Compared with adults, neonates tend to have stronger and more persistent biological perception of pain. They may have the memory for pain and the negative effects caused by pain may exist for a long time. Therefore, standardized pain management can reduce or prevent the adverse effect of pain on body and mind and promote the rehabilitation process. In order to further deepen the understanding of pain management and standardize the analgesic measures for neonates, the Neonatologist Branch of Chinese Medical Association and Editorial Board of Chinese Journal of Contemporary Pediatrics have developed an expert consensus based on the clinical evidence in China and overseas and with reference to clinical experience from the following aspects:evaluation of neonatal pain and methods and techniques of pain management. It is recommended to adopt a step-by-step analgesic management for neonates. For mild pain stimulation, it is effective to relieve the pain by gentle touch and non-nutritive sucking combined with sucrose feeding. For moderate pain, selection of appropriate trocar needle and skilled puncture are important to reduce the pain, and in addition, the application of local anesthetics at the site of puncture also has a good effect. For severe pain, intravenous sedative drugs are often required, but no consistent evidence has been obtained so far.

Objective To study the effect of gestational age at birth on the neurobehavioral development of preschool children. Methods A total of 25 254 preschool children from Ma'anshan of Anhui Province, Taizhou of Zhejiang Province, and Yangzhou of Jiangsu Province were enrolled. The preschool children were divided into three groups based on their gestational ages at birth:preterm group (2 760 cases; 28-36⁺⁶ weeks), early term group (6 005 cases; 37-38⁺⁶ weeks), and full term group (16 489 cases; ≥ 39 weeks). The Ages and Stages Questionnaires-Third Edition (ASQ-3) was employed to evaluate the children's neurobehavioral development. Results The preterm group had significantly lower scores of the five domains of ASQ-3, communication, gross motor, fine motor, problem solving, and personal-social, than the full term group (P < 0.05), and significantly lower scores of communication, gross motor, fine motor, and problem solving than the early term group (P < 0.05). There were no significant differences in the scores of the five domains of ASQ-3 between the early term and full term groups (P > 0.05). The multiple linear regression analysis indicated a significant positive correlation between gestational age and the five domains of ASQ-3 after adjustment for confounding factors including sex, age, body mass index, and parental education level (P < 0.01). Conclusions Children born preterm have poorer neurobehavioral development than those born full term and early term, whereas children born full term and early term have similar neurobehavioral development. Gestational age at birth is an independent influencing factor for neurobehavioral development in preschool children.

Objective To investigate whether there is a difference in cerebellar development between appropriate-for-gestational-age (AGA) infants and small-for-gestational-age (SGA) infants. Methods A total of 165 AGA infants and 105 SGA infants, with a gestational age of 26-40⁺⁶ weeks, were enrolled in this study. Within 24-48 hours after birth, ultrasound examination was performed to measure the transverse diameter of the cerebellum, the height of the vermis, the area of the vermis, the perimeter of the vermis, and the area and perimeter of the cerebellum on transverse section. A Pearson correlation analysis was used to investigate the correlation between cerebellar measurements and gestational age. Results In both AGA and SGA infants, all cerebellar measurements were positively correlated with gestational age (r=0.50-0.81, P < 0.05). In AGA and SGA infants, there were no significant differences in the measurements between the 25-27⁺⁶ weeks, 28-30⁺⁶ weeks, and 31-33⁺⁶ weeks of gestational age subgroups (P > 0.05), while in the 34-36⁺⁶ weeks and 37-40⁺⁶ weeks subgroups, the SGA infants had significantly lower measurements than the AGA infants (P < 0.05). Conclusions The SGA infants with a gestational age of <34 weeks have intrauterine cerebellar development similar to AGA infants, but those with a gestational age of ≥ 34 weeks have poorer intrauterine cerebellar development than AGA infants.

Objective To investigate the clinical features and outcome of neonatal acute respiratory distress syndrome (ARDS) in southwest Hubei, China. Methods According to the Montreux definition of neonatal ARDS, a retrospective clinical epidemiological investigation was performed on the medical data of neonates with ARDS who were admitted to Department of Neonatology/Pediatrics in 17 level 2 or level 3 hospitals in southwest Hubei from January to December, 2017. Results A total of 7 150 neonates were admitted to the 17 hospitals in southwest Hubei during 2017 and 66 (0.92%) were diagnosed with ARDS. Among the 66 neonates with ARDS, 23 (35%) had mild ARDS, 28 (42%) had moderate ARDS, and 15 (23%) had severe ARDS. The main primary diseases for neonatal ARDS were perinatal asphyxia in 23 neonates (35%), pneumonia in 18 neonates (27%), sepsis in 12 neonates (18%), and meconium aspiration syndrome in 10 neonates (15%). Among the 66 neonates with ARDS, 10 neonates (15%) were born to the mothers with an age of ≥ 35 years, 30 neonates (45%) suffered from intrauterine distress, 32 neonates (49%) had a 1-minute Apgar score of 0 to 7 points, 24 neonates (36%) had abnormal fetal heart monitoring results, and 21 neonates (32%) experienced meconium staining of amniotic fluid. Intraventricular hemorrhage was the most common comorbidity (12 neonates), followed by neonatal shock (9 neonates) and patent ductus arteriosus (8 neonates). All 66 neonates with ARDS were treated with mechanical ventilation in addition to the treatment for primary diseases. Among the 66 neonates with ARDS, 10 died, with a mortality rate of 15% (10/66), and 56 neonates were improved or cured, with a survival rate of 85% (56/66). Conclusions Neonatal ARDS in southwest Hubei is mostly mild or moderate. Perinatal asphyxia and infection may be the main causes of neonatal ARDS in this area. Intraventricular hemorrhage is the most common comorbidity. Neonates with ARDS tend to have a high survival rate after multimodality treatment.

Objective To investigate the current status of readmission of neonates with hyperbilirubinemia and risk factors for readmission. Methods From January 2017 to December 2019, a total of 85 infants who were readmitted due to hyperbilirubinemia were enrolled as the study group. A total of 170 neonates with hyperbilirubinemia but without readmission during the same period of time were randomly selected as the control group. The medical data were compared between the two groups. Multivariate logistic regression was used to assess the risk factors for readmission due to hyperbilirubinemia. Results The readmission rate was 2.30%, and the interval between readmission and initial admission was 5 days. Compared with the control group, the study group had significantly higher levels of total bilirubin and indirect bilirubin at discharge (P < 0.05) and a significantly longer duration of phototherapy during the first hospitalization (P < 0.05). The univariate analysis showed that compared with the control group, the study group had significantly lower birth weight, gestational age, and age on initial admission (P < 0.05) and a significantly higher proportion of infants with glucose-6-phosphate dehydrogenase (G-6-PD) deficiency or hemolytic disease (P < 0.05). The multivariate analysis showed that low gestational age (OR=1.792, P < 0.05), young age on initial admission (OR=1.415, P < 0.05), and G-6-PD deficiency (OR=2.829, P < 0.05) were independent risk factors for readmission of neonates with hyperbilirubinemia. Conclusions The infants with hyperbilirubinemia who have lower gestational age, younger age on initial admission, and G-6-PD deficiency have a higher risk of readmission due to hyperbilirubinemia. It is thus important to strengthen the management during hospitalization and after discharge for these infants to prevent the occurrence of readmission.

Objective To study the clinical effect of recombinant human interferon α1b assisting acyclovir on immune function, inflammatory factors, and myocardial zymogram in children with infectious mononucleosis (IM). Methods A total of 182 children with IM who were admitted to the hospital from January to December, 2018, were divided into an observation group with 91 children and a control group with 91 children using a random number table. The children in the control group were treated with intravenous drip of acyclovir, and those in the observation group were treated with inhalation of recombinant human interferon α1b in addition to the treatment in the control group. The two groups were compared in terms of clinical symptoms, immune function, inflammatory response, myocardial zymogram, and adverse reactions. Results Compared with the control group, the observation group had significantly shorter time to body temperature recovery and disappearance of isthmopyra, cervical lymph node enlargement, hepatomegaly, and splenomegaly (P < 0.05). After treatment, both groups had significant increases in CD4⁺, CD4⁺/CD8⁺, and CD19⁺, and the observation group had significantly higher levels of these markers than the control group (P < 0.05). After treatment, both groups had significant reductions in the levels of CD8⁺, tumor necrosis factor-α, interlukin-6, creatine kinase, and creatine kinase-MB, and the treatment group had significantly lower levels of these markers than the control group (P < 0.05). There was no significant difference in the incidence rate of adverse reactions between the two groups after treatment (P > 0.05). Conclusions For children with IM, recombinant human interferon α1b assisting acyclovir can effectively improve immune function, inhibit inflammatory reaction, reduce myocardial injury, and thus alleviate clinical symptoms.

Objective To study the role of microRNA-17-5p (miR-17-5p) in the pathogenesis of pediatric nephrotic syndrome (NS) and its effect on renal podocyte apoptosis via the activin A (ActA)/Smads pathway. Methods An analysis was performed on 55 children with NS (NS group) who were admitted from March 2018 to March 2019. Fifty healthy children who underwent physical examination during the same period of time were enrolled as the control group. The mRNA expression of miR-17-5p in peripheral blood was measured and compared between the two groups. Human renal podocytes were transfected with antisense oligonucleotide recombinant plasmid containing miR-17-5p (inhibition group) or control vector containing nonsense random sequence (negative control group), and untreated human renal podocytes were used as the blank group. These groups were compared in terms of cell apoptosis and the mRNA and protein expression of miR-17-5p, ActA, and Smads after transfection. Results The NS group had a significantly higher level of miR-17-5p in peripheral blood than the control group (P < 0.001). Compared with the blank and negative control groups, the inhibition group had significantly lower apoptosis rate and relative mRNA expression of miR-17-5p and significantly higher relative mRNA and protein expression of ActA, Smad2, and Smad3 (P < 0.001). Conclusions There is an increase in the content of miR-17-5p in peripheral blood in children with NS. Low expression of miR-17-5p can inhibit the apoptosis of human renal podocytes, which may be associated with the upregulation of the mRNA and protein expression of ActA, Smad2 and Smad3.

Objective To evaluate the efficacy and safety of tacrolimus in the treatment of children with myasthenia gravis (MG). Methods A total of 28 children with MG were treated with tacrolimus. MG-Activities of Daily Living (MG-ADL) scale was used to assess clinical outcome and safety after 1, 3, 6, 9, and 12 months of treatment. Results After tacrolimus treatment, the MG-ADL score at 1, 3, 6, 9 and 12 months was lower than that at baseline (P < 0.05), and the MG-ADL score showed a gradually decreasing trend. The response rates to tacrolimus treatment at 1, 3, 6, 9, and 12 months were 59%, 81%, 84%, 88%, and 88% respectively. At 6, 9, 12, and 18 months of treatment, 4, 13, 14, and 15 children respectively were withdrawn from prednisone. No recurrence was observed during treatment. Major adverse reactions/events were asymptomatic reduction in blood magnesium in 5 children and positive urine occult blood in 1 child, which turned negative without special treatment, and tacrolimus was not stopped due to such adverse reactions/events. One child was withdrawn from tacrolimus due to recurrent vomiting. According to CYP3A5 genotypes, all of the patients were divided into two groups:slow metabolic type (n=19) and non-slow metabolic type (fast metabolic type + intermediate type; n=9). The non-slow metabolism group received a higher dose of tacrolimus, but had a lower trough concentration of tacrolimus than the slow metabolism group (P < 0.05). The slow metabolism group had a higher response rates to tacrolimus treatment than the non-slow metabolism group (P < 0.05). Conclusions Tacrolimus appears to be effective and safe in the treatment of children with MG and is thus an option for immunosuppressive therapy. CYP3A5 genotyping has a certain guiding significance for determining the dosage of tacrolimus.

Objective To study the association of motor nerve conduction block (CB) with different subtypes of childhood Guillain-Barré syndrome (GBS). Methods A retrospective analysis was performed on the clinical and nerve electrophysiological data of 50 children with GBS. According to the results of nerve electrophysiology, the children were divided into an acute inflammatory demyelinating polyneuropathy (AIDP) group with 29 children and an acute motor axonal neuropathy (AMAN) group with 21 children. According to the presence or absence of motor nerve CB, the children with AMAN or AIDP were further divided into subgroups:group AMAN with or without motor nerve CB (n=10 and 11 respectively) and group AIDP with or without motor nerve CB group (n=19 and 10 respectively). The subgroups were compared in terms of age of onset, sex, Hughes Functional Grading Scale (HFGS) at nadir for the most severe involvement of motor function, and short-term prognosis based on HFGS score at 1 month after disease onset. Results Motor nerve CB was reversible in children with AMAN. AMAN children with motor nerve CB had a significantly lower HFGS score than those without motor nerve CB at 1 month after onset (P < 0.05). AIDP children with motor nerve CB had a significantly higher HFGS score than those with motor nerve CB at 1 month after onset (P < 0.05). Conclusions AMAN with reversible motor nerve CB suggests mild nerve fiber lesion and has better recovery than AMAN and AIDP without motor nerve CB in short term.

Objective To study the clinical features of aerophagia in children. Methods A retrospective analysis was performed on the medical data of 46 children with aerophagia who were diagnosed and treated in Children's Hospital Affiliated to Nanjing Medical University from October 2011 to September 2019. Results Among these 46 children, 15 (33%) had Tourette syndrome. Abdominal distension was the most common symptom and was observed in 45 children (98%). The 24-hour esophageal multichannel intraluminal impedance monitoring showed a mean number of 341 times of air swallowing and a mean number of 212 times of gas reflux, and 95% of gas refluxes occurred in the upright body position. Compared with those without Tourette syndrome, the children with Tourette syndrome had a significantly higher incidence rate of air swallowing symptoms (67% vs 6%, P < 0.001), but there were no significant differences in other symptoms and the results of 24-hour esophageal impedance. Dietary adjustment, psycho-behavioral therapy, and drug intervention significantly improved the scores of clinical symptoms and quality of life, among which psycho-behavioral therapy was an important intervention measure. Conclusions Some children with aerophagia may have Tourette syndrome, and such children are more likely to have air swallowing symptoms. Psycho-behavioral therapy is one of the most important treatment methods, and children with aerophagia tend to have a good prognosis after treatment.

Objective To study the expression level of cAMP response element-binding protein (CREB) in children with recurrent wheezing under three years of age and its effect on the expression of the serum orosomucoid 1-like protein 3 (ORMDL3) gene. Methods Thirty-six children with recurrent wheezing under three years of age who visited the hospital from June 2017 to June 2019 were selected as the recurrent wheezing group. Twenty-four healthy children from physical examination were selected as the control group. The CREB expression level in peripheral blood was measured by quantitative real-time PCR. Human bronchial epithelial cells (BEAS-2B) were cultured, and dual-luciferase reporter assay and quantitative real-time PCR were used to investigate the effects of overexpression and siRNA interference of CREB on the promoter activity and mRNA expression of the ORMDL3 gene in the BEAS-2B cells. Results The expression level of CREB in the recurrent wheezing group was significantly higher than that in the control group (P < 0.001). In BEAS-2B cells, overexpression of CREB significantly up-regulated the promoter activity and mRNA expression of the ORMDL3 gene (P < 0.05), while siRNA interference of CREB significantly reduced the promoter activity and mRNA expression of the ORMDL3 gene (P < 0.05). Conclusions The expression of CREB is increased in children with recurrent wheezing, and CREB may be involved in the pathogenesis of recurrent wheezing by regulating expression of the ORMDL3 gene.

Objective To study the influencing factors for the clinical effect of bronchoalveolar lavage (BAL) in children with Mycoplasma pneumoniae pneumonia (MPP) and atelectasis. Methods A total of 75 children with MPP and atelectasis were divided into a good response group with 51 children and a poor response group with 24 children according to the clinical effect of BAL treatment. LASSO logistic regression analysis was used to investigate the factors influencing the clinical effect of BAL treatment. The receiver operating characteristic (ROC) curve and restricted cubic spline model analysis were used to evaluate the value of the course of the disease at the time of BAL treatment in predicting the clinical effect of BAL treatment. Results Compared with the good response group, the poor response group had a significantly lower percentage of lymphocytes in bronchoalveolar lavage fluid, a significantly higher proportion of children with atelectasis of two or more lung lobes or stenosis of the bronchial cavity or opening caused by inflammation, and a significantly longer course of the disease at the time of BAL treatment and azithromycin treatment (P < 0.05). The LASSO logistic regression analysis showed that a prolonged course of the disease at the time of BAL treatment (OR=1.23), atelectasis of two or more lung lobes (OR=11.99), and stenosis of the bronchial cavity or opening caused by inflammation (OR=5.31) were independent risk factors for poor clinical effect of BAL treatment (P < 0.05). The ROC curve analysis showed that the course of disease of ≥ 11.5 days at the time of BAL treatment suggested a poor clinical effect of BAL treatment, with a sensitivity of 91.7% and a specificity of 54.9%. The restricted cubic spline model analysis showed that there was a non-linear dose-response relationship between the course of disease at the time of BAL treatment and the clinical effect of BAL treatment (P < 0.05). Conclusions Early BAL treatment may have a good clinical effect in children with MPP and atelectasis. Atelectasis of two or more lung lobes and inflammation-induced stenosis of the bronchial cavity or opening shown under bronchoscope may indicate a poor clinical effect of BAL treatment.

Objective To study the value of anti-neutrophil cytoplasmic antibody (ANCA) in assessing the severity of bronchiolitis obliterans (BO) in children. Methods A prospective analysis was performed on 59 children who were diagnosed with BO from June 2009 to October 2014. ELISA was used to measure the concentrations of myeloperoxidase (MPO)-ANCA and proteinase 3 (PR3)-ANCA in serum. According to the results of ELISA, the children were divided into three groups:double-negative ANCA (n=22), single-positive ANCA (n=17), and double-positive ANCA (n=20). The three groups were compared in terms of the scores of BO risk factors, clinical symptoms, chest high-resolution computed tomography (HRCT), and lung pathology on admission, as well as the changes in the expression level of ANCA and the scores of clinical symptoms and chest HRCT over time. Results Compared with the double-negative ANCA group, the double-positive ANCA group had a significantly higher score of BO risk factors (P < 0.05), and the single-positive ANCA group and the double-positive ANCA group had significantly higher scores of clinical symptoms, chest HRCT, and lung pathology (P < 0.05). The children were followed up for 6 months after discharge, and there were significant reductions in MPO-ANCA and PR3-ANCA titers from admission and discharge to the end of follow-up (P < 0.05), as well as a significant reduction in the score of clinical symptoms from admission to the end of follow-up (P < 0.05), while there was no significant change in the score of chest HRCT from admission to the end of follow-up (P > 0.05). The single-positive ANCA and double-positive ANCA groups still had a significantly higher score of clinical symptoms than the double-negative ANCA group (P < 0.05). Conclusions The expression level of ANCA is correlated with the severity of BO in children and thus has certain clinical significance in disease evaluation.

Objective To study the value of cleaved lymphocytes in peripheral blood smear in assisting the early diagnosis of pertussis. Methods Nasopharyngeal swabs and peripheral blood samples were collected from 107 children with pertussis-like disease. PCR-flow fluorescent hybridization was used to detect the nucleic acids of Bordetella pertussis. Based on the detection results, the children were divided into two groups:pertussis (n=52) and non-pertussis (n=55). According to age, the pertussis group was divided into two subgroups:<1 year old (n=42) and ≥ 1 year old (n=10). According to disease severity, the pertussis group was divided into another two subgroups:mild (n=45) and severe (n=7). An automatic blood cell analyzer was used to determine peripheral blood cell counts. Wright's staining and peroxidase staining were used to observe and count cleaved lymphocytes under a microscope. Results Cleaved lymphocytes in peripheral blood were round with small cytoplast, less cytoplasm and cleaved or lobulated nuclei. According to the negative peroxidase staining results, these cells were confirmed as lymphocytes. Compared with the non-pertussis group, the pertussis group had significantly higher leukocyte count, lymphocyte percentage, platelet count, and percentage of cleaved lymphocytes (P < 0.001). For the children with pertussis, the <1 year old subgroup had significantly higher lymphocyte percentage, platelet count, and percentage of cleaved lymphocytes than the ≥ 1 year old subgroup (P < 0.05). The severe subgroup had slightly higher leukocyte count, lymphocyte percentage, platelet count, and percentage of cleaved lymphocytes than the mild subgroup (P > 0.05). Conclusions The detection of cleaved lymphocytes combined with peripheral blood cell counts provides a new option for the early diagnosis of pertussis in children.

Objective To study the clinical screening and genetic diagnosis of children suspected of Prader-Willi syndrome (PWS), as well as the differences in the scores of clinical diagnostic criteria among the children with a confirmed diagnosis of PWS. Methods A total of 94 children suspected of PWS who were admitted from July 2016 to December 2018 were enrolled as subjects. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) was performed to confirm the diagnosis. For the children with a confirmed diagnosis of PWS, the scores of clinical diagnostic criteria were determined, and the perinatal characteristics were analyzed. Results A total of 11 children with PWS were confirmed by MS-MLPA, with a detection rate of 12%, among whom there were 7 boys and 4 girls, with a median age of 3 years and 4 months (range 25 days to 6 years and 8 months) at the time of confirmed diagnosis. Among the 11 children with PWS, only 5 children (45%) met the criteria for clinical diagnosis. The main perinatal characteristics of the children with PWS were decreased fetal movement (9 cases, 82%), cesarean section birth (11 cases, 100%), hypotonia (11 cases, 100%), feeding difficulties (11 cases, 100%), and weak crying (11 cases, 100%). Conclusions Gene testing should be performed as early as possible for children suspected of PWS by clinical screening. PWS may be missed if only based on the scores of clinical diagnostic criteria.

Objective To evaluate the value of capsule endoscopy in children with small intestinal diseases with hematochezia as the chief complaint. Methods A retrospective analysis was performed on the clinical data and capsule endoscopy findings of 93 children with hematochezia who were admitted to Children's Hospital of Fudan University from May 2015 to January 2019 and underwent capsule endoscopy. According to the capsule endoscopy findings of the jejunum and the ileum, they were divided into a positive lesion group with 39 patients and a negative lesion group with 54 patients. Related clinical data and the features of lesion on capsule endoscopy were analyzed for the two groups. Results There were no significant differences in age, sex, duration of capsule endoscopy, gastric transit time, and small intestinal transit time between the positive lesion and negative lesion groups (P > 0.05). The positive lesion group had a significantly lower level of hemoglobin than the negative lesion group (P < 0.05). Hemoglobin level was negatively correlated with the rate of positive lesions on capsule endoscopy (r=-0.342, P=0.001). Among the 39 patients with positive lesions on capsule endoscopy, the detection of Meckel's diverticulum was the highest (41%), followed by inflammatory bowel disease (21%). Conclusions Capsule endoscopy has a certain value in detecting small intestinal diseases, especially diseases in the jejunum and the ileum, in children with lower gastrointestinal hemorrhage.

To study the clinical effect of oral sirolimus in the treatment of children with blue rubber bleb nevus syndrome (BRBNS) in the gastrointestinal tract, a retrospective analysis was performed on the clinical data and follow-up results of two children with BRBNS treated by sirolimus. The two children with BRBNS had gastrointestinal bleeding and anemia and were treated with sirolimus at a dose of 1 mg/day as part of treatment. The plasma concentration of the drug was maintained between 2.5-12.0 ng/mL. The children showed disappearance of gastrointestinal bleeding and improvements in anemia and coagulation function, and blood transfusion could be stopped during treatment, with no obvious adverse drug reactions. PubMed, Wanfang Data, and CNKI were searched for related articles on sirolimus in the treatment of BRBNS. A total of 26 cases of children with BRBNS, aged 0-18 years, were obtained. With the addition of the 2 cases in this study, sirolimus treatment achieved a satisfactory clinical effect in all 28 cases. Sirolimus may be effective and safe in the treatment of children with BRBNS, and further prospective studies are needed to evaluate the long-term efficacy of this drug.

Objective To study the effect of advanced maternal age (AMA) on the development of hippocampal neural stem cells in offspring rats. Methods Ten 3-month-old and ten 12-month-old female Sprague-Dawley rats were housed individually with 3-month-old male rats (1:1, n=20), whose offspring rats were assigned to a control group and an AMA group. A total of 40 rats were randomly selected from each group. Immunofluorescence assay and Western blot were used to localize and determine the levels of protein expression of Nestin and doublecortin (DCX) on day 7 as well as neuronal nuclear antigen (NeuN) and glial fibrillary acidic protein (GFAP) on day 28 (n=8 for each marker). Immunofluorescence assay was also used to localize the hippocampal expression of polysialylated isoforms of neural cell adhesion molecule (PSA-NCAM) on day 14 (n=8 for each marker). Results According to the Western blot results, the AMA group had significantly lower protein expression of DCX than the control group (P < 0.05), while there were no significant differences in the protein expression of Nestin, NeuN, and GFAP between the two groups (P > 0.05). According to the results of immunofluorescence assay, the AMA group had significantly lower protein expression of Nestin, DCX, and PSA-NCAM in the hippocampal dentate gyrus (DG) region than the control group (P < 0.05), while there were no significant differences in the above indices in the hippocampal CA1 and CA3 regions between the two groups (P > 0.05). The AMA group had significantly higher expression of NeuN in the hippocampal CA1 region than the control group (P < 0.01), while there were no significant differences in the expression of NeuN in the hippocampal DG and CA3 regions between the two groups (P > 0.05). The AMA group had significantly lower expression of GFAP in the hippocampal CA1, CA3, and DG regions than the control group (P < 0.05). Conclusions AMA may cause inhibition of proliferation, survival, and migration of hippocampal neural stem cells. AMA may also affect their differentiation into neurons and astrocytes, which will eventually lead to developmental disorders of hippocampal neural stem cells in offspring rats.

Objective To study whether pyroptosis is involved in the bilirubin-induced injury of primary cultured rat cortical microglial cells. Methods Primary cultured rat cortical microglial cells were randomly administered with 30 μmol/L bilirubin (bilirubin group), 30 μmol/L bilirubin following 30 μmol/L VX-765 pretreatment (VX-765+bilirubin group), or an equal volume of dimethyl sulfoxide (control group). Modified MTT assay was used to measure the viability of microglial cells. Western blot was used to measure the expression of the pyroptosis-related proteins Caspase-1 and gasdermin D (GSDMD). Lactate dehydrogenase (LDH)-release assay was used to evaluate the cytotoxicity of microglial cells. EtBr/EthD2 with different molecular weights (394 Da/1 293 Da) was used to measure the size of plasma membrane pores. ELISA was used to measure the level of the inflammatory factor interleukin-1β (IL-1β) in culture supernatant. Results After bilirubin stimulation, the viability of microglial cells decreased and LDH release increased, both in a time-dependent manner. Compared with the control group, the bilirubin group had a significantly higher positive rate of small-molecule EtBr passing through the cell membrane (P < 0.001), while there was no significant difference in the pass rate of large-molecule EthD2 between groups (P > 0.05). The expression of activated Caspase-1 significantly increased at 0.5 hour after bilirubin stimulation (P < 0.05), and that of activated GSDMD significantly increased at 6 hours after bilirubin stimulation (P < 0.05). The release of IL-1β significantly increased at 6 hours after bilirubin stimulation and reached the peak at 24 hours (P < 0.001). Compared with the bilirubin group, the VX-765+bilirubin group had a significant increase in cell viability (P < 0.05) and significant reductions in the expression of activated GSDMD, the pass rate of EtBr, and the release of LDH and IL-1β (P < 0.05). Conclusions Pyroptosis is involved in bilirubin-induced injury of primary cultured microglial cells.

Objective To study the effect and related signaling pathways of ginsenoside Rb1 in the treatment of coronary artery lesion (CAL) in a mouse model of Kawasaki disease (KD). Methods BALB/c mice were randomly divided into a control group, a model group, an aspirin group, a low-dose ginsenoside Rb1 group (50 mg/kg), and a high-dose ginsenoside Rb1 group (100 mg/kg), with 12 mice in each group. All mice except those in the control group were given intermittent intraperitoneal injection of 10% bovine serum albumin to establish a mouse model of KD. The mice in the aspirin group, the low-dose ginsenoside Rb1 group, and the high-dose ginsenoside Rb1 group were given the corresponding drug by gavage for 20 days after modeling. Hematoxylin and eosin staining was used to observe the pathological changes of coronary artery tissue. ELISA was used to measure the levels of the inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in serum and coronary artery tissue. Western blot was used to measure the relative expression levels of proteins involved in the regulation of the AMPK/mTOR autophagy signaling pathway and the PI3K/Akt oxidative stress signaling pathway in coronary artery tissue. Results The observation of pathological sections showed that compared with the model group, the high-dose ginsenoside Rb1 group had significant improvement in the symptoms of vascular wall thickening, intimal edema, fiber rupture, and inflammatory infiltration of endothelial cells. Compared with the control group, the model and low-dose ginsenoside Rb1 groups had significant increases in the levels of TNF-α, IL-6, and IL-1β in serum and coronary artery tissue (P < 0.05); the model group had significant increases in the expression levels of P-AMPK/AMPK, P-mTOR/mTOR, and P-P70S6/P70S6 in coronary artery tissue (P < 0.05) and significant reductions in the expression levels of P-PI3K/PI3K, P-AKT/AKT, and P-GSK-3β/GSK-3β in coronary artery tissue (P < 0.05). Compared with the model group, the aspirin group and the high-dose ginsenoside Rb1 group had significant reductions in the levels of TNF-α, IL-6, and IL-1β (P < 0.05); the low- and high-dose ginsenoside Rb1 groups had significant reductions in the expression levels of P-AMPK/AMPK, P-mTOR/mTOR, and P-P70S6/P70S6 (P < 0.05) in a dose-dependent manner between the two groups (P < 0.05); the low-dose ginsenoside Rb1 group had no significant change in the expression level of P-PI3K/PI3K (P > 0.05) and had significant increases in the expression levels of P-AKT/AKT and P-GSK-3β/GSK-3β (P < 0.05), while the high-dose ginsenoside Rb1 group had significant increases in the relative protein expression levels of the above three proteins (P < 0.05). Compared with the low-dose ginsenoside Rb1 group, the aspirin group and the high-dose ginsenoside Rb1 group had significant reductions in the levels of TNF-α, IL-6, and IL-1β (P < 0.05); the high-dose ginsenoside Rb1 group had significant increases in the expression levels of P-PI3K/PI3K and P-AKT/AKT (P < 0.05). Conclusions Ginsenoside Rb1 can effectively alleviate CAL in a mouse model of KD in a dose-dependent manner, possibly by regulating the AMPK/mTOR/P70S6 autophagy signaling pathway to inhibit CAL inflammation and regulating the PI3K/AKT/GSK-3β oxidative stress signaling pathway to exert a biological activity of protection against coronary artery endothelial cell injury.

This article reports two children with hereditary hemorrhagic telangiectasia (HHT). Patient 1 was a boy aged 12 years and was admitted due to intermittent cough and wheezing for more than 10 years. This boy and his mother and grandmother had a history of epistaxis. The boy had a history of the rupture of cerebral arteriovenous malformations. Gene detection showed a heterozygous mutation, c.277C > T(p.Arg93^*), in the ENG gene. Patient 2 was a girl aged 13 years and was admitted due to cyanosis of lips for more than 1 year. The girl had a history of recurrent epistaxis and the manifestations of severe decline in pulmonary diffuse function, pulmonary hypertension, dilation of blood vessels at the distal end of lungs, and small arteriovenous communications in both lungs. Children with HHT often lack typical respiratory symptoms, which may lead to missed diagnosis and misdiagnosis in the early stage. Pulmonary computed tomography or right cardiac acoustic contrast can help with the diagnosis of HHT, and gene detection can improve the early diagnostic rate of this disease.

This article evaluates the potential influence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in pregnant women on the development of coronavirus disease 2019 in neonates and discusses the possibility of mother-to-child vertical transmission of SARS-CoV-2. With reference to related articles published up to now and the information on official websites, a retrospective review was performed for the clinical manifestations and laboratory examination results of the neonates born to the mothers with infection during pregnancy during the epidemics of severe acute respiratory syndrome and Middle East respiratory syndrome and after the outbreak of SARS-CoV-2 infection since December 2019. Based on the limited data, there is no conclusive evidence for mother-to-child vertical transmission of coronavirus disease 2019, and more cases need to be collected for further evaluation.