【Objective】 To explore the efficacy of Mathieu combined with tongue-shaped flap covering in children with distal hypospadias and micropenis head deformity. 【Methods】 The clinical data of 72 patients with distal hypospadias complicated with micropenis treated during Jan.2018 and Jun.2021 were retrospectively analyzed, including 35 patients who underwent modified Mathieu combined with tongue-shaped flap and external urethral orificium (study group), and 37 patients who underwent traditional Mathieu (control group). Urethral stricture, urethral fistula, urethral diverticulum and penile head cleft were compared between the two groups. 【Results】 Urinary fistula occurred in 2 cases (5.27%) and 9 cases (24.32%) in the study group and control group, respectively, the incidence being much lower in the study group (P=0.028). Urethral stricture occurred in 1 case (2.86%) and 1 case (2.70%) in the study group and control group, respectively; penile head dehiscence in 1 case (2.86%) and 3 cases (8.11%); urethral diverticulum in 0 case (0%) and 2 cases (5.41%). There were no significant differences between the two groups in the incidence of urethral stricture, urethral dehiscence and urethral diverticulum (P>0.05). 【Conclusion】 Mathieu combined with tongue-shaped flap covering to treat children with distal hypospadias with micropenis head deformity can reduce the incidence of postoperative complications, achieve high surgical satisfaction and appearance satisfaction in the first phase, reduce harm caused by surgery, and promote patients’ psychological health.

Objective:By exploring the source of children's total fluoride intake and the relationship between children's total fluoride intake and dental fluorosis prevalence, to calculate the benchmark dose (BMD) of children's total fluoride intake and its 95% confidence lower limit (BMDL), and to provide the basis for revision of "Total Fluoride Intake for Inhabitants" (WS/T 87-2016).Methods:The villages that had water improvement for 5 years and more in 6 provinces of Jiangsu, Shandong, Hebei, Anhui, Henan and Shaanxi were selected as survey sites in April 2014. The water fluoride content of these villages was 0.3 - 3.0 mg/L, tap water samples from the centralized water supply were collected, and fluoride content was detected by ion selective electrode method. Children aged 8 to 12 years were selected, children's dental fluorosis was checked by Dean's method. Children's dietary and drinking water volume were surveyed by duplicate portion study (measurement for 3 d), and dietary fluoride content was detected according to the "Method for Determination of Fluorine in Foods". The mean and standard deviation of drinking water fluoride intake, dietary fluoride intake, and total fluoride intake were measured. According to the dose-response relationship between children's total fluoride intake and the detection rate of dental fluorosis, BMD and BMDL were calculated, and the reference dose (RfD) was calculated based on BMDL.Results:The mean of water fluoride of all 29 villages was 1.26 mg/L (from 0.41 to 2.85 mg/L). Totally 3 043 children aged 8 to 12 years were checked, and the detection rate of dental fluorosis was 30.2% (919/3 034). The lowest detection rate of dental fluorosis was 2.0% (2/100) and the highest was 71.4% (30/42) in the 29 villages. The children's dietary and drinking water volume of 769 person-time aged 8 to 12 years were surveyed, the children's daily drinking water fluoride intake, dietary fluoride intake, and total fluoride intake were (0.83 ± 0.66), (1.13 ± 0.61) and (1.96 ± 0.89) mg/d, respectively. The BMD of children's daily total fluoride intake was 2.43 mg, the BMDL was 2.21 mg, and the RfD was 2.21 mg.Conclusion:The BMD of 8 to 12 years old children's daily total fluoride intake is the same as the allowable limit (2.4 mg) of the national standard "Total Fluoride Intake for Inhabitants" (WS/T 87-2016).

Objective: To analyze the effect of NCCN (2015) risk stratification on prognosis of patients with acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: Retrospective analysis of 258 patients with AML in CR (186 cases in CR₁, 72 cases in CR₂) who underwent allogeneic HSCT in our hospital between April 2012 and March 2015 according to NCCN (2015) risk stratification. Of them, 63 cases were classified as low risk, 112 cases intermediate risk and 83 cases high risk. Results: ①With the median follow up of 18 (5-41) months, two-year disease free surviva (DFS) in 258 patients was 78.0% (95% CI 60.4%-96.6%) . Two-year DFS in AML after transplantation was 78.6% (95% CI 61.0%-96.2%) in low risk, 76.0% (95% CI 84.0%-93.6%) in intermediate risk and 80.3% (95% CI 62.7%-97.9%) (P=0.886) in high risk groups respectively. ②Univariate analysis showed that DFS has no significant difference in patient age, the median disease course before HSCT, the WBC number at the beginning of the disease, blood routine and chromosomes examination before transplantation, extramedullary disease before transplantation, disease status before transplantation, conditioning regimen, donor type, donor and recipient sex, recipient blood type, transfused MNC number, transfused CD34⁺ cell number and transfused CD3⁺ cell number. DFS was significant lower in primary AML than that in secondary AML (P=0.006) and also lower in MRD positive than that in MRD negative (P=0.003) . The accumulative relapse was significant higher in CR₂ compared to that in CR₁ (P=0.046) . Accumulative non-relapse mortality (NRM) was significanlyt higher in secondary AML compared to that in primary AML (P=0.004) and also higher in MRD positive compared to that in MRD negative (P=0.010) . ③Multivariate analysis showed that MRD positive was the only significant factor in DFS and NRM. Conclusion Allo-HSCT treatment of AML CR patients could achieve a high efficacy, which is similar between CR₁ and CR₂ patients. There is no significant correlation between NCCN (2015) risk stratification and the prognosis of AML patients with allo-HSCT treatment. Pre-conditioning MRD status monitored by multiparameter flow cytometry was the only impact factor on DFS and NRM in allo-HSCT for CR-AML patients.

Objective To compare selenium content in hair samples of people in Kaschin-Beck disease (KBD) areas and non KBD areas two months after stopping selenium salt in Shaanxi Province,to provide a scientific basis for KBD control and prevention.Methods In September 2012,four historical KBD areas were selected,including Yongshou,Yuyang,Linyou and Nanzheng,four villages were selected as monitoring sites according to the four directions as east,west,south and north in each county.Meanwhile,four non KBD areas were selected,including Wugong,Mizhi,Qishan and Chenggu,which adjacent to the survey counties and were similar to the survey counties in environment and production and living conditions.Four villages were selected as monitoring sites according to the four directions as east,west,south and north in each county.Lianhu District in Xi'an city was selected as a urban non KBD area.Four communities were selected in the east,west,south and north as the monitoring sites.In each monitoring point,hair samples of 8 children aged 7-12 years old (gender balanced) and 8 adults over the age of 16 (gender balanced) were selected to determine the hair selenium.Samples were disposed by wet digestion method,the selenium content was determined by 2,3-diaminonaphthalene fluorescence method.Results A total of 256 hair samples were collected in the four KBD counties,256 hair samples in four non KBD counties,64 hair samples in one urban district.The average of hair selenium in each monitoring point was ≥0.25 mg/kg.Compared the hair selenium content in KBD areas,rural non KBD areas,and urban non KBD areas [(0.40 ±0.23),(0.42 ± 0.28),(0.37 ± 0.38) mg/kg],the differences were not statistically significant (F =0.045,P ＞ 0.05).In KBD areas,the hair selenium content of 37 people was ＜ 0.20 mg/kg,accounting for 14.45％;28 people was 0.20-＜ 0.25 mg/kg,accounting for 10.94％;127 people was 0.25-＜ 0.50 mg/kg,accounting for 49.61％;64 people was ≥ 0.50 mg/kg,accounting for 25.00％.In non KBD areas,the hair selenium content of 67 people was ＜ 0.20 mg/kg,accounting for 20.94％,28 people was 0.20-＜ 0.25 mg/kg,accounting for 8.75％;143 people was 0.25-＜ 0.50 mg/kg,accounting 44.69％;82 people was ≥0.50 mg/kg,accounting for 25.63％.Compared the hair selenium content of children aged 7-12 and adults in KBD areas,rural non KBD areas,and urban non KBD areas [children:(0.45 ± 0.29),(0.47 ± 0.31),(0.33 ± 0.12) mg/kg;adults:(0.41 ± 0.25),(0.37 ± 0.25),(0.40 ± 0.49) mg/kg],the differences were not statistically significant (F =0.007,0.024,all P ＞ 0.05).Compared the hair selenium content in different gender in KBD areas,rural non KBD areas,and urban non KBD areas [maile:(0.43 ± 0.23),(0.43 ± 0.26),(0.40 ± 0.51) mg/kg;female:(0.38 ± 0.22),(0.41 ± 0.31),(0.34 ± 0.18) mg/kg],the differences were not statistically significant (F =0.872,3.589,all P ＞ 0.05).Conclusion Two months after stopping to supply selenium salt in 2012 in Shaanxi Province,the hair selenium content of residents in KBD areas has not dropped significantly.Since this survey is carried out only 2 months after taking the measure,it is necessary to continue to monitor the selenium level in KBD areas.

Objective To investigate the population and role of IL-10+ CD19+ regulatory B cell (Breg) in patients with chronic hepatitis B.Methods Patients with acute hepatitis B (AHB) (n =28),chronic hepatitis B (CHB) (n =31) and normal subjects (n =25) were collected from Changshu No.2 People's Hospital between 2011 June and 2012 October.Peripheral blood mononuclear cells (PBMC) were isolated and stimulated with CpG ODN 2006 and PMA.Flow cytometry was used to analyze the population of IL-10-CD19 + Breg,CD4 + CD25high Treg,and ELISA was used to analyze the concentration of IL-10 in culture supernatant.Results The population of Breg in Peripheral blood of the CHB group [1.28％ (1.05％-2.20％)] was higher than that in the AHB group [0.87％(0.55％-1.22％)] and the HCs group [0.89％ (0.51％-1.37％)] (P =0.001,0.006),and the difference between the AHB group and the HCs group was not statistically significant (P=0.669).Breg in the CHB group [14.30％ (10.70％-16.70％)] was higher than that in the AHB group [10.30％ (7.05％-13.30％)] and the HCs group [10.40％(6.85％-12.60％)] (P =0.003,0.001),treg in the CHB group [5.80％ (4.20％-9.10％)] was also higher than that in the AHB group [4.05％ (2.53％-5.40％)] and the HCs group [4.50％ (2.55％-5.50％)] (P ＜0.001,P =0.005),and there was no significantly difference between the AHB group and the HCs group (Breg:P =0.796 ; Treg:P =0.227).Spearman correlation analysis showed that Breg was positively correlated with Treg in the CHB group (r =0.50,P =0.004),however there was no significantly correlation in the AHB group and the HCs group (r =-0.15,P =0.462; r =0.09,P =0.669).The concentration of IL-10 in the CHB group was higher than that in the AHB group and the HCs group (P ＜ 0.001),and the difference between the AHB group and the HCs group was not statistically significant (P=0.341).Spearman correlation analysis showed that IL-10 were positively correlated with the population of Breg in the CHB group (r =0.409,P =0.022).Conclusion The poluations of regulatory B cell and regulatory T cell increased in patients with chronic hepatitis B,and Breg cell might play the immune regulation role through secreting IL-10 in chronic HBV infection.

Objective To investigate the correlation between immune inflammation and overactivity of T helper cells in childhood asthma by cell proliferation assay and activation induced cell death in vitro.Methods Th1/Th2/Th17 cytokines were determined by cytometric bead array.Cell proliferation and activation induced cell death were detected when CD4+ T cells were purified by magnetic beads and stimulated by PHA and antiCD3.At last,mRNA of Fas,FasL and Bcl-2 were mesured by real-time PCR.Results Cytokines of IL-4(2.451± 1.052ng/L vs 1.796 ±0.615 ng/L,P =0.018),IL-10( 1.920 ±0.813ng/L vs 1.390 ±0.162ng/L,P =0.006)and TNF(5.112 ±5.842 ng/L vs 1.506 ±0.551 ng/L,P =0.009) in sera of asthma group were higher than those in control group.Compared to control group,proliferation ability of CD4 + T cells in asthma group was greater ( OD450:0.498 ± 0.052 vs 0.274 ± 0.032,P ＜ 0.001 ) and apoptosis rate was lower( 35.62 ± 0.05 ％ vs 65.28±3.85％,P ＜0.001 ).mRNA expression of Fas in asthma group was lower but Bcl-2 was higher than those in control group.Conclusion It is implicated that defective expression of Fas and over expression of Bcl-2 in CD4+ T cells may contribute to apoptosis inhibition and cell proliferation,which could explain overeactivity of CD4 + T cells and lvmphocvte infiltration in childhood asthma.

Objective To investigate the roles of TLR3 pathway activiated by polyI:C in proliferation and apoptosis of multiple myeloma (MM) RPMI8226 cell line.Methods RPMI8226 cells were cultured in RPMI 1640 with different dose of polyl:C.Cells were collected in different time.Proliferation and apoptosis were detected by CCK-8 kit and flow cytometry,separately.Results The proliferation of RPM18226 was inhibited by polyI:C,and it was dose and time dependent,24 h:12.30％ ±2.04％,22.50％±2.20％,37.90％ ±1.30％ ; 48 h:17.80％ ±1.52％,29.60％ ±0.85％,45.80％ ±1.68％ ;72 h:25.10％±1.01％,34.60％±1.27％,60.50％±2.08％,P＜0.05.RPMI8226 cells were incubated with 50 μg/ml,100 μg/ml and 200 μg/ml polyI:C for 48 h.Apoptotic rate were 5.60％ ±1.06％,8.71％ ±1.06％ and 13.93％ ±1.17％,P＜0.05.TLR3 and TRIF mRNA expression increased obviously and dose dependent,TLR3:1.41±0.10,2.24±0.16,4.08±0.13; TRIF:1.07±0.16,1.97±0.13,3.56±0.19,P＜0.05.Conclusion The proliferation of MM cells were inhibited by TLR3 pathway obviously,and apoptosis was induced by polyI:C.

Objective To investigate a new therapeutic pathway for primary biliary cirrhosis (PBC) by immune tolerance reestablishment in a PBC mouse model. Methods Spleenic cells from naive mice were incubated with M2 in the presence of ECDI and the ceils were injected into caudal vein of the mice which would be used for development of PBC model. Spleenic cells incubated with bovine serum albumin (BSA) were injected as controls. 16 weeks later, anti-mitoehondrial antibody (AMA) , alkaline phosphatase(AKP) and portal inflammation were assayed for evaluating the prevention effect. Results AMA positive rate in tolerance group was lower than that in BSA and PBC groups ( P = 0. 007, P = 0. 003 ). The difference between BSA and PBC was not significantly. Serum AKP levels in tolerance, BSA and PBC group were (80.5 ±9.8) U/L, (93.8 ±15.7) U/L and (92.5 ±17.7) U/L, separately. The level in tolerance group was lower than that in BSA and PBC groups (P =0.0095, P =0.029). The rates of portal areas with cell infiltration were 42. 67% ± 12. 30% , 57. 07% ± 11. 35% and 51. 53% ± 9. 96% , separately. The number of infiltrated portal tracts in tolerance group was less than that in PBC group (P = 0.039) and BSA group (P = 0. 0024). Conclusion PBC was prevented to some extent by reestablishing immune tolerance to M2 autoantigen. This provides clues for finding a better treatment proposal.

ObjectiveTo investigate mechanisms for IL-27 induced proliferation and differentiation of peripheral blood CD4+ T cells in primary biliary cirrhosis (PBC).MethodsPeriperal blood CD4+ T cells were isolated from patients with PBC,chonic hepatitis B (CHB) and health controls (HCs).After IL-27 stimulation,proliferation ability of CD4+ T cells was evaluated by CCK-8 kit,and cytokines were analyzed by ELISA.Real-time PCR was employed to assay mRNA expression of T-bet and GATA3 in CD4+ T cells.p-STAT-1 and pSTAT-3 expression in CD4+ T cells were detected by Western blot.ResultsEnhanced proliferation of CD4+ T cells was found in all subjects after IL-27 stimulation.However,the proliferation ability in patients with PBC was greater than that in CHB and HCs ( P＜0.001 ).Levels of IL-2 and IFN-γ in supernatant from IL-27-incubated PBC blood CD4+ T cells were higher than that from CHB and HCs (P＜0.001 ).In normal situation,T-bet mRNA of CD4+ T cells in PBC group was higher than that in CHB group (P=0.007).Furthermore,after IL-27 stimulation,elevated T-bet mRNA expression and GATA3 inhibition were found in patients with PBC.High expression of p-STAT-1 and p-STAT-3 in blood CD4+ T cells were found in PBC,CHB and HCs after stimulation by IL-27.But their expression in patients with PBC were higher than those in patients with CHB and HCs.ConclusionProliferation of blood CD4+ T cells could be induced by IL-27 in patients with PBC.The signaling pathways of p-STAT-1,p-STAT-3 were involved to induce Th1 immune response and related cytokines expression.This study implicated that IL-27 may play important roles in early inflammation damage in PBC.

Objective To investigate the relationship between cardiac vagus nerves and changes of connexins(Cx)and intracellular gap junction(GJ)distribution pattern in superior vena cava(SVC)myosleeve in dog with atrial fibrillation(AF).Methods Twenty four hybrid dogs were divided into sham operation group(Sham group,n=8),SVC-AO fat pad removed group(RM group,n=8)and SVC-AO fat pad reserved group(RS group,n=8).In RM group and RS group,right atrial pacing was performed at a frequency of 500～650/min for 6 weeks to establish AF model.AF was induced by programmed stimulation or burst stimulation of atrial pacing.The expression and distribution of Cx40 and Cx43 in the SVC myosleeve tissue in three groups were analyzed by immunofluorescence staining.Transmission electron microscopy was used to observe the uhrastructural organization of gap junction(GJ).Results The rate of inducing sustained AF(＞ 15 min)in RS group was significantly higher than that in RM group (P ＜ 0.01).The expression of Cx40 and Cx43 in the SVC myosleeve in sham group and RS group were significantly higher than that in RM group(P ＜ 0.05).Furthermore,the expression of Cx40 and Cx43 in RS group were obviously higher than that in sham group(P ＜0.05).The ratio of end-to-end to side-to-side in RS group was lower than that in Sham group and RM group.Comparing with RM group,the channel of GJ became shorter and wider in RS group(P ＜0.05).Sarcomere was dissolved and mitochondrion showed vacuole degeneration in RS group.Conclusion The remodeling of Cx40 and Gx43 in SVC myosleeve tissue may be mediated by vagus nerves.It is conducive to the maintenance and stability of AF.However,this effect can be weakened by removing SVC-AO fat pad of canine.