Multi-partners,unprotected sex,and alcohol and drug use are the main causes of HIV/STD transmission among MSM,and related interventions should be taken promptly.MSM are good candidates for studies and interventions as the retention rate is relatively high among this population group.

PUMA (p53 up-regulated modulator of apoptosis) is a recently discovered Bcl-2 family member which could be rapidly induced by p53 and has strong pro-apoptotic effects.PUMA has attracted much attention in the research of life science.PUMA expression results in potent growth suppression of some cancer cells through induction of apoptosis.PUMA can also significantly sensitize some cancer cells to chemotherapeutic agents and irradiation through induction of apoptosis.PUMA is potentially useful in gene therapy of tumor.But recently,researchers have also found that PUMA participates in the process of carcinogenesis and possessed important biological functions.

Objective To construct and identify the screening vector pUC18-EGFP,using EGFP as an indicator. Methods The EGFP gene was prepared by PCR and cloned into pUC18 resulting the vector pUC18-EGFP. Then DNA fragment was inserted into the MCS of pUC18-EGFP to test its practicability based on green fluorescence. Results The pUC18-EGFP was confirmed correctly by restriction enzyme analyses. The pUC18-EGFP was used to select recombinants. The green strains on the plate were confirmed by restriction enzyme and DNA analyses,which were E.coli harboring recombinants. Conclusion The screening vector PUCl8-EGFP was constructed successfully. Thus,we can select the positive clones on plates based on the green fluorescence of EGFP.

BACKGROUND: Critical closing pressure (CCP) is recently thought to play a key role in cerebral blood flow autoregulation as an effective downstream pressure of cerebral circulation and can objectively reflect the cerebrovascular tone, namely the vascular smooth muscle contraction and diastole, which is subjected to dynamic modulation.OBJECTIVE: To dynamically assess the hypertension-induced damage of the contraction function of cerebral microvascular smooth muscles and its correlation with morphological changes based on CCP evaluation.DESIGN: Randomized controlled experiment.SETTING: Institute of Neural Science of Second Hospital Affiliated to Guangzhou Medical College and Department of Neurology, First Hospital Affiliated to Sun Yet-san University.MATERIALS: The experiment was carried out at the Laboratory of Physiological Science of Sun Yet-san University between July 2002 and August 2003. Totally 160 health male SD rats were randomized into control group and hypertension group with 80 rats in each group. METHODS: Stroke-prone renovas cular hyp ortonsive rats were established in rats of the hypertension group by bilateral renal artery occlusion with two clips. The rats in the control group were not subjected to the occlusion with other treatments identical to those of the hypertension group. At the time points of 2, 4, 6, 8, 10, 12, 14 and 16 weeks after operation, respectively, 10 rats were randomly selected from each of the two groups for determination of arterial pressure and CCP. After the measurements the frontal-parietal lobe was obtained from the anaesthetized rats and cut into slices for quantitative analysis of the morphological changes in cerebral microvessels.different postoperative time points.mean arterial pressure in hypertension group obviously increased from the 6th postoperative week with significant difference from that of the control after operation to a level significantly higher than that of the control group at postoperative 14 and 16 weeks [(63.75±7.43) vs (37.28±3.68) mm Hg and (67.37±15.57) vs (38.39t7.41) mm Hg, respectively, P ＜ 0.05].significance from that of the control group at the 8th postoperative week (Paverage arterial pressure and cerebral arteriole tunica media (r=0.906 93,0.811 36, respectively, P ＜ 0.05). The changes in CCP was more obvious in the early and advanced stages of blood pressure elevation, but not so manifest during obvious blood pressure increment, displaying an inverted S-shaped curve of changes (R2=0.996 2, P ＜ 0.05).CONCLUSION: Contraction of the cerebrovascular smooth muscles is enhanced with the dynamic increment of arterial pressure after the development of hypertension. Vascular tone increase is more manifest during the early and advanced stages of hypertension.

Objective To observe the effects of PTD-hepatitis B core antigen (HBcAg) induced murine bone marrow-derived dendritic cells (DCs) maturation on T lymphocyte proliferation in vitro.Methods Bone marrow derived DCs isolated from BALB/c mice were cultured with recombinant granu|ocyte-macrophage colony-stimulating factor (rGM-CSF) and recombinant interleutin-4 (rIL-4)for 5 days.Tumor necrosis factor (TNF)-a,HBcAg and PTD-HBcAg were added to induce DCs maturation.The distribution and localization of intracellular immunofluorescence were observed by confocal microscopy,and DCs phenotypes were analyzed by flow cytometry.The level of IL-12 p70 in the supernatant was detected by enzyme linked immunosorhent assay (ELISA).The proliferation of T lymphocytes was performed by using cell counting kit-8 (CCK-8).All data were analyzed using t test.Results DCs were cultured and identified successfully.Recombinant PTD-HBcAg could penetrate into DCs cytoplasm while recombinant HBcAg was detected on the surface of cells.DCs surface molecules,such as CD80,CD86 and major histocompability complex (MHC) II were upregulated by PTDHBcAg;IL-12 p70 levels induced by 50 mg/L and 100 mg/L recombinant PTD-HBcAg were (142.50±18.31) ng/L and (124.30±15.12) ng/L,respectively,which were significantly higher than those induced by recombinant HBcAg [(42.31±4.21 ) ng/L,t = 9.234 and 9.045,respectively,P＜0.05].The proliferation of T lymphocytes induced by PTD-HBcAg was much higher than that in HBcAg group or positive control TNF-a group.Conclusions PTD-HBcAg could penetrate membrane of DCs and promote the differentiation and maturation of DCs.PTD-HBcAg could up-regulate the expressions of costimulatory molecules on cell surface of DCs,and enhance the ability of DCs on stimulating T lymphocytes proliferation and IL-12 p70 production.

Objective To investigate the effect and the mechanism of Sorcin (soluble resistancerelated calcium-binding protein )on development of drug-resistance in human gastric cancer cell line.Methods The full length Sorein cDNA was isolated by reverse transcription-polymerase chain reaction (RT-PCR).The FLAG-sorein-peDNA 3.1 plasmid was constructed by directed cloning of Sorein gene into the eukaryotie expression vector FLAG-peDNA 3.1,and was transfeeted into SGC7901 cells using liposome-mediated method.The expressions of Sorcin mRNA and protein in stable clone were detected by RT-PCR and Western blot. The intracellular concentration of Vineristine (VCR) in Sorein-transfected SGC7901 eells(SGC7901-F-Sor) and SGC7901 cells with or without Verapamil(VRP) was determined by high performance liquid chromatography(HPLC).Results The FLAG-Sorcin-peDNA3.1 plasmid vector was constructed successfully by DNA recombination and was transfected into SGC7901 cells.RT-PCR and Western blot analysis revealed that the expressions of Sorcin mRNA and the protein were up-regulated in SGC7901 F-Sor cells.Compared to the parent SGC7901 cells,the concentration of VCR in SGC7901-FSor cells was decreased by 76.89%,but it was increased by 2.41 times when treated with VRP.Conclusions Overexpression of Sorcin in SGC7901 cells results in decreasing concentration of VCR,which indicate that Sorcin may play a role in drug-resistance of SGC7901 cells by regulating the transference of chemotherapy drugs.The effect of Sorcin can be reversed by VRP.

Objective:To explore the wound-healing effect and the anti-inflammatory activity of the aqueous extracts from the fresh roots and rhizomes of A. calamus. Methods: The image analysis techniques and the histological analysis were used to determine the wound-healing effect in the excised wound test, and the real-time RT-PCR techniques was used to evaluate the anti-inflammatory activi-ty in the lipopolysaccharide-induced RAW 264. 7 cells test. Results:The aqueous extracts were given 3 times a day since the model was established. The skin wound area was reduced significantly in the aqueous extracts group when compared with that in the control group since the 3rd day, the wound area in the aqueous extracts group was only 15% of that in the control group on the 13th day, and the wound-healing rate was enhanced significantly by the extracts. Moreover, the mRNA expressions of the inflammatory mediators such as TNF-α, IL-1β and IL-6 in the inflammatory RAW 264. 7 cells induced by lipopolysaccharide were inhibited effectively by the ex-tracts in a dose-dependant manner. Conclusion:The results indicate that the aqueous extracts from the fresh roots and rhizomes of A. calamus have significant wound-healing activity in animal excised wound model and anti-inflammatory activity in vitro.

Objective To evaluate the plasma levels of adrenomedullin (ADM) and urotensin Ⅱ ( U-Ⅱ ) in children with capillary bronchiolitis, and their clinical significance. Methods One hundred and fifty three children with capillary bronchiolitis and 36 healthy children were recruited. Plasma levels of ADM and U- Ⅱ were measured at acute stage ( <7days) and convalescent stage (>14days) of airway inflammation. The relationship of plasma ADM and U-Ⅱ levels with symptom scores was evaluated. Results Plasma levels of ADM and U-Ⅱ in acute stage of capillary bronchiolitis were significantly higher than those in convalescent stage and healthy controls ( ADM: t = 20. 57 and 26. 26, P < 0. 01; U-Ⅱ: t = 14. 27 and 7. 61, P < 0. 01 ) , and there were significant differences among mild, moderate and severe capillary bronohiolitis (F = 245. 94 and 304. 79, P <0. 01). Plasma level of U-Ⅱ in convalescent stage of capillary bronchiolitis was lower than that of healthy controls (t = 6. 99, P <0. 01 ) , but ADM was still in a higher level ( t = 8. 98, P < 0. 01 ). In the convalescent stage, there was significant difference on U-Ⅱ levels among mild, moderate and severe capillary bronchiolitis ( F = 25.69, P < 0. 01 ) , but no significant difference was observed for ADM levels (F = 2. 25 , P > 0. 05 ) . Plasma levels of ADM and U- Ⅱ in acute stage showed positive correlation with symptom scores, and the regression coefficients were 0.884 (P =0. 000) for ADM and 0. 943 (P = 0. 000) for U-Ⅱ . Conclusion Plasma ADM and U-Ⅱ levels in children with capillary bronchiolitis are increased in acute stage and correlated with the symptom scores, which may serve as laboratory indicators for assessing the severity of the disease.

BACKGROUND: Tongbiling is the modified guizhi shaoyao zhimu tang (Decoction of Cinnamon Twig, Peony and Anemarrhena) recorded in Synopsis of Prescriptions of the Golden Chamber. It has been used for rheumatoid arthritis(RA) for more than ten years. Here is the experimental study on effects of tongbiling on interleukin 2(IL-2) and its α-chain, IL-2Rα(CD25), to explain the mechanism of tongbiling in regulating the cellular immunity and resisting RA From the aspects of in vivo and in vitro.OBJECTIVE: To study the effect of tongbiling on IL-2 in collagen introduced arthritis of rats and the influence of total base of tongbiling on expression of IL2-Rα (CD25).DESIGN: A totally randomized and controlled experimental study based on the experimental animalsSETTING: Department of Jinkui of a university and the center of organ transplantation and immunity of another university.MATERIALS: The experiment was carried out from April to July 2001 in the Central Laboratory of the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine. The 36 Wistar rats, clean degree, male,bought from the Animal Experimental Center of Medical College of Sun Yat-sen University, certificate No. 2000A050, weighted(110 ± 20) g, were divided into 6 groups with 6 in each: normal group, model group, methotrexate (MTX) high dosage group, MTX low dosage group, tongbiling high dosage group, and tongbiling small dosage group.METHODS: The experiment in vivo was done with the modeled rats of collagen-induced arthritis. The foot swelling was measured with volume method, the cells of primary synovium cultured, and the upper clear fluid of culture fluid tested with radioimmunoassay to observe the effect of tongbiling on synovial IL-2. The experiment in vitro was done in the way of separating the lymphocytes, giving extraneous stimulation with phorbol 12, 13-dibutyrate (PDB) or concanavalin A(ConA), using bicolor immunofluorescence labeling, testing with flow cytometer to observe the influence of total base of tongbiling on the expression of CD3 +, CD25 +.MAIN OUTCOME MEASURES: The influence of tongbiling on the foot swelling and synovial IL-2 of rats with collagen-induced arthritis.RESULTS: In the experiment in vivo, the high dosage of tongbiling could relieve obviously the foot swelling, compared with MTX, there was no remarkable difference ( P ＞ 0.05) . Both high and low dosage of tongbiling could inhibit the synthesis of synovial IL-2 ( P ＜ 0.01 ). In the experiment in vitro, the total base of tongbiling could extremely inhibit the expression of CD3 +, CD25 + stimulated by ConA, but not effective for that stimulated by PDB plus ionomycin.CONCLUSION: Tongbiling can inhibit the IL-2 synthesis and the signal transduction of IL-2Rα (CD25) activated by ConA, relieving the inflammation of local joint. It provides an experimental evidence for the application of tongbiling in the treatment of RA.

AIM:To study the effect of hypertensive arteriosclerosis on cerebral blood flow autoregulation (CBFA), and to introduce a new method to measure the lower limit. METHODS:The blood velocities and blood pressure was recorded simultaneously and the curves of CBFA were analyzed and classified into classical and non-classical pattern. The lower limit were determined by clinical closing pressure (CCP) and the curve CBFA. RESULTS:When the blood pressure was decreasing, the classical and non-classical pattern of the cerebral blood flow autoregulation were 25% and 75% respectively in normal SD rats, while they were 40.55% and 54.45% respectively in renovascular hypertensive rats (RHR). However, when the blood pressure was elevating, the classical and non-classical pattern were 76.47% and 23.53% respectively in SD rats, while they were all classical in RHR. Furthermore, in SD and RHR ras, the lower limits measured by CCP were well in accordance with that measured by CBFA. CONCLUSION:Hypertensive arteriosclerosis could influence the limits and the patterns of cerebral blood flow autoregulation. The lower limit of CBFA can be measured and analyzed by CCP.