In clinical practice,we have adopted the methods such as intensifying the knowledge of preclinical medicine,learning to collect case history,training medical check-up,understanding the significance of auxiliary examination and encouraging the students to form the clinical thinking ability,and thus achieved satisfactory effect and obtained the positive opinion of students.

Objective To screen single nucleotide polymorphism(SNP)of DNA methyltransferase 1 gene in patients with systemic lupus erythematosus(SLE).and to investigate the significance of abnormal expression of DNA methyltransferase in the pathogenesis of SLE.Methods Peripheral blood mononuclear cells(PBMCs)were obtained from 11 patients with SLE and 12 normal human controls.DNA was extracted from the PBMCs.and PCR was performed to amplify some exons(1 to 7)and corresponding introns followed by direct sequencing.Results There was a 21397T/C SNP in the intron 7 between exon 7 and exon 8 of DNA methyltransferase 1 gene in patients with SLE.The frequency of CC homozygote increased in patients with SLE compared with that in normal controls(5/11 vs 1/12,X~2=4.10,P＜0.05).Conclusions There is a 21397T/C SNP in DNA methyltransferase 1 gene in patients with SLE.which may be a component of genetic background underlying the abnormal expression Of DNA methyltransferase 1.

Purpose: To investigate the efficacy ofmassotherapy plus warming acupuncture for treatment of patellar malacia. Methods: Seventy-two patients with patellar malacia were randomly divided into observation and control groups, treated by massotherapy plus warming acupuncture and simple medication, respectively. Results: The effective rate was 83.3% in the observation group and 56.7% in the control group. There was a significant difference in curative effect between the two groups (P＜0.01). Conclusion: The curative effect of massotherapy plus warming acupuncture on patellar malacia is superior to that of simple medication.

Objective To investigate the effect of adenosine on pulmonary injury after extracorporeal circulation. Methods Twelve mongrel dogs were randomly divided into two groups, the test group and control group. The model of extracorporeal circulation was established. In the test group and control group, adenosine(50μg/kg*min) and normal saline were infused before extracorporeal circulation respectively. Hemodynamics, right cardiac function and arterial blood gas were assessed. Lung water and malondialdehyde content were assessed. Histological evaluation was also made. Results　There were no differences between the two groups with respect to heart rate, mean systemic artery pressure, left atrial pressure, and central venous pressure. In the test group, pulmonary vascular resistance was lower and right cardiac function improved remarkably; arterial oxygen pressure (PaO2) was also higher, less lung water was observed and the malondialdehyde content in the pulmonary tissue was lower (P＜0.05,P＜0.01). Histological evaluation of the test group revealed normal pulmonary architecture without abundant intra-alveolar neutrophils. Conclusion Adenosine can attenuate ischemia-reperfusion lung injury and right cardiac insufficiency caused by extracorporeal circulation. At a proper dose, there was no significant effect on systemic hemodynamics.

Post-stroke infection mainly includes stroke-associated pneumonia and urinary tract infection.It is one of the major causes of increasing disability and mortality in patients with stroke.Its occurrence and development is associated with the basal state,stroke location,infarct volume,severity,dysphasia,aphasia and other factors of the whole body system in patients with stroke.In recent years,stroke-induced immunosuppression is receiving increasing attention.

Objective To explore the possibility of bone marrow mesenchymal stem cells (MSCs) differentiating into active melanocytes in vitro.Methods Bone marrow stromal cells were harvested from femoral marrow of 6-week-old black male C57BL/6 mice,and subjected to a primary culture.After 6-passage subculture,an induction medium containing hydrocortisone,recombinant human insulin,transferrin and fibroblast growth factor was used to induce the differentiation of MSCs into melanocytes.Inverted light microscopy was applied to observe the process of cell differentiation,transmission electron microscopy to observe melanosome formation and maturation,and immunocytochemistry to determine the expression of melanocyte-associated epitopes,and flow cytometry to analyze cell cycles and yield of differentiated melanocytes.Results The total number of MSCs was close to 109 after 6 passages of subculture,and immunofluorescent studies showed an expression rate of 94.3％ for CD44 and 82.3％ for CD105 in these MSCs.After 180-day cultivation in the induction medium,the MSCs showed a morphological similarity to melanocytes with an increase in dendrites,formation of melanosome-like structures,and cell growth cycle was shortened to 3-4 days.Brown/black cell sediments were visualized by naked eyes.Electron microscopy revealed that intracellular melanosomes were mainly in Ⅳ phase.Immunofluorescence studies of the differentiated melanocytes showed a positive staining for tyrosinase related protein-1 (TRP-1),TRP-2,and microphthalmia-associated transcription factor (MITF).Flow cytometric analysis showed that most of the melanocytes differentiated from the MSCs were in G1 and S phases,and TRP-1-positive melanocytes amounted to 80％ of gate cells.Conclusions Bone marrow MSCs can be largely differentiated into melanocytes with a close similarity to normal melanocytes in morphology,ultrastructure and specific epitopes and a certain degree of proliferative activity.

Objective: To observe the clinical efficacy of combined tuina manipulations with Qi Zheng Qing Peng paste for herniation of the lumbar intervertebral disc. Method: All 568 cases were randomly allocated into an observation group (189 cases), control Ⅰ group (187 cases) and control Ⅱ group (192 cases). Combined tuina with Qi Zheng Qing Peng paste, combined tuina with diclofenac diethylamine emulsions and tuina alone were adopted in the above three groups respectively. The efficacies in the three groups were compared after 4 weeks of treatment. Results: The total effective rates in the observation group, control Ⅰ group and control Ⅱ group were 89.9%, 86.6% and 84.9% respectively, showing a significant difference (P ＜0.05) . Conclusion: Combined tuina manipulations with Qi Zheng Qing Peng paste works well for herniation of the lumbar intervertebral disc.

Objective: To observe the clinical effects of frozen shoulder treated by penetrating needling and manipulation Methods: 336 cases of frozen shoulder were randomly divided into treatment group, treated with needling Jianyu (LI 15) toward Jianqian (Ex) and Manipulation, and control group, treated with needling Jianyu (LI 15) toward Jianqian (Ex). The scores of shoulder movement and the changes of infrared image were observed before and after treatment. Results: The therapeutic effects were better in the treatment group than that in the control group (P<0.05). Conclusion: Penetrating needling combining with manipulation is effective in treating frozen shoulder.

Objective:To investigate the effects of epidermal growth factor (EGF)on cell cycle and cell cycle-related regulatory factors of human esophageal squamous cell carcinoma (ESCC) cell line Eca109.Methods: Serum starved Eca109 cells were treated with 20 ng/ml recombinant human EGF(rhEGF)for 24 h.The cell cycle phase distribution was detected by flow cytometry.The mRNA and protein expression levels of p21CIP1/WAF1(p21) and p27KIP1(p27) were detected by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot,respectively.Results: The proportions of G1 phase cells in EGF group and control group were ( 54.90 ±0.82 )% and ( 65.94 ±0.74 )%.The mRNA and protein expression levels of p 21 in EGF group was significantly higher ,and p27 was significantly lower than that in control group ( P<0.01 ) .Conclusion: EGF facilitates G1-S phase transition,and promotes the proliferation of Eca 109 cells,which may be associated with the up-regulation of p21 and down-regulation of p27.