Objective To discuss the influence of different doses budesonide aerosol inhalation on serum cytokines levels in children with severe asthma.Methods 96 children with severe asthma aged 4 to 14 years old in our hospital were chosen,and they were randomly divided into the observation group and the control group,48 cases in each group.All patients were given conventional treatment, and the control group was given 1mg/time budesonide treatment on the basis of conventional treatment (3 times a day),while the observation group was given 2mg/time budesonide treatment (3 times a day).Before and 1 week after treatment,the clinical symptoms of two groups were observed and compared,as well as the changes of IL-4,IFN-gamma,IL-10 and TNF-α.Results In the obser-vation group,wheezes,coughing,wheezy sound and rales disappearance time were (2.10 ±0.77)d,(5.45 ±1.20)d, (3.46 ±1.03)d,(5.55 ±1.35),which were significantly shorter than (2.98 ±1.02)d,(7.48 ±1.19)d,(5.43 ± 1.06)d,(7.56 ±1.67)d in the control group (t=4.77,8.32,9.23 and 8.32,all P<0.01).4 weeks after treat-ment,the total effective rate of the observation group was 89.6%,which was significantly higher than 72.9% of the control group (χ2 =4.376,P<0.05).After treatment,the IL-4,IL-10,TNF-alpha,IFN-gamma levels in the observation group were (4.06 ±1.77)pg/mL,(12.77 ±2.05)pg/mL,(4.15 ±1.11)ng/mL,(26.23 ±2.78)pg/mL, which had significant changes compared with (9.02 ±2.23)pg/mL,(10.21 ±1.30)ng/mL,(6.66 ±1.62)pg/mL, (17.33 ±2.31)pg/mL before treatment(t=12.07,24.56,16.20,17.25,all P<0.01).After treatment,the IL-4, IL-10,TNF-alpha,IFN-gamma levels in the control group were (9.11 ±2.05)pg/mL,(6.80 ±1.23)ng/mL, (9.88 ±2.20)pg/mL,(21.22 ±2.80)pg/mL,which had significant changes compared with (9.11 ±2.05)pg/mL, (10.38 ±1.37) ng/mL,(6.71 ±1.77) pg/mL,(17.30 ±2.05) pg/mL before treatment( t=5.36,13.47,7.77, 7.83,all P<0.01).But IL-4,TNF-alpha levels in the treatment group were significantly lower than those in the control group (t=7.32,11.08,all P<0.01),while IL-10 and IFN-gamma levels were significantly higher than those in the control group (t=6.65,8.80,all P<0.01).The incidence of adverse reactions of the two groups had no statistically significant difference (χ2 =0.771,P>0.771).Conclusion High doses of budesonide aerosol inhalation in the treatment of children with severe asthma has obvious clinical curative effects,which could significantly improve the patients'clinical symptoms,and also has low incidence of adverse reactions,which is worthy of clinical promotion.

Background Inflammation is one of the most popular aspects in the studies of diabetic retinopathy (DR) mechanisms.Researches showed that S100A8/A9 participate in the inflammatory procedure of many diseases,however,the relationship between S100A8/A9 complex and retinal inflammation of DR needs to be researched.Objective This study was to detect the serum S100A8/A9 level of diabetes mellitus (DM) and DR patients,and explore its role in DM an DR development.Methods A cases-controlled study was carried out.The DR patients,type 2 DM patients without retinal change and heathy controls were enrolled in Shanghai Xuhui Central Hospital from January to June 2014,and 30 patients for each group.The DR patients were subgrouped to non-proliferative DR (NPDR) group and proliferative DR (PDR) group.The periphery blood was collected to isolate the serum,and serum S100A8/A9 complex level was detected by ELISA.Serum high-sensitivity C-reactive protein (hsCRP) and glycosylated hemoglobin A1C (HbAlc) level was assayed by immunity turbidimetry and immune agglutination respectively.Results Serum S100A8/A9 complex levels in the DR group,DM group and normal control group were (9.74±0.59),(11.41 ±0.64) and (6.46 ±0.62) μg/L,respectively,and the serum S100A8/A9 complex level in the DM group and DR group was significantly higher than that in the normal control group,and the serum S100A8/A9 complex level in the DM group raised in compared with the DR group (all at P＜0.01).Serum hsCRP levels in the DR group,DM group and normal control group were (1.40±0.34),(1.27±0.13) and (1.11 ± 0.12)mg/L,respectively,with the highest value in the DR group and the lowest value in the normal control group (all at P=0.00).The serum HbAlc levels were higher in the DR group and DM group than those in the normal control group (both at P =0.00),while no significant difference was found in the serum HbAlc level between DR group and DM group (P =0.12).There was no significant differece in the serum S100A8/A9,hsCRP and HbAlc levels between NPDR group and PDR group (t=-0.10,P =0.92;t =-0.17,P =0.87;t =0.66,P =0.51).A weak positive correlation was seen between serum S100A8/A9 level and serum hsCRP level (r =0.36,P =0.00).Conclusions As an inflammatory marker,S100A8/A9 complex might play an important role in the pathogenesis and development of DR.Intensive control of glycemia can alleviate retinal inflammation in DM patients.

Objective To evaluate clinical significance of two real-time fluorescence quantitative PCR kits for quantitative detection of HBV DNA and detection performance at different viral load levels.Methods A series of calibrators with different concentrations(1×106,5×105,1×105,5×104,1×104,5×103,1×103,5×102,1×102,1×101 kIU/L) were prepared with AB-type sera using the second generation WHO international standard (NIBSC code:97/750). HBV viral load in the sera of 78 patients,30 healthy blood donors and 10 calibrators were detected by real-time fluorescence quantitative PCR HBV DNA test kit from PIJI Bio-Technical Development Company Ltd (PG kit) and Cobas AmpliPrep/Cobas TaqMan HBV test kit. The correlation of the two methods was evaluated, and the performance of the two kits different viral load levels was evaluated. The false negative rate was analyzed. Negative control, low positive control and high positive control were included in every batch. Results Both two kits showed the correct results for the 10 specimens from the WHO international standards. The lowest detection limit of HBV DNA for Roche Cobas AmpliPrep/Cobas TaqMan HBV test and PG kit were 2.00 (kIU/L, lg) and 3.00 (kIU/L,lg) ,respectively. There was linear correlation between the results from Roche Cobas AmpliPrep/Cobas TaqMan HBV test and PG kit ( R2=0.938 7, P < 0.01 ), the upper limit of Roche kit had coincided with theoretical value. The samples with HBV DNA level above the upper limit of detection were diluted and retested to obtain the precise result. The result form Roche Cobas AmpliPrep/Cabas TaqMan HBV test [(8.35±0.20) kIU/L, lg] was higher than that from PG kit [(7.73±0.42 ) kIU/L, lg] (t=3. 776, P <0.05) . The detection of 108 serum samples showed that the level of HBV DNA detected by Roche Cobas AmpliPrep/Cobas TaqMan HBV test [(5.88±1.64) kIU/L, lg] was higher than that by PG kit [(5.25±1.55 kIU/L,lg] (t=12. 297 ,P <0.01 ). The correlation coefficients were high in samples with high HBV viral load[HBV DNA(>5.00 and≤7.00) kIU/L,Ig,R2=0. 779 7, P <0.01 ;HBV DNA( >7.00 ands≤9.00) kIU/L,lg,R2=0.603 7, P <0.01]. The correlation coefficient was low in samples with low HBV viral load[HBV DNA ( > 3.00 and≤5.00) kIU/L, lg, R2=0. 417 3, P <0.01 )]. When HBV DNA ( >3.00 and≤4.00) kIU/L,lg,the false negative rate was 33.3% (5/15). When HBV DNA ( > 1.08and≤3.00) kIU/L,lg,none of positive samples was detected with PG kit. Conclusions PG kit is not as good as Cobas AmpliPrep/Cobas TaqMan HBV test . The linear correlation between the results from the two kits is good. The correlation between the results detected with PG kit and Cobas AmpliPrep/Cobas TaqMan HBV test is higher in the high viral load groups than in the low viral load group. It is suggested that PG kit had a narrower linear range.

Objective To determine the conversion equation for X(copies/ml, lg) quantity values of domestic HCV RNA quantitative fluorescence amplification assays approved by SFDA and Y( IU/ml, lg)reference values of standard substance. Methods The second generation WHO International Standard (NIBSC code:96/798) was mixed with human AB blood type serum to create 7 different dilutions which included 100 000, 50 000, 25 000, 10 000, 5 000, 2 500 and 1 000 IU/ml. Two different batches of each three domestic hepatitis C virus RNA real-time fluorescence quantitative PCR assays and 2 different batches of each assay were employed to detect the 7 different concentration samples with real-time PCR. Each test was performed 4 times repeatedly. Results The correlations between X( copies/ml,lg) values of domestic HCV RNA assays and Y(IU/ml,lg) reference values of standard substance were as follow,Assay A:Y =0. 902 0 X+0.284 9,R2 =0.953 3,P＜0. 01,n =56;Assay B: Y=0. 875 7 X +0.562 4,R2 =0.956 5,P＜0.01,n =56; Assay C: Y = 0. 843 8 X + 0. 560 5, R2 = 0. 945 8, P ＜ 0. 01, n = 56. Conclusions All the conversion equations are different among the quantity value of three assays and the reference values of standard substance, that suggests it is necessary to perform more stringent traceability analysis for the quantity values of 3 assays. Through standardizing the quantity values preliminarily, the conversion equation can enhance the comparability between the quantity values of different assays, and provide a standard of HCV RNA virus load detection for clinical diagnosis and treatment monitoring of HCV infection.

AIM: To study the effects of ultraviolet (UV) on mitochondrial functions and apoptosis in HaCaT cells.METHODS: After irradiation by UV at low dose(UVA 2 J/cm~2,UVB 10 mJ/cm~2) and high dose(UVA 6 J/cm~2,UVB(30 mJ/cm~2),) HaCaT cells were cultured for 15 hours.Flow cytometry was used to measure mitochondrial membrane potential,mitochondrial mass and apoptotic rate.Annexin V-FITC/PI staining of apoptotic cells was analyzed by laser confocal microscopy.RESULTS: After UV irradiation,cell proportion with low mitochondrial membrane potential increased with irradiation doses.The proportion of control group,low dose group and high dose group were 7.94%?1.02%,25.87%?4.55% and 39.27%?5.32%,respectively.Cells proportion with low mitochondrial mass increased with irradiation doses.The proportion of control group,low dose group and high dose group were 15.19%?1.58%,40.36%?4.41% and 68.79%?5.46%,respectively.The hypodiploid peaks of DNA content analysis represented the apoptotic rate of HaCaT cells.The apoptotic rate of control group,low dose group and high dose group were 1.82%?0.51%,30.16%?5.47% and 58.49%?5.98%,respectively.To analyze the cells apoptosis by staining with annexin V-FITC and PI,the results were consistent with those of DNA content analysis.Cells in control group showed almost no positive staining cells.Single annexin V-FITC positive cells in low dose group and double positive cells in high dose group were predominant,respectively.CONCLUSION: UV irradiation induces HaCaT cell mitochondrial depolarization,as well as mitochondrial mass loss.These changes are related to cell apoptosis.

In recent years, more and more people have recognized the importance of patients' family in the intensive care unit (ICU) in medical care, and advocated the use of patient- and family-centered care (PFCC) in the ICU. This article explains the content (family presence, family support, communication with family members, consultations and ICU team members, environmental issues) and significance of PFCC in the ICU, and provides guidance for the practice of PFCC in China.

Objective To determine the distribution of HCV genotypes in patients with chronic hepatitis C,study the distribution of genotype in different gender and the relationship between genotypes and serum HCV-RNA levels.Methods Two hundred and six cases of HCV RNA positive patients(all with relevant clinical data) receiving pegylated interferon therapy were collected from May to December 2010.HCV RNA was detected in 206 hepatitis C patients from 40 hospitals in China by Roche Cobas AmpliPrep/Cobas TaqMan HBV test,and genotype was determined by Abbott RealTime HCV G enotype Ⅱ .The distribution of genotypes in the gender was analyzed by chi-square test analysis.The relationship between genotypes and serum HCV RNA levels was detected by single factor analysis and two independent sample t test analysis.Results There were seven different subtypes of HCV in 206 samples,including genotype 1,7 cases(3.4% ,7/206); genotype 1a,2 cases(1.0%,2/206); genotype 1b,123 cases (59.7 %,123/206); genotype 2,32 cases(15.5 %,32/206); genotype 3,27 cases(13.1%,27/206); genotype 6,6 cases(2.9% ,6/206) ;genotype 1/6,5 cases(2.4% ,5/206) ;genotype 2/4,1 cases(0.5%,1/206).There was no significant difference between HCV genotype and gender in 132 cases with genotype 1 and 65 cases with non-genotype 1(genotype 2,3,6) (x2 = 0.000,P ＞ 0.05).There was significant association between quantity of HCV RNA and genotype in 188 patients with HCV(F = 3.371,P＜ 0.01).The 197 patients with HCV single genotype were divided into five groups in terms of region(East,South,West,North and Center).There was no significant difference between HCV genotype 1 and non-genotype 1 in the five groups(x2 = 5.840,P ＞ 0.05).Conclusions It is suggested that HCV 1 b is the most prevalent type in China,followed by HCV 2.There is no significant difference between HCV genotype and gender.The levels of HCV RNA with genotype 1b are significantly higher than those with genotype 3.The levels of HCV RNA with genotype 2 are significantly higher than those with genotype 3.The levels of HCV RNA with genotype 6 are significantly higher than those with genotype 3.

Objective To study the proliferation inhibition effect by silencing PLCε gene expression with RNA interference in BIU-87 cells. Methods The specific short hairpin RNA recombinant plasmids were constructed by gene clone technology.The expression level of PLCε protein and mRNA were detected by Western blot and RT-PCR respectively after transfected recombinant plasmids into BIU-87 cells.The influence on proliferation was check by MTT.The changes of proliferating cell nuclear antigen(PCNA)were analyzed by immunocytochemical method,and the distribution of cell cycle was analyzed using flow cytometry. Results After transfected with the specific recombinant plasmids,PCNA expression was decreased 33.08%,and the analysis of cell cycle indicated that cells of G0/G1 phase were increased comparision with(40.75±2.30)%and(40.00±1.76)0A,and its G2/M phase cells(8.16±0.51)%were decreased strikingly compared with group control(31.20±1.76)%and group NP(35.94±1.58)%.Cells were blocked at G0/G1 phase,the cell proliferation was inhibited obviously. Conclusion PLCε may play an important role in proliferation of bladder cancer cells,which could be a potential target of biological treatment on bladder cancer in the future.

ABSTRACT:Objective To explore the role of HIF‐1αin the pathogenesis of hepatopulmonary syndrome (HPS) and its relationship with GRP78 .Methods The HPS model in rats was induced by multiple pathogenic factor .The samples were assessed by using Western blotting analysis for HIF‐lα, GRP78 and VEGF164 . The expressions of VEGFR‐2 and CD105 were observed by using immunohistochemical staining .Results The protein level of HIF‐1αwas significantly increased in HPS group at week 8 compared with that at week 4 and 6 groups and corresponding normal control groups .With the development of HPS ,protein level of GRP78 was gradually increased at each time point significantly and reached the highest level at week 8 ;protein level of VEGF164 showed a similar change with GRP78 ,but the peak was at week 6 .Immunohistochemical results showed that the protein expressions of VEGFR‐2 and CD105 were gradually increased in lung tissue as HPS progressed .The protein level of GRP78 was positively correlated with HIF‐1α,VEGF164 ,VEGFR‐2 and CD105 ,respectively (P<0 .05) .Conclusion HIF‐1αis most likely together with GRP78 to play a critical role in promoting pulmonary microvascular remodeling in the pathogenesis of HPS in rats .

Objective: To investigate the changes of cardiovascular health indicator and arteriosclerosis in middle and elder population.
Methods: A total of 4190 subjects with the average age of (49.78 ± 9.74) years by 3 physical examinations in Kailuan group from 2006 to 2011 were randomly stratiifed for arm ankle arterial pulse wave velocity (baPWV) examination. According to 7 AHA cardiovascular health indicators of non-smoking, normal BMI, active excise, healthy diet, normal cholesterol, blood pressure and fasting blood glucose, each indicator had 3 conditions as ideal, general and poor by scores of 2, 1 and 0 respectively. Based on the 1st and 3rd physical examinations, the changes of cardiovascular health scores (△CHS), the subjects were divided into 8 groups as△CHS≤-4,-3,-2,-1, 0, 1, 2 and△CHS≥3, n=241, 368, 611, 855, 911, 647, 354 and 203 respectively. The impacts of△CHS on baPWV values were studied by liner and Logistic regression analyses.
Results: As△CHS increased by △CHS ≤ -4, -3,-2,-1,0,1, 2 and△CHS ≥ 3, the baPWV values were decreased accordingly by cm/s as (1590.78 ± 17.93), (1566.4 ± 14.5), (1552.83 ± 11.25), (1536.59 ± 9.51), (1508.85 ± 9.21), (1499.81 ± 10.93), (1485.92 ± 14.82) and (1475.85 ± 19.57) respectively. Multiple linear regression analysis showed that with adjusted confounding factors, as△CHS increasing 1 score, baPWV increasing 15.58 cm/s (B=15.58, P<0.001). Logistic regression analysis indicated that with adjusted confounding factors, as△CHS increasing 1 score, the risk for arteriosclerosis occurrence was decreased by 14%(OR=0.86, 95%CI 0.83-0.90).
Conclusion: △CHS was negatively related to baPWV in middle and elder subjects, improving cardiovascular health indicator may decrease arteriosclerosis occurrence.