Objective To evaluate intravesical instillation of epirubicin plus BCG to prevent recurrence of transitional cell cancer of bladder after surgical management. Methods A total of 44 cases of bladder cancer who underwent TURBT or partial cystectomy were divided into 2 groups.①Group 1 received intravesical instillation of epirubicin plus BCG (24 cases).Within 1 week following operation single dose of intravesical epirubicin instillation was given and from the second week regular intravesical BCG was instilled afterwards.②Group 2 received BCG alone (20 cases).Within 1 week after surgical management regular intravesical BCG instillation was given. All the patients were followed up for 24 months. Results In Group 2,3 cases had recurrence at 5,9,12 months individually,the incidence rate being 15% (3/20).No recurrence developed in Group 1.The recurrence rate was significantly different between the 2 groups (?2 test,P

Aim To construct human phage single-chain antibody library associated with esophageal cancer and to screen the specific scFv against Eca109 cells from the liberary. Methods Metastatic periesophageal lymph nodes of esophageal cancer patients were used as the B cells source, the total RNA of these B cells was extracted and prepared as the template of RT-PCR. First, we screened graticulely two pairs of primers of the heavy and light regions separately, then the V_H and V_L fragments were first amplified from the cDNA by the polymerase chain reaction (PCR). Second, the V_H-linker and V_L-linker were amplified from the V_H and V_L fragments. Last, the V_H-linker and V_L-linker were assembled into scFv gene fragments by SOE-PCR,and then Sfi I and Not I restriction site were inlet in it. ScFv gene was cloned into the pCANTAB-5E phagemid. Phagemids were introduced into E.coli TG1 by electrotransformation, followed by rescue of antibody-expressing phage using M13K07 helper-phage superinfection. Recombinant scFv phage library was constracted and PCR was used to identify the insert ratio of scFv antibodies library. Results of SfiI/Not I double digestion reaction positive insert clone were identified by 1.5% agarose gel electrophoresis. The phage library was panned with NHEEC and Eca109 cancer cells in suspension for four rounds. Strongly positive recombinant phage clones were used to infect E.coli HB2151. Expression of soluble scFv was induced by IPTG. Soluble scFv from periplasm were purified by affinity chromatography and identified by SDS-PAGE and Western blot. Cell ELISA , immunohistochemical staining and immunocytochemical staining were used to identify the activity of the soluble scFv. Results The result of agarose gel electrophoresis showed that total RNA of these B cells had two bands of 28 S and 18 S. The size of V_H fragment is about 450 bp,V_L fragment is about 350 bp and scFv is about 850 bp. The competence is 108 cfu??g-1 pUC18 DNA. Randomly digestive reac-tion showed that the positive insert ratio was 91.7% (22/24). After four rounds of panning, the fourth phage yield is 141 times as much as that of the first one. SDS-PAGE and Western blot showed that the MW of the soluble scFv was about 30 ku and the brand of 30 ku was stained. Immunohistochemical staining showed strong stainning of the tissue of esophageal cancer, but not the liver and gastric cancer tissue. Immunocytochemical staining showed significant staining of the esophageal cancer line Eca109. The result of cell ELISA assay revealed that soluble scFv had highly specific and could combined with Eca109 cells, but not with BGC-823 and NHEEC. Conclusion A human scFv phage display library associated with esophageal cancer has been constructed successfully and the specific scFv antibody against Eca109 has been identified from the liberary.

Objective To analyze quantitatively the histological components, and to observe the morphological changes of the transition zone in benign hyperplastic prostates (BPH) and adult normal prostates. Methods The quantitative morphometry of 40 BPH specimens obtained from suprapubic transvesical prostatectomy and 10 normal prostatic specimens (controls) were performed with computer-assisted image analysis system,and all the samples were stained with hematoxylin and eosin (HE) and the Masson trichrome.The area percentages of stromal (smooth muscle and connective tissue),glandular epithelial and luminal components,the average areas of acini,lumens and epithelia,and the heights of epithelial cells were quantitated,respectively. Results ①The mean area percentages of different components between control group and BPH group were as follows: smooth muscles,(23.83?8.53)% vs (35.35?8.33)% ( P 0.05);epithelia,(26.26? 7.45)% vs (17.76?4.61)% (P0.05).②The average areas of acini, epithelia and lumens of control group and BPH group were ( 0.087?0.028)mm 2 vs (0.062?0.030)mm 2 (P

Objective:To study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs.Methods:BABL/c mice were randomly divided into VLPs experimental group, alum adjuvant experimental group, PBS control group and alum adjuvant control group,the experimental group mice were intramuscular immunization with HBoV1 VP2 VLPs and HBoV1 VP2 VLPs added alum,control group mice were immunization with alum or PBS buffer,then to study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs by cellular and humoral immune strength.Results: Alum adjuvant decreased cellular immune response induced by HBoV1 VP2 VLPs(P<0.001),enhance the HBoV1 VP2 VLPs immuned serum IgG titer(P<0.05)and IgG activity(P<0.01).Conclusion:Alum adjuvant can enhance humoral immune response induced by HBoV1 VP2 VLPs,but weaken cellular immune response induced by HBoV1 VP2 VLPs,when HBoV1 VP2 VLPs used as a prophylactic vaccine it should add alum adjuvant,when used as a therapeutic vaccine,it should not add alum adjuvant.

Objective To observe the changes in CD4 + CD25 + regulatory T cells(Treg) and T helper 17 cells (Th17) cells' proportions in the peripheral blood in children with Kawasaki disease(KD) before and after the treatment,and to analyze the role of Treg/Th17 cell imbalance in the pathogenesis of KD.Methods Fifty-two children with acute KD(KD group) and 34 age-matched healthy children(healthy control group) were selected at Jiangxi Provincial Children's Hospital from April to December of 2014.Morning peripheral vein blood was collected from 2 groups:one before the treatment by Immunoglobulin and Aspirin,and the other 3 days after defervescence treatment.Flow cytometry was used to detect proportions of Treg cells and Th17 cells in the peripheral blood.The enzyme linked immunosorbent assay was used to detect the levels of interleukin(IL)-6,IL-10,IL-17,IL-23 and transforming growth factor(TGF)-β.Results Proportion of Treg cells in the acute KD group was remarkably lower than that in the healthy control group [(1.48 ± 0.21) ％ vs.(5.13-± 0.32) ％,t =28.41,P ＜ 0.05],but it was significantly increased after treatment,and there was a significant difference [(4.71 ± 0.36) ％ vs.(1.48 ± 0.21) ％,t =-23.32,P ＜ 0.05].Proportion of Th17 cells in the acute KD group was markedly higher than that in the healthy control group [(8.06 ± 0.48) ％ vs.(2.65 ± 0.50) ％,t =-23.47,P ＜ 0.05],which was significantly decreased after treatment [(3.04 ±0.35) ％ vs.(8.06 ± 0.48) ％,t =25.55,P ＜ 0.05].Compared with the healthy control group,the levels of serum IL-6,IL-17,IL-23 in the acute KD group were significantly increased before treatment,and there were significant differences [(34.53 ± 0.53) ng/L vs.(10.88 ± 0.83) ng/L,t =-72.36;(57.05 ± 0.78) ng/L vs.(14.29 ± 0.98)ng/L,t =-55.29;(45.18 ± 1.52) ng/L vs.(18.25 ± 1.08) ng/L,t =-43.27;all P ＜ 0.05],but after treatment the levels were significantly decreased [(14.94 ± 1.06) ng/L vs.(34.53 ± 0.53) ng/L,t =49.63;(27.64 ± 0.91)ng/Lvs.(57.05±0.78) ng/L,t =26.49;(24.50-±1.13) ng/L vs.(45.18-±1.52) ng/L,t =32.17;allP＜0.05].The levels of serum IL-10,TGF-β in the acute KD group significantly decreased than those of the healthy control group,and there were significant differences [(14.29-± 0.64) ng/L vs.(29.57 ± 0.87) ng/L,t =42.24;(16.88 ±-0.90) ng/L vs.(38.83 ±0.84) ng/L,t =53.51;all P ＜0.05],but after treatment the levels were significantly increased,and there were significant differences [(23.01-± 0.61) ng/L vs.(14.29-± 0.64) ng/L,t =-29.54;(33.47±-0.82) ng/Lvs.(16.88±-0.90) ng/L,t=-40.68;allP＜0.05].Conclusion Imbalance betweenTreg cells and Th17 cells may be an important cause for the immune disorder of KD,the changes in related cytokines are involved in the pathogenesis of KD.

Objective To investigate the viral prevalence of acute lower respiratory tract infection (ALRTI)in children. Methods Totally 1165 children with clinical diagnosis of ALRTI during the period from August 2007 to September 2008 were involved in our study. The nasopharyngeal aspirate specimen was collected from each patient. RT-PCRs were performed to detect common respiratory tract viruses including respiratory syncytial virus (RSV), rhinovirus (HRV), parainfluenza virus (PIV, type 1 -3 ), influzenza virus type A and B (IFA,IFB), and PCR was used to detect adenovirus (ADV). Results 783 patients were identified to have at least one kind of viral pathogens and the overall positive rate was 67.2%. The most common virus was RSV (27%), followed by HRV ( 17.4% ) and PIV3 ( 13. 9% ). The peak infection seasons were winter and autumn. The etiological spectrum of ALRTI varied in different age groups. Two or more viruses were identified in 284 out of 783 cases ( 36. 3% ). The mixed infection rate was high in infants under 1 year old (63.7%) while it decreased to 8. 5% in children older than 3 years of age. Conclusion RSV, HRV and PIV3 were the most predominant pathogens in children less than 1 year old. The peak infection seasons were winter and autumn. The infection rate and mixed infection rate in infants under 1 year old were highest. The most common style was RSV and HRV mixed infection.

Objective To explore the efficacy and safety of Tocilizumab in the treatment of systemic juvenile idiopathic arthritis (sJIA)in children.Methods Twenty-four sJIA patients were collected who were hospitalized at the Department of Rheumatology and Immunity,Jiangxi Provincial Children's Hospital from October 2015 to May 2016,and they received Tocilizumab combined with Methotrexate (MTX) treatment for 12 weeks.The clinical laboratory and physiological indices,including routine blood,liver and kidney function tests,number of joints with active arthritis,number of joints with limited range of motion,physicians and patients assessment of disease activity,childhood health questionnaire,erythrocyte sedimentation rate(ESR),C-reactive protein (CRP),and compliance rates of Pediatrics of American College of Rheumatology(ACR Ped) 30,50,70 were observed after 4,8 and 12 weeks of treatment,and the adverse reactions were recorded.Results After 4 weeks of treatment,the levels of white blood cells,platelet,ESR and CRP in 24 cases of sJIA significantly decreased compared with those of the patients before treatment [(15.1 ± 2.7) × 109/L vs.(24.2 ±3.5) × 109/L,(277 ±73) × 109/L vs.(368 ± 62) × 109/L,(25 ± 12) mm/1 h vs.(75 ± 15) mm/1 h,(20 ± 13) mg/L vs.(64 ± 1) mg/L],and the differences were statistically significant (t =10.08,4.65,70.71,26.78,all P ＜0.05);the hemoglobin was increased dramatically[(110 ± 12) g/L vs.(98 ± 10) g/L],and the difference was statistically significant(t =-3.76,P ＜ 0.05).The compliance rates of ACR Ped 30,50,70 after 4 weeks of treatment were 82％,74％,68％,and they were continuously improved after 8 weeks of treatment (90％,82％,78％)and 12 weeks of treatment (98％,93％,92％),and the differences were all statistically significant (F =7.11,7.29,8.86,all P ＜0.05).The levels of IL-6 after 12 weeks of treatment had no significant change compared with those of the patients pre-treatment [(10.8 ±2.5) ng/L vs.(12.7 ±3.0) ng/L,t =1.96,P ＞0.05].Conclusion Tocilizumab is effective and safe in the treatment of sJIA patients,which can improve the symptoms,signs and laboratory inflammatory activity indexes of sJIA in a short time.

Objective To investigate the characteristics and liver function of the population with occupational exposure to hepatotoxicants. Methods A total of 17 093 workers with occupational hepatotoxicants exposure who underwent occupational medical examination during their employment in a occupational medical examination institution of Shanghai in 2021 were selected as the research subjects by judgement sampling method. Occupational medical examination data were collected, and the prevalence of abnormal liver function and fatty liver were analyzed. The association between hepatotoxicants exposure and abnormal liver function were analyzed. Results The median and the 0th-100th percentiles of the duration of exposure to hepatotoxicants was 6.5(1.0-42.0) years. The prevalence of fatty liver was 48.4% and the incidence of abnormal liver function was 23.7%. Among the workers with fatty liver, the prevalence of abnormal liver function was higher in workers exposed to metals, metalloids and their compounds than in unexposed workers (33.9% vs 30.0%, P<0.01). The results of multivariate logistic regression analysis demonstrated that the risk of abnormal liver function increased with the number of different hepatotoxicants mixed exposures (all P<0.01), after correcting for confounding factors including gender, age, years of exposure, marital status, drinking, hypertension, fatty liver and blood sugar. Conclusion Exposure to hepatotoxicants is a risk factor for abnormal liver function. The more diverse types of hepatotoxicants an individual is exposed to, the stronger the association with this risk.

Objective To establish the method for detecting human Rhinovirus (HRV) subtypes A,B,and C by reverse real-time fluorescent quantitative polymerase chain reaction (RT-PCR) and to assay HRV nasopharyngeal samples of children with acute respiratory tract infections.Methods The specific primers and Taqman probes of targeted HRV gene were designed,RNA standards were prepared to establish a standard curve,the sensitivity,specificity and reproducibility has been tested,222 clinical respiratory specimens were retrospectively detected.Results The linear ranges of human Rhinovirus A,B,and C subtypes were 109-10copies/μl,109-10 copies/μl,109-103copies/μl respectively,correlation coefficients were 0.999,0.997,0.999.Variation within the group was less than 3％.The detection rate was 27.02％ by RT-PCR,the sensitivity and specificity were 100％ and 98.18％.Conclusion The RT-PCR method can simultaneously detect rhinovirus subtypes A,B,and C with good sensitivity,specificity and reproducibility.

Objective To investigate the epidemiological features and the types of human adenovirus (HAdV) in hospitalized children with acute lower respiratory tract infection (ALRTI) in Changsha from 2013 to 2014.Methods Six hundreds and three nasopharyngeal aspirates were collected from the children with ALRTI,who were hospitalized at the pediatric of People' s Hospital of Hunan Province from April 2013 to March 2014.Eleven kinds of respiratory virus were firstly detected by Real-time PCR,the HAdV-positive samples are typed by Nested-PCR based on the sequence of hexon gene.The sequences were analyzed and compared with GenBank data.Results Adenovirus was detected in one hundreds and nineteen samples from six hundreds and three samples and the rate is 19.73％.Among the eighty eight samples that successful classified,twenty seven samples for HAdV-7 (30.68％),seventeen for HAdV-2 (19.32％),fourteen for HAdV-3 (15.91％),fourteen for HAdV-1 (15.91％),six for HAdV-5 (6.82％),3 for HAdV-6(3.41％),four for HAdV-4(4.55％),two for HAdV-57(2.27％),one for HAdV-14 (1.14％).HAdV infection could occur in any season,and the peak season was summer.The vulnerable groups for HAdV were younger than seven years old.Conclusions HAdV is one of the most important pathogens of acute lower respiratory infection in children in Changsha,the most prevalent types of adenovirus were HAdV-7 and HAdV-2.The pathogenicity of HAdV-3 and HAdV-7 in B species was stronger than other types in C and E species.