Background There is a lack of research evidence on the association between sugar-sweetened beverage (SSB) consumption and gestational diabetes mellitus (GDM) in China. Objective To explore the association between frequency of SSB consumption before pregnancy and risk of GDM in pregnant women in Shaanxi Province, and to provide a scientific basis for targeted interventions to control maternal blood glucose. Methods The recruitment to the China Birth Cohort study started in October 2020. Pregnant women at 6-16 weeks who had their first prenatal examination at five hospitals in Shaanxi Province were recruited. A maternal health questionnaire was used to collect basic information about pregnant women. A semi-quantitative food frequency questionnaire was used to collect the consumption of carbonated beverages, fruit and vegetable juice beverages, coffee beverages, and milk tea beverages in one year before pregnancy, which were summed to obtain the SSB consumption. Pregnant women were divided into three groups according to SSB consumption, namely <1 serving·week⁻¹, 1-4 servings·week⁻¹, and ≥5 servings·week⁻¹. GDM was confirmed by oral glucose tolerance test (OGTT) between 24-28 weeks of gestation. A binary logistic regression model was applied to explore the association between SSB consumption and risk of GDM. Multiple linear regression was applied to investigate the associations between SSB consumption (per 1-serving·d⁻¹ increase) and OGTT fasting plasma glucose, 1-hour glucose, and 2-hour glucose. Results A total of 3811 pregnant women were finally enrolled in this study, of which 752 developed GDM, with an incidence rate of 19.7%. The incidence rates of GDM in pregnant women with SSB consumption frequency of <1 serving·week⁻¹, 1-4 servings·week⁻¹, and ≥5 servings·week⁻¹ were 18.0%, 21.1%, and 26.8%, respectively. After adjusting for maternal age, pre-pregnancy body mass index (BMI), education, number of children born, family history of diabetes, smoking, alcohol consumption, physical activity level, and total energy intake, the risk of GDM increased by 26% (OR=1.26, 95%CI: 1.05, 1.50) in the 1-4 servings·week⁻¹ group and by 76% (OR=1.76, 95%CI: 1.31, 2.38) in the ≥5 servings·week⁻¹ group compared to the <1 serving·week⁻¹ SSB consumption group, respectively. Further stratified analysis revealed no interaction effect (P_interaction>0.05) between SSB consumption and maternal age, pre-pregnancy BMI, or first labor or not. For each additional SSB consumption per day, the risk of GDM increased by 94% (OR=1.94, 95%CI: 1.37, 2.75); and the maternal OGTT 1-hour glucose and 2-hour glucose increased by 0.33 mmol·L⁻¹ and 0.18 mmol·L⁻¹, respectively (P<0.05), and no significant increase in fasting plasma glucose was found (P>0.05). Conclusion Higher SSB consumption before pregnancy increases the risk of GDM in pregnant women.

【Objective】 To study the genotypes of ABO ambiguous blood group samples(n=20) and identify their molecular biological characteristics. 【Methods】 The serological phenotype of the samples was analyzed by serological techniques. Seven exons of ABO gene were amplified by polymerase chain reaction (PCR) and the PCR products were directly sequenced; the genotypes and sequences of ABO subtypes were analyzed. 【Results】 The serological phenotypes of 20 samples presenting ABO ambiguous blood group were as follows: weak A antigen (n=5), weak A antigen combined with anti-A1 antibody (n=5), normal A antigen combined with anti-A1 antibody (n=2), weak B antigen (n=8). The genotypes of them were as follows: Ax02/O01 (n=3), Ael07/O01 (n=2), B313/O01 (n=2), A204/O02 (n=1), A220/O01 (n=1), Ael07/O02 (n=1), Ael02/O01 (n=1), Ael02/O02 (n=1), Ax03/O01 (n=1), Ax03/O02 (n=1), B313/O02 (n=1), B302/O01 (n=1), B302/O02 (n=1), Bw19/O02 (n=1), A102/B313 (n=1) and A101/Bw37 (n=1). 【Conclusion】 ABO genotyping technology can accurately identify the ambiguous blood group of samples, provide definite genetic information of blood group and ensure the safety of clinical transfusion.

Objective:To investigate the clinical phenotype and gene mutation in a child with developmental disorders caused by CTNNB1 gene mutation. Methods:Clinical data of a child with CTNNB1 gene mutation who was admitted to Xiamen Hospital of Fudan University Affiliated Pediatric Hospital in May 2017 were collected, whole exome sequencing technology was applied to verify the family lineage of the child, and the pathogenicity of mutation site was analyzed. Results:The patient was a 6 years and 1 month old male, with a clinical phenotype including mental retardation, motor developmental disorders, speech disorders, visual disorders (internal strabismus), microcephaly, and behavioral problems (social withdrawal, overdependence, etc.), as well as panic syndrome (i.e., sudden shrieking in response to auditory and visual stimuli, extensional rigidity of the body, etc., followed by short periods of general extensional rigidity). The whole exome sequencing results showed the presence of a de novo mutation c.283(exon4)C>T in the CTNNB1 gene, and the c.283(exon4)C>T mutation was interpreted as pathogenic (PVS1+PS2+PS1+PM2+PM) according to the American College of Medical Genetics and Genomics variant classification criteria and guidelines. No relevant genetic variants were found in the parental family verification. Conclusion:CTNNB1 gene mutation c.283(exon4)C>T can cause neurodevelopmental disorders, including mental retardation, motor developmental disorders, speech disorders, visual disorders, microcephaly and behavioral abnormalities.

Objective:To deeply analyze differences in characteristics of neurosyphilis between male and female patients with neurosyphilis, as well as between patients with symptomatic neurosyphilis and those with asymptomatic neurosyphilis, and to provide reference for the prevention and control, clinical diagnosis and treatment of neurosyphilis.Methods:A total of 131 inpatients with neurosyphilis were collected from Department of Dermatology and Venereology, the First Affiliated Hospital of Anhui Medical University from June 2015 to December 2019, and their clinical manifestations and laboratory findings were retrospectively analyzed. These patients were grouped according to gender and neurological/psychiatric symptoms. Measurement data were compared by using two-independent-sample t test or Mann-Whitney U test, and enumeration data were compared by using chi-square test and Fisher′s exact test, to analyze differences in clinical characteristics and laboratory indicators between different groups. Results:Among the 131 patients, there were 72 with asymptomatic neurosyphilis (asymptomatic group) and 59 with symptomatic neurosyphilis (symptomatic group). The proportion of patients receiving syphilis treatment was significantly lower in the symptomatic group (10.17%) than in the asymptomatic group (98.61%, OR = 0.002, P < 0.001). The misdiagnosis rate at the first clinical visit was significantly higher in the male patients (50.00%) than in the female patients (24.49%, OR = 3.08, P = 0.004), as well as in the symptomatic patients (89.83%) than in the asymptomatic patients (0, OR = 13.00, P < 0.001). The proportion of symptomatic patients was significantly higher in male patients (57.32%) than in female patients (14.64%, OR = 4.14, P = 0.003). Compared with the female patients, the male patients showed significantly increased positive rates of toluidine red unheated serum test (TRUST) in the cerebrospinal fluid samples (52.44% vs. 26.54%, OR = 3.05, P = 0.004), increased proportions of patients with elevated levels of total protein (> 0.5 g/L) in cerebrospinal fluids (79.27% vs. 59.18%, OR = 2.64, P = 0.01), increased total protein levels in cerebrospinal fluids (0.76 ± 0.41 g/L vs. 0.56 ± 0.25 g/L, P = 0.002), and increased detection rates of brain magnetic resonance imaging abnormalities (72.22% vs. 44.90%, OR = 2.13, P = 0.039). The age at diagnosis of the symptomatic female patients (50.82 ± 9.31 years) was significantly higher than that of the asymptomatic female patients (42.30 ± 12.18 years, P = 0.038). The positive rate of TRUST in the cerebrospinal fluid samples was significantly higher in the patients with symptomatic neurosyphilis (55.93%) than in those with asymptomatic neurosyphilis (31.94%, OR = 2.70, P = 0.006), and so was the total protein level in cerebrospinal fluids (0.79 ± 0.46 g/L vs. 0.60 ± 0.24 g/L, P = 0.003) . Conclusion:The misdiagnosis rate of neurosyphilis is high at the first clinic visit; the condition of male patients is more serious than that of female patients; anti-syphilitic treatment history, gender and age may play some role in the development of neurosyphilis.

Jaundice is a common clinical problem in neonatal period.Phototherapy is a common treatment for neonatal jaundice, but it also has side effects such as fever, diarrhea, rash and so on.In recent years, probiotics have been widely used in neonates with jaundice because they are beneficial to the health of the host, especially when they are treated with light and probiotics are added at the same time, which is more conducive to the elimination of jaundice.Studies have shown that abnormal bilirubin metabolism is closely related to microecology.This article reviews the mechanism and clinical application of probiotics in adjuvant treatment of neonatal jaundice.

OBJECTIVE: To explore the genetic basis for a fetus featuring growth restriction and validate the effectiveness of a novel noninvasive prenatal testing (NIPT) technique for the detection of chromosomal microdeletions. METHODS: Next-generation sequencing(NGS) and fluorescence in situ hybridization(FISH) were used to analyze the DNA of the fetus. Conventional G-banding was used to analyze the karyotypes of the fetus and its parents. High-throughput sequencing was used to analyze free fetal DNA. RESULTS: NGS analysis has revealed a 4.88 Mb deletion at 15q11.2-q13.1 region in the fetus, which has a 99% overlap with the critical region of Prader-Willi syndrome (Type 2) and Angelman syndrome (Type 2) and encompassed critical genes including SNRPN and UBE3A. NIPT also revealed a 4.6 Mb deletion at 15q12, which was consistent with the results of fetal cord blood and amniotic DNA testing. FISH assay has confirmed the result of NGS. By karyotying, all subjects showed a normal karyotypes at a level of 320~400 bands. CONCLUSION It is quite necessary to carry out genetic testing on fetuses showing growth restriction. NIPT for fetal chromosomal microdeletions/microduplication syndromes is highly accurate for the diagnosis of Prader-Willi/Angelman syndrome.
Angelman Syndrome ; Chromosome Banding ; Chromosomes, Human, Pair 15 ; Female ; Fetus ; Humans ; In Situ Hybridization, Fluorescence ; Prader-Willi Syndrome ; Pregnancy

Objective To study the relationships between differences in tumorigenicity and immu-nogenetic backgrounds of nude mice. Methods According to the Chinese Pharmacopoeia, positive and neg-ative groups were set up in both Laboratory A and B with ten nude mice in each group. Organ tissues were collected for clinicopathological analysis. Blood samples were collected and detected using flow cytometry. DNA was extracted and analyzed with 23 STR markers. Results The positive group of Laboratory B was in-valid (7/10 tumor formation). The two laboratories showed no significant difference in the results of patho-logical analysis, but had significant differences in CD25, CD8, CD4, Th1 and Th2. There were 13 and 18 polymorphic sites respectively found in nude mice of Laboratory A and B. Further analysis of the non-tumor-bearing nude mice in Laboratory B positive group revealed that CD25, Th2, D3Mit29 and D5Mit48 were the specific indexes. Conclusion Differences in tumorigenicity might be related to the diversity of immunoge-netic backgrounds of nude mice.

Objective To optimize and compare four in vitro cytotoxicity assays in order to find a relatively rapid assay that can replace the traditional 51Cr release assay to evaluate the cytotoxicity of immune cells prepared for immunotherapy.Methods Four assays including BATDA, CAM (calcein acetoxymethyl ester), CytoTox-Glo and PKH were optimized and used to measure the in vitro cytotoxicity of NK-92 cells to K562 cell line.Intra-and inter-assay reproducibility of these assays and their correlation with 51Cr release assay were analyzed.Results After optimization, all of the four cytotoxicity assays showed good correlation with effector to target (E/T) ratio in a certain range.Compared with the other three assays, CytoTox-Glo assay showed obvious hook effect at a high E/T ratio of 40∶1.BATDA assay could detect the significant cytotoxicity of NK-92 cells to K562 cells after incubating them for one hour and that was the shortest time taken by the four assays to detect in vitro cytotoxicity.Both CAM and PKH assays took about four hours and CytoTox-Glo assay took six to eight hours to detect the significant cytotoxic potency.All of the four assays, especially BATDA and CAM assays, showed good intra-and inter-assay reproducibility.Among these assays, BATDA assay showed the best correlation with the traditional 51Cr release assay.BATDA assay, as compared with the other three assays, could be used to detect the cytotoxicity of Caov3 cells, an adherent cell line, and showed good results in evaluating the cytotoxic potency of autologous primary NK cells and CD19-CAR T cells.Conclusion Compared with the other three assays, BATDA assay shows the best linear correlation with 51Cr release assay and has the advantages of time-saving and good reproducibility.Besides, it is a better assay for detecting the cytotoxicity of adherent cells.BATDA assay is a promising substitute for 51Cr release assay in evaluating the in vitro cytotoxic potency of NK cells and other immune cells.

Objective:To study the metabolism of pedunculoside treated with rat intestinal bacteria in vitro. Methods:Pedunculo-side and rat intestinal bacteria were incubated in vitro for 0, 4, 8, 24 and 48 hours under anaerobic condition. After extracted repeat-edly by ethyl acetate, the metabolites in the incubation media were qualitatively and quantitatively analyzed by HPLC. Results:Totally 90. 8% of pedunculoside was transformed to M2 after incubated with rat intestinal bacteria in vitro for 48 hours, and a detailed compari-son of HPLC profiles between M2 and rotundic acid showed M2 was rotundic acid. Conclusion: Pedunculoside can be metabolized to rotundic acid by rat intestinal bacteria in vitro.

Objective To observe the attachment of Treponema pallidum to human brain microvascular endothelial cells (HBMECs) in vitro.Methods Some primary cultured HBMECs were inoculated into in 24-well plates to be cocultured with the suspension of T.pallidum at a concentration of 1.6 × 107 treponemes/ml.After 0.5,2 and 4 hours of co-culture,scanning electron microscopy was conducted to observe the attachment of T.pallidum to HBMECs.Some HBMECs were cocultured with the presence of T.pallidum suspensions at different concentrations (4 × 106,8 × 106,1.6 × 107 treponemes/ml) for 2,4,6 and 16 hours,then,dark-field microscopy was performed to count the number of treponemes that attached to single HBMECs.Statistical analysis was carried out by using repeated-measures analysis of variance.Results As scanning electron microscopy showed,treponemes gathered at some regions on the surface of HBMECs when they attached to HBMECs.In addition,T.pallidum partly merged with the membrane of HBMECs at the site of attachment.After co-culture with T.pallidum suspensions,the number of treponemes that attached to single HBMECs was significantly different among different time points (F =387.72,P ＜ 0.001) and among different concentrations of T.pallidum suspensions (F =593.23,P ＜ 0.001),with an interaction effect between the concentration of T.pallidum suspensions and incubation period (F =98.74,P ＜ 0.001).Concretely speaking,the number of treponemes that attached to single HBMECs increased over time until 6 hours after the start of coculture,then showed a decreasing trend,and reached the nadir value at 16 hours.Conclusion T.pallidum can adhere to cultured HBMECs in vitro,likely by the merger of its end with the membrane of HBMECs at some regions.