Objective:To investigate the value of vasoactive inotropic score (VIS) in assessing adverse outcomes in neonates with early-onset sepsis (EOS).Methods:A retrospective study was conducted on 110 neonates with EOS admitted to the Affiliated Suzhou Hospital of Nanjing Medical University from January 2020 to March 2024. The patients were divided into a survival group ( n=88) and a death group ( n=22). Perinatal factor, and complications were compared between the two groups using t test, Mann-Whitney U test, and Chi-square test. Logistic regression analysis was used to identify the risk factors for death in patients with EOS, and receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive value of VIS for death in EOS patients. Results:The gestational age and birth weight of the death group were lower than those of the survival group [29.4 (26.8-35.3) weeks vs. 32.8 (30.1-37.2) weeks, 1 050.0 (737.5-2 162.5) g vs. 1 700.0 (1 212.5-2 587.5) g, Z values were－2.16 and－2.30, both P<0.05]. Compared with the survival group, the proportion of asphyxia [45.5% (10/22) vs. 21.6% (19/88)], low temperature [27.3% (6/22) vs. 8.0% (7/88)], mechanical ventilation [100.0% (22/22) vs. 54.5% (48/88)], disseminated intravascular coagulation [22.7% (5/22) vs. 3.4% (3/88)], persistent pulmonary hypertension [36.4% (8/22) vs. 11.4% (10/88)] and pulmonary hemorrhage [40.9% (9/22) vs. 8.0% (7/88)] were higher in the death group ( χ2=5.16, 4.59, 15.71, 7.09, 6.32 and 12.84, all P<0.05). The VIS values at 24 hours and 48 hours after admission in the death group were 15.0 (10.0-18.1) and 18.8 (12.8-30.0), respectively, which were higher than those in the survival group [10.0 (7.5-10.0) and 10.0 (7.5-10.0), Z values were－4.60 and－4.94, respectively, both P<0.05].Logistic regression analysis showed that 24-h VIS value ( OR=1.163, 95% CI: 1.018-1.328), 48-h VIS value ( OR=1.114, 95% CI: 1.031-1.204), birth asphyxia ( OR=3.815, 95% CI: 1.017-14.310), and pulmonary hemorrhage ( OR=4.470, 95% CI: 1.174-17.017) were independent risk factors for death (all P<0.05). ROC curve analysis showed that the optimal cutoff value for predicting death was 11 for 24-h VIS, with an area under the curve (AUC) of 0.807, Youden's index of 0.466, sensitivity of 68.2%, and specificity of 78.4%, and 12.5 for 48-h VIS, with an AUC of 0.851, Youden's index of 0.659, sensitivity of 95.5%, and specificity of 70.5%. Conclusions:The VIS value after the onset of EOS is closely related to the outcome. A 48-h VIS value exceeding 12.5 is associated with a high risk of death.

This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

Objective:To investigate the value of vasoactive inotropic score (VIS) in assessing adverse outcomes in neonates with early-onset sepsis (EOS).Methods:A retrospective study was conducted on 110 neonates with EOS admitted to the Affiliated Suzhou Hospital of Nanjing Medical University from January 2020 to March 2024. The patients were divided into a survival group ( n=88) and a death group ( n=22). Perinatal factor, and complications were compared between the two groups using t test, Mann-Whitney U test, and Chi-square test. Logistic regression analysis was used to identify the risk factors for death in patients with EOS, and receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive value of VIS for death in EOS patients. Results:The gestational age and birth weight of the death group were lower than those of the survival group [29.4 (26.8-35.3) weeks vs. 32.8 (30.1-37.2) weeks, 1 050.0 (737.5-2 162.5) g vs. 1 700.0 (1 212.5-2 587.5) g, Z values were－2.16 and－2.30, both P<0.05]. Compared with the survival group, the proportion of asphyxia [45.5% (10/22) vs. 21.6% (19/88)], low temperature [27.3% (6/22) vs. 8.0% (7/88)], mechanical ventilation [100.0% (22/22) vs. 54.5% (48/88)], disseminated intravascular coagulation [22.7% (5/22) vs. 3.4% (3/88)], persistent pulmonary hypertension [36.4% (8/22) vs. 11.4% (10/88)] and pulmonary hemorrhage [40.9% (9/22) vs. 8.0% (7/88)] were higher in the death group ( χ2=5.16, 4.59, 15.71, 7.09, 6.32 and 12.84, all P<0.05). The VIS values at 24 hours and 48 hours after admission in the death group were 15.0 (10.0-18.1) and 18.8 (12.8-30.0), respectively, which were higher than those in the survival group [10.0 (7.5-10.0) and 10.0 (7.5-10.0), Z values were－4.60 and－4.94, respectively, both P<0.05].Logistic regression analysis showed that 24-h VIS value ( OR=1.163, 95% CI: 1.018-1.328), 48-h VIS value ( OR=1.114, 95% CI: 1.031-1.204), birth asphyxia ( OR=3.815, 95% CI: 1.017-14.310), and pulmonary hemorrhage ( OR=4.470, 95% CI: 1.174-17.017) were independent risk factors for death (all P<0.05). ROC curve analysis showed that the optimal cutoff value for predicting death was 11 for 24-h VIS, with an area under the curve (AUC) of 0.807, Youden's index of 0.466, sensitivity of 68.2%, and specificity of 78.4%, and 12.5 for 48-h VIS, with an AUC of 0.851, Youden's index of 0.659, sensitivity of 95.5%, and specificity of 70.5%. Conclusions:The VIS value after the onset of EOS is closely related to the outcome. A 48-h VIS value exceeding 12.5 is associated with a high risk of death.

BACKGROUND: Antenatal corticosteroids (ACS) can significantly improve the outcomes of preterm infants. This study aimed to describe the ACS use rates among preterm infants admitted to Chinese neonatal intensive care units (NICU) and to explore perinatal factors associated with ACS use, using the largest contemporary cohort of very preterm infants in China. METHODS: This cross-sectional study enrolled all infants born at 24 +0 to 31 +6 weeks and admitted to 57 NICUs of the Chinese Neonatal Network from January 1st, 2019 to December 30th, 2019. The ACS administration was defined as at least one dose of dexamethasone and betamethasone given before delivery. Multiple logistic regressions were applied to determine the association between perinatal factors and ACS usage. RESULTS: A total of 7828 infants were enrolled, among which 6103 (78.0%) infants received ACS. ACS use rates increased with increasing gestational age (GA), from 177/259 (68.3%) at 24 to 25 weeks' gestation to 3120/3960 (78.8%) at 30 to 31 weeks' gestation. Among infants exposed to ACS, 2999 of 6103 (49.1%) infants received a single complete course, and 33.4% (2039/6103) infants received a partial course. ACS use rates varied from 30.2% to 100% among different hospitals. Multivariate regression showed that increasing GA, born in hospital (inborn), increasing maternal age, maternal hypertension and premature rupture of membranes were associated with higher likelihood to receive ACS. CONCLUSIONS The use rate of ACS remained low for infants at 24 to 31 weeks' gestation admitted to Chinese NICUs, with fewer infants receiving a complete course. The use rates varied significantly among different hospitals. Efforts are urgently needed to propose improvement measures and thus improve the usage of ACS.
Humans ; Infant, Newborn ; Infant ; Pregnancy ; Female ; Gestational Age ; Infant, Premature ; Intensive Care Units, Neonatal ; Cross-Sectional Studies ; Adrenal Cortex Hormones/therapeutic use*

Objective:To study the clinical features and genotypes of neonatal Glanzmann thrombasthenia(NGT).Methods:A male neonate with NGT admitted to the Department of Neonatology of our hospital was retrospectively reviewed. CNKI, Wangfang database, VIP, the Chinese Medical Journal Full Text database, PubMed and Embase database were searched using key words '(neonate OR newborn) AND (Glanzmann thrombasthenia)' both in English and Chinese. The clinical features and genotypes of NGT were summarized and analyzed.Results:A male full-term neonate was admitted to our hospital for mass on the forehead and ecchymosis and petechiae on the body within half an hour after birth. He gradually developed subgaleal hemorrhage and severe anemia. Platelet count, mean platelet volume and coagulation functions were normal. The platelet aggregation test indicated decreased platelet aggregation rate induced by arachidonic acid and adenosine diphosphate. Genetic testing revealed two heterozygous mutations in the patient's ITGA2B gene: NM_000419.4: c.886G>A(p.Gly296Arg) and NM_000419.4: c.2855dup(p.Phe953Valfs*83). A total of 42 literature involving 44 patients (our case included) with NGT were retrieved. 33 cases (75.0%) of NGT showed ecchymosis or petechiae on the first day after birth. For 13 cases with detailed information, 5 cases with severe anemia were given erythrocyte and plasma transfusion and platelet transfusion was given in 1 case. 4 cases had homozygous variants and 4 cases showed compound heterozygous variants. 10 cases had follow-up records, including 2 cases without any bleeding and 8 cases with varying degrees of bleeding during follow-up. No deaths were reported.Conclusions:Neonates with ecchymosis and petechiae in the early postnatal period should be suspected of NGT. Blood transfusion is preferred when the indication for transfusion is met.

Objective:To analyze the risk factors of bronchopulmonary dysplasia(BPD)in very preterm infants(VPI), and to provide scientific basis for the prevention and treatment of BPD in VPI.Methods:A prospective multicenter study was designed to collect the clinical data of VPI in department of neonatology of 28 hospitals in 7 regions from September 2019 to December 2020.According to the continuous oxygen dependence at 28 days after birth, VPI were divided into non BPD group and BPD group, and the risk factors of BPD in VPI were analyzed.Results:A total of 2 514 cases of VPI including 1 364 cases without BPD and 1 150 cases with BPD were enrolled.The incidence of BPD was 45.7%.The smaller the gestational age and weight, the higher the incidence of BPD( P<0.001). Compared with non BPD group, the average birth age, weight and cesarean section rate in BPD group were lower, and the incidence of male infants, small for gestational age and 5-minute apgar score≤7 were higher( P<0.01). In BPD group, the incidences of neonatal respiratory distress syndrome(NRDS), hemodynamically significant patent ductus arteriosus, retinopathy of prematurity, feeding intolerance, extrauterine growth restriction, grade Ⅲ~Ⅳ intracranial hemorrhage, anemia, early-onset and late-onset sepsis, nosocomial infection, parenteral nutrition-associated cholestasis were higher( P<0.05), the use of pulmonary surfactant(PS), postnatal hormone exposure, anemia and blood transfusion were also higher, and the time of invasive and non-invasive mechanical ventilation, oxygen use and total hospital stay were longer( P<0.001). The time of starting enteral nutrition, cumulative fasting days, days of reaching total enteral nutrition, days of continuous parenteral nutrition, days of reaching 110 kcal/(kg·d) total calorie, days of reaching 110 kcal/(kg·d) oral calorie were longer and the breastfeeding rate was lower in BPD group than those in non BPD group( P<0.001). The cumulative doses of amino acid and fat emulsion during the first week of hospitalization were higher in BPD group( P<0.001). Multivariate Logistic regression analysis showed that NRDS, invasive mechanical ventilation, age of reaching total enteral nutrition, anemia and blood transfusion were the independent risk factors for BPD in VPI, and older gestational age was the protective factor for BPD. Conclusion:Strengthening perinatal management, avoiding premature delivery and severe NRDS, shortening the time of invasive mechanical ventilation, paying attention to enteral nutrition management, reaching whole intestinal feeding as soon as possible, and strictly mastering the indications of blood transfusion are very important to reduce the incidence of BPD in VPI.

Objective:To establish a fluorescent reporter human induced pluripotent stem cell line (hiPSCs) for monitoring the expression of visual system homeobox 2 ( VSX2). Methods:VSX2_small guide RNA (sgRNA) was inserted into vector PX459 to construct knockout plasmid, and the P2A-eGFP knock-in donor plasmid was conducted at the same time.The two plasmids were transfected into BC1-hiPSCs.Single cell clones were generated after treatment of puromycin.Correct insertion was confirmed by PCR and Sanger sequencing.The isogenicity of the parental and the reporter hiPSCs was confirmed by STR analysis and karyotyping.Pluripotency capacity of the reporter hiPSCs was analysed by reverse trascription PCR and immunofluorescence.Three-germ-layer formation experiment was carried out to analyse the multi-lineage differentiation ability of the reporter hiPSCs.The reporter hiPSCs were further differentiated to obtain three-dimension (3D) retinal organoids, and immunofluorescence was used to identify the co-localization of the enhanced green fluorescent protein (eGFP) and VSX2.Results:A VSX2 eGFP reporter hiPSC clone was successfully obtained by CRISPR/Cas9 technology, which was consistent with the parental hiPSCs (BC1-hiPSCs) in morphology, without any chromosomal aberrations or cell line cross-contamination.Reverse transcription PCR assay and immunofluorescence showed obvious positive expressions of iPSCs markers in BC1- VSX2 eGFP-iPSCs, including NANOG, OCT4, SOX2, DNMT3B and GDF3 mRNA as well as NANOG, OCT4, SSEA4 and TRA-1-60 protein.The α-fetoprotein (AFP), α-smooth muscle actin (α-SMA) and neuronal class Ⅲ β-tubulin (TUJ1) were expressed in endoderm, mesoderm and ectoderm, respeetively, derived from BC1- VSX2 eGFP-iPSCs, and eGFP and VSX2 were co-stained in the neural retinal layer of 3D retinal organoids derived from BC1- VSX2 eGFP-iPSCs by immunofluorescence. Conclusions:VSX2 fluorescent reporter hiPSCs is successfully generated, which can monitor the temporal and spatial expression changes of VSX2 protein in real time, providing a powerful tool for evaluation of retina development mechanism and cell therapy.

Objective To evaluate the effect of endoscopic management of foreign bodies in the upper digestive tract. Methods Clinical data and endoscopic treatment methods of 41 patients were retrospectively analyzed from October 2014 to May 2016. Patients with incomplete medical records were excluded. Results Foreign bodies in the upper digestive tract occurred high frequency in elderly. 53.6% of the foreign bodies were located in the esophagus. Date stones was the main type of foreign bodies (56.1%). 41 cases with foreign bodies in digestive tract were successfully extracted, while 1 case occurred perforation. Conclusion Endoscopic management of gastrointestinal foreign bodies is safe and effective.

Objective To investigate the effect of brain ischemic preconditioning (IP) combined with traditional Chinese medicine three seven three alcohol saponin (PTS) on proliferation of endogenous neural stem cells and the mRNA expressions of delta opioid receptor (DOR),Bax,Bcl-2 in hippocampus at 7d post middle cerebral artery occlusion (MCAO).Methods The focal-focal ischemic tolerance models were established with twice suture method.80 SD rats were included and randomly divided into 5 groups:sham group,MCAO group,sham+ MCAO group,IP+ MCAO group,PTS+MCAO group (n=16 each).We chose 10 SD rats from each group to evaluate their neurological status,and made BrdU fluorescent immunolabeling.In addition,we chose the other 6 SD rats to detect the expression levels of DOR,Bax and Bcl-2 mRNA in ischemic region in hippocampusby using RT-PCR.Animals were given one set of BrdU injections (on day 6,three times,4h apart,50mg/kg) to label the proliferating cells.The neurological status was assessed by using Zea Longa neurological deficit scores at 7 days following cerebral infarction.Results Zea longa neurologic deficit scores in MCAO group and sham+ MCAO) group had significantly differences with IP+ MCAO group and PTS+ MCAO group respectively at 7d post MCAO(P＜0.01).There was no significant differeuce in Zea-longa neurologic deficit scores between MCAO group versus sham+ MCAO group,and IP+ MCAO group versus PTS+ MCAO group(P＞0.05).The number of BrdU+ ceils in hippocampus had significant differences between IP+ MCAO and PTS+ MCAO groups at 7d post MCAOand three groups of sham,MCAO and sham+ MCAO respectively (P＜0.01).There was no difference in the number of BrdU+ cells between MCAO versus Sham + MCAO groups and IP + MCAO versus PTS+MCAO groups(P＞0.05).DOR and Bcl-2 mRNA expression levels were higher and Bax mRNA expression level was lower in IP+ MCAO group than in MCAO,Sham+ MCAO and PTS+MCAO groups (P＜0.01).There were no significant differences in DOR,Bcl-2 and Bax mRNA expressions among MCAO,Sham + MCAO and PTS + MCAO groups (P＞ 0.05).Conclusions Acute cerebral infarction can induce the proliferation of endogenous neural stem cells in hippocampus in SD rats.IPC can facilitate the proliferation of endogenous neural stem cells in hippocampus afteracute cerebral infarction,improve the symptoms of neurologic dysfunction,increase DOR and Bcl 2 mRNA expressions,and reduce Bax mRNA expression in SD rats.PTS can facilitate the proliferation of endogenous neural stem cells in hippocampus after acute cerebral infarction in SD rats,and improve the symptoms of neurologic dysfunction,but it has no influence on the expressions of DOR,Bcl-2 and Bax mRNA.