Teaching in out-patient clinic is a new and unique teaching mode,which is characteristic of one-to-one tutoring experiences.Students are put in the key positions while mentors serve as assistants in the whole process.By means of repeated practicing,summing-ups and perceiving,students improve their ability to combine theory with practice,enhance capabilities of communication,enrich their social experience and increase their talent of independent working.Meanwhile,further trainings may endow our medical students with better abilities of summarization.Through practice,teaching clinic has been proved to be a reasonable teaching mode which deserves promotion.

Objective To establish the method to detect the expression of apolipoprotein E gene in human adipose cells. Methods Apolipoprotein E gene expression was determined by competitive reverse transcription-PCR in adipose cells of healthy children. Results Apolipoprotein E gene expressed in the adipose cells of healthy children. The mean expression quantity of apolipoprotein E mRNA in adipose cells was (0.41?0.12 ) in healthy children. Conclusions Competitive reverse transcription-PCR is a rapid, simple, sensitive and accurate method to analyze expression of apolipoprotein E gene.

BACKGROUND:At present, the development of reprogramming technology provides a wide prospect for stem cellresearch. Through the ectopic co-expression of reprogramming factors, the somatic cells can be reprogrammed to a pluripotent state, termed as induced pluripotent stem cells, which can avoid the ethical controversy faced in the research and application of embryonic stem cells. Also, we can generate patient-specific and disease-specific induced pluripotent stem cells, which significantly decrease immuno-rejection. However, reprogramming technology faces some chal enges, such as low efficiency and safety.
OBJECTIVE:Based on the characteristics of induced pluripotent stem cells and the principles of reprogramming, to detail the progress in reprogramming technology from five aspects, including cellresources, carriers, transcription factors, microRNA and signal transduction pathway. 
METHODS:A computer-based online retrieval was performed to search papers published form January 1990 to April 2013 in VIP periodical ful-text database, Wanfang periodical ful-text database, CNKI periodical ful-text database, PubMed database and Springer database with key words of“reprogramming, induced pluripotent stem cell, signal transduction pathway, epigenetics, microRNA, transcription factor, vector, somatic cell, smal molecule compound, safety”both in Chinese and English. After excluding objective-independent papers, 67 papers were included for further analysis.
RESULTS AND CONCLUSION:By exploring different cellresources, different carriers, various combination of transcription factors, microRNAs or inhibition of the signal transduction pathways, the reprogramming efficiency and safety have been improved greatly. However, currently, induced pluripotent stem cells stil could not meet the requirement of clinical application. To achieve the clinical application of induced pluripotent stem cells, it is urgent to explore the mechanism of reprogramming, and to optimize the programming strategy.

Objective To investigate effects of oxidative stress, expression change of transforming growth factor-β1 and Nestin in re-nal injury of hyperthyroidism rats. Methods The model of hyperthyroidism rat was made by giving them levothyroxine (L-T4). The activity of SOD, CAT, GSH-Px and the level of MDA on the 25th day, the 45th day, and the 60th day were determined by chromatometry in renal cor-tex.. The expressions of TGF-β1 and Nestin on the 25th day, the 45th day, and the 60th day were determined by immunohistochemistry. Results Along with the course of hyperthyroidism, it showed activity of SOD, CAT, GSH-Px and the expression of Nestin reduced, while the level of MDA and the expression of TGF-β1 increased. Conclusion Oxidative stress can increas the expression of TGF-β1 and podocyte in-jury may played an important role in hyperthyroidism renal injury.

OBJECTIVE: To study the role of P-JNK and P-c-Jun of JNK (c-Jun N-terminal kinase) on nasal mucosa remodeling in allergic rhinitis rats. METHOD: Sixty male Wistar rats (weighing about 200-250 g) were randomly divided into AR group (A group) and B group(control group). The rats in A group were sensitized for inducing AR by intraperitoneal injection of ovalbumin and Al(OH)₃. Rats in group A were randomized into A4, A8 and A12 group (each had 10 rats). Ovalbumin was dropped in each nasal cavity of every rat for 4,8,12 weeks, respectively. Rats in group B were sensitized by saline instead of OVA, and were also divided into B4, B8 and B12 group. Each group had 10 rats. Pathological changes of nasal mucosa in each period were observed by hematoxylin and eosin stain dyeing. The phosphorylation of JNK and c-Jun were tested by immunohistochemistry. RESULT: In A8 group, mucosal congestion and edema thickening with inflammatory cells infiltration of eosinophils were observed in the eighth week, and the inflammatory changes were significantly increased as time went on. The mean absorbance values of P-JNK and P-c-Jun in A group were significantly higher than those in the corresponding B group (all P < 0.01). Moreover, the mean absorbance values of A12 group were significantly higher than A4 group and A8 group (all P < 0.01 ). CONCLUSION The expression of P-JNK and P-c-Jun in the process of nasal mucosa remodeling in allergic rhinitis rats were increased, which suggested that P-JNK and P-c-Jun played important roles in nasal mucosa remodeling of the allergic rhinitis rats.
Airway Remodeling ; Animals ; Disease Models, Animal ; Eosinophils ; cytology ; Injections, Intraperitoneal ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Male ; Nasal Mucosa ; metabolism ; Ovalbumin ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Rhinitis, Allergic ; metabolism ; Signal Transduction

BACKGROUND:Induced pluripotent stem cel technology have solved the contradiction between the ethics and immune rejection, and this high-efficient and safe technique is becoming the mainstream of today’s research. OBJECTIVE:To comprehensively review the safety and application of induced pluripotent stem cel s.METHODS:A computer-based online retrieval of PubMed and CNKI was performed to search relevant papers published from January 2006 to April 2016, with the key words of“induced pluripotent stem cel , reprogramming, clinical application, safety, transcription factor, disease mode”in English and Chinese, respectively. RESULTS AND CONCLUSION:In recent years, research on induced pluripotent stem cel s has attracted much attention from the scientific community and the medical community, and this technique has successful y gained induced pluripotent stem cel s and overcome the problems of immunity and ethics. However, it is limited to the theoretical and laboratory research due to the inability to solve the safety, efficiency and re-differentiation mechanism of induced pluripotent stem cel s. Therefore, we are faced with enormous difficulties and chal enges, which involve al aspects of basic research, including how to safely and effectively induce the differentiation of induced pluripotent stem cel s into the desired cel type and how to establish a suitable disease model as wel as a high-throughput drug screening platform.

BACKGROUND: Beta thalassaemia is a monogenic disease, which lacks effective clinical treatments. Hematopoieticstem cell transplantation currently is the only radical treatment for beta thalassaemia, but the limits of suitable donor and costs minimize its clinical application. Given the technology of reprogramming using somatic cells is well established,gene therapy using induced pluripotent stem cells has become the new direction of beta thalassaemia treatment.OBJECTIVE: To put forward the advantages of CRISPR/Cas9 technology in gene therapy of beta thalassaemia in thefuture by summarizing the mechanisms of three kinds of gene editing technologies and the preliminary experimentalresults in animal models.METHODS: In order to search relevant articles about beta thalassaemia, the first author retrieved PubMed database andCNKI (from 1989 to 2015) using the key words of beta thalassemia, genetic therapy, genome editing, homologousrecombination, iPSCs in English and Chinese, respectively. After eliminating literatures which were irrelevant toresearch purpose or containing a similar content, 67 articles were chosen for further analysis.RESULTS AND CONCLUSION: Gene editing technology has made considerable progress and three kinds of directedgene editing technologies have been developed, including ZFNs, TALENs, CRISPR/Cas technology. By targeting inducedpluripotent stem cells from thalassemi patients, these three kinds of gene editing technologies have been expected tocorrect pathogenic genes of thalassemia. The CRISPR/Cas system is more simple, rapid, safe and efficient than the others.The CRISPR/Cas9 system is expected to repair β-globin genes in the induced pluripotent stem cells, germ cells, fertilizedeggs and embryos from beta thalassaemia patients, laying the foundation for future clinical application.

Objective To evaluate the safety and efficacy of radical antithrombotic therapy(aspirin,clopidogrel and tirofiban) in the treatment of senile (≥ 75 years) non-ST-elevation acute coronary syndrome (NSTEACS).Methods A total of 146 senile patients with NSTEACS were divided into observation group (70 cases) and control group(76 cases) by random digits table method.The control group was given aspirin,low molecular weight heparin and clopidogrel,and the observation group was added tirofiban (intravenous loading dosage:0.4 μ g/ (kg· min) for 30 min and then maintaining 0.1 μ g/ (kg· min) for 48-72 h).The occurrence of major adverse cardiovascular events (MACE) within 30 d after treatment,7 d angina pectoris control rate and adverse reactions were observed in two groups.Results The occurrence rate of MACE within 30 d after treatment was 5.7％ (4/70) in observation group,and 14.5％ (11/76) in control group,and there was significant difference (P ＜ 0.05).The 7 d angina pectoris control rate was 92.9％ (65/70) and 76.3 ％ (58/76) in observation group and control group,and there was significant difference (P ＜ 0.05).The incidence of bleeding was 10.0 ％ (7/70) and 7.9 ％ (6/76) in observation group and control group,and there was no significant difference (P ＞ 0.05).There were no major bleeding events in two groups.One case had thrombocytopenia in observation group,but there was no significant difference compared with that in control group(P ＞ 0.05).Conclusion Triofiban on the basis of aspirin and clopidogrel in the treatment of senile NSTEACS is effective and safe.

Objective To explore the impact of blockade of SDF-1/CXCR4 signaling pathway by T140 on degradation of typeⅡ collagen in human articular cartilage and to define the mechanism of action of T140. Methods 144 pieces of cartilage (Mankin score of 0 or 1) were obtained from osteoarthritis patients undergoing total knee replacement ( OA cartilage group) and 144 pieces of cartilage (Mankin score of 0 or 1) were obtained from patients receiving traumatic amputation (normal cartilage group). Each group was treated with SDF-1 of 100 ng/mL, then divided into three subgroups A, B, and C. The cartilage tissue in each group was cultured in the nutrient solution containing of T140, MAB310, or SDF-1 (1 000 nmol/L) for 48 or 96 hours. RT-PCR was used to detect expression of typeⅡcollagen mRNA in the cartilage tissue. Results Level of type Ⅱcollagen mRNA was markedly higher in subgroup A than in subgroup B and subgroup C at the same group and the same time (P <0.05). The expression level of type Ⅱcollagen mRNA at the same time and in the same subgroup of OA cartilage group were lower than that in normal cartilage group (P < 0.05). Conclusions SDF-1 induces degradation of typeⅡcollagen in human articular cartilage thruogh the SDF-1/CXCR4 signaling pathway. T140 can block the SDF-1/CXCR4 signaling pathway and reduce the degradation of type II collagen.

Objective To establish an assay for detecting α2-macroglobulin activity in Cohn fraction Ⅳ.Methodsα2-M reacted with trypsin to form α2-M-trypsin complex.After the chromogenic substrate Na-benzoyl-DL-arginine 4-nitroanilide hydrochloride ( BAPNA) was added, absorption at 410 nm was detected with the microplate reader .α2-M activity in Cohn fractionⅣwas quantitatively detected according to the established standard curve of plasma α2-M activity. Result Several critical parameters in this assay were optimized .A standard curve of plasma α2-M activity was established . According to this standard curve ,α2-M activity in Cohn fraction Ⅳsample was detected to be 1.578 PU/ml.Conclusion Using normal human plasma as the reference material , theα2-M activity in Cohn fractionⅣcan be detected through chro-mogenic substrate assay.This study provides a simple method to detect α2-M activity during the purification process of α2-M from Cohn fraction Ⅳ.