Objective To study the expressions of cysteinyl aspartate specific proteinase 3 ( Caspase 3 ) and proliferating cell nuclear antigen ( PCNA ) in intestinal tissue of neonatal rats with necrotizing enterocolitis ( NEC ) , and the protective effect of glutamine ( Gln ) on NEC. MethodsThirty-six neonatal Sprague-Dawley ( SD) rats were randomly assigned into 3 groups at 48 h after birth (12 in each group). The control group were fed with milk replacer. The NEC group were fed with milk replacer and experiencing cold exposure after hypoxic-reoxygenation twice a day for 3 days, The Gln+NEC group were fed with milk replacer plus Gln and experiencing cold exposure after hypoxic-reoxygenation twice a day for 3 days. All the rats were sacrificed and intestinal tissues obtained at day 3 of the establishment of model. The histological changes of ileal tissues were studied using hematoxy lin-eosin ( HE ) staining. The expressions of Caspase 3 and PCNA were detected using immunohistochemical(IHC)method.Results Caspase3expressioninNECgroup(77.3±8.6)℅was significantly higher than the control group (18. 9 ± 3. 4)℅ and Gln+NEC group (50. 3 ± 6. 2)℅ ( P<0. 05). Also, Caspase 3 in Gln+NEC group was significantly higher than the control group (P<0. 05). PCNA expression in the NEC group ( 15. 0 ± 1. 9 )℅ was significantly lower than the control group (34. 2 ± 5. 8)℅ and the Gln +NEC group ( 24. 0 ± 3. 9 )℅ ( P <0. 05 ) . PCNA expression in the Gln+NEC group was significantly lower than the control group ( P<0. 05). The pathological score of the intestinal tissues was significantly correlated with Caspase 3 expression ( r = 0. 769, P = 0. 005 ), Caspase3/PCNA ratio (r=0. 835,P=0. 002) and PCNA expression (r= -0. 698, P=0. 014) in the NECgroup.Conclusions Up regulation of Cas pase3 and down regulation of PCNA might be correlated with the process of NEC. Gln might be effective in prevention and healing of NEC by inhibiting apoptosis and promoting cell proliferation.

Objectives To understand the effects of different doses of vitamin D supplementation on serum calcium,phosphorus,alkaline phosphatase and 25-hydroxyvitamin D levels in very low birth weight infants (VLBWI) and to provide guidance for early prevention of metabolic bone disease in VLBWI.Methods A total of 90 VLBWI hospitalized in the Department of Neonatology,Huzhou Maternal and Child Healthcare Hospital between January 2014 and January 2016 were enrolled and randomly divided into two groups:highdose group and low dose group.High-dose group was given vitamin D 900 U/d orally and low-dose group was given 400 U/d since the eighth day after birth.Serum calcium,phosphorus and alkaline phosphatase levels were detected at 1,7,21 and 42 days of age and serum 25-hydroxyvitamin D levels were detected at 7,21and 42 days of age.Two-sample t-test,Chi-square test,one-way analysis of variance and LSD or Dunnett's T3 test were used for statistical analysis.Results No significant differences in serum calcium,phosphorus and alkaline phosphatase levels were found between the two groups at 1 and 7 days of age,nor were found in serum 25-hydroxyvitamin D level at 7 days of age (all P＞0.05).At 21 days of age,high dose group had higher serum calcium,phosphorus and 25-hydroxyvitamin D levels than low-dose group [(2.38 ± 0.09) vs (2.04 ± 0.15) mmol/L,t=2.421;(1.80±0.50) vs (1.71 ±0.60) mmol/L,t-0.637;(45.58± 18.43) vs (42.53± 16.33) nmol/L,t=0.421],but lower alkaline phosphatase level [(505.12± 185.61) vs (588.32± 168.72) U/L,t=5.314] (all P＜0.05).The same trends were found at 42 days of age.In high-dose group,serum calcium level increased at 7,21 and 42 days of age as compared with that at 1 day of age [(2.43±0.13),(2.38±0.09),(2.39±0.08) vs (2.06±0.57) mmol/L];serum phosphorus level at 7 days of age was lower than that at 1,21 and 42 days of age [(1.31 ±0.26) vs (1.89±0.39),(1.80±0.50),(1.98±0.30) mmol/L];serum alkaline phosphatase level at 7,21 and 42 days of age was higher than that at 1 day of age [(475.18± 133.73),(505.12± 185.61),(538.43 ± 168.16) vs (296.15 ± 99.41) U/L] and a significant increase was observed at 42 days of age as compared with that at 7 days of age;serum 25 hydroxyvitamin D level at 21 days of age was higher than that at 7 days of age,and that at 42 days of age was higher than that at 7 and 21 days of age [(73.55±23.65) vs (30.63± 12.66) and (45.58 ± 18.43) nmol/L];the differences were all statistically significant (LSD or Dunnett's T3 test,all P＜0.05).Conclusions Vitamin D supplementation from the eighth day after birth can improve calcium and phosphorus metabolism in VLBWI and the dose of 900 U/d is more effective than 400 U/d.

ObjectiveTo investigate the effect of Glutamine (Gln) on the expression of PCNA in intestinal tissue of neo-natal rats with necrotizing enterocolitis (NEC), and to explore the protective mechanism of Gln in intestinal mucosa.Methods Forty-eight neonatal rats at the age of 48 hours were selected, and divided into 4 groups, control group, Gln group, NEC group, NECGln group. Each group had 12 rats. Control group were fed mice milk substitutes; Gln group were fed mice milk substitutes mixed with Gln; NEC group were fed mice milk substitutes and had cold/ hypoxia exposure twice a day for 3 days; NECGln group were exposed to cold stress, hypoxia and treated with Gln mixed in the milk. The expression of PCNA was detected using immunohistochemical method.Results Compared with control group were and Gln group, the general condition was worse, and the weight was decreased in NEC and NECGln group. The inifltrated inlfammatory cells, congestion, edema, intrinsic layer separation were observed in intestinal mucosa in NEC and NECGln group. The intestinal villus was lost in severe in NEC and NECGln group. The PCNA index was 34.17±5.78, 34.42±5.38, 15.00±1.94, 30.67±3.14 in control, Gln, NEC and NECGln group respectively, with signiifcant difference between each groups (H=24.32,P=0.000). The expression of PCNA in NEC group was lower than that in normal, Gln, and NECGln group (P<0.008). The expression of PCNA had no signiifcant difference among normal, Gln, and NECGln group (P>0.008).Conclusions The expression of PCNA in intestinal mucosa was decreased in NEC rats. Gln supplement could raise the expression of PCNA in intestinal mucosa of NEC rats, and accelerate the speed of intestinal mucosa repair.

OBJECTIVETo investigate the HLA-DRB1, DQB1 allele polymorphism in Kunming Yi nationality population.

METHODSHLA-DRB1, DQB1 DNA types in 70 healthy children of Yi nationality in Kunming were analyzed by polymerase chain reaction with sequence specific primer (PCR-SSP).

RESULTSTwelve alleles at HLA-DRB1 locus were observed in the 70 children: the alleles with gene frequencies higher than 10% were HLA-DRB1*12(33.57%), DRB1*0901(11.43%), DRB1*04(11.43%); the alleles with gene frequencies between 10% and 5% were HLA-DRB1*01(8.57%), DRB1*11(7.86%), DRB1*14(7.14%), DRB1*15(7.14%), DRB1*08(5%); the alleles with gene frequencies lower than 5% were HLA-DRB1*03(2.86%), DRB1*13(2.14%), DRB1*07(1.43%), DRB1*16(1.43%). Seven alleles at HLA-DQB1 locus were observed in the 70 children: the alleles with gene frequencies higher than 10% were HLA-DQB1*0301(45%), DQB1*05(22.14%), DQB1*0303(12.14%); the alleles with gene frequencies between 10% and 5% were HLA-DQB1*04(6.43%), DQB1*06(6.43%); the alleles with gene frequencies lower than 5% were HLA-DQB1*0201(4.29%) and DQB1*0302(3.57%).

CONCLUSIONThe distribution of HLA-DRB1, DQB1 allele polymorphism in the Kunming Yi nationality population is distinctive. It is neither like that in the South Han population nor like that in the North Han population.

Alleles ; China ; HLA-DQ Antigens ; genetics ; HLA-DQ beta-Chains ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Polymorphism, Genetic

Objective To investigate the clinical features and antimicrobial susceptibility of neonatal Listeria septicemia.Methods Eleven cases of neonatal Listeria septicemia that were treated in the Huzhou Maternity and Children Health Hospital from March 2013 to March 2017 were enrolled in this study.Clinical data including the results of bacterial culture,antimicrobial susceptibility test and antibiotic treatment were collected and retrospectively analyzed.Moreover,maternal history of Listeria monocytogenes (LM) infection before delivery was retrieved.Results All of the 11 mothers had fever before delivery and nine of them showed different grades of amniotic fluid contamination during delivery.Clinical symptoms of the 11 neonates included dyspnea (11 cases),fever (ten cases),apnea (nine cases),slow response and feeding difficulty (nine cases),convulsion (six cases),vomiting and abdominal distension (two cases),pulmonary hemorrhage (one case),progressive jaundice (one case) and systemic skin bleeding point (one case).All cases showed abnormal results of blood routine test and increased calcitonin and C-reactive protein.Ten cases received cercbrospinal fluid examination,seven of which were abnormal.Altogether 17 strains of LM were isolated from various specimens.These strains were all sensitive to piperacillin-tazobactam,ampicillin-sulbactam,meropenem,vancomycin,cotrimoxazole,ciprofloxacin and gentamycin,but resistant to oxacillin.Strains those were sensitive to penicillin,ampicillin,erythromycin and clindamycin accounted for 10/17,11/17,9/17 and 8/17,respectively.The 11 neonates were treated with piperacillin-tazobactam,meropenem or vancomycin.All of them improved (11/11)and ten were cured (10/11).All cases were followed up through phone calls at one week and one month after discharge.Two were lost to follow-up,while thc others were all in good condition.Conclusions Neonatal Listeria septicemia is usually a severe disease characterized by rapid progression and mainly presented with dyspnea and fever,besides there is a high possibility of purulent meningitis.Some LM strains are resistant to single-agent penicillin antibiotics.However,antibiotics such as piperacillin-tazobactam,meropenem and vancomycin are effective in the treatment of neonatal Listeria septicemia.

Objective:To investigate the effect of high flow nasal catheter ventilation (HFNC) in mechanical ventilation withdrawal of premature infants and its influence on blood gas analysis.Methods:The clinical data of 60 premature infants with mechanical ventilation from January 2017 to December 2018 in Huzhou Maternal and Child Health Hospital were analyzed retrospectively. After withdrawal, 30 premature infants treated with nasal continuous positive airway pressure (NCPAP) were as NCPAP group, and 30 premature infants treated with HFNC were as HFNC group. The ventilation time, total oxygen use time, complications, and arterial partial pressure of oxygen (PaO 2) and arterial partial pressure of carbon dioxide (PaCO 2) 12 and 24 h after withdrawal were compared between 2 groups. Results:There were no statistical differences in ventilation time and total oxygen use time between 2 groups ( P>0.05). The incidence of complications in HFNC group was significantly lower than that in NCPAP group: 13.33% (4/30) vs. 46.67% (14/30), and there was statistical difference ( P<0.05). The PaO 2 12 and 24 h after withdrawal in HFNC group was significantly higher than that in NCPAP group: (68.83 ± 2.76) mmHg (1 mmHg=0.133 kPa) vs. (64.79 ± 2.31) mmHg and (78.46 ± 3.32) mmHg vs. (74.72 ± 2.18) mmHg, the PaCO 2 was significantly lower than that in NCPAP group: (48.93 ± 2.51) mmHg vs. (52.31 ± 3.18) mmHg and (38.78 ± 4.23) mmHg vs. (43.67 ± 3.65) mmHg, and there were statistical differences ( P<0.05). Conclusions:HFNC is effective in mechanical ventilation withdrawal of premature infants, which can reduce complications and improve blood gas analysis.