Objective To study the changes of Th17 cell, regulatory T cell (Treg) and interleukin (IL)-6 in the peripheral blood of patients with systemic lupus erythematosus (SLE) and their relationship with disease activity. Methods Percentage of Th17 and Treg in the peripheral blood of 103 patients with SLE and 28 healthy volunteers were detected by flow cytometry. The concentration of IL-6 in SLE patients and healthy volunteers was detected by cytometric bead array (CBA). The disease activity of SLE was measured by SLEDAI. SLE patients were divided into two groups: stable SLE (SLEDAI≤ 9, n=37) and active SLE (SLEDAI＞9, n= 66). The change of Th17, Treg, IL -6 and their relationship with disease activity were analyzed. Nonparamentric tests, t -test and spearman correlation were used for statistical analysis. Results The percentage of Th17 cells and the concentration of IL-6 in the peripheral blood in patients with SLE was higher than that in normal controls [respectively for (1.2±1.1)%, (35±92) pg/ml and (0.6±0.4)%, (6±3) pg/ml, P＜0.05]. However, the percentage of Treg in patients with SLE was lower than that in normal controls [respectively for (1.6±1.2)%,(2.6±1.8)%, P＜0.05]. The percentage of Th17, Th17/Treg IL-6 level in active SLE patients was higher than those in inactive SLE and those in normal controls (P＜0.05). However, the percentage of Treg in active SLE was lower than that in stable SLE patients and that in normal controls (P＜ 0.05). The percentage of Th17, Th17/Treg and concentration of IL-6 was positively correlated to disease activity(P＜0.05). But the percentage of Treg had negative correlation with the percentage of Th17 and disease activity (P＜0.05). Conclusion Th17, Treg and serum IL-6 in SLE patients are abnormal and they maybe contribute to the pathogenesis of SLE.

Objective To explore the correlation between Th1,Th2,Th17,and Treg cells differentiation and related cytokines in patients with rheumatoid arthritis (RA).Methods Seventy-one patients with active RA were enrolled in this study.They were divided into low,moderate and high disease activity groups according to disease activity score (DAS28).The frequencies of Th1,Th2,Th17,and Treg cells in the peripheral blood of RA patients group (n=71) and healthy conlrol group (n=18) were determined by flow cytometry.T test was used for statistical analysis.Results Significant difference could be detected between the proportions of Th1 [ (6.2±4.5)％],Th17 [ (1.1±0.9)％] and Treg [(1.8±1.2)％] cells in the peripheral blood of RA patients and the control group (P＜0.05),and there was correlation between proportions of these three kinds of cell and the DAS28.Conclusion Th1 and Th 17 cells may promote the development of RA disease,but Th2 and Treg cells could prevent further development of RA disease.

Objective To access the expression and clinical significance of CD19+CD5+B cells and interleukin (IL)-10 in patients with systemic lupus erythematosus (SLE). Methods Forty-six SLE patients and ten healthy controls were recruited in the Second Affiliated Hospital of Shanxi Medical University. CD19+CD5+B cells subsets were detected with flow cytometry. IL-10 level in serum were detected with enzyme linked immunosorbent assay (ELISA). The correlation between the expression of CD19+CD5+B cells and serum level of IL-10 with ESR, systemic lupus erythematosus disease activity index (SLEDAI) score and complement was analyzed. Pair-wise comparison of means of groups was conducted with one-way ANOVA. Comparison between the two groups was conducted by LSD-t test. Correlations between variables were carried out using Spearman's rank correlation test. Results The percentage of CD19+CD5+B cells in peripheral blood of SLE patients [(5.7±2.1)%] was significantly lower than those in healthy control group [(19.1±2.9)%](t=2.431, P=0.005), and it had a negative correlation with SLEDAI score (r=-0.292, P=0.049). The IL-10 serum level of SLE patients [(18.8±13.5) pg/ml] was significantly higher than the healthy control group [(8.3±2.9) pg/ml] (t=3.021, P=0.003), and it had a positive correlation with SLEDAI score (r=0.322, P=0.029). Conclusion CD19+CD5+B cells and IL-10 may participate in the occurrence and development of SLE. The changes of CD19+CD5+B cells and IL-10 in the peripheral blood might contribute to the pathogenesis and correlate with the disease activity of SLE.

Objective To improve the rheumatologists' understanding of noncirrhotic portal hypertension.MethodsA case of systemic sclerosis complicated by noncirrhotic portal hypertension was reported and the related literatures were reviewed.Results A 51-year-old female who had been diagnosed as systemic sclerosis presented clinically with an chronic onset of portal hypertension.She also had pancytopenia,splenomegaly,and significant esophageal varices.Liver function tests were normal.Hepatitis viral serology was negative.Ultrasound scan of liver revealed no focal lesion.ACT scan confirmed the absence of portal vein thrombosis.Taking into account the above evaluation we concluded that the patient had systemic sclerosis and noncirrhotic portal hypertension.The patient was on prednisolone and immunosuppressant and the condition was well.Conclusion Noncirrhotic portal hypertension complicated by autoimmune disease,especially SSC,is very poor,characteriged by significant portal hypertension as well as histological evidence that cirrhosis is absent.Rheumatilogist should pay attention to it.

OBJECTIVETo investigate the effect of IL-12 on IFN-gamma and IL-10 production of peripheral blood mononuclear cells (PBMC) in chronic hepatitis B virus infection patients during IFN-alpha treatment.

METHODSBefore and after IFN-alpha treatment of 3 months and 6 months, PBMC of 20 patients with chronic hepatitis B virus infection were collected and cultured in vitro in the culture fluid containing PHA (100 microg/ml), HBcAg (1 microg/ml), or HBeAg (1 microg/ml) for 48 h under the condition of the presence or absence of IL-12 (10 ng/ml). Then the levels of IFN-gamma and IL-10 were determined by ELISA.

RESULTSThere were 12 responders and 8 nonresponders to IFN-alpha treatment. In the responders, the enhancing effect of IL-12 on IFN-gamma production was significantly greater after IFN-alpha treatment than before the treatment. The production of IL-10 was suppressed in the presence of IL-12 after 3 months and 6 months of IFN-alpha treatment. In the same treatment time, the level of IFN-gamma in the presence of IL-12 was significantly higher than that in the absence of IL-12. To the nonresponders, the enhancing effect of IL-12 on IFN-gamma production was also significantly increased after IFN-alpha treatment. Moreover, in the same treatment time, the level of IFN-gamma in the presence of IL-12 was significantly higher than that in the absence of IL-12.

CONCLUSIONSThe enhancing effect of IL-12 on IFN-gamma production of PBMC in patients with chronic hepatitis B virus infection is increased during IFN-alpha treatment. IFN-alpha and IL-12 may enhance the efficacy for the treatment of chronic hepatitis B virus infection.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-12 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; drug effects ; metabolism ; Male ; Middle Aged ; Time Factors

【Objective】 To construct the Rb luciferase reporter gene assay system and detect the activation ability of Rb gene for screening the targeted drugs. 【Methods】 The synthetic Rb gene sequence was annealed to form a double-stranded DNA structure and then inserted into the polyclonal site of pGL6-TA. The junction product was transformed into E.coli DH5α competent cells for expanded culture, and the constructed pGL6-Rb-Luc plasmid and pGL6-TA plasmid were transfected into HEK293 cells. The monoclonal cell line HEK293-Rb-Luc with stable expression was screened by G418, and the activation and inhibition of Rb in HEK293-Rb-Luc were tested by serum and CDK4/6 inhibitor Palbociclib. 【Results】 The sequence of Rb reaction elements in pGL6-Rb-Luc was completely correct. The recovery of serum culture significantly increased the luciferase activity in HEK293-Rb-Luc (P<0.001). Compared with 0 nmol/L, 25, 50, 75 and 100 nmol/L, CDK4/6 inhibitor Palbociclib made the inhibition rate of Rb activity rise to 6.90%, 40.23%, 50.57% and 52.07%, respectively (P<0.05). 【Conclusion】 The Rb luciferase reporter gene detection system HEK293-Rb-Luc was successfully constructed, which can effectively detect the activation level of Rb.

【Objective】 To clone the full-length of human kidney and brain protein (KIBRA) coding sequence in eukaryotic expression vector and provide a model for studying the biological function of KIBRA in breast cancer cells. 【Methods】 Total RNA of human breast cancer cell line MCF7 was extracted. After reverse transcription, the full length of KIBRA (NM_001161661.2) coding region was amplified by PCR, and cloned into eukaryotic expression vector pCMV-Blank. After identification, it was defined officially as pCMV-KIBRA. Then it was transfected into MCF7 cells, and the expression of KIBRA was detected by real-time PCR and Western blotting after 48 hours. The primary, secondary and tertiary structures and post-transcriptional modification sites of KIBRA were analyzed with bioinformatics software. 【Results】 Bacterial PCR, double enzyme digestion and DNA sequencing results showed that the correct sequence of KIBRA was inserted into the vector pCMV-KIBRA. The mRNA and protein expressions of KIBRA were significantly increased in MCF7 cells transfected with pCMV-KIBRA. Bioinformatics analysis showed that KIBRA was composed of 1119 amino acids. There were 52 phosphorylation sites, 1 acetylation site and 5 ubiquitination sites, and the protein structure was mainly α-helix and random coil. 【Conclusion】 The eukaryotic expression vector of full-length of human KIBRA coding sequence was successfully constructed and overexpressed in breast cancer cell line MCF7, which can lay a foundation for studying the biological function of KIBRA in breast cancer.