OBJECTIVETo express and purify recombinant human interleukin-35[IL-35-IgG1 (Fc) in eukaryotic expression system and to study the interaction of IL-35 with gp130 protein.

METHODSA mammalian expression vector, pSTEP2-IL35-LFc, was constructed and transfected into HEK293T cells. Then rhIL-35-IgG1 (Fc) was expressed and purified with protein A affinity chromatography, and was examined with SDS-PAGE and Western blot analysis. The binding of IL-35 to its receptor gp130 was investigated using enzyme-linked immunosorbent assay (ELISA). The biological effect of IL-35 on gp130 was explored in M1 myeloid leukemia cells.

RESULTSrhIL-35-Fc with high purity on reduced SDS-PAGE was obtained. ELISA confirmed that IL-35-Fc was bound to gp130 and neutralized the function of gp130 in M1 myeloid leukemic cells.

CONCLUSIONSHigh-purity and biologically active rhIL-35-Fc protein successfully produced in this study. IL-35 binds to gp130 and neutralizes its activity of in M1 myeloid leukemic cells.

OBJECTIVE：To investigate the contents of 5 kinds of heavy metal as copper （Cu），arsenic（As），cadmium （Cd），mercury（Hg）and lead （Pb）in Yougui pills ，and to evaluate its safety risk. METHODS ：Using yttrium （89Y），indium （115In）and bismuth （209bi）as internal standard ，the contents of each element were determined by ICP-MS. ICP-MS condition included that atomization gas flow rate was 0.95 L/min，auxiliary gas flow rate was 1.2 L/min，plasma gas （argon）flow rate was 18 L/min，pump speed was 30 r/min. RF power of inductively coupled plasma was 1 200 W，the voltage in simulation stage was 1 750 V，the voltage in pulse stage was 1 300 V，the voltage of deflection device was －12 V，and the detector was in analog and pulse dual-mode. The determination methods of various elements were investigated ，and 45 batches of marketed Yougui pills were determined. Hazard index （HI）was used to analyze the non-carcinogenic risk of each element and calculate the maximum residual limit（MRL）of each element. RESULTS ：The linear range of Cu ，As，Cd，Hg and Pb ranged from 10-200，1-50，0.4-30，0.2-6 and 2-100 μg/L（all r＞0.999 0），respectively. The limits of quantitation were 0.67，0.23，0.20，0.07，0.27 μg/L. The limits of detection were 0.20，0.07，0.06，0.02，0.08 μg/L. RSDs of precision，stability and reproducibility tests were all lower than 3.5％ （n＝6 or n＝5）. Average recoveries were 92.96％-100.89％（RSD＝2.23％-3.62％，n＝3）. Average contents of Cu ，As，Cd，Hg and Pb in 45 batches of Yougui pills were 2.72，0.28，0.07，0.05，0.62 mg/kg，and superimposed HI of each element was less than 1. The contents of 5 kinds of heavy metals in Yougui pills were lower than the proposed MRL （MRL of Cu ，As，Cd，Hg and Pb were 20，2，1，0.2，5 mg/kg or 111.11，4.44，2.22，1.48，8.89 mg/kg respectively ）. CONCLUSIONS ：Established method can be used for the determination of content of 5 kinds of heavy metal in Yougui pills ；the heavy metal pollution rate of marketed Yougui pills is low and the safety risk is small.

To study the effect of serum containing Xihuang pill on the proliferation of human breast cancer cell lines MDA-MB-435 and MCF-7 and the gene and protein expressions of Bcl-2, Bax, TP53, in order to explore the effect and mechanism of Xihuang pill in resisting breast cancer. The serum of the rats was prepared by the method of MTT assay. The expressions of Bcl-2 and Bax were detected by RT-PCR. The serum levels of Bcl-2 and Bax and the mRNA expression of TP53 were detected by immunofluorescence. The rats with serum containing Xihuang pill could inhibit the proliferation of MDA-MB-435 cells and MCF-7 cells (<0.05). The serum containing Xihuang pill increased TP53 and Bax in MDA-MB-435 cells (<0.05), and the ratio of Bcl-2/Bax was decreased (<0.05). Meanwhile, the serum containing Xihuang pill could up-regulate the mRNA expression of Bax in MCF-7 cells and decrease the expression of Bcl (<0.05), but there was no significant difference between the expression of TP53mRNA and Bax protein expressions after the treatment of MCF-7 cells with Xihuang pill serum. Serum containing Xihuang pill can induce the apoptosis of human breast cancer cells, and the mechanism of estrogen receptor-negative breast cancer cell apoptosis may be induced by up-regulating the mRNA expression of TP53, which can induce the expression of Bax and promote the metastasis of Bax to mitochondria, and ultimately play the role of inducing apoptosis.

Objective To develop neutralizing monoclonal antibodies (MAbs) against H10N8 avian influenza virus hemagglutinin and to identify the binding sites. Methods MAbs against hemagglutinin of H10N8 avian influenza virus were developed by genetic engineering. Neutralizing MAbs were screened by microneutralization assay,and then tested by enzyme-linked immunosorbent assay and Western blot to identity the binding sites.The homology modeling process was performed using Discovery Studio 3.5 software,while the binding epitopes were analyzed by BioEdit software. Results One MAb that could neutralize the H10N8 pseudovirus was obtained and characterized. Analysis about epitopes suggested that the antibody could bind to the HA1 region of hemagglutinin,while the epitopes on antigen were conserved in H10 subtypes.Conclusions One neutralizing antibody was obtained by this research.The MAb may potentially be further developed as a pre-clinical candidate to treat avian influenza H10N8 virus infection.
Antibodies, Monoclonal ; immunology ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; immunology ; Hemagglutinin Glycoproteins, Influenza Virus ; immunology ; Influenza A Virus, H10N8 Subtype ; Neutralization Tests

Animals ; Antibodies, Monoclonal ; immunology ; Humans ; Receptors, Interleukin-17 ; immunology

OBJECTIVE：To prepare Imperatorin ultradeformable liposomes gel （IMP-UDLs-Gel），and to evaluate its quality. METHODS：Based on single factor test ，using 12 h accumulative penetration amount （Q12h）as evaluation index ，the proportion of carbomer 940，glycerol and propyl glycol in formulation of IMP-UDLs-Gel were investigated by orthogonal test. The optimal formulation was screened. The quality of IMP-UDLs-Gel prepared with the optimal formulation was evaluated. RESULTS ：The optimal formulation of IMP-UDLs-Gel included carbomer 940 proportion of 1％，glycerol proportion of 15％ and propyl glycol proportion of 10％. Q12 h of IMP-UDLs-Gel was （11.543±0.241）μg/cm2；the appearance was milky white and translucent ；the particle size was （93.13±1.68）nm，PDI was 0.268±0.012，Zeta potential was （－24.96±1.99）mV；pH was 7.32±0.03； viscosity was （45.37±1.27）g·s；steady flow was （0.727±0.002）μg·h/cm2，lag time was （4.358±0.175）h，apparent permeability coefficient was 1.392×10－3 cm/h，and it has good physical and optical stability. CONCLUSIONS ：The preparation method is stable and feasible ，and the prepared IMP-UDLs-Gel has good adhesion ，stability and transdermal property.

A mangiferin aglycon derivative J99745 has been identified as a potent xanthine oxidase (XOD) inhibitor by previous study. This study aimed to evaluate the hypouricemic effects of J99745 in experimental hyperuricemia mice, and explore the underlying mechanisms. Mice were orally administered 600 mg/kg xanthine once daily for 7 days and intraperitoneally injected 250 mg/kg oxonic acid on the 7th day to induce hyperuricemia. Meanwhile, J99745 (3, 10, and 30 mg/kg), allopurinol (20 mg/kg) or benzbromarone (20 mg/kg) were orally administered to mice for 7 days. On the 7th day, uric acid and creatinine in serum and urine, blood urea nitrogen (BUN), malondialdehyde (MDA) content and XOD activities in serum and liver were determined. Morphological changes in kidney were observed using hematoxylin and eosin (H&E) staining. Hepatic XOD, renal urate transporter 1 (URAT1), glucose transporter type 9 (GLUT9), organic anion transporter 1 (OAT1) and ATP-binding cassette transporter G2 (ABCG2) were detected by Western blot and real time polymerase chain reaction (PCR). The results showed that J99745 at doses of 10 and 30 mg/kg significantly reduced serum urate, and enhanced fractional excretion of uric acid (FEUA). H&E staining confirmed that J99745 provided greater nephroprotective effects than allopurinol and benzbromarone. Moreover, serum and hepatic XOD activities and renal URAT1 expression declined in J99745-treated hyperuricemia mice. In consistence with the ability to inhibit XOD, J99745 lowered serum MDA content in hyperuricemia mice. Our results suggest that J99745 exerts urate-lowering effect by inhibiting XOD activity and URAT1 expression, thus representing a promising candidate as an anti-hyperuricemia agent.

The process of globalization increases the risk of global transmission of infectious diseases, resulting in pressure for country's prevention and control of imported infectious disease. Based on the risk assessment of disease importation and local transmission, a strategy that conducting importation prevention and routine prevention and control before the importation of disease and taking emergency control measures after the importation of disease was developed. In addition, it is important to take part in global infectious disease response action, aid the countries with outbreak or epidemic to actively decrease the risk of disease importation.
Communicable Diseases ; Communicable Diseases, Imported/transmission* ; Disease Outbreaks/prevention & control* ; Epidemics ; Global Health ; Humans ; Risk Assessment ; Travel

Objective: To explore the effect of BMI and waist circumference on diabetes of adults. Methods: After excluding participants with heart disease, stroke, cancer and diabetes at baseline study, 53 916 people aged 30-79 in the China Kadoorie Biobank (CKB) study from Tongxiang city of Zhejiang province were recruited. Cox regression model was used to estimate the hazards ratios (HR) for the associations of baseline BMI and waist circumference with incident diabetes. Results: Among 391 512 person-years of the follow-up program between 2004 and 2013 (median 7.26 years), a total of 944 men and 1 643 women were diagnosed as having diabetes. Compared to those with normal weight, after adjusting for known or potential factors, HR of both overweight and obesity in men for incident diabetes appeared as 2.72 (95%CI: 2.47-2.99) and 6.27 (95%CI: 5.33-7.36), respectively. The corresponding figures in women were 2.19 (95%CI: 2.04-2.36) and 3.78 (95%CI: 3.36-4.26). Compared to those with normal waist circumference, after adjusting for known or potential factors, HR of Ⅰgrade andⅡgrade in men for diabetes were 2.56 (95%CI: 2.22-2.95) and 4.66 (95%CI: 4.14-5.24), respectively. The corresponding figures in women were 1.99 (95%CI: 1.80-2.21) and 3.16 (95%CI: 2.90-3.44), respectively. Conclusions: Overweight, obesity and central obesity were all associated with the increased incident of diabetes. Strategies on diabetes prevention should include not only losing weight, but reducing waist circumference as well.
Adult ; Aged ; Body Mass Index ; China/epidemiology* ; Diabetes Mellitus/ethnology* ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Obesity/epidemiology* ; Obesity, Abdominal/epidemiology* ; Overweight/epidemiology* ; Proportional Hazards Models ; Prospective Studies ; Risk Factors ; Waist Circumference