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.

The knock out vector pHK was constructed with E coli vector pET 28a and shuttle vector pHY300PL, by using denatured DNA and homologous recombination technique, the kanamycin resistance gene ( Kan r) from integrated alkaline protease gene engineering strain BP071 was knocked out successfully, and the 11 positive clones were obtained The yield of the best positive clone BP0715 was stable as same as BP071 The methods provided the good experience for the industrial microbiology research, and it was foundation for studying on the safety of genetically modified organisms

Objective To investigate the routine methods of α-amylase (AMY) test in scrum which meets the requirements of ISO 15189.Methods Fifty human serum samples with different concentrations of AMY (40- 750 U/L) were collected from March to December in 2008,to form the patients' frozen serum group.Four AMY measurement systems including Roche,Wako,MINDRAY and MAKER were used.The frozen standard materials with concentrations of ( 70.1 ± 3.7 ) U/L and ( 418.3 ± 22.1 ) U/L and the patients' frozen serum group were measured simultaneously by using IFCC reference method and 4 AMY measurement systcms based on 7170A automatic biochemistry analyzer.Thc linear regression analysis was made between the measurement results of each system and IFCC reference method.The equivalence,agreement and trueness were also evaluated by using the file EP9-A2 method.Bland-Altman GraphicalAnalysis and the improved Bland-Altman Graphical-Analysis of MVS1.80 software.Results Judging by the standards of IFCC reference method,the measurement results of 4 measurement systems were obviously different. ( 1 ) When measuring standard materials the results were 66.4,70.6,69.4 and 49.2 U/L respectively and 394.0,456.4,406.7,302.4 U/L respectively.The measurement results of MINDRAY were in agreement with that of IFCC reference method.( 2 ) When mcasuring the patients' scrum group by 4 measurement systems and IFCC reference method,the slopes of the linear regression equations were 0.934,1.070,0.930 and 0.731.respectively.And the intercepts were 0.886,6.249,5.388 and 3.574,respectively.According to the EP9-A2 method,the measurement results of Roche Wako,MINDRAY were equivalent to that of IFCC reference method.According to Bland-Altman Graphical-Analysis, the measurement results of Roche and MINDRAY were in agreement with that of IFCC reference method.The average biases of each measurement system were - 6.11％ ( Average bias ± 2s were 2.81％ and -9.40％ ),1.99％ ( Average bias ± 2s were 10.35％ and - 6.36％ ),- 2.70％ (Average bias ± 2s were 2.37％ and -7.77％ ) and -34.72％ ( Average hias ±2s were -24.20％ and -45.24％ ),respectively.According to the improved Bland-Altman Graphical-Analysis, the measurement results of MINDRAY are correct.The average biases of each measurement system were - 5.92％ (Average bias ± 2s were -2.81％ and -9.03％),2.10％ (Average bias ±2s were 10.74％ and -6.53％),-2.64％ ( Average bias ± 2s were2.24％ and -7.51％ ) and - 29.51％ ( Average bias ± 2s were 21.82％ and - 37.21％ ),respectively.Conclusions ( 1 ) The measurement results of different measurement systems do not necessarily have crreet results though they have claimed to have traceability.(2) The trueness of measurement results using the same system may not come to the same conclusion when evaluated by different methods.So laboratories should select and establish a procedure to evaluate trueness of routine methods and adopt those meeting the trueness requirements of ISO 15189.

Objective To investigate the effects of proanthocyanidins on depressant-like behaviors and the structure of adrenal gland in chronic unpredictable mild stress (CUMS) rats. Methods Rats were randomly divided into 6 groups:control group, stressed group (CUMS + vehicle), three treatment groups (CUMS + proanthocyanidins 25,50,100 mg·kg-1,respectively) ,and imipramine group (CUMS + imipramine 10 mg·kg-1). Used the CUMS model in rats to investigate the effects of chronic oral administration (21 days) of proanthocyanidins and imipramine (ip) on the open-field;and forced swimming and sucrose consumption tests and the ratio of adrenal gland/body weight,and its thickness were examined by HE stain. Results Compared with control group, rats subjected to CUMS exhibited increased ratio of adrenal gland /body weight ( P < 0. 01), less sucrose consumption( P<0.01) and inhibited in the open-field test( P<0.01) as well as more despair time in the forced swimming test( P<0.01). While compared with stressed group,treatments with proanthocyanidins (25,50,100 mg·kg-1, po ,21 days) could significantly improve the activities in open-field test ((39.6±3.4) vs (49±4.5), (52.6±3.7),(54.1±1.8) ;all P<0.01) and sucrose consumption( (5.8±2.5)ml vs (8.1±3.3)ml,(8.5±4.1) ml, (9.2±2.6) ml; P < 0.05, P < 0.05, P < 0.01 respectively); Meanwhile, it could reduce the duration time in forced swimming test significantly( (103.5±10.2)s vs (83.7±8.8)s,(75.8±5.9)s,(67.2±6.5)s; all P<0.01) as well as thickness of the adrenal gland(P<0.05, P<0.01).Conclusions This study suggests that the proanthocyanidins (25,50,100 mg·kg-1) has an antidepressant-like effects in CUMS rats. The antidepressant actions of proanthocyanidins, in some degrees, may be related with the regulation of the adrenal gland's structure.

Objective To evaluate the system measurement procedure effects on the analytic precision of clinical chemistry analytes.Methods In June 2009, June 2010 and September 2010 respectively,the National Center for Clinical Laboratories of China and the Organization of Five Hospitals in Fukuoka Japan organized comparison activities of 26 clinical chemistry analytes which were ALT,AST,GGT, ALP,CK,LDH,AMY,ChE,TG,TC,HDL-C,LDL-C,Glu,Cr,BUN,UA,K,Na,Cl,Ca,TP,Alb,TBil,DBil, P,Fe.In this paper, we investigated 26 analytes of three sets in Beijing Aerospace General Hospital as follows.(1)The precision of different reconstitution methods was observed by using three kinds of pipetting tools, such as measuring pipette, pipette and dispenser.(2)The experiments were carried out in three stages by testing the dried powder control samples of two concentration levels(101-Ⅰ,101-Ⅱ)provided by Hitachi Japan.They were measured on 28 consecutive days at each stage in order to observe the precision of 26 clinical chemistry analytes.In the first stage,we used the former measurement procedure to measure the control samples;in the second stage we added three conditions of the measurement procedure.The first was two calibration modes,which were once-a--day calibration and twice-a--day calibration.The second was the calibration standard and the last was the conditions of the freeze-thaw samples.In the third stage, we used the twice-a-day calibration only for GGT,ALP,ChE,TG,Cr,Na,K,CL,ALB.(3)JSCC and Health Industry Standard quality objectives were implemented to evaluate whether the precision of the improved measurement procedure met the requirements.(4)Paired T test were used to compare the precision of measurement between the second stage and the first stage, and between the third stage and the second stage of the measurement procedure.Results (1)The precision of three kinds of pipetting tools were 0.56%,0.10%, 0.01%.(2)The ranges of precision of ALT,AST,GGT,ALP,CK,LDH,AMY,ChE,TG,TC,HDL-C,LDL-C,Glu,Cr,BUN,UA,K,Na,Cl,Ca,TP,Alb,TBil,DBil,P,Fe were 0.99%-10.5% about 101-Ⅰ and 0.91%-7.03%about 101-Ⅱin the first stage.The ranges of precision were 0.66%-8.81%of 101-Ⅰand 0.66%-4.28%of 101-Ⅱin the second stage.The ranges of precisions were 0.60%-3.91%of 101-Ⅰand 0.73%-3.39%of 101-Ⅱin the third stage.(3)73%/80%of the samples met the standard of JSCC about 101-Ⅰand 101-Ⅱand 80%/88%of the samples met the standard of Health Industry Standard in the first stage.88%/100% of the samples met the standard of JSCC about 101-Ⅰand 101-Ⅱ and 100%/100%samples met the standard of Health Industry Standard in the second stage.The ratio of samples meeting the standard of JSCC about 101-Ⅰand 101-Ⅱwere 96%/100% and that of Health Industry Standard were 100%/100%in the third stage.(4)Precision of 101-Ⅰand 101-Ⅱwas statistically significant between the measurement procedures of second stage and the first stage,and there was no significant difference between the third stage and the second stage.Conclusion (1)The precision of samples using dispenser to reconstitute is higher than that of the other two pipetting methods.(2)Improving the calibration mode and reconstitution of samples increase the precision of 26 clinical chemistry analytes by over 50%.

Objective:To investigate the relationship between polymorphisms and haplotypes of cyclin-dependent kinase inhibitor 2B antisense RNA 1 ( CDKN2 B- AS1) gene and the risk of ulcerative colitis (UC). Methods:From January 2012 to January 2021, a total of 534 UC patients diagnosed at the Department of Gastroenterology, the Second Affiliated Hospital of Wenzhou Medical University (Yuying Children′s Hospital) and during the same period 560 gender- and age-matched healthy controls were selected. Genotypes of CDKN2 B- AS1 (rs1063192, rs10757274, rs10757278, rs1333048, rs2383207) in venous blood were determined by matrix assisted laser desorption ionization time-of-flight mass spectrometry technique. Unconditional logistic regression was used to analyze the difference in the distribution of CDKN2 B- AS1 gene polymorphisms between UC patients and healthy controls, as well as the influence on the clinicopathologic characteristics of UC patients. Software Haploview 4.2 was used to analyze the linkage disequilibrium and haplotype. Chi-square test was used for statistical analysis. Results:The frequencies of variant genotype (AG+ GG) and variant allele (G) of rs1063192 in UC patients were higher than those in healthy controls (32.4%, 173/534 vs. 24.8%, 139/560; 18.1%, 193/1 068 vs. 13.7%, 153/1 120), and the differences were statistically significant ( OR=1.45 and 1.40, 95% confidence interval(95% CI) 1.12 to 1.89 and 1.11 to 1.77, P=0.006 and 0.004, corrected P=0.030 and 0.020). The frequency of variant allele (G) of rs10757274 in UC patients was lower than that in healthy controls (34.7%, 371/1 068 vs. 39.5%, 442/1 120), and the difference was statistically significant ( OR=0.82, 95% CI 0.69 to 0.98, P=0.025). However, the difference was not significant after Bonferroni correction (corrected P>0.05). According to the Montreal classification, the frequency of homozygous variant genotype (GG) of rs1063192 in the patients with extensive colitis was higher than that in patients with proctitis plus left-sided colitis (6.6%, 14/211 vs. 1.9%, 6/323), and the difference was statistically significant ( OR=3.92, 95% CI 1.47 to 10.42, P=0.006, corrected P=0.030). There was linkage disequilibrium among rs10757274, rs2383207, rs10757278 and rs1333048 of CDKN2 B- AS1 gene. The frequency of haplotype GGGC in UC patients was lower than that in healthy controls (33.3%, 355.5/1 068 vs. 37.8%, 423.4/1 120), and the frequency of haplotype AGGC in UC patients was higher than that in healthy controls (6.7%, 71.7/1 068 vs. 3.6%, 40.3/1 120), and the differences were statistically significant ( χ2=4.81 and 11.16, P=0.028 and<0.001). Conclusions:The variation of rs1063192 in CDKN2 B- AS1 gene may increase the risk of UC. The risk of extensive colitis in patients carrying homozygous variant genotype (GG) of rs1063192 may rise. Among the haplotypes composed of rs10757274, rs2383207, rs10757278 and rs1333048, the risk of UC may decrease in the individuals carrying haplotype GGGC. However, the risk of UC may increase in the individuals carrying haplotype AGGC. The correlation between the variation of 10757274 and the risk of UC still needs to be further verified by expanding the sample size.

Objective:To assess the association between the polymorphisms of integral protein α4 ( ITGA4) and intercellular adhesion molecule 1 ( ICAM-1) genesand the risk and clinicopathological characteristics of Crohn′s disease (CD) among Chinese patients. Methods:From January 2010 to January 2021, a total of 215 CD patients and 529 gender- and age-matched healthy controls were enrolled from the Second Affiliated Hospital of Wenzhou Medical University as the study subjects. Genotypes of ITGA4 (rs6740847, rs7562325) and ICAM-1 (rs5498) were determined by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Harvey-Bradshaw Index (HBI) was applied to assess the disease activity of CD, and the patients were further divided into subgroups based on the Montreal Classification Criteria of CD. Unconditional logistic regression was employed to analyze the distribution of ITGA4 (rs6740847, rs7562325) and ICAM-1 (rs5498) polymorphisms between the patients and healthy controls and their association with the clinicopathological characteristics of the patients. Results:The frequencies of T allele and CT+ TT genotypes of ITGA4 (rs7562325) were higher in CD patients than the healthy controls (40.70% vs. 31.57%, P=0.001; 62.79% vs. 54.36%, P=0.042). The G variant and AG+ GG genotypes of ITGA4 (rs6740847) were less common in patients with moderately to severely active CD compared with those with mildly active CD (31.18% vs. 51.72%, P=0.002; 55.91% vs. 75.86%, P=0.042). However, the opposite conclusion was drawn for the G allele (G) and AG+ GG genotypes of ICAM-1 (rs5498) (31.45% vs. 17.24%, P=0.027; 54.30% vs. 31.04%, P=0.020). Compared with patients with terminal ileal or ileocolic CD, G allele and AG+ GG genotypes of ITGA4 (rs6740847) were more prevalent in patients with colonic CD (55.26% vs. 29.38%, P<0.000 1; 84.21% vs. 53.11%, P<0.000 1). The same conclusion could also be drawn for the G allele and AG+ GG genotypes of ICAM-1 (rs5498) (42.11% vs. 26.84%, P=0.008; 73.69% vs. 46.33%, P=0.002). The frequency of homozygous GG genotype of ICAM-1 (rs5498) was lower in patients with stricturing and penetrating CD than those with non-stricturing and non-penetrating CD (0.00% vs. 12.32%, P=0.001). The G allele and AG+ GG genotypes of the ITGA4 (rs6740847) were more common in patients with perianal lesions than those without (40.28% vs. 30.77%, P=0.049; 72.22% vs. 51.75%, P=0.004). Conclusion:Variants of the ITGA4 (rs7562325) may be a risk factor for CD, whilst those of the ITGA4 (rs6740847) may be associated with the decline of disease activity and risk for colon involvement and perianal lesions. Variants of the ICAM-1 (rs5498) may increase the risk of disease activity and colonic involvement in CD patients, however, it may be a protective factor for stenosis and penetration. In addition, variants of the ITGA4 (rs6740847) and ICAM-1 (rs5498) may be associated with the early onset of CD.

Objective:To exploring the application effect of doctors-nurses-patients theory of goal attainment combined with StMarys cluster intervention mode in patients with tracheotomy after stroke.Methods:Using convenient sampling method, a total of 130 stroke patients with tracheostomy who were admitted to Shangqiu First People's Hospital from August 2020 to February 2022 were selected as the research objects. The patients were divided into the control group and the observation group using the random number table method, with 65 cases in each group. The control group received routine nursing care, while the observation group applied doctors-nurses-patients theory of goal attainment combined with StMarys cluster nursing intervention mode on the basis of the control group. After intervention, scores of Self Rating Anxiety Scale (SAS) and Self Rating Depression Scale (SDS) and patient nursing satisfaction were compared between the two groups.Results:After intervention, the SAS and SDS scores of the observation group were lower than those of the control group, and the nursing satisfaction was higher than that of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The doctors-nurses-patients theory of goal attainment combined with StMarys cluster intervention mode can help improve the psychological state of patients and increase nursing satisfaction.