BACKGROUND: Both animal experiments and clinical practice have confirmed that mild or moderate hypothermia is effective in reducing secondary brain injury, but its effect on homeostasis is not very clear.OBJECTIVE: To investigate the effect of a combined therapy of mild hypothermia and hibernation on the homeostasis of patients with severe brain injury.DESIGN: A randomized controlled study.SETTING: Neurosurgical Institute of Beijing; Neurosurgical Department of the First Clinical Medical College Affiliated to Harbin Medical University;and Neurological Department of the Second Clinical Medical College Affiliated to Harbin Medical University.PARTICIPANTS: The study was conducted at the Department of Neurosurgery, the First Affiliated Hospital of Harbin Medical University, from June to December 2002. Totally 24 patients (aged 35-60 years) with severe cerebral hemorrhage or brain injury were randomly divided into combined therapy group and normothermia group. Their Glasgow Coma Scale scores ranged from 3 to 8. The subjects signed the informed consent.METHODS: Within 10 hours of their injury, patients in hypothermia and hibernation combination group were given half dosage of No. 1 hibernation cocktail (chlorpromazine 25 mg, pethidine hydrochloride 50 mg, and promethazine 25 mg), and were cooled by cooling blankets to make their body temperature dropped to 32-34 ℃ (rectal temperature). Their temperature was kept within this range for 5 days, at 35 ℃ for 24 hours, and then was slowly increased to their normal level. The body temperature of patients in normothermia group was maintained at 37-38 ℃. The mean arterial pressure and heart rate of all patients were measured continuously by HP monitor. On the 3"d and 7th days of hospitalization, intracranial pressure and creatine phosphate kinase were measured via lumbar puncture.Femoral artery puncture was performed to check the partial pressure of arterial O2 and CO2. Platelets count and blood electrolytes K+ and Na+ concentration of each patient were measured, too. On the 7th day Glasgow Outcome Scale scores of each patient and mortality of each group were recorded.activity of creatine phosphokinase, platelets count, blood K+ and Na+ conand CO2 of patients in combined therapy group on the 3rd and 7th days of hospitalization.intracranial pressure, creatine phosphokinase and platelets count: The decreased values of intracranial pressure, creatine phosphokinase and platelet number in combined therapy group were all significantly higher than those in normothermia group [(104.09±54.90), (58.75±25.33) mm H2O; (26.95±19.22), (10.17±7.18) μkat/L; (89.82±46.36)×109/L, (48.83±44.59)×109/L,the mean arterial pressure, blood electrolytes, and partial pressure of artewas significantly lower than that of normothermia group (25.0%, 66.6%,P ＜0.05).CONCLUSION: This combined therapy of hypothermia and hibernation can effectively decrease intracranial pressure and creatine phosphokinase,but has no significant effects on the mean arterial pressure, blood electrolytes concentration, and partial pressure of arterial O2 and CO2. It has the risk of disturbing the patients' hematopoiesis.

Objective:To analyze the clinical effect and safety of Lingdan tablets and Wuling capsules in the treatment of alopecia areate. Methods:Totally 84 patients with alopecia areate were divided into the observation group and the control group with 42 ones in each. Minoxidil tincture was used for external in the two groups. The control group were treated with 0. 9 g Lindan tablets, tid, for 4 months and the treatment group was additionally treated with 0. 99 g Wuling capsules,po,tid, for 4 months. The changes in the area and position of trichomadesis, light pull test, the tissue growth score and Pittsburgh Sleep Quality Index ( PSQI) score were compared before and after the treatment. Results:The efficacy of the observation group was better than that of the control group(P<0. 05). Af-ter the treatment, the evaluated indices in the two groups were lower than those before the treatment(P<0. 01), and the reduction of the observation group was more significant than that of the control group(P<0. 01) . There was no notable adverse reaction appeared in the observation group. Conclusion:Lingdan tablets and Wuling capsules can improve sleeping quality, promote the growth of halr and enhance the curative effect.

Aim To investigate effects of ZX-5 on the expression and activity of nitric oxide synthase ( NOS) in endothelial cells,and to confirm which kind of NOS increases NO production and promote choroidal blood flow in ZX-5 -induced HUVECs. Methods HUVECs ( human umbilical vein endothelial cells) were used to determine the expression of eNOS,iNOS and nNOS by Western blot; the activities of NOS were investigated by the corresponding kit. Results ( R,R) ZX-5 upregulated eNOS expression and increased NO production; ( S,S) ZX-5 upregulated iNOS expression and slightly increased NO release. Conclusions ( R,R) ZX-5 promotes choroidal blood flow via upregulating eNOS expression and activity and promoting NO production; the compound may be used for the prevention and treatment of age-related macular degeneration in the elderly.

Objective:To study the inhibition of astragaloside on the proliferation of human keloid fibroblasts. Methods: Com-pared with that of normal skin, the expression of transforming growth factor-β( TGF-β) and its transduction factors Smad in the human keloid fibroblasts was detected. The optimal concentration was screened by MTT after HFF-1 human skin fibroblast was infected with astragaloside at different concentrations. The mRNA expression of Smad2, Smad3, Smad4 and Smad7 in the fibroblasts was studied by using real-time. The protein expression of TGF-βRⅡ, Smad2, Smad3, Smad4 and Smad7 in the fibroblasts was detected by using Western blot. Results: Compared with that of normal skin tissue, the expression of Smad protein was significantly increased ( P <0. 05) in the human keloid fibroblasts, and there was no significant difference in the TGF-βRⅡ expression (P>0. 05). The optimal concentration of astragaloside was 0. 5μg·ml-1 . The expression level of Smad2 protein in the two groups was significantly increased, and the level of Smad3 expression was significantly decreased (P<0. 05). Conclusion:Astragaloside can inhibit the formation of fi-broblast possibly through Smad2 over-expression and Smad3 inhibition in the TGF-β/Smad signal transduction pathway.

The plasma level and the regulation of cortisol in type 2 diabetic patients were invesligated.Plasma and urinary cortisol levels were measured, and dexamethasonc suppression test and oral cortisone test in vivo were performed. Compared with controls, diabetic patients had higher urinary cortisol level. The activity of hepatic 11 β-hydroxy-steroid dehydrogenase-1 (11β-HSD1) in type 2 diabetic patients was decreased, suggesting that the elevated basal cortisol in type 2 diabetic patiens may due to impaired hepatic degradation of cortisol.

Diabetic rat model was induced by high fat diet combined with streptozotocin (STZ). After the model was established, blood samples were taken from jugular veins to examine plasma adrenocorticotropic hormone (ACTH) and corticosterone, and hypothalamus and pituitary were removed for real-time PCR. There were no significant differences in basal plasma ACTH and corticosterone level among control, obese, and obese diabetic rats (P=0.07). The corticosterone rhythm in obese and obese diabetic rats was impaired. Hypothalamus glucocorticoid receptors (GR) mRNA expressions yielded similar results in the groups, but 11β-HSD1 mRNA expression in obese diabetic rats was up-regulated ( vs control rats, P<0.05 ). The expressions of GR and 11β-HSD1 in pituitary of obese diabetic and obese rats were significantly down-regulated (both P<0.05). In the obese diabetic rats, the impaired glucocorticoid negative feedback was partly due to down-regulation of 11 β-HSD1 and GR expressions in pituitary.

Objective To investigate the function of the hypothalamic-pituitary-adrenal (HPA) axis in postmenopausal women with type 2 diabetes and visceral obesity. Methods Subjects were divided into three groups:control group(group C),type 2 diabetes mellitus with non-obesity group (group DM) and type 2 diabetes mellitus with visceral obesity group (group DM + OB). General clinical characteristics, morning blood cortisol concentrations and 24 h urine free cortisol of three groups were compared. Serum cortisol levels were also compared after 0.25 mg dexamethasone suppression test and followed by oral intake of 25 mg cortisone acetate. Results (1) There were no significant differences in basal cortisol levels, but after inhibition with dexamethasone the group DM + OB showed significantly higher cortisol level than that in the control group (P < 0.01). (2) Conversion of oral cortisone to plasma cortisol differed significantly between the group C (lower) and group DM + OB (P < 0.05). (3) Plasma LH and FSH concerntrations were significantly lower in group DM + OB compared with group C (P < 0.01). Conclusion In the postmenopausal women with type 2 diabetes mellitus, the negative feedback mechanism and hepatic 11β-HSD-1 activity were impaired, especially in those with visceral obesity.

Objective To investigate the effects of miRNA-146a on the differentiations and functions of CD4+ T lymphocytes in patients with psoriasis vulgaris.Metbods Thirty patients with psoriasis vulgaris and twenty heathy subjects were enrolled in this study.Quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of miRNA-146a in CD4+ T lymphocytes isolated from the peripheral blood samples.The levels of IFN-γ and IL-4 in serum samples were determined by using enzyme-linked immunosorbent assay (ELISA).Mononuclear cells were isolated from human peripheral blood samples by using Ficoll-Hypaque density-gradients centrifugation,from which the CD4+ T lymphocytes were separated by magnetic-activated cell sorting.The CD4+ T cells (2× 106/ml) were seeded in culture plates with 6 wells.The CD4+ T lymphocytes were divided into 3 groups including the control group,miRNA-146a group and miRNA-146a inhibitor group.The numbers of Th1 and Th2 cells were measured by flow cytometry analysis (FACS).The expression of IFN-γRα,T-bet and GATA-3 at mRNA and protein levels were measured by using RT-PCR and Western blot assay,respectively.The levels of IFN-γ and IL-4 in culture supernatants of CD4+ T lymphocytes were detected by using ELISA.Results In comparison with the normal control group,there were significant increases in the expression of miRNA-146a in CD4+ T lymphocytes and the level of IFN-γin serum samples from patients with psoriasis vulgaris [(2.43±0.94) vs (1.05±0.23),(27.69±7.64) ng/L vs (9.75±2.81) ng/L,all P＜0.01].A positive correlation between the expression of miRNA-146a and the level of IFN-γ in serum was observed (r=0.837,P＜0.01).Results of the in vitro culture of CD4+ T lymphocytes showed that the number of Th1 cells,the expression of T-bet at mRNA and protein levels and the level of IFN-γin culture supernatant were significantly increased,while the expression of IFN-γRα protein was decreased in the miRNA-146a group in comparison with those of the control group (all P＜0.01).No significant differences in the number of Th2 cells,the expression of GATA-3 protein,the expression of GATA-3 and IFN-γRα at mRNA level and the level of IL-4 in culture supernatants were found between the control and miRNA-146a groups (all P＞0.05).The miRNA-146a inhibitor could effectively attenuate the effects of miRNA-146a on Th1 cells.Conclusion The miRNA-146a could promote the differentiation and enhance the function of Th1 cells.It might play an important role in the pathogenesis of psoriasis vulgaris.

Objective To evaluate changes of Rho kinase (ROK)activity in peripheral blood T lymphocytes from patients with atopic dermatitis (AD), and to analyze their clinical significance. Methods Eight milliliters of heparin-anticoagulated blood samples were collected from 60 patients with AD and 60 healthy human controls followed by separation of T lymphocytes and sera from these blood samples as well as culture of isolated T lymphocytes with 10% fetal bovine serum for 24 hours. Both patient- and control-derived T lymphocytes were classified into two groups to be cultured with patient- or control-derived sera. In addition, some patient-derived T lymphocytes were classified into 4 groups:Y27632 group treated with the Rho kinase-specific inhibitor Y2763, CD3/CD28 group treated with anti-CD3/anti-CD28 monoclonal antibodies, Y27632 + CD3/CD28 group treated with Y27632 and anti-CD3/anti-CD28 monoclonal antibodies, and control group treated with patient-derived sera. Subsequently, Western-blot analysis was performed to evaluate ROK activity in cells, methyl thiazolyl tetrazolium(MTT)assay to evaluate proliferative activity of T lymphocytes, and ELISA to measure interleukin 6 (IL-6)and IL-10 levels in supernatants of T lymphocytes. Results ROK activity was significantly lower in fresh T lymphocytes from patients than in those from healthy controls (2.47% ± 0.89% vs. 0.65% ± 0.35%, t =2.729, P < 0.05). After 24-hour culture with 10% fetal bovine serum in vitro, ROK activity was significantly decreased in patient-derived T lymphocytes compared with those before culture (0.70% ± 0.38% vs. 2.47% ± 0.89%, t = 2.658, P <0.05), but no significant difference was observed between patient- and control-derived T lymphocytes(0.70% ± 0.38% vs. 0.63% ± 0.32%, t = 1.010, P > 0.05). Compared with T lymphocytes cultured with control-derived sera, those cultured with patient-derived sera showed significantly increased ROK activity (F = 8.22, P < 0.001). Concretely speaking, ROK activity was significantly higher in patient-derived T lymphocytes cultured with patient-derived sera than in those cultured with control-derived sera (2.41% ± 0.87% vs. 0.76% ± 0.41%, P < 0.05), and higher in control-derived T lymphocytes cultured with patient-derived sera than in those cultured with control-derived sera(2.17% ± 0.85% vs. 0.64% ± 0.33%, P< 0.05)at 24 hours. Y27632 could significantly inhibit the proliferation of as well as secretion of IL-6 (F = 18.68, 22.95, respectively, both P < 0.001)by patient-derived T lymphocytes, but had insignificant effects on secretion of IL-10. The cellular proliferative activity and IL-6 supernatant level were significantly lower in the Y27632 group than in the control group, and lower in the Y27632 + CD3/CD28 group than in the CD3/CD28 group (all P < 0.05). Conclusion Aberrant activation of ROK exists in T lymphocytes from patients with AD, which may play a certain role in the pathogenesis of AD.

Objective To evaluate regulatory effects of miRNA-146a on peripheral blood CD4 + T lymphocytes from patients with psoriasis vulgaris, and to investigate the role of miRNA-146a in the pathogenesis of psoriasis vulgaris. Methods Totally, 30 patients with psoriasis vulgaris and 30 healthy human controls were enrolled into this study. Venous blood samples were obtained from these subjects, and CD4 + T lymphocytes were isolated from these samples by using magnetic activated cell sorting (MACS). Real-time quantitative PCR (RT-PCR)was performed to measure the expression of miRNA-146a in peripheral blood CD4+ T lymphocytes, and enzyme-linked immunosorbent assay(ELISA)to determine plasma levels of interferon-γ(IFN-γ)and interleukin 4(IL-4). Some CD4+ T lymphocytes were divided into 3 groups to be transfected with 50 nmol/L negative control miRNA (control group), miRNA-146a mimics(miRNA-146a group)or miRNA-146a inhibitor (miRNA-146a inhibitor group). After 24-hour additional culture, flow cytometry was conducted to determine the number of Th1 and Th2 cells, Western-blot analysis and RT-PCR were performed to measure the protein and mRNA expressions of IFN-γ receptor α (IFN-γRα), T-box expressed in T cells (T-bet)and GATA-binding protein-3 (GATA-3)respectively, and ELISA was carried out to determine the levels of IFN-γ and IL-4 in supernatants of CD4 + T lymphocytes. Results Compared with the healthy control group, the patient group showed significantly increased miRNA-146a expression in peripheral blood CD4 + T lymphocytes (243.81% ± 94.32% vs. 105.74% ± 22.93%, t = 6.653, P < 0.01)and plasma IFN-γ level (27.69 ± 7.64 ng/L vs. 9.75 ± 2.81 ng/L, t = 4.237, P <0.01). Moreover, miRNA-146a expression was positively correlated with plasma IFN-γ level in the patients(r = 0.837, P <0.01). After 24-hour culture in vitro, there was a significant increase in the number of Th1 cells, protein and mRNA expressions of T-bet, and supernatant level of IFN-γ, but a significant decrease in the protein expression of IFN-γRα in the miRNA-146a group compared with the control group (all P < 0.01). However, no significant differences were observed in the number of Th2 cells, mRNA or protein expressions of GATA-3, or supernatant level of IL-4 among the control group,miRNA-146a group and miRNA-146a inhibitor group (all P > 0.05). Conclusion miRNA-146a may play an important role in the pathogenesis of psoriasis vulgaris by participating in the regulation of peripheral blood CD4+ T lymphocytes via affecting Th1 cell differentiation and function.