardiography. Marrow stromal cells were isolated from cell-transplanted group rabbits and were culture-expanded on the 8th week. On the 10th week, cells were labeled with 4,6-diamidino-2-phenylindole (DAPI), and then injected into the myocardium of the same rabbits. The results showed that viable cells labeled with DAPI could be identified in myocardium at 2nd week after transplantation. Histological findings showed the injury of the myocardium around the injection site was relieved with less apoptosis and more expression of bcl-2. The echocardiography found the improvement of local tissue movement from (2.12+/-0.51) cm/s to (3.81+/-0.47) cm/s (P<0.05) around the inject site, but no improvement of heart function as whole. It was concluded bone marrow stromal cells transplantation for dilated cardiomyopathy was feasibe. The management of cells in vitro, the quantity and the pattern of the cells transplantation and the action mechanism still need further research.

During growth and progression, the microenvironment of tumors suffers a series of abnormal characteristics, which include hypoxia, acid pH, increased oxidative stress, excess glutathione (GSH), as well as certain overexpressed enzymes. Although affect or limit the cancer therapeutic outcomes, these factors provide possible approaches to strategies for cancer detection and novel therapy at the same time. Recently, based on these properties of the tumor microenvironment (TME), various kinds of responsive nano-platforms have been continuously developed and applied in cancer theranostics preliminarily. Thus, this review would introduce the typical features of TME firstly, then detailly summarize the design principles and research progress of corresponding hypoxia-responsive, pH-responsive, redox-responsive, enzyme-responsive, dual-responsive and multi-responsive nano-platforms. Finally, the challenges and the perspectives of the TME-responsive nano-platforms are briefly discussed.

Objectives To construct a cDNA subtractive library of dermal papilla cells (DPCs) in anagen with suppression subtractive hybridization (SSH) and clone differentially expressed genes related to DPCs in anagen. Methods Total RNA was isolated from DPC of anagen and telogen follicles. Then ds cDNAs were synthesized in turn using SMART cDNA synthesis technique. After cDNAs from anagen and telogen follicle DPCs were hybridized with each other twice and underwent two rounds of nested PCR, PCR products were ligated with arms of T/A plasmid vectors to set up the subtractive library. Selected clones were verified by reverse Nothern blot and DNA sequencing, and the acquired sequences were analyzed for homology based on Genbank nucleotide database. Results cDNA subtractive library of DPCs in anagen follicle was set up successfully with high subtractive efficiency. Thirty-five genes were identified with 22 known functional genes and 13 unknown functional genes. Conclusions These results demonstrate the effectiveness and sensitivity of SSH in detecting differentially expressed genes from a small amount of clinical samples. Information about such alterations in gene expression might be useful for elucidating the genetic events in hair follicle growth regulation.

Objective To explore the expression of PIWIL1 protein and DICER enzyme in hepa tocellular carcinoma (HCC) and their significance.Methods Immunohistochemical method was used to detect the expression of PIWIL1 and DICER in 47 cases of HCC and the adjacent HCC tissues.Western blot method was used to detect the expression of PIWIL1 and DICER in 31 cases of fresh HCC tissues and their adjacent HCC tissues.The relationship between PIWIL1 and DICER and their relationships were analysed with clinical features.12 cases of normal liver tissues were used as control group.Results The expression of PIWIL1 was high in HCC but low in normal liver tissues (P＜ 0.05).The expression of DICER was high in normal liver tissues but low in HCC (P＜0.05).The expression of PIWIL1 was positively correlated with invasion to adjacent tissues and histological differentiation (P＜0.05).The expression of DICER was negatively correlated with invasion to the adjacent tissues and histological differentiation (P＜0.05).There was a negative correlation between PIWIL1 and DICER (P＜ 0.05).Conclusions High expression of PIWIL1 and low/missing expression of DICER was related to pathological differentiation and invasion of adjacent tissues.