OBJECTIVE:To determine plasma concentration of caffeine,dapsone and chlorzoxazone in rats,and to calculate pharmacokinetic parameters. METHODS:6 rats were given the mixture of caffeine,dapsone and chlorzoxazone intragastrically, 1.5,2 and 3 mg/kg,respectively. 0.2-0.3 ml blood were collected before medication and 0.5,1,2,3,4,6,8,12,24 h after medication.The plasma sample was treated with solid phase extraction. The plasma concentration of caffeine,dapsone and chlorzoxa-zone were determined by HPLC using N-(2-Hydroxyethyl) phthalimide as internal standard. The pharmacokinetic parameters were calculated using DAS 2.0 software. RESULTS:The linear ranges of caffeine,dapsone and chlorzoxazone were all 0.2-30 μg/ml (r were 0.996 4,0.996 1,0.998 8,respectively). The limit of quantitation were 0.2 μg/ml. The recoveries of low-concentration, medium-concentration and high concentration were(84.8±3.6)%-(111.4±10.2)%(RSD were 4.3%-9.8%,n=3),(107.0±13.3)%-(113.5±8.1)%(RSD were 7.1%-14.0%,n=3),(104.2±10.8)%-(111.1±12.2)%(RSD were 8.0%-11.0%,n=3). Pharmacoki-netic parameters were as follows as tmax(1.70±0.99),(1.50±1.00),(1.92±0.80)h;t1/2(0.73±0.22),(2.77±1.35),(2.78±2.34) h;cmax (2.60 ± 0.50),(5.78 ± 1.19),(9.76 ± 1.37) mg/L;AUC0-t (8.43 ± 0.79),(20.68 ± 1.91),(26.71 ± 2.45) mg·h/L(n=6). CONCLUSIONS:The method is simple,sensitive and accurate,and can be used for the plasma concentration determination and pharmacokinetic study of caffeine,dapson and chlorzoxazone.

[Objective] To assess the role of imbalance between regulatory T (Treg) cells and T helper 17 (Th17) cells in the pathogenesis of atopic dermatitis (AD).[Methods] Peripheral blood was obtained from 41 patients with AD and 38 age- and sex-matched healthy controls.Flow cytometry was performed to determine the percentage of Treg cells (CD4+CD25+Foxp3+ T cells) and Thl7 cells (CD4+ILl7+ T cells),real-time quantitative reverse transcription (RT)-PCR to detect the mRNA expressions of Foxp3 and RORγt,which are the specific transcription factors of Treg and Th17 cells respectively.Serum concentrations of transforming growth factor (TGF)-β,IL-17 and IL-23 were measured by enzyme linked immunosorbent assay(ELISA).Data were statistically assessed by independent-samples t test and Pearson correlation analysis.[Results] The patients with AD showed an obvious decrease in Treg cell percentage,transcription factor Foxp3 mRNA level and Treg/Th17 ratio (2.01％ ± 0.57％ vs.5.04％ ± 1.44％,t =12.47,P＜ 0.01; 0.65 ± 0.19 vs.1.71 ± 0.69,t=9.47,P＜0.01; 1.26 ± 0.61 vs.14.53 ± 5.77,t =14.11,P ＜ 0.01),but a significant increase in peripheral Th17 cell percentage and transcription factor RORγt mRNA level (1.77％ ± 0.55％ vs.0.39％ ± 0.15％,t =14.82,P ＜0.01; 5.97 ± 1.45 vs.1.49 ± 0.57,t =17.78,P ＜ 0.01 ) compared with the healthy controls.Further comparison revealed that Treg/Th17 ratio was significantly lower in patients with acute AD than in those with subacute AD (0.88 ± 0.04 vs.1.29 ± 0.11,t =4.02,P ＜ 0.01 ) and those with chronic AD (2.05 ± 0.24,t =4.83,P ＜ 0.01 ),statistically different between patients with subacute AD and chronic AD (t =2.89,P ＜ 0.05).There was no significant difference in the serum concentration of TGF-β between patients with AD and healthy controls ((15.28 ± 2.34) μg/L vs.(16.56 ± 3.27) μg/L,t =1.96,P＞ 0.05).A significant increase was observed in the serum levels of IL-17 and IL-23 in patients with AD compared with those in the healthy controls( (33.24 ± 7.06)ng/L vs.(11.68 ± 2.67) ng/L,t =17.96,P＜ 0.01; (56.35 ± 12.16) ng/L vs.(18.43 ± 3.90) ng/L,t =18.36,P＜ 0.01).In patients with moderate and severe AD,SCORing atopic dermatitis (SCORAD) index was negatively correlated with the percentage of Treg ceils (r =-0.40,P＜ 0.05 ),but positively correlated with that of Th17 cells (r =0.42,P ＜ 0.05 ).[Conclusion]s There exists a change in Treg/Th 17 ratio,mRN A expressions of RORγt and Foxp3,and serum levels of relevant cytokines in patients with AD,which may lead to immune imbalance and subsequently contribute to the development of AD.

ObjectiveTo detect the expression levels of macrophage migration inhibitory factor (MIF) in peripheral blood mononuclear cells (PBMCs),sera and skin tissue fluid from patients with vitiligo vulgaris,and to investigate their clinical significance.MethodsThirty-nine patients with vitiligo vulgaris and 31 age- and sex-matched normal human controls were recruited in this study.Real-time reverse transcription-PCR was employed to assess the expressions of MIF mRNA in PBMCs,enzyme-linked immunosorbent assay (ELISA) to detect the concentrations of MIF in sera and skin tissue fluid from these subjects.ResultsPatients with vitiligo vulgaris showed a significantly higher level of MIF mRNA in PBMCs (6.70 (2.64 - 8.65) vs.1.67 (1.24 - 2.45),Z=5.895,P＜ 0.05),MIF protein in sera (32.76 (10.67 - 40.98) μg/L vs.7.89 (6.13 - 9.54) μg/L,Z=5.936,P ＜ 0.05 ) and skin tissue fluid ( 167.80 ( 107.40 - 219.60) μg/L vs.42.44 (32.29 - 49.74) μg/L,Z =4.715,P ＜ 0.05) compared with the normal human controls.The expression levels of MIF mRNA in PBMCs,and MIF protein in sera and skin tissue fluid were also higher in patients with progressive vitiligo than in those with stable vitiligo (7.89 (3.89 - 9.12) vs.5.62 (2.23 - 7.29),Z=2.213,P＜ 0.05; 37.80 (29.50 - 45.70) μg/L vs.22.70 (9.36 - 37.78) μg/L,Z=2.141,P＜ 0.05; 211.50 (131.70 - 248.75) μg/L vs.144.65 (89.13 - 167.30) μg/L,Z =2.100,P ＜ 0.05).In addition,the vitiligo area severity index (VASI) score was positively correlated with the expression levels of MIF mRNA in PBMCs (r =0.486,P ＜ 0.05)and MIF protein in sera (r =0.453,P ＜ 0.05).ConclusionMIF might play a certain role in the pathogenesis of vitiligo vulgaris.

Objective To investigate the inhibitory effect of immunosuppressive agent FRY720 on hepatocellular carcinoma Hepal-6. Methods Hepal-6 cells were cultured, and divided into 4 groups, namely control group and 0.1 μg/ml, 10 μg/ml, 100 μg/ml quality concentration groups. The cells were treated by the drugs for 24 to 48 hours respectively. The inhibitory rate of the cells was measured by MTT assay, and cell cycle and cell apoptotic rate were detected by flow cytometry (FCM). Results The ability of tumor cell growth were inhibited by FTY720 after 48 h. The maximal inhibition rate was 62.10 %, The apoptosis ratio was increased when FTY720 was 0.1-100 μg/ml, and it was 4.07 %, 8.16 %, 19.84 % respectively. FTY720 significantly prolonged cell G1 phase. Conclusion FTY720 could inhibit the growth of hepatocellular carcinoma, arrest the cell in G1 phase, and increase apoptosis.

OBJECTIVE:To establish a method for the content determination of total flavonoids in Morus alba. METHODS:UV-visible spectrophometry was performed with Al(NO2)3-NaNO2-NaOH color-test at the wavelength of 510 nm with the reference of rutin. RESULTS:The linear range of rutin was 0.031 2-0.156 mg/ml(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recovery was 95.7%-101.0%(RSD=2.1%,n=6). CONCLUSIONS:The method is simple,sta-ble and reproducible,and can be used for the content determination of total flavonoids in M. alba.

OBJECTIVE:To establish a method for the determination of residuals of petroleum ether,ethanol,xylene and ace-tic acid in ecabet sodium crude drug. METHODS:Capillary GC was performed on the column of PGE-20M capillary column at the flow rate of 1.7 ml/min,detector was hydrogen flame ionization detector,carrier gas was nitrogen with high purity,column temper-ature was 45 ℃,maintaining 4 min,it increased to 80 ℃ with rate of 10 ℃/min,then increased to 135 ℃ with rate of 30 ℃/min,maintaining 3 min,the injection mode was direct injection,inlet temperature was 250 ℃,and the volume injection was 1.0 μl. RESULTS:The mass concentration was 0.050-1.952 g/L for petroleum ether,0.050-1.941 g/L for ethanol,0.024-0.948 g/L for xy-lene and 0.050-1.947g/L for acetic acid(r=0.999 1-0.999 7);RSDs of precision,stability and reproducibility tests were no more than 1.7%;recoveries were 99.3%-101.0%(RSD=0.7%,n=9),102.3%-103.7%(RSD=0.4%,n=9),101.2%-102.1%(RSD=0.3%,n=9) and 100.3%-102.2%(RSD=0.6%,n=9),respectively. CONCLUSIONS:The method is simple and accurate,and can be used for the control of residual of organic solvents in ecabet sodium crude drug.

ObjectiveTo investigate the epidemiology of senile dementia in Shanghai.Methods15158 residents aged 65 or over were selected by stratified random cluster sampling from 3 districts in Shanghai. Two stages was carried to ascertain dementia: in the first stage, Mini Mental State Examination (MMSE) and Activity of Daily Living Scale (ADL) were used as screening tools; in the second stage, DSM-Ⅳ criteria for dementia was applied to diagnosis.ResultsScreening positive rate of MMSE and ADL are both 9.9%. 1271 cases of elderly with dementia were identified. The crude and age-standardized prevalence in residents aged 65 or over was 8.1% and 6.1%. The female, older age, lower education and worse marriage seemed to be more susceptible to senile dementia(P<0.01).ConclusionSenile dementia has become one of major diseases harmful to elderly's health.

ObjectiveTo evaluate probability and severity of behavioral and psychological symptoms in dementia (BPSD) in elderly with dementia in Shanghai.MethodsOn the basis surveyed before, total 1271 elderly with dementia were investigated with the international neuropsychiatric inventory (NPI) to evaluate BPSD.ResultsThe proportion of elderly with dementia that had BPSD was 50.95% , it was 32.17% for clinical significant disturbance (CSD). The symptoms had greatest impact on the caregivers were paranoia (2.32±1.48), followed by abnormal motor behavior (2.30±1.31) and hallucinations (2.16±1.25). The severity rating included mild 620 cases, moderate 238 cases, and severe 406 cases.ConclusionThe incidence rate of BPSD in elderly with dementia was high. The standard of severity grading should be selected with purposes.

ObjectiveTo analyze burden and tolerance of caregivers of elders with dementia in Shanghai.MethodsOn the basis of surveyed before, total 1271 caregivers of elders with dementia were investigated with the questionnaire that concerned the demographic characteristics, influence on body, psychology, life and tolerance.ResultsNearly 60% of the caregivers felt stress, nearly 40% of the caregivers thought that mental pressure had a negative impact on their health as a result of caring patients. Most of the caregivers responded with tolerance to non-adaptive behavior.ConclusionDementia caregivers are old, the burden is heavy. It is necessary to establish a social support system for elderly with dementia as soon as possible.

Objective To investigate the presence and significance of Th9 cells and the related transcription factor ( PU.1 ) and cytokine ( IL-9 ) in peripheral blood of patents with hashimoto thyroiditis ( HT) .Methods Thirty patients with HT and thirty age/gender matched healthy subjects were recruited in this study.The peripheral blood and serum samples were collected from each subject.The percentages of Th9 cells and the transcriptional levels of PU.1 in peripheral blood mononuclear cells ( PBMCs) were meas-ured by flow cytometry analysis and real-time RT-PCR.The concentrations of IL-9, the functions of thyroid and the titers of thyroid-specific autoantibodies ( TPOAb and TgAb) in serum samples were detected by using enzyme-linked immunosorbent assay ( ELISA ) and electrochemiluminescence immunoassay analysis ( ECLIA) .Results Compared with healthy subjects, the percentages of Th9 cells and the expression of PU.1 at mRNA level in PBMCs and the concentrations of IL-9 in serum samples were all significantly in-creased in patients with HT [(1.49±0.68)%vs (0.87±0.24)%], 4.91±2.14 vs 1.66±0.52, (26.90± 7.74) pg/ml vs (16.71±5.87) pg/ml, all P<0.01).Serum concentrations of IL-9 were positively correla-ted with the percentages of Th9 cells (r=0.419, P=0.021).Moreover, the percentages of Th9 cells were positively correlated with the titers of TPOAb and TgAb in serum samples (r=0.394, P=0.032;r=0.457, P=0.011) of patients with HT.Conclusion The levels of Th9 cells and the related cytokine IL-9 were in-creased in the peripheral blood of patients with HT.A positive correlation was found between the percentage of Th9 cells and the titers of thyroid-specific autoantibodies.This study indicated that Th9 cells might be in-volved in the pathogenesis of autoimmune damage in thyroid.