Objective To explore the perinatal risk factors and clinical characteristics of complications of early term neonates.Methods Data of 5 468 live term newborns and their mothers hospitalized in the Third Affiliated Hospital of Zhengzhou University from January 2013 to December 2013 were analyzed.Background information,morbidity and complications of the mothers were compared among early,full and late term groups (n=l 933,3 013,412,respectively).And background information and incidence of complications were also investigated among neonates of early,full and late term groups (n=2 033,3 023,412,respectively),and neonates born between 37-37+6 (n=695) and 38-38+6 weeks (n=1 338).One-way analysis of variance,LSD-t test,logistic regression analysis,Chi-square or Fisher exact test,Pearson Chi-square test,corrected Chi-square test were used for statistical analysis.Results 1.Comparison among the early,full and late term group showed that higher proportions of elder gravida [21.1％(407/1 933),10.5％(317/3 013),6.8％(28/412),x2=127.690],multipara [43.7％(844/1 933),23.1％(697/3 013),15.0％(62/412),x2=287.765],scarred uterus [27.9％(539/1 933),8.9％(267/3013),1.5％(6/412),x2=396.521],higher incidence of cesarean section [75.2％(1 453/1 933),56.2％(1 693/3 013),54.1％(223/412),x2=196.348],hypertensive disorder complicating pregnancy [9.2％(178/1 933),3.5％(105/3 013),2.9％(12/412),x2=79.915],multiple pregnancy[5.1％(99/1 933),0.3％(9/3 013),0.0％(0/412),x2=147.860],gravidity＞1[63.1％(1 220/1 933),47.3％(1 425/3 013),39.6％(163/412),x2=147.668],premature rupture of fetal membranes[20.6％(398/l 933),14.2％(428/3 013),10.2％(42/412),x2=47.217],abnormal amniotic fluid[17.8％(344/1 933),12.3％(370/3 013),11.2％(46/412),x2=32.777],gestational diabetes mellitus[11.5％(223/1 933),5.9％(178/3 013),5.1％(21/412),x2=56.169],abnormal presentation [9.5％(184/1 933),5.1％(155/3 013),2.9％(12/412),x2=43.511],abnormal placenta [7.6％(146/1 933),3.1％(92/3 013),2.7％(11/412),x2=57.739],hysteromyoma[4.9％(94/1 933),2.3％(68/3 013),0.7％(3/412),x2=35.062] in the early term group than in the full and late term group,respectively (all P＜0.016).Multivariate logistic analysis showed that multiple pregnancy (OR=21.736,95％CI:10.785-43.806),scarred uterus (OR=3.302,95％CI:2.679-4.071) and hypertensive disorder complicating pregnancy(OR=2.658,95％CI:2.040-3.465) were the leading three perinatal risk factors for early term delivery.2.The incidence of the following neonatal conditions were different among early,full and late term infants (all P＜0.05):hyperbilirubinemia [12.5％(255/2 033),3.9％(119/3 023),4.9％(20/412),x2=138.343],infectious diseases [4.3％(88/2 033),2.0％(59/3 023),1.7％(7/412),x2=27.122],asphyxia[3.0％(60/2 033,1.4％(42/3 023),1.0％(4/412),x2=17.795],brain damage [2.3％(46/2 033),0.5％(15/3,023),10.％(4/412)],respiratory distress syndrome [1.1％(23/2 033),0.2％(7/3 023),0.0％(0/412)],feeding problems [2.0％(41/2 033),0.3％(10/3 023),1.0％ (4/412)],surgical diseases[2.0％(41/2 033),0.9％(28/3 023),1.5％(6/412),x2=0.709],intracranial hemorrhage [1.9％(39/2 033),0.9％(26/3 023),0.5％(2/412),x2=13.263],wet lung [0.9％(19/2 033),0.4％(11/3 023),0.5％(2/412)].Incidences of the above complications in the early term infants were all higher than in the full term infants,but when compared with the later term infants,only that of hyperbilirubinemia and infectious diseases was higher (all P＜0.016).Incidence of admission ot NICU [24.5％(170/695) vs 11.5％(153/1 338),x2=57.729],hyperbilirubinemia [19.0％(132/695) vs 9.2％(123/1 338),x2=40.046],infectious diseases[6.2％(43/695) vs 3.4％(45/1 338),x2=8.807],brain damage[4.0％(28/695) vs 1.3％(18/1 338),x2=14.828],and NRDS[2.0％(14/695) vs 0.5％(9/1 338),x2=7.329],feeding problems [3.2％(22/695) vs 1.5％(20/1 338),x2=6.271],intracranial hemorrhage [3.2％(22/695) vs 1.3％(17/1 338),x2=8.684],wet lung [1.7％(12/695) vs 0.5％(7/1 338),x2=7.049] of the early term infants born at 37-37+6 weeks were all higher than those born at 38-38+6 weeks(all P＜0.05).Conclusions Multiple pregnancy,scarred uterus and hypertensive disorder of pregnancy are the three leading perinatal risk factors of early term delivery.The incidence of neonatal complications among early term infants are higher than those among full term infants,and early term infants are more likely to stay in NICU.We should take preventive measures to decrease the rate of early term delivery and improve the follow-up management of early term infants.

ObjectiveTo investigate the effects of caffeine citrate (CC) on the neuronal proliferation and apoptosis and long-term learning ability in neonatal rats with hypoxia-ischemic brain damage (HIBD).MethodsForty-eight 7-day-old Sprague-Dawley neonatal rats were randomly divided into 3 groups: sham operation group (n=16), HIBD group (n=16), HIBD + caffeine citrate group (CC group,n=16). Rats in HIBD and CC groups received ligation of left common carotid artery followed by 2 hours of hypoxia to establish HIBD model. Rats in CC group were injected intraperitoneally with CC (20 mg/kg) before and at 0 min, 24 h, 48 h, and 72 h after hypoxia-ischemic (HI), and rats in the other two groups were injected intraperitoneally with an equal volume of normal saline at the corresponding time. Meanwhile, from postnatal day 10, each rat was injected intraperitoneally with 5-bromo-2’-de-oxyuridine (BrdU) (50 mg/kg) for 5 consecutive times, once every 12 h. On postnatal day 12, BrdU in the hippocampal dentate gyrus and cleaved caspase-3 in the hippocampal CA1 area were detected by immunohistochemistry, and neuronal apoptosis in hippocampal CA1 area were detected by TUNEL staining. On postnatal day 28, long-term learning and memory ability of rats was tested by Y maze.ResultsThere was signiifcant difference in the number of BrdU-positive cells in brain tissues of rats among three groups (F=101.38,P<0.01). The BrdU-positive cells in HIBD group and CC group were signiifcantly more than those in sham operation group (P<0.05). There was signiifcant difference in the number of cleaved caspase-3-positive cells in hippocampal CA1 area among three groups (F=379.77,P<0.01). The cleaved caspase-3-positive cells in CC group were signiif-cantly fewer than those in HIBD group but signiifcantly more than those in sham operation group (P<0.05). The TUNEL-pos-itive cells in hippocampal CA1 area were signiifcantly different among three groups (F=505.92,P<0.01) which was most in HIBD group and fewest in sham operation group and signiifcant difference was found through multiple comparison (P<0.05). The total learning number of avoiding electric shock tested by Y maze was signiifcantly different among three groups (F=32.05, P<0.01) which was most in HIBD group. Correct response rate was significantly different among three groups (F=24.99, P<0.01) which was lowest in HIBD group.ConclusionsCaffeine citrate can improve the ability of long-term learning and memory in neonatal rats with hypoxia-ischemic brain damage, the mechanism of which may be related to reducing the neuronal apoptosis after hypoxia ischemia.

Objective To investigate the protective effect of caffeine citrate on white matter damage in 2-day-old neonatal rats induced by postnatal infection.Methods Forty-eight 2-day-old SD rats were randomly divided into the control group (group A,n =16),lipopolysaccharide(LPS) infection group (group B,n =16),and caffeine citrate intervention group (group C,n =16) according to the random table method.The newborn rats of group B and C were continuously injected LPS 0.6 mg/kg intraperitoneally for 5 days from 2 days old,and the newborn rats of group A were continuously injected by an equal volume of 9 g/L saline intraperitoneally.Group C was continuously injected by caffeine citrate 10 mg/kg intraperitoneally for 7 days from 4 days old;equal volume of 9 g/L saline was injected into group A and B for 7 days continuously.At 12 days old,8 rats of each group were sacrificed randomly to evaluate the expression of myelin basic protein(MBP) subcortical white matter by immunohistochemical method.Both sides of hippocampus of the rest 8 rats of each group were taken out in ice surface rapidly.The left hippocampus was used to detect the expression of MBP and Caspase-3 by Western blot method,and the right hippocampus was used to evaluate the MBP and Caspase-3 protein level by quantitative real-time PCR(qPCR) method.Results The mean integral optical density (IOD) of subcortical MBP positive expression in group A,group B and group C were 132.64 ± 1.94,102.43 ± 2.12,114.25 ± 2.04,and the difference was statistically significant among 3 groups (F =22.912,P ＜ 0.05).The relative expression levels of MBP mRNA of 3 groups in hippocampus were 0.79 ± 0.01,0.39 ± 0.03,0.55 ± 0.02,and the difference was statistically significant among 3 groups (F =18.584,P ＜ 0.05).The relative expression levels of MBP protein of 3 groups in hippocampus were 0.64 ± 0.03,0.31 ± 0.03,0.51 ± 0.05,and the difference was statistically significant among 3 groups (F =25.780,P ＜ 0.05).The relative expression levels of Caspase-3 mRNA of 3 groups in hippocampus were 0.34 ± 0.02,0.74 ± 0.03,0.57 ± 0.04,and the difference was statistically significant among 3 groups (F =6.105,P ＜ 0.05).The relative expression of Caspase-3 protein of 3 groups in hippocampus were 0.11 ± 0.03,0.36 ± 0.02,0.23 ± 0.03,and the difference was statistically significant among 3 groups (F =40.541,P ＜ 0.05).Conclusions Caffeine citrate has showed protective effect on white matter damage in neonatal rats of 2 days old induced by postnatal infection.The mechanism may be related to inhibition of inflammatory response and reducing apoptosis.

Objective In order to study the biological characteristics of macrophages and provide the materials to study the survival mechanism of intracellular parasites, we conducted this study to establish a high-purity alveolar macrophage isolation and culture method.Methods Goat lungs were lavaged with normal saline in sterile environment several times, and cells were collected and then goat alveolar macrophages were purified by density gradient centrifugation using peripheral blood mononuclear cells (PBMC) solution.The isolated goat alveolar macrophages were cultured in cell culture medium containing 10% fetal bovine serum and cell morphology was observed under an inverted microscope every day,and the phagocytic activity of the cells was detected by chicken red blood cell phagocytosis test.Flow cytometry was used to detect CD14, a characteristic monocyte-macrophage surface marker.Results The adherent cells were characterized by typical macrophage morphology, pseudopodia and protrusions, showing round and irregular shape, rich cytoplasm, and large cell body.Of the cultured macrophages, 54.5% could phagocytize chicken erythrocytes and showed good phagocytic activity.After one month of in vitro culture, 93.7% of the cells were able to express CD14 antigen, which had a macrophage-specific immunophenotype.Conclusions The alveolar macrophages obtained in this study have high purity and good bioactivity, thus provide a cell model for studying the immune mechanism of intracellular parasites.

AIM:To investigate the effects of Arg-Gly-Asp-Ser (RGDS) tetrapeptide on proliferation,apoptosis and caspase 3 expression in FN-stimulated HSCs in vitro. METHODS:[ 3H]-thymidine incorporation,Annexin-V/Propidium Iodide double-labeled flow cytometry(FCM),TUNEL,scanning electron microscope and transmission electron microscopy were employed to estimate the influence of RGDS on proliferation and apoptosis of HSCs. The adhesion rates were observed by toluidine blue colorimetric assay. The expression of caspase-3 protein was detected by FCM. RESULTS:①Compared with control and FN groups,RGDS tetrapeptide at concentrations of 25 mg?L -1 ,50 mg?L -1 and 100 mg?L2 1 inhibited the proliferation of HSCs ( P

Interleukin-21 is a type I cytokine mainly produced by activated CD4+ T cells that acts as a regulator of immune system. In this work, hIL-21cDNA was amplified from human peripheral blood lymphocytes by RT-PCR, and then inserted into pPIC9K. The recombinant vector pPIC9K-hIL21cDNA was linearized by Sac I, and transformed into Pichia pastoris strain GS115 by electroporation. Transformants were selected by G418 and confirmed by PCR. The recombinant protein was expressed and secreted into the supernatant after inducing by methanol. SDS-PAGE analysis indicated the molecular weight of rhIL-21 was about 16 kD. ELISA results show that the yield of rhIL-21 reach 229.28 mg/L, rhIL-21 was purified from culture supernatants, and it was purified to about 95% purity with ion-exchange chromatography. When co-stimulate with Con A, rhIL-21 can promote the proliferation of human lymphocytes. This is the first expression of bio-active rhIL-21 in Pichia pastoris. It lays a foundation for further research in immunotherapy and cancer therapy.
Electroporation ; Genetic Vectors ; genetics ; Humans ; Interleukins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; analysis ; biosynthesis ; genetics

OBJECTIVETo evaluate the potential role of insulin-like growth factor-1 receptor mRNA (IGF-IR mRNA) in the onset and development of retinopathy in diabetic rats.

METHODSA diabetic model was duplicated in Wistar rats. The early changes in the retina were examined using light and transmission electron microscopy. Expression of IGF-IR mRNA was analyzed using in situ hybridization.

RESULTSWeak expression of IGF-IR mRNA (5%) was found in retinas of normal rats, but was significantly increased (15% and 18%) in the retinas of diabetic rats after 3 and 6 months of diabetes (P < 0.01). In situ hybridization and morphological study demonstrated that there was a positive correlation between IGF-IR mRNA expression and retinal changes at various stages.

CONCLUSIONIncreased IGF-IR mRNA might play an important role in the onset and development of diabetic retinopathy.

Animals ; Blood Glucose ; analysis ; Diabetic Retinopathy ; etiology ; Glycated Hemoglobin A ; analysis ; Insulin-Like Growth Factor I ; physiology ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Receptor, IGF Type 1 ; genetics ; Retina ; metabolism ; pathology

Promoter-trap strategy for enriching targeted colonies has been usually used to elevate the gene targeting efficiency in somatic cells. Knocking out Prnp in animals by gene targeting can render them resistant to Prion diseases. We constructed a bovine Prnp promoter-less targeting vector BoPrPneo, then transfected the linearized vector into the bovine fetal fibroblasts BFF through electroporation. After selecting in cell culture medium with 250 microg/mL G418, we obtained 99 drug-resistant cell colonies, 4 of them were positive for targeted events after PCR screening, and the targeted colonies were further confirmed by sequencing and Southern blotting. This suggests that one allele of Prnp has been successfully knocked out in bovine fetal fibroblasts. This research supplies a simple, safe and effective method to targeting bovine Prnp.
Animals ; Cattle ; Electroporation ; Fetus ; Fibroblasts ; metabolism ; Gene Knockout Techniques ; methods ; Prion Diseases ; genetics ; prevention & control ; Prions ; genetics ; Promoter Regions, Genetic ; genetics ; Transfection

The clinical data, laboratory testing, genetic testing results, diagnosis and treatment process of a child with PERCHING syndrome diagnosed and treated in the Department of Neonatology, the Third Affiliated Hospital of Zhengzhou University in June 2022 were retrospectively analyzed, and the relevant literatures were reviewed.The proband mainly presented with dyspnea and feeding difficulties after delivery, facial nevus flammeus, protrusion of eyes, small fissure of eyes, wide nasal root, limited opening of mouth, slightly high palatal arch, special posture, cryptorchid, hypospadias, and high muscle tone of limbs.Magnetic resonance imaging of the brain suggested possible agenesis of corpus callosum.Genetic testing showed complex heterozygous variations in the KLHL7 gene, and the two mutation sites have not been previously reported.A case of PERCHING syndrome caused by the KLHL7 gene mutation in China was reported for the first time, which provided new ideas for the diagnosis and treatment of children with PERCHING syndrome and reliable genetic evidence for family reproduction.

Objective:To investigate the clinical characteristics and prognosis of bloodstream infection caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) in preterm infants, and to provide basis for early clinical diagnosis and infection control. Methods:The clinical data of infants with CRKP bloodstream infection admitted to the Preterm Infants Ward of Children′s Hospital Affiliated to Zhengzhou University from January 2015 to December 2022 were retrospectively analyzed. The risk factors for death in preterm infants caused by CRKP bloodstream infection were explored through multivariate logistic regression analysis, and the receiver operating characteristic (ROC) curve was used to analyze the clinical value of each factor on evaluating prognosis. The area under curves (AUC) of each factor in different ROC curve were compared by Delong′s test.Results:A total of 96 preterm infants with CRKP bloodstream infection were included, including 70 in the survival group and 26 in the death group. The first onset symptoms of CRKP bloodstream infection in preterm infants were persistent tachycardia (heart rate>180 per minute) (69 cases, 71.9%), fever (61 cases, 63.5%), and apnea (59 cases, 61.5%). There were 88(91.7%) cases of infection combined with septic shock, and 91(94.8%) cases required vasoactive drug support. Multivariate logistic regression analysis showed that the maximum vasoactive-inotropic score (VIS) within 48 hours of onset (odds ratio ( OR)=1.058, 95% confidence interval (95% CI) 1.022 to 1.095, P=0.001), concurrent purulent meningitis ( OR=8.029, 95% CI 1.344 to 47.972, P=0.022), and concurrent necrotizing enterocolitis (NEC) ( OR=10.881, 95% CI 1.566 to 75.580, P=0.016) were independent risk factors for death in preterm infants with CRKP bloodstream infection. The ROC curve showed that the AUCs for evaluating the prognosis of preterm infants with NEC and purulent meningitis were 0.784 and 0.711, respectively. The AUC for evaluating the prognosis of preterm infants with a maximum VIS ≥52.5 points within 48 hours of onset was 0.840, and the AUC for combining the three factors was 0.931. Compared with NEC and purulent meningitis, the AUC for combining factors was higher, the differences were statistically significant ( P=0.002, P<0.001). Conclusions:Preterm infants with CRKP bloodstream infection who have a maximum VIS ≥52.5 points within 48 hours of onset, with NEC and purulent meningitis have a higher risk of death.