[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.

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.

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 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 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 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.

Objective To investigate the expression and clinical features of amplified in breast cancer 1 (AIB1) and epithelial mesenchymal transition (EMT) markers in human hepatocellular carcinoma and to observe the effect of AIB1 silencing by RNA interference (RNAi) on expression of EMT markers and invasiveness of HepG2 cells.Methods In this study,expression of AIB1,E-cadherin,Vimentin,ZO-1,and N-cadherin protein in 81 hepatocellular carcinomas were assessed through immunohistochemistry and clinicopathological significance was analyzed.After the lentiviral vector of AIB1 RNA interference was transfected into HepG2 cells,the expression of AIB1 and EMT markers was detected by real-time PCR and Western blot.The invasion and metastasis was evaluated by Transwell analysis.Results The expression of AIB1 protein was significantly up-regulated in the hepatocellular carcinoma tissue compared to the normal tumor adjacent tissue.The frequency of AIB1 overexpression in hepatocellular carcinomas with lymph node metastasis is 63％ (P ＜ 0.05).Correlation analysis demonstrated that the AIB1 protein expression was inversely correlated with E-cadherin,and positively correlated with Vimentin in hepatocellular carcinomas.After transfection with AIB1 targeting siRNA,the expression of AIB1 mRNA and protein decreased significantly (P ＜ 0.05).Knockdown of AIB1 expression increased the expression of E-cadherin and inhibited the expression of Vimentin.In addition,the invasion of HepG2 cells silenced AIB1 were significantly descented.Conclusion Above data suggests that overexpression of AIB1 might promote invasiveness and metastasis of cancer cells through regulation of E-cadherin and Vimentin expression in hepatocellular carcinomas.

Objective To reveal the chromosome abnormalities and their relationship with the clinical phenotype of neonates with congenital malformation.Methods Karyotype analysis of peripheral blood lymphocytes was performed on 396 newborns with congenital malformation,who were recruited at the Children's Hospital Affiliated to Soochow University from Jan.2006 to May 2012,chromosome karyotypes were prepared with neonatal peripheral lymphocytes by conventional G-banding technique.Results 1.Of 396 newborns,159 (40.2％) cases were detected to have chromosomal abnormalities,including karyotype first reported domestically and internationally in 3 cases.2.Trisomy-21 (Down's syndrome),which was the most common abnormal karyotype,was seen in 130 cases,accounting for 81.8％,of whom 119 cases show the standard type,10 cases accompanied by the Robertsonian translocation involving group D or group G,and 1 case accompanied by sexual chromosomal abnormality:inv(Y) (p1 1 q 1 1),+ 21.3.Other common karyotype abnormalities were as follows:del (5) (p 1 2-14) (cats cry syndrome) in 4 cases,trisomy-18 (Edwards syndrome)in 4 cases,45,XO (Turner' s syndrome) in 4 cases,inv (9) (p1 1 q1 2-21) in 4 cases,trisomy-X (super female syndrome) in 1 case,rob(13;14) in 1 case,trisomy-8 in 1 case and del(18) (q22) in 1 case.4.Special faces were seen in 147 cases (92.5 ％),congenital heart disease in 97 cases (61.0％),low birth weight in 72 cases (45.3 ％),congenital anal atresia in 13 cases(8.1％),multiple malformations in 11 cases (6.8％),intestinal malformations in 10 cases (6.2％),extrinsic genital abnormalities in 9 cases(5.7％),meow-like cry in 4 cases(2.5％),limb edema in 4 cases (2.5％),fingers and toe abnormalities in 6 cases(3.6％),congenital brain dysplasia in 6 cases (3.6％),webbed neck in 5 cases(3.1％) and cleft lip and palate in 3 cases(1.8％).Conclusions Chromosomal abnormality is an important factor leading to neonatal birth defects,of which special face,congenital heart disease,low birth weight,and multiple malformations are the main clinical manifestations of chromosomal diseases.

Objective To investigate the expression of BAMBI in pancreatic carcinoma tissue and its clinical value.Methods Expression of BAMBI was detected by immunohistochemistry in 69 cases of pancreatic cancer and 15 normal pancreatic samples.The survival and its association with clinicopathologic parameters were analyzed.Results The expression of BAMBI in the cancer tissue was much higher than that in the normal tissues (56.5％ vs 16.7％,P ＜0.01).The expression of BAMBI was not associated with age,sex and tumor size,differentiation,but expression of BAMBI in patient with lymph node metastasis was higher than that in patient without lymph node metastasis (67.4％ vs 38.5％,P =0.019),the expression of BAMBI in TNM stage Ⅲ was higher than that in stage Ⅰ + Ⅱ (75.0％ vs 31.0％,P =0.001).The median survival of BAMBI-positive group (6 months) was lower than that of BAMBI-negative group (10 months).Conclusions BAMBI is over-expressed in pancreatic carcinoma,and its expression is associated with lymph node metastasis,TNM staging,differentiation and prognosis.

Objective To investigate the pathogenesis,diagnosis,treatment and prognosis of aort-oesophageal fistula(AEF).Methods Retropective analysis was performed on 6 patients presenting with AEF between January 2002 and December 2014,and relative literature was reviewed on its pathogenesis,di-agnosis,prognosis and treatment.Results Five men and 1 woman with a mean age of 49 (range,27-71 years)were recruited to the study.One case of AEF was caused by esophageal foreign body,2 cases were caused by aneurysm while the other 3 patients presented AEF after aortic surgery.All 6 patients showed he-matemesis,among whom 3 presented sentinel hemorrhage,1 presented exsanguination after sentinel hemor-rhage,2 presented sudden exsanguination.Among 4 patients with sentinel hemorrhage,2 accompanied with chest pain,1 with dysphagia and 1 with fever.Two patients had a history of hypertension.Diagnostic rate was nearly 100% by gastroscopy or CT/CTA.Four patients died from hemorrhagic shock and 2 patients re-covered from surgery.Conclusion AEF should be seriously considered for patients with a history of hyper-tension,aortic disease or esophageal foreign body presenting sentinel hemorrhage,chest pain,dysphagia,fa-tal exsanguination followed by symptom-free interval.Prompt examinations and aggressive surgery are of great significance for survival.