Objective To investigate whether allocardiac graft acceptance in the specific NF-κB impaired mice is due to regulatory T cell(Tr) and Th17 cells.Methods Mice abdominal heterotopic cardiac transplantation was performed and then divided in to control group(BALB/c→C57BL/6) and experimental group(BALB/c→IκBα/△N-Tg).Pretransplant and at day 7,30,100 posttransplant,spleens were harvested from the IκBα△ N-Tg mice,and then the Tr were detected by the fluorescence activated cell sorter.At day 5 posttransplant,the CD4 + Th17 cells from the spleens of the two groups were examined by the FACS.Additionally,at day 3 and 5 posttransplant,IL-17 expressed in the cardiac allograft was detected by Western blot.Results In the IκBα/ N-Tg mice group,the cardiac allografts were survived more than day 100,and without obviously lymphocytes infiltration.At the day 7 and 30 posttransplant,the Tr was obviously increased(21.23 ± 3.95,23.17 ± 4.11 vs 11.64 ± 1.96,P ＜ 0.05); however,the Tr decreased at the day 100 posttransplant,and had no difference with before transplant(10.79 ±2.48 vs 11.64 ± 1.96,P ＞0.05).Compared with the control group,at day 5 posttransplant,CD4+ Th17 cells in the IκBα/N-Tg mice and IL-17 expression of the cardiac allograft were both decreased.Conclusion In the early stage after transplantation,specific T cell NF-κB impaired could abrogate the balance of the Tr and Th17 cells,and induce the T cells differentiated into Tr and inhibit the Th17 cells differentiation,and then induce tolerance.

Objective To investigate the alterations of Treg cells , Th17 cells and related cyto-kines in peripheral blood of patients during the early stage of hand-foot-mouth disease ( HFMD) .Methods Flow cytometry was performed to analyze the percentages of Treg cells ( CD4+CD25+Foxp3 T cells) and Th17 cells ( CD3+CD8-IL-17+T cells) in peripheral blood samples collected from 49 patients with severe HFMD , 26 patients with common HFMD and 30 healthy children.The levels of IL-6, IL-10, IL-17, IL-23 and TGF-β1 in serum samples were measured by ELISA .Results The percentages of Treg cells , ratios of Treg/Th17 cells, serum levels of TGF-β1 and IL-10 in patients with HFMD were significantly decreased as compared with those of control group (F=5.580, 6.205, 0.000, 0.014, respectively, P<0.05).Patients with se-vere HFMD showed a significantly increased Th17 cells (F=3.189 P<0.05) and a tendency of enhanced IL-17 expression , but no significant differences with the levels of IL-17 were observed .No significant differ-ences with the expression of IL-23 in the patients among each group were detected (P>0.05).The levels of IL-6 in serum samples from severe disease group were obviously increased as compared with those of common HFMD group and control group (F=7.318, P<0.05).Conclusion The results of this study demonstrated that the levels of Treg , Th17 cells and some related cytokines were varied in peripheral blood of patients dur-ing the early stage of HFMD .Inflammatory responses were enhanced to promote anti-virus activities by sup-pressing Treg cells and stimulating Th17 cells.

OBJECTIVETo observe the expression of bcl-2 and bcl-xL in rat brain neuron after acute brain trauma.

METHODSThe rat model of mild and sever brain trauma were made by diffused brain injury. The expression of bcl-2 and bcl-xL in rat brain neuron was examined by immunohistochemical staining. Rat neuron apoptosis was detected by TUNEL method.

RESULTSThere were few bcl-2 (cortex: 4.40 +/- 1.67, hippocampal: 3.20 +/- 1.30) and lots of bcl-xL (cortex: 45.60 +/- 4.34, hippocampal: 50.20 +/- 3.50) expression in normal controls. After impact, the expression of bcl-2 in rat brain neuron increased, most distinctly on day 1(cortex: 30.0 +/- 4.3, hippocampal: 46.6 +/- 3.2), in mild group. A negative correlation was seen between bcl-2 expression and neuronal apoptosis (-1 < r < -0.847, P< 0.01, n = 10). No change was seen in bcl-xL expression.

CONCLUSIONSbcl-2 expression increased after acute brain trauma but bcl-xL did not change much. Both bcl-2 and bcl-xL are concerned with anti-apoptosis in neuron after acute brain trauma.

Acute Disease ; Animals ; Apoptosis ; Brain Injuries ; metabolism ; pathology ; Immunohistochemistry ; Male ; Neurons ; chemistry ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Rats ; Rats, Wistar ; bcl-X Protein

OBJECTIVETo analyze the role of Caspase 3 in neuronal apoptosis after acute brain injury.

METHODSExperiments were carried out with rat diffuse brain trauma model. The neuronal DNA injury in cortex and hippocampus was observed by TUNEL stain. The mRNA and protein expressions and enzyme activation of Caspase 3 were observed by Northern blot, in situ hybridization, immunohistochemistry stain and Western blot, respectively. Special Caspase 3 enzyme inhibitor was used to observe the therapeutic effect.

RESULTSTUNEL positive neurons appeared 2 hours after severe trauma, peaked at 1 day and lasted for 7 days. Northern blot showed that the Caspase 3 mRNA expression was increased and peaked at 1 day, about twice higher than the control. In the area of cortex and hippocampus, positive mRNA staining neurons appeared most distinct on one day. With the antibody for Caspase 3 P20 subunit, the active Caspase 3 expression peaked at 1-3 days. The electrophoresis band of PARP degradation would be seen by Western blot. Caspase 3 enzyme inhibitor could reduce apoptotic neuronal death without any effect on Caspase 3 P20 subunit expression.

CONCLUSIONSAfter brain trauma, Caspase 3 mRNA and protein expressions and enzyme activation are enhanced in combination with neuronal apoptosis. Special Caspase 3 enzyme inhibitor can apparently decrease the neuronal apoptosis.

Acute Disease ; Animals ; Apoptosis ; physiology ; Brain Injuries ; enzymology ; physiopathology ; Caspase 3 ; Caspases ; metabolism ; Enzyme Activation ; Enzyme Inhibitors ; pharmacology ; Nervous System ; physiopathology ; Neurons ; enzymology ; physiology ; RNA, Messenger ; metabolism ; Rats ; Rats, Wistar

Objective To investigate the frequency and distribution characteristics of gene mutations in children with phenylalanine hydroxylase(PAH)deficiency in Xinjiang.Methods A total of 230 children diag-nosed with PAH deficiency in Urumqi Maternal and Child Health Care Hospital from January 1st,2015 to February 28th,2023 were enrolled in the study.The variation of PAH gene was analyzed and the variation sites of PAH gene in children with different phenotypes were compared.Results A total of 441 PAH gene va-riants were detected in 230 children with PAH deficiency in Xinjiang,with a total detection rate of 95.87％.A-mong them,2 variants were detected in 227 cases,only 1 variant was detected in 2 cases,and 3 variants were detected in 1 case.217 cases were complex heterozygous variants,and 10 cases were homozygous variants.The high-frequency variant loci were c.158G＞A[23.39％(102/441)],c.728G＞A[11.70％(51/441)],c.688G＞A[5.05％(22/441)],c.721C＞T[3.90％(17/441)],c.611A＞G[3.67％(16/441)],c.1238G＞C[3.21％(14/441)].The high-frequency variant loci for classic PKU were c.728G＞A,c.331C＞T,and c.782G＞A;the high-frequency variant loci for mild PKU were c.721C＞T,c.1068C＞A,and c.1301C＞A;the high-frequency variant loci for children with mild HPA were c.158G＞A and c.688G＞A.There were significant differences in the frequency of high frequency mutations among the above three phenotypes(P＜0.05).Conclusion Mild HPA predominates in children with PAH deficiency in Xinjiang.The hotspot loci of the PAH gene in Xinjiang have been clarified,and specific PAH gene loci have been observed in the three different phenotypes,which can provide theoretical basis for prenatal diagnosis and clinical genetic counselling.

OBJECTIVE: To summarize the genetic diagnosis, low-depth copy number variation sequencing (CNV-seq) and prenatal finding in 7 fetuses with 2p16.3 deletions only involving the NRXN1 gene. METHODS: The 7 fetuses have all been found to have loss of heterozygosity at 2p16.3 by CNV-seq, which were verified by quantitative real-time PCR (qPCR). Specific regions of NRXN1 gene deletions were identified, and the CNVs were verified in their parents. Outcome of the pregnancies were followed up. RESULTS: Among 16 502 prenatal samples, 7 fetuses were found to harbor a 120 kb ~ 900 kb microdeletion in the 2p16.3 region, which yielded a prevalence of 0.424‰. The deleted region mainly involved 50 200 000-51 880 000 positions of chromosome 2 and involved only the NRXN1 gene. All of the 7 fetal CNVs were confirmed by qPCR, including 2 cases with heterozygous deletion of exons 1 to 6, 1 with heterozygous deletion of exons 1 to 19, 1 with heterozygous deletion of exons 19 to 22, and 3 with heterozygous deletion of introns 6 to 7 of the NRXN1 gene. Verification in the parents had found that one deletion was inherited from the father, 1 was from the mother, 2 cases were de novo in origin, whilst the remaining 3 had refused parental verification. After genetic counseling, one couple had elected induced abortion, 1 case has not been born yet, whilst the other 5 cases were born healthy. Follow up had identified no mental abnormalities among the children. CONCLUSION Seven fetuses with heterozygous 2p16.3 deletions only involving the NRXN1 gene were detected by CNV-seq. The specific deletion of the NRXN1 gene was verified by qPCR. Prenatal genetic counseling and fertility guidance has been provided to the particular family by combining the results of CNV testing, pedigree analysis and pregnancy outcome.
Female ; Humans ; Pregnancy ; Calcium-Binding Proteins/genetics* ; Cell Adhesion Molecules, Neuronal/genetics* ; DNA Copy Number Variations ; Nerve Tissue Proteins/genetics* ; Neural Cell Adhesion Molecules/genetics* ; Prenatal Diagnosis ; Real-Time Polymerase Chain Reaction ; Infant, Newborn

Objective:To study the genetic profiles of phenylalanine hydroxylase (PAH) gene mutations in neonates with phenylketonuria (PKU) in Xinjiang.Methods:From January 2015 to December 2021,neonates born and genetically diagnosed with PKU in our region were retrospectively included. The genetic profiles of different ethnic groups were analyzed and compared with PKU patients from central, northwest and northern regions of China.Results:A total of 131 neonates with PKU were enrolled, including 82 Han, 25 Hui and 20 Uyghur patients, 4 cases of other ethnic groups. 46, 20 and 14 types of pathogenic variants were detected in each ethnic group with detection rates of 95.1% (156/164), 66.0% (33/50), and 60.0% (24/40), respectively. The variants were mainly missense mutations and located in exons 2, 3, 6,7 and 11. The most common loci in Hui patients were c.158G>A (18.2%), c.728G>A (18.2%) and c.898G>T (9.1%). The most common loci in Uyghur patients were c.158G>A (33.3%), c.355C>T (12.5%) and c.1068C>A (8.3%). c. 898G>T might be most unique in Hui patients and c.355C>T most unique in Uyghur patients in Xinjiang. A novel variant of PAH gene, c.828G>C (p.M276I) in exon 7 was identified. Compared with northern, central and northwestern regions of China, PKU patients in Xinjiang had significantly higher incidence of c.158G>A mutation and lower incidence of c.728G>A mutation ( P<0.05). Conclusions:Missense mutations of PAH gene are common in some regions of Xinjiang. The compositions of PAH gene variations are similar to northwest and northern China with significant differences in hotspots of mutations.

【Objective】 To investigate the influencing factors behind the follow-up compliance of patients with low/no phenylketonuria (PKU) for special medical use, in order to provide a basis for regulating the follow-up of PKU patients and ensuring the effectiveness of special diet treatment. 【Methods】 A survey was conducted on PKU patients treated in Urumqi Maternal and Child Health Hospital for over 1 year, from January 2010 to December 2020. Interviews and questionnaires were conducted with their caregivers to collect and analyze the current status of PKU patients undergoing special diet treatment, and to identify the influencing factors behind their compliance with follow-up treatment. 【Results】 Patients who had received neonatal disease screening, neonatal gene diagnosis, and maternal Down′s screening during pregnancy had better compliance, with statistically significant differences (χ²=5.753, 10.993, 9.189, P<0.05). PKU children with parents who had a college education or above showed significantly higher adherence to special diet treatment (χ²=8.321, 7.415, P<0.05). PKU children with parents having a fixed occupation also showed higher compliance, with a statistically significant difference (χ²=20.626, 7.895, P<0.05). Patient age, interval of buying special diet, number of blood samples sent and enrollment of normal age, all had a significant impact on the follow-up compliance of PKU patients with special diet (χ²=19.443, 8.090, 69.482, 12.001, P<0.05). 【Conclusions】 PKU is a treatable genetic metabolic disease. Strengthening health education, formulating standardized follow-up plans and procedures, and improving follow-up treatment compliance are crucial in enhancing the treatment and follow-up effectiveness of PKU patients.

Objective:To explore the characteristics of patients with Alagille syndrome(ALGS), so as to provide evidence for early clinical diagnosis.Methods:The clinical data of 13 children diagnosed with ALGS in Children′s Hospital of Chongqing Medical University from February 2014 to November 2018 was retrospectively analyzed, and some children were followed up.Results:Among the 13 patients, 12 cases(92.3%) had cholestasis, 8 cases(61.5%) with heart malformation, 5 cases(38.5%) with characteristic facial features, 5 cases(38.5%) with pruritus, and 1 case(7.7%) with a positive family history.Among the 7 pediatric ophthalmologists, 2 patients suffered from ocular lesions.Seven patients underwent spine radiography, and 1 patient had typical butterfly vertebra.In the 9 cases with hepatic pathology, 4 cases had partial hepatic sinus pressure or occlusion, 1 case had no small bile duct in part of the portal area, 4 cases had small bile duct hyperplasia, and 5 cases underwent biliary tract exploration.Genetic testing of 12 children with ALGS showed JAG1 gene mutation in 8 cases and NOTCH2 gene mutation in 4 cases.Among the 11 followed up patients, 8 cases were in stable condition, 1 patient with progressive cirrhosis was registered for liver transplantation, 1 patient died of liver failure, and 1 patient developed with drug-induced liver injury at the age of 4 months had progressive liver failure, and the jaundice was gradually subsided after liver transplantation. Conclusions:ALGS may appear on multiple systems involvement, and it demonstrates variable clinical expressivities and incomplete penetrance, thus bring certain difficulties to clinical diagnoses.It′s easily misdiagnosed as biliary atresia in infancy.Genetic testing is integral in the diagnosis of ALGS.