Objective To investigate the protective effect of human amniotic mesenchymal stem cells(hAMSCs)on pulmonary microvascular endothelial cell(PMVECs)through observing the effects of prolifer-ation,apoptosis and inflammatory response after PMVECs injury induced by pine sawdust smoke solution.Methods HAMSCs and rat PMVECs were isolated and cultured.The flow cytometry and immunofluores-cence were used to identify hAMSCs and PMVECs respectively.The experimental grouping:control group(normal cultured PMVECs),smoke group(PMVECs injury induced by pine sawdust smoke solution),smoke+hAMSCs group(after PMVECs was injured by smoke solution,hAMSCs and PMVECs were co-cul-tured in Transwell culture system).The proliferative activity of PMVECs after co-culture for 12,24 h was measured by cell counting kit-8(CCK-8),the apoptosis of PMVECs was measured by flow cytometry,and the ex-pression levels of TNF-α and IL-6 were detected by enzyme-linked immunosorbent assay(ELISA).Results hAMSCs and PMVECs were successfully isolated and cultured,and the hAMSCs surface markers CD105(95.4％),CD73(99.8％)and CD90(99.8％)were identified as strongly positive expression,while CD34,CD45,CD14,CD19 and HLA-DR were weakly expressed(1.96％in total).The vascular endothelial cell mark-er CD34 in PMVECs was positively expressed,moreover its combination with aggulutinin BSI was also posi-tive.At the observation time point of 12,24 h co-culture,compared with the Control group,the proliferation activity of PMVECs in the Smoke group was inhibited(P＜0.05),and the cellular apoptosis was increased(P＜0.05),the TNF-α and IL-6 expression levels were up-regulated(P＜0.05);the phenomena of PMVECs proliferation activity inhibition,apoptosis increase and inflammatory factor expression level up-regulation in the Smoke+hAMSCs group were reversed compared with the Smoke group(P＜0.05).Conclusion After PMVECs are injured by smoke solution,hAMSCs could decrease the PMVECs inflammatory factors expres-sion,promote its proliferation activity and inhibit its apoptosis,thus play the protective effect on PMVECs.

Objective: To investigate the prevalence of glucose-6-phosphate dehydrogenase (G6PD) gene mutation in newborns of Hainan Province, and to provide reference for prevention and treatment of G6PD deficiency in this region. Methods: Neonatal disease screening filter paper dry blood spots from all midwifery agencies were collected in 18 cities (counties) in Hainan Province from January to June 2016, G6PD activity screening test was conducted at Neonatal Disease Screening Center, Hainan Women and Children's Medical Center, and the enzyme activity was normal when fluorescence value > 1 600, and enzyme deficiency when fluorescence value ≤1 600. Referred to "China's Major Birth Defects and Genetic Diseases Survey and Biological Resources Collection", women newborns (3 371 cases) with fluorescence values ≤2 200 and men newborns (1 620 cases) with fluorescence values ≤1 700 were included in the study. Molecular screening was performed using multicolor melting curve analysis (MMCA), and the distribution and mutation characteristics of G6PD gene mutants were analyzed. Results: Among the 4 991 newborns, 2 111 cases with G6PD gene mutation, and the detection rate was 42.30% (2 111/4 991). A total of 27 G6PD gene mutants were found, including 14 single mutants and 13 composite mutants. The mutation detection rates of c.1376 G>T (21.42%, 1 069/4 991), c.1388 G>A (11.54%, 576/4 991), c.95 A>G (2.40%, 120/4 991) and c.871 G>A (2.24%, 112/4 991) were higher. The mutation detection rate (61.54%, 136/221) of Baisha County (inland area) was the highest, and the mutation detection rate (22.15%, 64/289) of Wenchang City (coastal area) was the lowest. The mutation detection rate (49.88%, 628/1 259) of the Li nationality was the highest. A total of 873 female heterozygotes were detected, including 13 gene mutants, of which 44.67% (390/873) with normal enzyme activity. The c.86 C>T heterozygous mutation was first detected in a sample of Li female with normal enzyme activity. Conclusions Hainan Province is the high incidence area of G6PD gene mutation, the most common mutations are c.1376 G>T, c.1388 G>A, c.95 A>G and c.871 G>A in newborns. Inland area is the high incidence area, and Li nationality is the high incidence population.

Objective To investigate the treatment,outcomes and disease burden of severe hand-foot-and mouth diseases(HFMD),and to evaluate the compliance to the 2011 guideline for treatment in regions with a high in-cidence of HFMD.Methods A collaborative study group was established including Children's Hospital of Fudan Uni-versity,Jiangxi Provincial Children's Hospital,Anhui Provincial Children's Hospital,Linyi People's Hospital and the Second People's Hospital of Fuyang City.Clinical manifestation,treatment,prognosis and other data of severe HFMD pa-tients were prospectively collected by filling out a survey form in real time from April 2014 to October 2016. Results Two hundred and twenty-six severe HFMD cases were collected during the research time,including 114 ca-ses in stage 2,75 cases in stage 3,and 37 cases in stage 4.Two hundred and twenty-one cases(97.8%)were given mannitol,with a mortality of 6.2%;91 cases(40.3%)were given mannitol and glycerol fructose,with a mortality of 3.3%;the combined use of mannitol and glycerol fructose might have a better result than the single use of mannitol. One hundred and ninety-eight cases(87.6%)were given intravenous immunoglobulin(IVIG).One hundred and ninety cases(84.1%)were given antiviral drugs.One hundred and forty-five cases(64.2%)were given hormone therapy,and the use of hormone could reduce temperature,but did not reduce the mortality.One hundred and fifty cases (66.4%)needed vasoactive agent,including milrinone,dobutamine,phentolamine and sodium nitroprusside. The vasoactive agent use in stage 3 and 4 were 88.0%(66/75 cases)and 91.9%(34/37 cases),respectively.Sixty-nine cases(30.5%)received continuous positive airway pressure(CPAP),91 cases(40.3%)with mechanical ventila-tion,peak inspiratory pressure(PIP)≥20 cmH2O(1 cmH2O=0.098 kPa)accounted for 61.6%(53/86 cases),po-sitive end-expiratory pressure(PEEP)≥10 cmH2O accounted for 36.3%(33/91 cases).The mean mechanical ven-tilation time was(125.9 ± 101.8)h.Eleven cases(4.86%)died or died after giving up treatment,in which the mortality in stage 3 was 6.7%(5/75 cases),and the mortality in stage 4 was 16.2%(6/37 cases).The death causes were respiratory failure,pulmonary hemorrhage,and circulatory failure. The average time from onset to death was (5.91 ± 5.26)(1-15)d.Length of stay in hospital was(9.18 ± 5.16)(1-37)d in which length of stay in hospi-tal in stage 3 and 4 were(11.3 ± 6.35)d,(11.4 ± 6.62)d,respectively,which were significantly longer than that in stage 2[(7.50 ± 3.04)d],and the difference was statistically significant(P <0.05). The cost was(19 136 ± 12 556)CNY,of which the cost in stage 3 and 4 was(23 121 ± 13 846)CNY,(29 849 ± 16 015)CNY,respectively, which were significantly higher than that in stage 2[(12 961 ± 4 272)CNY],and the difference was statistically sig-nificant(P<0.05). Multivariate analysis found that respiratory rhythm abnormality,capillary refill time more than 3 seconds were risk factors for the deaths in the severe HFMD.Conclusion The 2011 edition of guidelines for treat-ment of children with severe HFMD was well executed.Hormone,IVIG,antiviral drugs did not significantly reduce the mortality of severe HFMD in children.

Objective To investigate the clinical features of severe hand,foot and mouth disease (HFMD) in recent three years,and to analyse the risk factors of severe HFMD.Methods A multicenter collaborative research group was set up,including five children's hospitals(pediatric department)with high incidence of HFMD.Prospective epidemiologic approaches were adopted.After training,staffs from the col-laborative center executed the study by filling up the record forms.Results We collected 114 HFMD cases in stage 2,75 cases of HFMD in stage 3,37 cases of HFMD in stage 4 from April 2014 to October 2016.The age ranged from 2 months to 13 years old,the median age was 2 years old,younger than 3 years accounted for 86.3%.Fever was observed in all the severe HFMD,mean temperature was (39.2 ± 0.7) ℃,fever lasted for (4.54 ± 2.89)d.The mean heart rates in stage 2,3,4 were (128.2 ± 13.3,176.1 ± 22.2,184.2 ± 27.5) times/min,respectively.The mean arterial pressure was (84.4 ± 14.6) mmHg(1 mmHg=0.133 kPa),the mean respiratory rate was (39.0 ± 8.4)times/min,the mean respiratory rates in stage 2,3,4 were (37.8 ± 7.36,38.7 ± 8.13,43.4 ± 10.7) times/min,respectively. Respiratory rhythm abnormality in stage 2,3,4 were 9.6%,14.9% and 56.8%,respectively.The blood glucose increased gradually in 2,3 and 4 stages,the mean blood glucose in stage 4 was(12.4 ± 4.74)mmol/L.The incidence of coma in stage 4 was higher than those in stage 2 and 3. Multivariate Logistic regression analysis found that tachycardia,drowsiness,coma, respiratory rate increase,respiratory rhythm abnormality,capillary refilling time more than 3 seconds were risk factors for severe HFMD.Conclusion The incidence of severe HFMD is still high in children under 3 years of age in last three years.The severe patients have obvious changes in the nervous,respiratory and circulatory system. Tachycardia,drowsiness,coma,respiratory rate increase,respiratoy rhythm abnormality,capillary refilling time more than 3 seconds are risk factors for severe HFMD.

Objective To study the levels of plasma catecholamine( norepinephrine,epinephrine and dopamine) in children with severe hand,foot and mouth disease( HFMD) ,and to assess the influence of cate-cholamine on the pathogenesis of severe HFMD. Methods A collaborative study group was established, including Children′s Hospital of Fudan University, Jiangxi Provincial Children′s Hospital, Anhui Provincial Children′s Hospital,Linyi People′s Hospital and No. 2 People′s Hospital of Fuyang City. Blood samples from children with severe HFMD were collected from April 2014 to October 2016. The levels of blood epineph-rine,norepinephrine,dopamine were measured at 2 h,24 h and 48 h after diagnosis for HFMD. Results The level of epinephrine at 24 h after diagnosis was ( 213. 0 ± 139. 8 ) ng/L in children with pulmonary edema, which was significantly higher than that of children without pulmonary edema[(137. 8 ± 45. 5)ng/L)](P=0. 022). The level of epinephrine at 24 h after diagnosis was (373. 2 ± 298. 1)ng/L in the dead children,and was (144. 2 ± 42. 5)ng/L in the survival children. The concentration of norepinephrine at 24 h after diagnosis was (1935. 7 ± 1824. 1)ng/L in the dead children,and was (858. 3 ± 212. 7)ng/L in the survival children. The levels of epinephrine and norepinephrine of those who died from HFMD were significantly higher than those of survival children at 24 h after diagnosis. There were no significant differences in catecholamine con-centrations among stage 2,3,4 HFMDs at 2 h,24 h and 48 h after diagnosis. Sex and enterovirus 71 virus infection had no significant influences on plasma catecholamine concentrations in children with severe HFMD. Conclusion Plasma epinephrine levels increase in children with HFMD complicated with pulmonary edema. Epinephrine and norepinephrine may play an important role in the death of children with severe HFMD.

Objective:To compare two flexible embryo catheters and determine whether clinical out-come differs in the in vitro fertilization-embryo transfer (IVF-ET)cycles.Methods:This prospective control study was conducted by one doctor between July 2012 and November 2013.In the study,2 064 patients undergoing fresh embryo transfer by using IVF-ET/intracytoplasmic sperm injection (ICSI)-ET in Reproductive Medical Center of Peking University Third Hospital were recruited.The subjects were di-vided into two groups.Cook Sydney IVF embryo transfer catheters (product model:K-JETS-7019-SIVF) were used for embryo transfer in group 1 (n =949),and FrydmanCCD catheters (product model:131230301)were used in group 2 (n =1 115).Pregnancy outcomes were compared between these two groups.Results:There was no significant difference in age,diagnosis for infertility and stimulation proto-col used between the two groups.In addition,there was no difference in the number of oocytes collected and in the number and score of embryos transferred.The significantly higher implantation rate,clinical pregnancy rate,and live birth rate (34.40% vs.26.92%,51.21% vs.41.52%,42.57% vs. 33.09%,P <0.05)were observed in group 1 compared with group 2.The abortion rate was not signifi-cantly different between the two groups (11.93% vs.15.98%,P >0.05).The proportion of difficult transfer was higher in group 1 than that in group 2 (5.27% vs.3.41%,P <0.05 ).There was no difference in the clinical pregnancy rate and live birth rate between the two difficult transfer cycles.Con-clusion:The type of embryo transfer catheter affects the clinical outcome in IVF.Good clinical outcome can be obtained by using Cook Sydney IVF catheter,which is worthy of clinical promotion.

Objective to provide the evidence for inducing the SAP model in rats with proper concentration of sodium taurocholate.Methods 60 SD rats were divided into sham operated group, group of 1.5% in concentration, group of 3.5% in concentration and group of 5% in concentration randomly, while the SAP model was induced by the sodium taurocholate concentration of 1.5%,3.5% and 5% with the method of retrograde injection into the biliopancreatic duct.To calculate the mortality of different groups, measure the serum amylase, tumor necrosis factor -α(TNF -α) and interleukin -6 (IL -6),and to observe the pancreatic pathological scores of HE staining in rats.Results The mortality in group of 5% in concentration has a significant ascending compared with group of 1.5% in concentration, while the serum amylase, tumor necrosis factor -α(TNF -α) , interleukin -6( IL -6), pathological score of hemorrhage and acinar necrosis in group of 5% in concentration have a significant ascending compared with group of 1.5% in concentration and group of 3.5% in concentration.Conclusions A better SAP model may be induced by sodium taurocholate with the concentration of 5% by the method of retrograde injection into the biliopancreatic duct, which may accord with the physiological and pathological manifestation of SAP.

Subclinical hypothyroidism during pregnancy is associated with some adverse outcomes during maternal pregnancy.The present study investigated thyroid function parameters measured by electroehemiluminescence (ECL) immunoassays in subclinical hypothyroid women treated with levothyroxine (L-T4) during pregnancy.The results showed that in evaluating thyroid function with ECL immunoassays during replacement with L-T4,determination of serum TT4 appears to have a closer correlation with TSH and may better reflect the effìcacy of treatment.

Objective To study the levothyroxine doses and related factors in the treatment of pregnant women with subclinical hypothyroidism (SCH).Methods Fifty-six pregnant women with SCH (diagnosed before 12 weeks of gestation) were recruited and divided into 2 groups according to the baseline TSH levels,SCH group 1 (2.5 mIU/L ≤ TSH ≤ 5.0 mIU/L,n =24) and SCH group 2 (TSH＞5.0 mIU/L,n =32).Thyroid autoantibodies [thyroid peroxidase antibody(TPOAb) and thyroglobulin antibody(TGAb)] were detected.All the subjects were treated with levothyroxine and the doses were adjusted according to the TSH level.The therapeutic target was to keep the TSH levels under control,0.3 to 2.5 mIU/L for the first trimester and 0.3 to 3.0 mIU/L for the second and third trimesters.Results There was a positive correlation between the levothyroxine doses and baseline TSH levels (r =0.533,P＜0.01) in pregnant women with SCH.A significant difference in the levothyroxine doses between SCH group 1 and SCH group 2 was found [(0.583 ± 0.341) vs (0.961 ± 0.405) μg/kg,t =-3.695,P＜ 0.01].The levothyroxine doses in SCH group 2 were 64.84％ higher than those in group 1.There was a significant difference in the levothyroxine doses between thyroid autoantibody negative and positive subjects [(0.680 ± 0.370) vs (0.918 ±0.440) μg/kg,t =-2.197,P =0.032].The levothyroxine doses in thyroid autoantibody positive subjects were 35 ％ higher than those in the negative subjects.In addition,there was a significant difference in the levothyroxine doses between subjects with negative and positive thyroid autoantibody [(0.421 ± 0.192) vs (0.720 ± 0.385)μg/kg,t =-2.331,P =0.029] in SCH group 1.While in SCH group 2,the difference did not reach statistical significance.Conclusion The baseline TSH levels and status of thyroid autoantibodies may affect the levothyroxine dosage in pregnant women with SCH.