As a patient's right,informed consent has been acknowledged universally.Both hospital and patients attach great importance to normal informed consent during clinic.But the subjects' right in clinical trial didn't draw enough attention,which leads to encroachment of the subjects' right and even the clinical disputes.The law and rule,based on the ethics principle,can enhance the protection of the subject's right of informed consent.In this article,the problem on the legal protection against the infringement of the subject's right and the corresponding counter-measures are discussed.

Objective To clone and express the extracellular portion of mouse Fcγreceptor Ⅱ-b ( FcγRⅡb) and to analyze the functions of the expressed proteins in a mouse model of systemic lupus ery-thematosus ( SLE) .Methods The gene fragment encoding FcγRⅡb was amplified by PCR, and then in-serted into the prokaryotic expression vector pET-32a(+) to construct the recombinant expression plasmid pET-FcγRⅡb.The expression plasmid was identified with restriction enzymes and then sent to the Shanghai Bio-Engineering Co.LTD for further sequencing analysis.The transformed Escherichia coli ( E.coli) BL21 ( DE3) strains carrying the recombinant expression plasmid pET-FcγRⅡb were induced by isopropylβ-D-1-thiogalactoside ( IPTG) .The expressed fusion proteins were analyzed by Western blot assay and purified with purification kits.The immune complex ( IC)-binding ability of FcγRⅡb was measured by ELISA.MRL/lpr mice with SLE in both the prevention (12 weeks old, n=40) and the treatment groups (19 weeks old, n=40) were randomly divided into four groups including 60 μl (4.8 μg) treatment group, 120 μl (9.6 μg) treatment group, 180μl (14.4μg) treatment group and PBS treatment group with 10 mouse in each group. The MRL/lpr mice with SLE were injected with the fusion protein through tail vein once a week for four con-secutive weeks.Serum samples were collected from each mouse after one week of observation.The levels of FcγRⅡb in soluble form in mice form both the prevention and treatment groups as well as the levels of anti-double stranded DNA antibodies were detected by ELISA.Results The gene encoding FcγRⅡb was ampli-fied and the recombinant expression plasmid pET-FcγRⅡb was successfully constructed.The recombinant proteins were expressed in the prokaryotic expression system, and then successfully purified.The recombi-nant proteins could bind to IC.Compared with the corresponding PBS control group, the levels of FcγRⅡb in soluble form were increased in mice from both prevention and treatment groups after treating with various concentrations of the recombinant protein (P<0.05).Significant differences in the levels of FcγRⅡb were found among mice of the same age after treating with different concentrations of the recombinant protein ( P<0.05).Compared with the corresponding PBS control group, the levels of anti-double stranded DNA anti-bodies were decreased in mice from both prevention and treatment groups after treating with various concen-trations of the recombinant protein (P<0.05).The levels of anti-double stranded DNA antibodies were grad-ually decreased along with the increasing dosage of protein (P<0.05).Conclusion The extracellular por-tion of murine FcγRⅡb in soluble form was successfully expressed.The recombinant proteins played a cer-tain role in the prevention and treatment of SLE in a mouse model.

Objective To construct an eukaryotic vector expressing short hairpin RNA(shRNA) of HIF-1α,and to observe its effects on location and metastasis of HeLa cells under hypoxic condition.Methods shRNA templates was developed based on HIF-1α gene sequence and then cloned into pSilencer2.1-U6-neo vector.The resultant plasmid was transfected into HeLa cells with Lipofectamine 2000.The cells were incubated in hypoxic condition.The HIF-1α protein and mRNA were detected by Western blot and RT-PCR.The colony formation assay and Transwell cabin assay were performed to measure the colony formation and metastasis.Results The plasmid pSilencer2.1-U6-neo-HIF-1α was successfully constructed and transfected into HeLa cells.The expression of HIF-1α in HeLa cells decreased,and the number of colony formation in soft agar and cells penetrating matrigel also decreased under hypoxic condition.Conclusion The shRNA expressing plasmid targeting at HIF-1α may suppress the location and metastasis of cervical carcinoma cells under hypoxic condition.

Medical Ethics Committee founded in medical orgnizations,universities,academic periodicals and hygiene institutes is an international regulation in the filed of medicine.Although we have done a large amount of work to be in line with international standards,there are still some problems such as no systemic roles,function deficiencies etc.Therefore,we should study its establishing background and main function and strengthen its construction.

Objective To observe the expression of Toll-like receptor(TLR) on peripheral blood dendritic cells(DC) in children with Henoch-Schtinlein purpura(HSP),and to investigate the pathogenesis of the abnormal expression of TLR in children with HSP.Methods Twenty hospitalized children with HSP in the Affiliated Hospital of Qingdao University Medical College from Dec.2011 to Jul.2012 were enrolled in the study(HSP group).Twenty agemetched healthy children were selected as a healthy control group.Peripheral venous blood was sampled under aseptic condition,peripheral blood mononuclear cells (PBMC) were isolated from density gradient centrifugation,and DC were generated by recombinat human granulocyte-macrophage colony-stimulating factor(GM-CSF),interleukin-4(IL-4) and tumor necrosis factor-α(TNF-α) in vitro.Expressions of CD83,CD86 and TLR2,TLR3,TLR4 in peripheral blood DC were examined by fluorescent activated cell sorter (FACS).Results 1.No significant distinction was found in the expression of the C Ds3 on peripheral blood DC between HSP group and healthy control group(t =0.80,P ＞ 0.05) ;in HSP group had remarkably increased expression of the CD86 on peripheral blood DC than that of the healthy control group (t =9.56,P ＜ 0.01).2.Expression rates of TLR2,TLR3,TLR4 on peripheral blood DC in the HSP group were higher than those in the healthy control group(t =1 1.79,13.29,9.45,all P ＜ 0.01).3.Expression rates of TLR2,TLR3 and TLR4 in HSP group had positive correlation with expression rates of CD86 (r =0.84,P ＜ 0.01 ; r =0.53,P ＜ 0.05 ; r =0.66,P ＜ 0.05).Conclusions Expressions of TLR2,TLR3 and TLR4 on peripheral blood DC significantly increased and were positively correlated with expression of CD86.This implies that TLR and co-stimulatory molecules might participate in the pathogenesis of HSP by mediating signal transduction,leading to abnormity of cytokines,then inducing Th1/Th2 immune imbalance by showing the advantage of Th2 function.

Objective To determine the prevalence of penicillinase-producing Neisseria gonorrhoeae(PPNG) and the distribution of blaTEM-135 gene variants in PPNG at several gonococcal antimicrobial surveillance sites in China, to compare N. gonorrhoeae multi-antigen sequence typing(NG-MAST)types of PPNG and its blaTEM-135 gene variants, and to assess the difference and association in NG-MAST types of blaTEM-135 gene variants among different regions. Methods A total of 572 N. gonorrhoeae isolates were collected at 6 gonococcal antimicrobial surveillance sites from Jiangsu, Shanghai, Zhejiang, Tianjin, Guangdong and Guangxi in 2012. After isolation, purification, and identification, cefalotin paper discs were used for detection of PPNG. DNA was extracted by QIAxtractor DX kits after cultivation of the PPNG strains. Then, mismatch amplification mutation assay (MAMA) PCR was performed to identify blaTEM-135 variants, and NG-MAST analysis to determine N. gonorrhoeae genotypes. Results Among the 572 N. gonorrhoeae strains, 38.1%(218/572) were identified as PPNG, and of the PPNG strains, 52.3% (114/218) were blaTEM-135 variants. The detection rate of PPNG at these surveillance sites from high to low was as follows: 51.7% (45/87, Zhejiang), 45.6%(36/79, Shanghai), 38.0% (78/205, Guangdong), 37.5% (12/32, Guangxi), 31.2% (24/77, Jiangsu) and 25.0%(23/92, Tianjin), and that of blaTEM-135 variants was as follows: 68.9%(31/45, Zhejiang), 58.3%(14/24, Jiangsu), 50.0%(39/78, Guangdong), 47.2%(17/36, Shanghai), 39.1%(9/23, Tianjin)and 33.3%(4/12, Guangxi). NG-MAST analysis showed that the ST2318, ST1768, ST1866, ST1053 and ST8726 types predominated among these bla TEM-135 variants, and a strong correlation was found between blaTEM-135 variants and some NG-MAST types, such as ST1768, ST1053 and ST8726 types. The distribution of NG-MAST types was significantly different between the surveillance site in Tianjin (in the Northern part of China) and the other sites (in the Southern part of China), but highly similar among the surveillance sites in Jiangsu, Zhejiang and Shanghai regions. Conclusions There is a high prevalence of PPNG and its blaTEM-135 variants at several gonococcal antimicrobial surveillance sites in China, with significant differences in NG-MAST genotype distribution of PPNG and its blaTEM-135 variants among different regions.

Objective After implementation of new standard iodized salt,to comprehensively assess the iodine nutrition levels of different populations in Dali City of Yunnan Province.Methods From 2012 to 2015,in Dali City,there were 5 districts divided into east,west,south,north and middle,each district selected 1 township (town),and each township (town) selected 4 administrative villages,15 households for edible salt in each administrative village were sampled,and the salt iodine content was measured by "General Test Method in Salt Industry Determination of Iodine" (GB/T 13025.7-2012).In 2014,in the five districts of east,west,south,north and middle of Dali City,one township (town) was selected,and 20 pregnant women in the early,middle and late stages,respectively,20 lactating women,20 ordinary healthy adults and 20 children aged 0 to 4 were selected from each township (town);one primary school in each township (town) was selected in each district,and 40 students aged 8-10 years old were selected from each primary school as the survey objects.The urine samples of the survey objects were collected,and the urinary iodine content was measured by "Method for Determination of Iodine in Urine by As3+-Ce4+ Catalytic Spectrophotometry" (WS/T 107-2006).In 2015,in each administrative village of Dali,a water source with the largest number of drinking people was investigated,and water iodine was detected by the "Method of Water Iodine Detection Suitable for Iodine Deficiency and High Iodine Areas".Through questionnaires,the sources of iodine supplementation for pregnant and lactating women were investigated.Results The qualified iodized salt consumption rate of residents was higher than 90％ per year from 2012 to 2015,and median of salt iodine decreased from 29.38 mg/kg (2012) to 24.96 mg/kg (2015).The medians of urinary iodine in different populations were 136.85 μg/L for pregnant women (n =356),102.63 μg/L for lactating women (n =111),164.03 μg/L for adults (n =163),209.61 μg/L for 8-10 years old children (n =200),157.27 μg/L for children aged 0-＜ 2 years old (n =57),and 134.08 μg/L for 2-4 years old children (n =50).The median of iodine content of drinking water (n =142) in Dali was 0.62 μg/L,the range of iodine content was 0.00-9.92 μg/L.The average intake frequencies of iodine-rich seaweed for pregment women and lacting women were 0.99,1.07 time/month,respectively,only 1.99％ (9/453) of the population supplemented iodine through multivitamin and minerals tablets.Conclusions After reduction of salt iodine content,the iodine nutrition of populations in Dali City (a low water iodine region) is generally at an appropriate level.Maintaining a higher level of qualified iodized salt consumption rate,strengthening the monitoring of different populations and promotion of healthy behaviors are key steps in prevention and control of the disease in the future.

To investigate the leukemogenic potential of PML-RARalpha fusion protein in vivo, hCG-PML-RARalpha transgene was constructed using molecular cloning technique and hCG-PML-RARalpha transgenic mice were generated. The genotype and phenotype of hCG-PML-RARalpha transgenic mice were analyzed by PCR, RT-PCR, morphology of peripheral blood and bone marrow cells, and pathological examination of spleen, liver and bone marrow. As a result, acute promyelocytic leukemia was developed in 3 hCG-PML-RARalpha transgenic mice in 1 - 5 months. The results demonstrated that PML-RARalpha fusion protein plays a crucial role in leukemogenesis.

Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine