Objective] To sum up Pro. Cheng Zhiqing’s clinical experience in treating hypertension with insomnia in menopause. [Method] Select 3 classical cases of the disease above treated by Pro. Cheng Zhiqing for analysis and discussion, and summarize her differentiation thought and treatment features. [Result] In her view, such disease is mostly concerned with liver, as well as related to heart, kidney, etc., the pathological course concerns mother-organ disorder involving its child-organ, so the therapy shall center on stretching liver, clearing liver, smoothing liver and tonifying liver, also purging child-organ in excess, nourishing mother-organ in deficiency, treating multi-organ in the same time, therefore to get better clinical effect. [Conclusion] For the hypertension with insomnia in menopause, Pro. Cheng Zhiqing is good at grasping disease mechanism, revising famous and recipe decoctions in generations, with close compatibility of medicines, definite cure effect, worth reference.

[Objective] Using gene chip to detect the influence on gene expression due to replicate Heart-Qi Deficiency Syndrome on Wistar Rat caused by complex factor including slimming, forced swimming and large dosage of propanolol. [Methods] Every animal in experiment group took food according to 5g/100g body weight, and was forced to swim by taking weight according to its 5% body weight until sank beyond 10 seconds. From 17th day ig propanolol 24mg/Kg and lasted for 4 days. At 21st day we took out hearts and picked up general DNA to prepare probe, then cross with gene chip. [Results] Compared with control group, expression of gene NM-017329,NM-016997 and NM-030850 showed down regulation obviously, expression of gene NM-031345 and NM-024349 showed up-regulation significantly. [Conclusion] Repression of immunological function and damage of vascular endothelium may be the main mechanism resulting in Heart-Qi Deficiency Syndrome.

OBJECTIVE To ensure clinical application of standard procedure in the management of dental handpieces collection and sterilization.METHODS To establish the center of sterilization and supply for dental handpieces;to equip with the device for depuration and sterilization;to standardize the procedure for depuration,retrieval,pre-disposal treatment,cleaning,drying,lubrication,envelopment and sterilization.RESULTS The management of collection and sterilization for dental handpieces ensured the clinical application of standard procedure,avoided the exogenous infection and guaranteed the patients safety.CONCLUSIONS The clinical management of collection and sterilization for dental handpieces is an effective method.

OBJECTIVE: To investigate part mechanisms of Th cell differentiation, and to observe the interference effect of Qingxin-II Recipe in the chronic stage of viral myocarditis (VMC), so as to provide some experimental evidences for illuminating the pathogenesis of VMC and treatment mechanisms of Qingxin-II Recipe. METHODS: According to 20%-40% death rate of experiment in advance, 100 BALB/c male mice (4 weeks old and weighing 12-15 g) were used. Twenty mice were randomly assigned to normal control group, and the other 80 mice were intraperitoneally injected with 0.1 ml normal saline containing coxsackie virus B3 at the 1st, 4th and 28th day (the virus densities were 1:2000, 1:1000 and 1:500 respectively) to induce chronic VMC. At the 42nd day, the surviving mice were randomly divided into untreated group and treatment group, with 20 mice in each group. Mice in the treatment group were orally administered with 0.2 ml Qingxin-II Recipe every day, while mice in the normal control group and the untreated group were administered with 0.2 ml normal saline. All the mice were sacrificed after 45 days, and the sera, heart and spleen cells were collected. Then the myocardial pathological changes were observed by using a light microscope, and the levels of IL-4, IL-10 and IFN-gamma in serum were detected by enzyme linked immunosorbent assay (ELISA). And the Th cell differentiation was observed by flow cytometry. RESULTS: No obvious myocardial pathological changes were observed in mice of the normal control group. Myocardial pathological changes in the treatment group were slighter than those in the untreated group. The difference of serum IL-10 level between the normal control group and the untreated group showed no significance (P>0.05), and the levels of IFN-gamma and IL-4 of the untreated group were higher than those of the normal control group (P<0.05 or P<0.01). There was no statistical difference in IL-10 level between the treatment group and the untreated group (P>0.05), while the serum levels of IL-4 and IFN-gamma of the treatment group were lower than those of the untreated group (P<0.05 or P<0.01). There was no significant difference of the Th1 cell responder between the treatment group and the untreated group (P>0.05), while the Th2 cell responder was inhibited significantly in the treatment group (P<0.05). CONCLUSION: Qingxin-II Recipe can restore the balance of Thl and Th2 cells through inhibiting the reaction of Th2.

Objective To establish the magnetic activated cell sorting system(MACS)for isolation and incubation of human ovarian carcinoma-derived microvascular endothelial cells(ODMECs) and to observe their ability to form tubule-like structure(TLS) in vitro.Methods MACS was used to purify ODMECs with antiCD31 antibody and ODMECs were identified according to their morphological,biochemical and phenotypic characteristics.The ability of ODMECs to intake acetylated-low-density lipoprotein(DIL-acLDL) and Ulex europeaus agglutinin-1(UEA-Ⅰ) was assayed to observe the formation of TLS in vitro.Results Flow cytometry showed that the purity of ODMECs was up to 99.6%.The ODMECs were characterized by cobble stones,contact inhibition,expressed CD31,CD105 and FⅧ-Rag,and exhibited the ability to bind to UEA-1,intake DIL-acLDL and form TLS during incubation.Conclusion The MACS we established can be used to isolate and incubate human ODMECs and study the formation characteristics of TLS.

Objective:To investigate the changes of Th cell differentiation in the VMC and interference effect of Qingxin capsule on them.Methods:BALB/c mouses with different courses of VMC were established, after treated with Qingxin capsule, the cardiac pathological changes were observed by light microscope and transmisson electron microscope,levels of IL-2,IL-4,IL-10 and IFN-? in blood serum were detected by ELISA.Results: Whatever in acute stage or recovery stage of VMC, the myocard pathological changes of mouses were lighter after treated with Qingxin capsule;moreover, levels of IFN-? and IL-2 in blood serum of mouses with VMC decreased significantly(P0.05).Conclusion:Qingxin capsule can restore the balance of Th1 and Th2 cells through inhibiting reaction of Th1 and enhancing reaction of Th2.

Objective:To conjugate IVIG and MTX to produce a specific cytotoxicity upon phagocytes.Methods :MTX was conjugated with IVIG by indirect conjugating methods. HSA was used as an intermedi-ary to conjugate MTX with IVIG. The indirect immunofluorescence was adopted to test the binding abilityof Fc fragment. MTT assay was used to measure the cytotoxicity of conjugation on phagocytes. Results:Conjugation showed stronger cytotoxicity upon target cells than free MTX,and it showed only less cyto-toxic effect on Fc receptor negative cells compared with the positive ones. The specific cytotoxicity of IVIG-HSA-MTX was significantly stronger than that of MTX. Conclusion: In vitro the conjugation showed ahighly specific cytotoxicity upon phagocytes.

Aim To study the expression of human VEGF165 cDNA in Pichia pastoris and to obtain high-level expressed recombinant human VEGF165 with good biological activity. Methods Amplifying human VEGF165 cDNA by PCR, after confirmed by DNA sequence analysis, the gene was inserted into the Pichia pastoris expression vector pPIC9K containing AOX1 promoter and α secreting signal peptides, the recombinant expression plasmids pPIC9K/hVEGF165 was constructed and transformed into KM71. The multiple insert transformants were screened， fermented in flasks and induced by 10 mL/L methanol. Results After 4 days of methanol induction, the expressed hVEGF165 reached up to 60％ of total proteins in supernatant by SDS-PAGE. Western blot assay proved the expressed hVEGF165 having good antigenicity and high specificity. The recombinant protein was further purified with Heparin-Sepharose CL6B affinity chromatography, and was proved to have good biological activity in stimulating HUVEC proliferation. Conclusion High-level expression of secreted hVEGF165 has been successfully achieved in Pichia pastoris expression system.

Objective To observe the curative effect of electro-acupuncture plus Tuina on prostatic hypertrophy.Method Fifty-six cases of prostatic hypertrophy were treated according to the principle of taking the kidney as the main aspect. Points Guanyuan (CV 4), Qugu ( CV2), Shenshu(BL 23), Ciliao(BL32) and Sanyinjiao( SP6) were selected and electro-acupuncture were used together with Tuina. Those who took the tablet Finasteride orally were treated as the control group. Result The total effective rate was 94.6% in treating group, while that was 86.7% in the control group, P＞0.05. Conclusion It is suggested that electro-acupuncture plus Tuina has a certain effect on prostatic hypertrophy. Compared with western medicine, it is lower in price and has no side effect.

Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.