Objective To explore the effects of fasudil on inhibiting vascular smooth muscle cells (VSMCs) proliferation in vitro,increasing cell apoptosis,and inhibiting the Ras-MEK 1/2-ERK 1/2 pathway.Methods Healthy male SD rats (80～100 g) were selected.VSMCs were separated by the thoracoabdominal aortic vascular membrane dissection.Cultured VSMCs were randomly divided into 5 groups:serum-free group; serum group; serum + 1 μmol/L fasudil intervention group; serum + 10 μmol/L fasudil intervention group; serum + 100 μmol/L fasudil intervention group.The proliferation and migration of VSMCs were detected by MTT method and wound healing assay.Cell cycle and apoptosis of VSMCs were examined by flow cytometric analysis.The mRNA expressions of pro-apoptotic protein (Bax) and anti-apoptotic protein(Bcl-2) were determined by RT-PCR method,and the ratio of Bax/Bcl 2 was calculated.Western blot were performed to detect the protein expressions of Ras,MEK1/2,ERK1/2 and Akt in VSMCs.Results Fasudil inhibited rat VSMCs proliferation and migration,and blocked FBS-induced progression from the G0/G1 phase to S phase in a dose-dependent manner.Fasudil inhibited the early and late apoptosis in VSMCs,increased Bax mRNA expression and inhibited Bcl 2 mRNA expression.Fasudil significantly inhibited the protein expressions of FBS-stimulated intracellular Ras,phosphorylated MEK1/2,ERK1/2 in a dosedependent manner,but did not affect the protein expression of phosphorylated Akt.Conclusions Fasudil can attenuate VMSCs proliferation by blocking Ras-MEK1/2-ERK1/2 pathway and increasing cell apoptosis.

Objective:To evaluate the efficacy of individualized cognitive behavioral therapy for unprotected sex and sexual attitude of middle school students having unprotected sex.Methods:A target sample of 68 adolescents having unprotected sex was recruited from 4 secondary schools in Changsha,Hunan [the unprotected sex (US) score of Health-Risk Behavior Inventory for Chinese Adolescents (HBICA) ≥ 1].Subjects were randomized assigned to cognitive behavioral therapy group (CBT group) and control group.Each group had 34 subjects.The CBT group was giving one-on-one counseling for 6 weeks (50 to 60 minutes weekly).The control group didn't receive intervention by counselors.The US and Attitudes Toward Sexuality Scale (ATSS) were selected as criterion measurements.Outcome assessments were made at baseline and at 1-and 3-month follow-up.Results:The reduction rate of US scores showed that the response rate of therapy was over 80％.Mixed linear model analysis showed that there were significant group effect,time effect and group × time effect in scores of US and ATSS (Ps ＜ 0.05).Simple effect analysis indicated that the scores of US and ATSS of CBT group were significant lower than those of baseline from 1-month follow-up [(2.2 ± 2.9) vs.(4.7 ± 3.1),(3.2 ± 1.6) vs.(4.7 ± 3.1);(38.2 ± 4.9) vs.(40.9 ±5.1),(37.2 ±5.4) vs.(40.9 ±5.1),Ps ＜0.01],whereas the scores of those in the control group did not show any significant difference (Ps ＞0.05).At l-month and 3 month follow-up,moderate effect sizes were found for the CBT and control groups on all the outcome measures (Cohen's d =0.50-0.70).Conclusion:The individualized cognitive behavioral therapy could effectively reduce the level of unprotected sex and sexual attitude of adolescents having unprotected sex.

Objective The aim of this study was to determine the content of phthalate in disposable plastic tableware sold on Chengdu market,and to provide primary data for safety evaluation.Methods Sample selection was based on stratified sampling.Sixteen phthalate compounds were investigated in 60 disposable plastic tableware,divided into seven groups.The analysis was performed by gas chromatography-mass spectrometry (GC-MS).Results In this survey,diethyl phthalate,diisobutyl phthalate,dibutyl phthalate and diethylhexyl phthalate were detected,while the other 12 phthalate compound were not.The positive rates of the four detected phthalate were 6.7％ (4/60),10.0％ (6/60),46.7％ (28/60) and 28.3％ (17/60) respectively,and the highest concentrations were 10.3,6.4,7.2 and 65.6 mg/kg,respectively.Conclusion The observed level of detection rates and maximum concentrations were relatively high in this survey.In addition,some subgroups of PAEs that were not allowed to use in food contact materials were detected.Therefore,the migration in different food simulant would be analyzed in the next step for further health outcome assessment.

OBJECTIVE: To observe the effects of psoralen plus ultraviolet-A light (PUVA) on K562 cells and the relative mechanism. METHODS: The effects of psoralen, ultraviolet-A light and PUVA on K562 cells were assayed by monotetrazolium test (MTT). DNA content was analyzed by flow cytometry (FCM). The apoptotic rates of K562 cells treated with 40 and 80 microg/ml psoralen for 24 and 48 hours were assayed by Annexin-V-FITC/PI reagent kit on FCM respectively. The ultrastructures of apoptotic cells were observed by a transmission electron microscope (TEM). RESULTS: Either single psoralen therapy or single ultraviolet-A irradiation had inhibiting effect on K562 cells. The inhibiting effect of PUVA on K562 cells was stronger than that of the single psoralen therapy or single ultraviolet-A light irradiation (P<0.05). Apoptotic peak (AP) was detected by FCM. TEM test showed that K562 cells treated with PUVA were smaller, having condensed cell nucleus, assembled chromatin, disintegrated nucleus body and the majority of the cells appeared to be apoptotic conformation. CONCLUSION: Psoralen has inhibiting effect on K562 cells, and the effect of PUVA is more significant. It is suggested that 10 min irradiation and 40 microg/ml terminal concentration of psoralen be probably the best choice for PUVA. The inhibiting effect of PUVA is due to apoptosis.

Objective:To investigate the effect of silica gel column separation component of Artemisia asiatica (AEM-SC) on the maturation and immune function of mouse dendritic cells (DCs). Methods:Artemisia asiatica components were prepared by macroporous resin eluted with 70% ethanol, and then isolated by silica gel column to obtain AEM-SC. The contents of polysaccharides, flavonoids and triterpenes were quantified. Flow cytometry was used to detect the expression level of DCs surface molecules and antigen phagocytosis ability and to stimulate the proliferation of allogeneic T cells. ELISA method was used to detect the effect of DCs on cytokine secretion. Results:The contents of polysaccharides, flavonoids and triterpenes in AEM-SC were 10.12%, 5.7% and 3.62%, respectively. Functional tests showed that AEM-SC significantly reduced the expression levels of LPS-induced DCs surface molecules CD40, CD86 and MHC-II, reduced the expression levels of inflammatory cytokines IL-12p40, TNF-α and IL-6 (all P<0.05), improve the ability of phygocytosis ( P<0.01), and reduce the ability of DCs to stimulate the proliferation of CD4 +T and CD8 +T lymphocytes in the spleen of mice (all P<0.001). In the inflammatory mouse model experiment, AEM-SC significantly reduced the expression levels of DCs surface molecules CD40, CD86, CD80 (all P<0.001), and the expression levels of inflammatory cytokines TNF-α, IL-12p40 in serum (all P<0.01). Conclusions:AEM-SC can inhibit the maturation of DCs-induced LPS both in vitro and in vivo, indicating that AEM-SC has the immunosuppressive effect.

Objective To explore the efficiency of transcervical video-assisted mini-incision thyroidectomy for thyroid tumor. Methods 53 patients with thyroid tumors who underwent the video-assisted mini-incision thyroidectomy from Jan. 2006 to Dec. 2009 in the Department of Thyroid Surgery, Yantai Yuhuangding Hospital,were retrospectively studied. Results All of the patients, except 5 cases converting to open thyroidectomy, underwent the video-assisted mini-incision thyroidectomy successfully. The temporary recurrent laryngeal nerve injury occurred in 1 patients. The subcutaneous fluid accumulation occurred in 3 patients. The skin burns around the incisions occurred in 3 patients. There was no complication of hematoma, hypocalcemia, or superior laryngeal nerve injuries. The follow-up period was from 3 months to 35 months. There was no evidence of recurrence and metastasis by postoperative ultrasonic and CT scan and radioiodine scintigraphy. Conclusions The video-assisted mini-incision thyroidectomy is feasible, safe, and with the advantage of cosmetic results. For thyroid tumor,especially thyroid cancer, longer follow-up is necessary to draw definitive conclusion on recurrence and survival rate.

In this paper is recommended a highly sensitive and reagent-safe method to determine plasma heamoglobin (FHb) in viscacha hemolytic test. The 2,4-dichlorophenol method (2,4-DCP) of Trinder reaction has been improved. The performance of 2,4-DCP is verified. The sensitivity of 2,4-DCP is 2.39 times that of phenol method. It is well used with run precision and day-to-day precision. The reaction color is stable. The reference value FHb is 1-36.7 mg/L. Sodium citric is an excellent anticoagulant liquid to keep erythrocyte. The 2,4-DCP method is neither carcinogenic nor poisonous;it is suitable for viscacha hemolytic test in clinical and biomedical engineering.
Chlorophenols ; Coombs Test ; methods ; Hemoglobins ; analysis ; Hemolysis ; Humans ; Sensitivity and Specificity

Purpose/Significance To investigate the status quo and influencing factors of self-management level of elderly diabetic mellitus patients in Dongcheng District of Beijing,and to provide references for further development of community diabetes health educa-tion and health management.Method/Process The survey is conducted on 1 962 cases of elderly patients with type 2 diabetes mellitus who are randomly selected from community-level health service institutions in Dongcheng District of Beijing and are investigated by self-made questionnaires and the summary of diabetes self-care activities.Multiple stepwise regression and logistic regression are used to analyze the influencing factors of patients'self-management level.Result/Conclusion The self-management level of elderly diabetic mellitus patients is affected by many factors,among which the awareness of disease-related knowledge,the form of medical security and the use of health-related software to manage chronic diseases are the main influencing factors(P＜0.05).Local management depart-ments can take targeted health interventions to further improve the self-management ability of elderly diabetic mellitus patients.

Objective To investigate the effects of Glycyrrhiza uralensis Fisch L. crude polysac-charides (GUCP) as an adjuvant on the immunity of mice and the immune responses induced by human pap-illoma virus ( HPV)-DNA vaccine. Methods ICR mice were injected with different concentrations of GUCP by different ways to detect the influences of GUCP on body weight, organ indexes and the numbers of immune cells in spleen. C57BL/ 6 mice were co-immunized with HPV-DNA vaccine and GUCP to detect the adjuvant efficacy on antigen-specific cellular and humoral immune responses. Results GUCP in all injected groups had no side effect on mouse body weight and liver, heart, lung and kidney indexes, but intraperitone-al injection of GUCP significantly increased spleen and thymus indexes and the numbers of B cells, CD4+ T cells, macrophages and dendritic cells in spleen. Subcutaneous injection of GUCP significantly increased the numbers of B cells and macrophages in spleen and intragastric administration significantly increased the num-bers of CD4+ and CD8+ T cells in spleen. Furthermore, GUCP as an adjuvant enhanced the antigen-specific CD4+ and CD8+ T cell responses and the levels of IgG, IgG1 and IgG2a induced by HPV-DNA vaccine at a certain degree. Conclusion GUCP enhanced the immunity of mice and the antigen-specific cellular and hu-moral immune responses induced by HPV-DNA vaccine. These results suggested that GUCP might be used as an adjuvant for DNA vaccine.

Objective:To investigate the effects of 30% ethanol elution fraction of Artemisia absinthium extract with macroporous resin (AAEM-30%) on the dendritic cell (DC) and immunity of mice. Methods:AAEM-30% was obtained from the alcoholic extracts of A. absinthium by AB-8 macroporous resin, and its polysaccharide, flavonoid, and terpenoid contents were determined. The expressions of AAEM-30% on DC surface molecular cluster of differentiation (CD) 40, CD80 and CD86 were detected in vitro by flow cytometry, and the expressions of DC cytokines IL-6 and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). The effect of AAEM-30% on the immune function of ICR mice was measured in vivo with different doses (50 and 100 mg/kg) and different administration methods (subcutaneous injection, intraperitoneal injection, and gavage). Results:The contents of polysaccharides, flavonoids, and terpenoids in AAEM-30% were 24.30%, 22.50% and 28.19%, respectively. AAEM-30% significantly enhanced the expression of CD40, and CD86 and the secretion of IL-6 and TNF-α (all P<0.001). Compared with the control group, no statistically significant differences were found in the body mass of mice compared with the three administration methods (all P>0.05). The thymus index in the 50 and 100 mg/kg AAEM-30% intraperitoneal injection groups and the spleen index in the 50 mg/kg AAEM-30% gavage group were increased (all P<0.05). CD19 + cells increased in the 100 mg/kg AAEM-30% intraperitoneal injection group ( P<0.01) and in the 50 mg/kg AAEM-30% gavage group ( P<0.05). The CD11b + and CD11c + counts increased in the 100 mg/kg AAEM-30% gavage group ( P<0.05). The number of CD4 + and CD8 + T lymphocytes was increased by both gavage and intraperitoneal administration (all P<0.05). Conclusions:AAEM-30% can promote the maturation of DC and enhanced the immunity of mice without obvious side effects.