1.Protective effects of valproic acid on gut barrier function after major burn injury and its mechanism
Hongmin LUO ; Sen HU ; Huining BIAN ; Shaoyi ZHENG ; Bing XIONG ; Zhifeng HUANG ; Zu'an LIU ; Chuanwei SUN ; Lianghua MA ; Hanhua LI ; Wen YU ; Minghua DU ; Huada CHEN ; Wen LAI
Chinese Critical Care Medicine 2017;29(3):221-227
Objective To investigate the potential protective effects of valproic acid (VPA) on gut barrier function after major burn injury in rats and its mechanism.Methods Forty male Sprague-Dawley (SD) rats were divided into sham + normal saline (NS),sham + VPA,scald + NS,and scald + VPA groups,with 10 rats in each group.Rat with 55% total body surface area (TBSA) third-degree severe-bums model was reproduced by immersing into 80 ℃ water,and the rats in sham groups were given sham-bums by immersing into 37 ℃ water.The rats after severebums were immediately treated with 0.25 mL of 300 mg/kg VPA or NS by subcutaneous injection.Rats were sacrificed at 2 hours and 6 hours after injury,and abdominal aortic blood and ileal tissue were harvested.The levels of vascular endothelial growth factor (VEGF) were determined by enzyme-linked immunosorbent assay (ELISA).The intestinal permeability was evaluated by fluorescein isothiocyanate-dextran (FITC-dextran) determination.The histomorphological changes in gut barrier were evaluated by Chiu grading system.Levels of acetylated lysine at the ninth position of histone 3 protein (Ac-H3K9),hypoxia-inducible factor 1α (HIF-1α),zona occludens 1 (ZO-1) and myosin light chain kinase (MLCK) were determined by immunofluorescence staining and Western Blot.Results Compared with sham + NS group,rats in scald + NS group showed intestinal mucosal damage 2 hours after bum injury,as well as increased mucosal permeability,protein expression levels of HIF-1 α,VEGF,MLCK,and lowered levels of AC-H3K9 and ZO-1.These changes were much more prominent at 6 hours after injury.VPA treatment significantly attenuated the bum-induced intestinal damage.Compared with scald + NS group,the protective effects in scald + VPA group was not evident at 2 hours after injury;however,intestinal damage was much less severe at 6 hours after injury (Chiu score:2.03 ± 0.27 vs.3.12 ± 0.15),intestinal permeability was significantly decreased [FITC-dextran (μg/L):709 ± 76 vs.1138 ± 75],histone acetylation was enhanced [Ac-H3K9 (gray value):1.55 ± 0.12 vs.0.48±0.12],ZO-1 degradation was significantly inhibited (gray value:0.69 ± 0.12 vs.0.43 ± 0.16),the protein expression levels of VEGF and MLCK were significantly down-regulated [VEGF (ng/mg):51.7±3.7 vs.71.2±4.3,MLCK (gray value):1.98±0.20 vs.2.80±0.24],while the HIF-1 α protein expression levels were significantly reduced at both 2 hours and 6 hours after injury (gray value:2.50±0.39 vs.3.88±0.42 at 2 hours,1.83±0.42 vs.4.42±0.41 at 6 hours,all P < 0.05).Conclusions Severe bum injury can induce histone deacetylation,ZO-1 degradation and intestinal barrier dysfunction.VPA can improve the levels of histone acetylation and ZO-1,and protect intestinal epithelial barrier function.These may probably be mediated through inhibiting HIF-1α and its downstream gene VEGF and MLCK.
2.Inhibitory effects of Salvia miltiorrhiza injection coordinated with dexamethasone on interleukin-13 and eotaxin expression in lung of asthmatic rats.
Dan LI ; Sheng-dao XIONG ; De-bing DU
Chinese Journal of Integrated Traditional and Western Medicine 2006;26(11):1007-1010
OBJECTIVETo investigate the molecular mechanism of inhibitory effect of Salvia miltiorrhiza Injection (SMI) coordinated with dexamethasone (DXM) on allergic airway inflammation in asthmatic rats.
METHODSForty SD rats were randomly divided into 5 groups equally: the normal group, the asthma model group, the DXM group, the SMI group and the DXM + SMI group, they were treated with correspondant herbal medicines. Pathologic changes of lung tissue were obseved with HE stain, count of WBC and eosinophil (Eos) in bronchoalveolar lavage fluid (BALF) were estimated and the expressions of interleukin-13 (IL-13) and Eotaxin in lung tissue were measured by RT-PCR and SP method of immunohistochemistry assay.
RESULTSThere was moderate inflammation in lung tissue in the SMI group, and mild inflammation in the DXM + SMI and the DXM group, which was similar to that in the normal group. Compared with the asthma model group, Eos and WBC count in BALF and the expression of IL-13 and Eotaxin in the lung tissue were significantly lower in the three treated groups (P < 0.05), particularly in the DXM + SMI group, showing a significant difference as compared with the other two groups (P < 0.05 or P < 0.01). Additionally, IL-13 expression was positively correlated with Eotaxin expression (r = 0.92, P < 0.01).
CONCLUSIONSMI could inhibit the expression of IL-13 and Eotaxin in the lung of asthmatic rats, showing inhibitory effects synergistic with DXM on airway inflammation.
Animals ; Anti-Inflammatory Agents ; administration & dosage ; pharmacology ; Asthma ; drug therapy ; genetics ; metabolism ; Chemokine CCL11 ; biosynthesis ; genetics ; Dexamethasone ; administration & dosage ; pharmacology ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Eosinophils ; drug effects ; metabolism ; Immunohistochemistry ; Injections, Intraperitoneal ; Interleukin-13 ; biosynthesis ; genetics ; Lung ; drug effects ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Salvia miltiorrhiza ; chemistry
3.Evolution and prognosis of the acute flaccid paralysis associated with enterovirus 71 infection evaluated through a clinical and magnetic resonance imaging follow-up study.
Bing-wei PENG ; Zhi-hong DU ; Xiao-jing LI ; Hai-sheng LIN ; Hong-sheng LIU ; Wen-xiong CHEN ; Jian-ning MAI ; Hui-ci LIANG
Chinese Journal of Pediatrics 2012;50(4):255-260
OBJECTIVETo explore the development and prognosis of the acute flaccid paralysis (AFP) associated with enterovirus 71 (EV71) infection through clinical follow-up study for clinical and magnetic resonance imaging (MRI) features based on the research progress of virology and pathology.
METHODSixteen children with HFMD associated with AFP in hospital from May 1, 2011 to August 31, 2011 were investigated and the patients received intensive rehabilitation training. The 16 cases were divided into two groups (the recovery or the sequela) by if the muscle strength recovered to level 4 after intensive rehabilitation. The MRI findings of 15 children were analyzed and among them, 6 patients were reexamined after one month. The clinical markers were compared between groups including course of disease, WBC, WBC in cerebrospinal fluid (CSF), ventilator support, therapy, the worst muscle strength, the initial tendon reflex, the muscle atrophy, and multi-limb paralysis. The data were analyzed by t test and χ2 test with SPSS10.0.
RESULTAll the 16 children were infected with enterovirus 71 (EV71). The myodynamia of 7 children were level 0, 4 children had serious upper limbs paralysis. The neck muscle in 3 cases and the brain stem motor ruckus in 4 cases were involved. The ankle clonus of non-completely paralyzed limbs in 14 cases occurred during rehabilitation. Eight children had the better prognosis, the other 8 children had sequela. 0 level muscle strength (0 case vs. 7 cases, χ2=12.4), the initial tendon reflex (2 cases vs. 8 cases, χ2=9.6), obvious muscle atrophy (0 case vs. 8 cases, χ2=16), were significantly different in the children with the recovery when compared to the sequela (P<0.01). The severe upper limbs paralysis had the worse prognosis than the severe lower limbs paralysis. MR imaging showed signs of spinal nerve root inflammation and the bilateral hyperintense lesions, symmetrical in the posterior portions of the medulla, pons, and asymmetrical in the ventral horns of cervical spinal cord. Signal enhancement occurred only in the early MRI examination.
CONCLUSIONIn the evolution of AFP due to EV71 infection, the upper motor neuron damage is common, the prognosis is related with the severity of early paralysis and neuron damage. MR imaging is helpful to understand the pathological mechanism of AFP.
Child, Preschool ; Enterovirus A, Human ; pathogenicity ; Female ; Follow-Up Studies ; Hand, Foot and Mouth Disease ; diagnosis ; pathology ; virology ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Paralysis ; diagnosis ; pathology ; virology ; Prognosis
4.Protective effects of 3'-methoxy-puerarin on rat brain suffering from ischemia.
Yi-Bing ZHANG ; Gui-You DU ; Yu-Lan XIONG ; Yong ZHAO ; Hai-Feng CUI ; Chun-Yu CAO ; Sha LIU
China Journal of Chinese Materia Medica 2008;33(5):537-540
OBJECTIVETo research on the protective effect of 3'-methoxy-puerarin (3'-mo-pue) on rat brain suffering from ischemia and to offer the experimental basis for widening the applied areas of Radix Puerariae.
METHODAll rats were randomly divided into sham operation group, control group, Pue group and 3'-mo-pue group. The influence of different dosage on relevant indexes were measured through cerebral artery occlusion (MCAO) model rat covered by Fecl3, and cerebral ischemia-reperfusion model rat after 2VO.
RESULTThe result showed that 3'-mo-Pue could reduce the scores of neurological deficit, cerebral infarcted zone to varying degrees and the water content of brain tissue in MCAO model. It could obviously increase the activity of CAT and GPx in the hippocampus and also increase the activity of CAT and GPx in the cortex in the ischemia field. Moreover, 3'-mo-Pue could decrease the content of LPO, LD in the brain tissue of cerebral ischemia-reperfusion model rat.
CONCLUSION3'-mo-Pue has favorable protective effect on brain suffering from ischemia. The mechanism is possibly related to its effect of anti-oxidation.
Animals ; Brain ; drug effects ; enzymology ; metabolism ; Brain Ischemia ; drug therapy ; metabolism ; prevention & control ; Catalase ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Enzyme Activation ; drug effects ; Glutathione Peroxidase ; metabolism ; Isoflavones ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar
5.Autoantibodies against alpha1 adrenergic receptor related with cardiac remodeling in hypertensive patients by clinical observation.
Zheng-zai LI ; Yu-hua LIAO ; Zi-hua ZHOU ; Yu-miao WEI ; Min WANG ; Feng ZHU ; Ming CHEN ; Xiong-bing DU ; Yan-xiang SUN ; Hai-yan ZHANG
Chinese Journal of Cardiology 2006;34(7):602-604
OBJECTIVETo investigate the effects of autoantibodies against alpha(-) adrenergic receptor on cardiac remodeling in patients with hypertension.
METHODSFive hundred and fifty three patients with hypertension in our hospital were selected. The autoantibodies against alpha(1) adrenergic receptor in sera of donor were detected by ELISA, and the results of echocardiography were recorded. By multiple logistic regressions, the risk factors were analyzed on left ventricular enlargement of hypertension.
RESULTSThe percentage of autoantibodies against alpha(1) adrenergic receptor positive was 32.3% (179/553). There were significant difference between the positive group and negative group on the ratio of left atrial enlargement (53.6%, 44.3%, respectively; P < 0.05) and left ventricular enlargement (12.8%, 6.1%, respectively; P < 0.01). The result of regression analysis demonstrated that 4 risk factors were related to left ventricular enlargement, including male, course of disease, heart rate (HR) and autoantibodies against alpha(1) adrenergic receptor in the serum (all P < 0.05).
CONCLUSIONSThe autoantibodies against alpha(1) adrenergic receptor have a relationship with left ventricular enlargement of hypertension. Patients with the activity of autoantibodies against alpha(1) adrenergic might contribute to predict cardiac remodeling.
Adult ; Aged ; Autoantibodies ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Hypertension ; immunology ; physiopathology ; Logistic Models ; Male ; Middle Aged ; Receptors, Adrenergic, alpha-1 ; immunology ; Ventricular Remodeling ; immunology
6.Humoral immune responses in rabbits induced by an experimental inactivated severe acute respiratory syndrome coronavirus vaccine prepared from F69 strain.
Chuan-Hai ZHANG ; Zhong-Min GUO ; Huan-Ying ZHENG ; Jia-Hai LU ; Yi-Fei WANG ; Xin-Ge YAN ; Yong ZHAO ; Xiong-Wei DU ; Xin ZHANG ; Ling FANG ; Wen-Hua LING ; Shu-Yuan QI ; Xin-Bing YU ; Nan-Shan ZHONG
Chinese Medical Journal 2004;117(11):1625-1629
BACKGROUNDThe etiologic agent of severe acute respiratory syndrome (SARS) has been confirmed to be a novel coronavirus (CoV), namely SARS-CoV. Developing safe and effective SARS-CoV vaccines is essential for us to prevent the possible reemergence of its epidemic. Previous experiences indicate that inactivated vaccine is conventional and more hopeful to be successfully developed. Immunogenicity evaluation of an experimental inactivated SARS-CoV vaccine in rabbits was conducted and reported in this paper.
METHODSThe large-scale cultured SARS-CoV F69 strain was inactivated with 0.4% formaldehyde and purified, then used as the immunogen combined with Freund's adjuvant. Eight adult New Zealand rabbits were immunized four times with this experimental inactivated vaccine. Twelve sets of rabbit serum were sampled from the third day to the seventy-fourth day after the first vaccination. The titers of specific anti-SARS-CoV IgG antibody were determined by indirect enzyme-linked immunosorbent assay, and the neutralizing antibody titers were detected with micro-cytopathic effect neutralization test.
RESULTSRapid and potent humoral immune responses were induced by the inactivated SARS-CoV vaccine in all the eight test rabbits. Titers of both specific IgG antibody and neutralizing antibody peaked at about six weeks after first vaccination, with the maximum value of 1:81 920 and 1:20 480, respectively. After that, serum antibody levels remained at a plateau or had a slight decrease, though two boosters were given in the succedent 4 to 5 weeks. Cross neutralization response existed between SARS-CoV F69 strain and Z2-Y3 strain.
CONCLUSIONSThe inactivated SARS-CoV vaccine made from F69 strain owns strong immunogenicity, and the cross neutralization response between the two different SARS-CoV strains gives a hint of the similar neutralizing epitopes, which provide stable bases for the development of inactivated SARS-CoV vaccines.
Animals ; Antibodies, Viral ; blood ; Immunoglobulin G ; blood ; Neutralization Tests ; Rabbits ; SARS Virus ; immunology ; Vaccines, Inactivated ; immunology ; Viral Vaccines ; immunology
7.Progressive increase of serum circulating immune complexes and its significance in patients during the progression from chronic hepatitis B to hepatocellular carcinoma.
Jun DU ; Ping TIAN ; Tao-yang CHEN ; Jian-xiong WU ; Jin-bing WANG ; Yong WEI ; Li-ming WANG ; Li-guo LIU ; Wan-sheng JI ; Chun-feng QU
Chinese Journal of Oncology 2011;33(12):905-910
OBJECTIVETo investigate the significance of increasing circulating immune complex (CIC) in patients during the progression from chronic hepatitis B to hepatocellular carcinoma (HCC).
METHODSSerum levels of CIC from 20 hospitalized patients diagnosed by pathology with primary HCC, and 13 with hepatic hemangioma, and from 45 subjects with chronic HBV infection who finally developed into HCC (45 cases), and age- and gender-matched 45 subjects who kept the chronic HBV infection after consecutively followed up for 10 - 13 years by June of 2009 were quantified by ELISA. The serum levels of anti liver-kidney microsomal (anti LKM-1) antibodies were also measured by ELISA, and that of HBV-DNA were quantified by Taqman probe-based real time PCR in the followed up chronic HBV infection subjects. In the 45 chronic HBV subjects who finally developed into HCC and the 45 controls, serum samples were collected and determined at 3 time points: the baseline when the subjects were recruited, the middle point during the follow-up, and the end of follow-up.
RESULTSThe serum level of CIC was significantly higher in the 20 HCC patients than that in the 13 hemangioma cases (P < 0.001). When HCC was diagnosed, the CIC concentration was significantly higher than that in the baselines (P < 0.001) in the 45 chronic HBV subjects who finally developed into HCC after the consecutively follow-up for 5 - 13 years. Of them, 36 patients (80.0%) showed progressively increased CIC during the follow-up (P < 0.001). In the controls, the CIC levels were kept relatively stable during the follow-up. Among them, 17 patients (37.8%) showed CIC slightly increased (P = 0.046). Kaplan-Meier survival analysis indicated that elevated serum CIC during the follow-up increased cumulative HCC incidence (HR = 2.77, 95%CI 1.47 - 5.22). In addition, the serum levels of anti-LKM-1 and HBV-DNA were also significantly higher in the patients who finally progressed into HCC than that in the controls and maintained at a high level during the follow-up tested at all the 3 time points. Further analysis indicated that the serum level of CIC was correlated with that of serum HBV-DNA only when HCC was diagnosed (r = 0.344, P = 0.026).
CONCLUSIONProgressive increase of serum CIC level may be one of risk factors reflecting HCC development from chronic HBV infection.
Antigen-Antibody Complex ; blood ; Autoantibodies ; blood ; Carcinoma, Hepatocellular ; immunology ; virology ; DNA, Viral ; blood ; Disease Progression ; Female ; Follow-Up Studies ; Hemangioma ; immunology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; complications ; immunology ; Humans ; Liver Neoplasms ; immunology ; virology ; Male ; Middle Aged ; Risk Factors
8. Synchronous lymphoma and carcinoma-clinical analyses of 17 patients
Yuanyuan XIONG ; Zhijuan LIU ; Lin CHEN ; Fangfang YUAN ; Qingsong YIN ; Ruihua MI ; Bing ZHANG ; Jianwei DU ; Qinglan ZHANG ; Quande LIN ; Li’na ZHANG ; Xue GAO ; Lihua DONG ; Yufu LI ; Yongping SONG ; Xudong WEI
Chinese Journal of Hematology 2018;39(4):277-280
Objective:
To explore the clinical features of patients with synchronous lymphoma and carcinoma.
Methods:
The clinical data of 17 patients with Synchronous lymphoma and carcinoma from February 2012 to October 2017 were analyzed retrospectively.
Results:
Among 17 patients of lymphoma, 1 case HL, 2 cases B-NHL, 6 cases MZBL, 3 cases DLBCL, 1 case mantle cell lymphoma (MCL) , 3 cases NK/T- cell lymphoma, 1 case anaplastic large cell lymphoma(ALCL). In terms of 17 patients with carcinoma, 3 cases esophageal carcinoma, 3 cases gastric carcinoma, 2 cases colorectal carcinoma, 7 cases thyroid carcinoma, 1 case hepatocellular carcinoma and lung cancer. Up to 15 patients received operation, and some of them combined with chemotherapy, radiotherapy and autologous transplant. Follow-up analysis showed that 3 cases was undergoing treatment, 2 cases lost follow-up, 4 cases died, 3 cases achieved CR, 3 cases remained to be at SD, and 2 cases assessed for progression or recurrence.
Conclusion
The relationship between lymphoma and carcinoma was under discussion, patients with synchronous lymphoma and carcinoma were not unusual. We herein should raise awareness to avoid misdiagnosis.
9.Synchronous lymphoma and carcinoma-clinical analyses of 17 patients.
Yuan Yuan XIONG ; Zhi Juan LIU ; Lin CHEN ; Fang Fang YUAN ; Qing Song YIN ; Rui Hua MI ; Bing ZHANG ; Jian Wei DU ; Qing Lan ZHANG ; Quan De LIN ; Li Na ZHANG ; Xue GAO ; Li Hua DONG ; Yu Fu LI ; Yong Ping SONG ; Xu Dong WEI
Chinese Journal of Hematology 2018;39(4):277-280
Objective: To explore the clinical features of patients with synchronous lymphoma and carcinoma. Methods: The clinical data of 17 patients with Synchronous lymphoma and carcinoma from February 2012 to October 2017 were analyzed retrospectively. Results: Among 17 patients of lymphoma, 1 case HL, 2 cases B-NHL, 6 cases MZBL, 3 cases DLBCL, 1 case mantle cell lymphoma (MCL) , 3 cases NK/T- cell lymphoma, 1 case anaplastic large cell lymphoma(ALCL). In terms of 17 patients with carcinoma, 3 cases esophageal carcinoma, 3 cases gastric carcinoma, 2 cases colorectal carcinoma, 7 cases thyroid carcinoma, 1 case hepatocellular carcinoma and lung cancer. Up to 15 patients received operation, and some of them combined with chemotherapy, radiotherapy and autologous transplant. Follow-up analysis showed that 3 cases was undergoing treatment, 2 cases lost follow-up, 4 cases died, 3 cases achieved CR, 3 cases remained to be at SD, and 2 cases assessed for progression or recurrence. Conclusion: The relationship between lymphoma and carcinoma was under discussion, patients with synchronous lymphoma and carcinoma were not unusual. We herein should raise awareness to avoid misdiagnosis.
Humans
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Lymphoma
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Neoplasm Recurrence, Local
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Neoplasms
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Neoplasms, Multiple Primary
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Retrospective Studies
10.Mortality and Morbidity of Extremely Low Birth Weight Infants in the Mainland of China: A Multi-center Study.
Hui-Jia LIN ; Li-Zhong DU ; Xiao-Lu MA ; Li-Ping SHI ; Jia-Hua PAN ; Xiao-Mei TONG ; Qiu-Ping LI ; Jian-Guo ZHOU ; Bing YI ; Ling LIU ; Yun-Bing CHEN ; Qiu-Fen WEI ; Hui-Qing WU ; Mei LI ; Cui-Qing LIU ; Xi-Rong GAO ; Shi-Wen XIA ; Wen-Bin LI ; Chao-Ying YAN ; Ling HE ; Kun LIANG ; Xiao-Yu ZHOU ; Shu-Ping HAN ; Qin LYU ; Yin-Ping QIU ; Wen LI ; Dong-Mei CHEN ; Hong-Ru LU ; Xiao-Hong LIU ; Hong LIU ; Zhen-Lang LIN ; Li LIU ; Jia-Jun ZHU ; Hong XIONG ; Shao-Jie YUE ; Si-Qi ZHUANG
Chinese Medical Journal 2015;128(20):2743-2750
BACKGROUNDWith the progress of perinatal medicine and neonatal technology, more and more extremely low birth weight (ELBW) survived all over the world. This study was designed to investigate the short-term outcomes of ELBW infants during their Neonatal Intensive Care Unit (NICU) stay in the mainland of China.
METHODSAll infants admitted to 26 NICUs with a birth weight (BW) < l000 g were included between January l, 2011 and December 31, 2011. All the data were collected retrospectively from clinical records by a prospectively designed questionnaire. The data collected from each NICU transmitted to the main institution where the results were aggregated and analyzed. Categorical variables were performed with Pearson Chi-square test. Binary Logistic regression analysis was used to detect risk factors.
RESULTSA total of 258 ELBW infants were admitted to 26 NICUs, of whom the mean gestational age (GA) was 28.1 ± 2.2 weeks, and the mean BW was 868 ± 97 g. The overall survival rate at discharge was 50.0%. Despite aggressive treatment 60 infants (23.3%) died and another 69 infants (26.7%) died after medical care withdrawal. Furthermore, the survival rate was significantly higher in coastal areas than inland areas (53.6% vs. 35.3%, P = 0.019). BW < 750 g and GA < 28 weeks were the largest risk factors, and being small for gestational age was a protective factor related to mortality. Respiratory distress syndrome was the most common complication. The incidence of patent ductus arteriosus, intraventricular hemorrhage, periventricular leukomalacia, bronchopulmonary dysplasia, retinopathy of prematurity was 26.2%, 33.7%, 6.7%, 48.1%, and 41.4%, respectively. Ventilator associated pneumonia was the most common hospital acquired infection during hospitalization.
CONCLUSIONSOur study was the first survey that revealed the present status of ELBW infants in the mainland of China. The mortality and morbidity of ELBW infants remained high as compared to other developed countries.
China ; Female ; Humans ; Infant ; Infant Mortality ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; statistics & numerical data ; Male ; Morbidity ; Respiratory Distress Syndrome, Newborn ; mortality ; Retrospective Studies ; Surveys and Questionnaires