Objective To investigate the effects of homocysteine (Hcy) on methylation status and mRNA expression of TNF superfamily member 4 (TNFSF4) gene in THP-1 macrophages. Methods Cultured THP-1 monocytes were induced to macrophages by 0.1 ?mol/L PMA treatment for 72 h, then the differentiated THP-1 macrophages were incubated with homocysteine (50-1 000 ?mol/L) for 24, 48 or 72 h. The status of methylation of TNFSF4 gene in THP-1 macrophages was analyzed by methylation specific polymerase chain reaction (MS-PCR).The expression level of TNFSF4 mRNA was determined by RT-PCR. Results The MS-PCR results showed that the unmethylated bands gradually became stronger, and the methylated bands gradually became weaker along with the prolongation of treatment time and the increased Hcy concentrations. The 72-hour treatment with Hcy induced a complete demethylation in the promotor region of TNFSF4 gene, where left the only unmethylated bands. Meanwhile, the TNFSF4 mRNA expression was also increased in a dose-dependent manner. Conclusion Hcy may contribute to atherogenesis by inducing demethylation in the promotion region of the TNFSF4 gene and increasing TNFSF4 expression in THP-1 macrophages.

BACKGROUND: Inflammatory response is one of potential cellular mechanisms by which hyperhomocysteinemia accelerates atherosclerosis. There are still many details need to be illuminated. TNFSF4 gene and interleukin (IL)-10 are two inflammatory factors related to atherosclerosis. But their correlation with hyperhomocysteinemia is seldom reported. OBJECTIVE: To investigate the effects of homocysteine (Hcy) on expression of TNFSF4 gene and IL-10 in the monocytes cultured in vitro. DESIGN, TIME AND SETTING: A randomized control experiment in vitro was conducted from July to December in 2007 in the Pathophysiology Laboratory of West China School of Preclinical and Forensic Medical Sciences of Sichuan University. MATERIALS: THP-1 monocytes were offered by the Pathophysiology Laboratory of West China School of Preclinical and Forensic Medical Sciences of Sichuan University); Hcy, dexamethasone (Dex) and vitamin D3 (Vit D3) were all purchased from Sigma. METHODS: Cultured THP-1 monocytes were induced to macrophages by 0.1 ?mol/L phorbol myristate acetate, and the differentiated THP-1 macrophages were incubated with 200 ?mol/L Hcy for 24, 48 and 72 hours respectively. Then cell supernatant and lysate were collected as condition medium. Any other differentiated THP-1 macrophages were incubated with a combination of Dex and Vit D3 for 6 days, and then the condition mediums were added to incubate together for 24 hours. Cells were divided into blank control group, 200 ?mol/L Hcy group, Dex+Vit D3 group, 24-hour Hcy+Dex+Vit D3 group, 48-hour Hcy+Dex+Vit D3 group, and 72-hour Hcy+ Dex+Vit D3 group. MAIN OUTCOME MEASURES: The expression level of TNFSF4 mRNA was determined by reverse transcription polymerase chain reaction. The expression level of IL-10 protein was determined by ELISA. RESULTS: Compared with blank control group, no significant difference of TNFSF4 mRNA expression was found in the 24-hour Hcy group, while the expression level was significantly higher in 48-hour and 72-hour Hcy groups. Compared with Dex+Vit D3 group, the IL-10 expression of all Hcy+Dex+Vit D3 groups was significantly lower than that in the Dex+Vit D3 group. CONCLUSION: Hcy may increase TNFSF4 mRNA expression and decrease IL-10 expression in THP-1 macrophages.

Objective To explore, the levels of serum highsensitive C-reactive protein (hs-CRP) in different subtypes of acute ischemic stroke (AIS) and its significance. Methods According to TOAST criteria, 124 cases of AIS were divided into four groups: large-artery atherosclerosis (LAA,47 cases), small-artery occlusion (SVO,37 cases), cardiogenic cerebral embolism (CE,21 cases), acute stroke of other determined etiology and stroke of other undetermined etiology (ODE and UE, 19 cases). The serum concentration of hs-CRP was measured with the immunoturbidimetry in every patient of each group at 72 hours, 7 and 14 days after admitted to hospital. Meanwhile 81 healthy persons were involved in control group. Results The serum concentrations of hs-CRP at 72 hours, 7 and 14 days after onset were (10.77 ± 4.27),(16.41±5.61), (7.63±3.59) mg/L in LAA group;(3.99± 1.56), (6.45±3.25), (4.22±3.21) mg/L in SVO group; (11.60±4.85), (25.14±7.12), (9.24±4.61) mg/L in CE group; (6.09±2.43), (9.65 ±4.65), (5.89 ± 2.68) mg/L in ODE and UE group. The serum concentrations of hs-CRP in LAA group, CE group, ODE and UE group were significantly higher than those in control group [(4.26 ± 1.34) mg/L] (P <0.05) except for that in SVO group, the level of serum hs-CRP in the three groups showed significant difference among the different time (P< 0.05). Conclusions The level of serum hs-CRP in AIS patients is increased and positively correlated with the disease severity. The changes of hs-CRP level are performed in various subtypes of TOAST, and show time-dependent effects. Inflammatory response may play an important role in the pathological mechanism of AIS.

Objective To understand the characteristic of value orientation during occupational choice and occupational development of nursing students with different background. Methods The study was non-experimental and descriptive with questionnaires. Results The correlation coefficients on 15 work value scales in different background students ranged from 0.561 to 0.940, P value ranged from 0.01 to 0.05. And the coefficients were significant. But there were significant differences on some scales in 15 work value of students who were different in grade, sex, family or areas and so on. Conclusions Nursing stu-dents with different background place emphasis on the same work value and ignore the same others because of the same social-cultural background. But there are some differences in recognition degree of some work val-ue in nursing students with different background.

Objective To exPlore the theraPeutic effect of cycloVirobuxine_D ( CVB_D) on heart failure by obserVing the influences of CVB_D on cardiac function,myocardial morPhology and structure in rats with heart failure. Methods The rat model of heart failure was established by ligating left anterior descending coronary artery,and then the rats were randomly diVided into 5 grouPs,Plus the sham grouP. EchocardiograPhy was used to assess cardiac function,including EF,LVSF,SV,HR,CO and so on. Myocardial tissue section was obtained. Change in the myocardial tissue was obserVed by HE staining. Myocardial ultra_structure was obserVed by using transmission electron microscoPe. Results As comPared with the model grouP,EF and LVSF were significantly increased in medium and high dose CVB_D grouPs (P<0. 05 or P<0. 01),SV was increased in low and medium dose CVB_D grouPs (P<0. 05),CO was increased in low,medium and high dose CVB_D grouPs (P<0. 05),LVIDs was decreased and LVPWs was increased in high dose CVB_D grouP (P<0. 05). Under light microscoPe,changes of myocardial structures were obserVed in different CVB_D dose grouPs: myocardial fiber gradually became tidy, striPes became clear, and contour became clear. The changes of myocardial ultra_structure were obserVed under transmission electron microscoPe: the myocardial fiber breakage was reduced and the number of mitochondria was increased,swelling degeneration was alleViated. Conclusion CVB_D significantly imProVed cardiac contraction and blood PumPing function of rats with heart failure, exerted mitigatiVe effect on damage of cardiac muscle fiber and mitochondria. It suggests that CVB_D may be effectiVe in the treatment of heart failure.

In order to have a better basic research of needle sticking, reports regarding basic research of needle sticking in recent years are arranged and summarized, including the concepts of needle sticking, the history origin, manipulation methods, precautions and selection requirements of needles. In the meanwhile, the reinforcing and reducing manipulation in sticking of the needle is preliminarily analyzed, and based on analysis of ancient records, three hypotheses are proposed: (1) needle sticking partly belongs to reinforcing method; (2) needle sticking partly belongs to reducing method; (3) needle sticking can perform reinforcing and reducing effects according to different manipulations. It is also believed that the needle sticking in modern clinical research is mostly used for reducing effects. However reinforcing and reducing manipulation in sticking of the needle still lacks of the support from ancient literature theory and modern clinical application, which needs to be improved.
Acupuncture Therapy ; instrumentation ; methods ; Humans ; Medicine in Literature ; Needles

AIM: To select proper hydrolysis time to determine Ginkgo biloba flavonoids content. METHODS: Sample dissolved in 25mL MeOH-25%∶HCl(4∶1) mixture and was heated to hydrolyze for various time, then diluted. Flavonoids'content was determined by HPLC. The conditions were developed by Shimpack CLC-ODS(150mm?6mm) column using 0.4% phosphoric acid solution-Methanol(50∶50), the detective wavelength was at 360nm with quercetin、kaempferol and isorhamnetin as standards, temperature being 30℃. RESULTS: Contents of flavonoids of Ginkgo biloba leaves,Ginkgo biloba crude extracts and Ginkgo biloba refined extracts were highest in the hydrolysis time of 2.0h. Ginkgo biloba flavonoids can be completely hydrolyzed in 2.0h. CONCLUSION: The hydrolysis time of 2.0h is best.

OBJECTIVE To search for the reasons of unqualified level of bacteria in exit-entrance fluid of hemodialyzer and draft the measures of adjustment to meet the evaluation criteria for exit-entrance fluid of hemodialyzer.METHODS According to MOH′s Hospital Infection Management Standards to detect level and species of bacteria.RESULTS The average numbers of bacteria in exit-entrance fluid of hemodialyzer decreased from 2152.95?826.45 CFU/ml before adjustment to 579?541.04 CFU/ml after adjustment,checking with chi square test,the entrance fluid of hemodialyzer was ?2 =15.92,P

AIM:To observe the characteristics of estrous cycle of female ICR mice and ovariectomized mice that had reached sexual maturity stage .METHODS:ICR mice were randomly divided into 3 groups, normal group, sham operation group and ovariectomized group .After ovariectomy and sham operating , at the 8th day, the mice were taken vagi-nal smears twice daily (09:00 and 16:00) for 12 consecutive days.The feature of vulva in the mice and vaginal smear cy-tology, occurrence rate of regular estrous cycle , duration of estrous cycle and each stage duration of estrous cycle were re -corded and observed .RESULTS:The characteristics of estrous cycle in normal group and sham operation group were simi-lar.Nearly 85%of the ICR mice in sexual maturity stage showed regular estrous cycle .The majority manifested 6-day cy-cle (83%), and only a small number exhibited 5-day cycle (17%).Compared with 5-day group, the duration of oestrus was significantly increased in 6-day group (P<0.05).The number and kind of the cells in vaginal smear were a little dif-ferent between 5-day and 6-day groups.The ovariectomized mice which showed irregular estrous cycle (76%) were charac-terized by continuous diestrus .CONCLUSION:Female ICR mice have the strain characteristics of estrus cycle .

Objective To construct 2 recombinant plasmids carrying cassette chromosome recombinase C(ccrC)and ccrAB respectively and introduce the plasmids into methicillin-resistant Staphlococcus aureus(MRSA)strain 81/0432,and to observe the precise excision of Staphylococcal cassette chromosome mec(SCCmec)island from bacterial chromosome.Methods ccrC and ccrAB genes were amplified with chromosomal DNAs isolated from MRSA strains 81/0342 and N31S as PCR templates.We constructed recombinant plasmids pSR5C and pSR2AB by cloning ccrC and ccrAB genes into temperature-sensitive plasmid pYI3,after introducing them into MRSA strains 81/0432 and N315 by electroporation.PCR was performed to identify SCCmec excision from the bacterial chromosome.The transformants were serial passaged for 10 days,and then the drug resistance of these rransformants was detected by replica experiment.Results The fragment length of ccrC gene was 1.9 kb,smaller than the fragment length of ccrAB from N315.The recombinant plasmids of pSR5C and pSR2AB were successfully constructed.After these 2 recombinant plasmids were introduced into MRSA strain 81/0342,type-V SCCmec island was excised from the chromosome and formed a closed circular structure in the bacteria.However,type-Ⅱ SCCmec island could be excised only in N31S strain after the expression of pSR2AB.Replica experiment verified that transformed strains of 0342(pSR2AB),0342(pSR5C),and N315 (pSR2AB)were mostly susceptible to ceftizoxim or tobramycin after the excision of SCCmec island.Conclusion cciC could serve as a recombinase as ccrAB,which could mediate precise excision of SCCmec island from the chromosome of type-V MRSA strain.This study shows that type-V SCCmec island is widely disseminated between Staphylococcus aweus strains in community setting.