OBJECTIVE: To explore the clinical characteristics and genetic etiology of a child with Aicardi-Goutières syndrome 3 (AGS3). METHODS: Trio whole exome sequencing was carried out for the child and his parents, and candidate variants were verified by Sanger sequencing. To further clarify their pathogenicity, the crystal structure of the variants was simulated and analyzed, and the plasmid of variants was expressed in vitro. A literature search was also carried out to summarize the phenotypic and genetic characteristics of AGS3. RESULTS: The child was found to harbor novel compound heterozygous variants of the RNASEH2C gene, namely c.434G>T (p.Arg145Leu) and c.494G>C (p.Ter165Ser), which were inherited from his mother and father, respectively. Analysis of protein crystal structure suggested that the c.434G>T (p.Arg145Leu) variant may affect the stability of local structure, and in vitro experiments showed that this variant can lead to protein degradation. The c.494G>C (p.Ter165Ser) variant has destroyed the stop codon, resulting in prolonged variant. CONCLUSION The novel compound heterozygous variants of the RNASEH2C gene probably underlay the AGS3 in this child, which has enriched the phenotypic and mutational spectrum of this disorder.
Humans ; Child ; Mutation ; Autoimmune Diseases of the Nervous System/genetics* ; Nervous System Malformations/genetics*

Background: A small shift in high-sensitivity cardiac troponin T (hs-cTnT) assays can lead to different result interpretation and consequent patient management. We explored whether a small bias could be detected using conventional internal quality control (QC) procedures, evaluated the performance of moving average (MA)-based QC procedures, and proposed a new QC procedure based on the moving rate (MR) of positive patient results of hs-cTnT assays. Methods: The ability of conventional QC to detect a 5 ng/L bias was examined using the 1 3s/ 22s/R4s multi-rule procedure as deviation rules.We developed MA and MR procedures for the hs-cTnT assay using eight months of patient data. The performance of different MA or MR procedures was investigated by calculating the median number of patient samples affected until a bias introduced into the dataset was detected (MNPed). After comparing the MNPed across different procedures, we selected an optimal MA or MR procedure for validation. Validation graphs were plotted using the minimum, median, and maximum number of results affected until bias detection. Results: Our conventional QC procedures could not detect a positive bias of 5 ng/L. When a positive bias was introduced, MNPed was much higher using MA than using MR, with cut-off values of 5 ng/L and 14 ng/L, respectively. MR validation charts for optimal procedures provided insight into the MR performance. Conclusions The MR procedure could detect different errors with few false alarms. In the hs-cTnT assay, the MR procedure with a smaller cut-off value outperformed MA and conventional QC procedures for small bias detection.

Objective:To analyze the causes, influencing factors and trends of dead on arrival cases in children′s Hospital in the past 5 years, aiming to provide direction and basis for reducing the dead on arrival cases of children.Methods:We collected the dead on arrival cases in the department of emergency at Shanghai Children′s Hospital from January 2015 to December 2019, classifed and analysed the gender, age, native place, death season, time of death, and possible causes of death, and then studied the correlation between above factors and the cases.Results:A total of 151 dead on arrival cases were collected.The annual number decreased year by year, and boys were more than girls in gender.Most of them were infants under 1 year old, and nonlocal children were more than Shanghai native.The above differences were statistically significant, but there was no significant difference in the distribution of death season and death time.In terms of the cause of death, perinatal diseases accounting for 33.8%(51/151), those accompanied with severe underlying diseases accounting for 39.1%(59/151), accidental death accounting for 14.6%(22/151), unexplained deaths accounting for 12.6%(19/151). Those distribution differences were statistically significant( χ2=32.497, P<0.001). Meanwhile, there were statistic differences in gender and age of the cases with severe underlying diseases( χ2=4.898, P=0.027; χ2=32.169, P<0.001), and the year and age distributions of the accidental death cases also had significant differences( χ2=16.636, P=0.002; χ2=14.727, P=0.002). Conclusion:To reduce dead on arrival cases of children, we should do a good job in perinatal health care and screening, reduce premature birth and birth defects, actively conduct propaganda to prevent children′s accidental injuries, popularize medical first aid knowledge, and strengthen children′s transport system.

Objective:To investigate the effects of Astragaloside Ⅳ on the secretion of exosomes in human endothelial progenitor cells (EPCs) and the expression of microRNA (miRNA)-126 in exosomes.Methods:The umbilical cord blood from one healthy full-term newborn from the Department of Obstetrics and Gynecology of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine in 2019 was harvested for isolating mononuclear cells by density gradient centrifugation and cultured for 7 days. Morphological observation was performed during this period. Cells of the third passage were collected for identification by CD31 immunomagnetic bead sorting and double fluorescence staining. According to the random number table, the identified EPCs were divided into Astragaloside Ⅳ group and phosphate buffer solution (PBS) group. The cells in Astragaloside Ⅳ group were cultured with Astragaloside Ⅳ in final mass concentration of 100 mg/L for 24 hours, and the cells in PBS group were cultured with the same volume of PBS for 24 hours. After culture, the exosomes from the cell culture supernatant of the two groups were collected, and the expressions of characteristic markers of exosomes CD9, CD63, and CD81 were detected by Western blotting, the morphology of EPC exosomes (EPC-Exos) was observed under transmission electron microscope, and the particle size of EPC-Exos was detected by nanoparticle tracking analysis technique. The concentration of EPC-Exos was determined by dioctyl butyric acid method (the sample number was 3), and the expressions of miRNA-126-3p and miRNA-126-5p related to angiogenesis in EPC-Exos were determined by reverse transcription polymerase chain reaction (the sample number was 3). Data were statistically analyzed with independent sample t test. Results:(1) On the 4th day of culture, the cells began to adhere to the wall, and the multi-forms such as circle, fusiform, and strip appeared at the same time. On the 7th day of culture, the edge of the cells was clear and arranged like a paving stone, the central cells were round, and the surrounding cells were fusiform. (2) CD31 immunomagnetic beads sorting method identification showed that the membrane was stained with green fluorescence and the nucleus was stained with blue fluorescence. Double fluorescence staining method showed that the cells were orange-yellow. The cells were identified as EPCs. (3) After 24 hours of culture, the expressions of CD9, CD63, and CD81 in EPC-Exos were all positive, confirming that EPC-Exos were extracted successfully in this experiment. (4) After 24 hours of culture, the EPC-Exos of the two groups showed round membrane vesicles, and there was no significant difference in morphology. (5) After 24 hours of culture, the particle size of 98.7% EPC-Exos in Astragaloside Ⅳ group was 84.7 to 143.1 nm, and that of 98.0% EPC-Exos in PBS group was 88.7 to 123.5 nm. (6) After 24 hours of culture, the mass concentration of EPC-Exos in Astragaloside Ⅳ group was (310±5) μg/mL, which was significantly higher than (257±5) μg/mL in PBS group, t=13.369, P<0.01. (7) After 24 hours of culture, there were more miRNA-126-3p ( t=16.062, P<0.01) and miRNA-126-5p ( t=3.252, P<0.05) in EPC-Exos of Astragaloside Ⅳ group than in PBS group. Conclusions:Astragaloside Ⅳ can improve the function of human EPC secretory exosomes, and the secreted exosomes are loaded with miRNA-126.

Objective To explore the effect of rat nerve growth factor combined with early rehabilitation intervention in the treatment of hearing injury caused by cytomegalovirus (CMV) infection.Methods 106 cases of hearing impairment caused by cytomegalovirus infection diagnosed in our hospital from January 2012 to February 2018 were randomly divided into observation group and control group,53 cases in each group.The control group were treated only with ganciclovir and comprehensive rehabilitation therapy,while the treatment group was treated with nerve growth factor on the basis of the control group.Before and after treatment,the children in both groups underwent multi-frequency steady-state evoked potential test,cytomegalovirus antibody DNA fluorescence quantification,and their mothers underwent cytomegalovirus antibody test.The total effective rate of the two groups and the effective rate of the observation group with different degrees of hearing impairment were compared,and the therapeutic effect of the observation group with congenital and acquired CMV infection was compared.Results The total effective rate was 80.7％ in the observation group,which was higher than 65.9％ in the control group (P ＜ 0.05).The effective rate of mild to moderate hearing loss in observation group was higher than that in severe hearing impairment (P ＜0.05).The efficacy of the rat nerve growth factor in the treatment of hearing loss after CMV infection was higher than that of hearing loss caused by congenital cytomegalovirus infection (P ＜ 0.05).Conclusions Rat nerve growth factor combined with early comprehensive rehabilitation intervention is an effective method to treat acoustic nerve injury caused by CMV infection.The effect of rat nerve growth factor on mild and moderate hearing impairment was better than that of severe hearing impairment.The therapeutic effect of acquired CMV infection on hearing damage was better than that of by congenital infection.

OBJECTIVE:To investigate the risk factors for secondary pulmonary fungal infection in patients with severe craniocerebral trauma after tracheotomy,and to provide reference for clinical prevention and treatment. METHODS:In retrospective study,87 severe craniocerebral trauma patients with secondary pulmonary fungal infection after tracheotomy were selected from Ezhou Municipal Central Hospital(called"our hospital"for short)during Jan. 2014-Jun. 2017 as observation group;87 severe craniocerebral trauma inpatients without secondary pulmonary fungal infection after tracheotomy were selected as control group. The distribution and drug resistance of infected fungal in observation group were analyzed. χ2 test and binary Logistic analysis were adopted to investigate risk factors of secondary pulmonary fungal infection in patients with severe craniocerebral trauma after tracheotomy. RESULTS:Totally 174 clinical specimens were detected in observation group of our hospital;7 kinds of fungus were detected and isolated from 87 strains,and the fungi with high detection rate were Candida albicans(41 strains,47.13%)and Candida glabrata(23 strains,26.44%). The resistance rates of C. albicans and Candida tropicalis to commonly used antifungal agents as fluconazole,itraconazole and fluoncytosine were lower than 20%;resistance rates of C. glabrata to fluconazole,itraconazole and fluoncytosine were more than 25%,to amphotericin B and nystatin were lower than 20%. χ 2test and binary Logistic analysis showed that independent risk factors of secondary pulmonary fungal infection included hypoproteinemia,Glasgow coma score(GCS,<8 points)at admission,serum creatinine clearance(<30 mL/min)at admission,tracheal incision ventilation time(≥7 days),the time of antibiotics use(≥14 days),combined use of antibiotics,the use of carbapenems and systemic glucocorticoid [odd ratios were 3.02,2.98,2.21, 2.05,2.48,2.35,4.74,5.97;95%CI were(1.59,5.74),(1.58,5.63),(1.18,4.41),(1.11,3.78),(1.34,4.59),(1.27,4.34), (2.49,8.35),(3.08,11.49),P<0.05]. CONCLUSIONS:The fungus of secondary pulmonary fungal infection in patients with severe craniocerebral trauma after tracheotomy in our hospital are mainly C. albicans and C. glabrata,which are sensitive to commonly used antifungal agents. Hypoproteinemia,GCS at admission,serum creatinine clearance rate at admission,tracheal incision ventilation time,the time of antibiotics use,combined use of antibiotics,the use of carbapenems and systemic glucocorticoid are independent risk factors of secondary fungal infection in patients with severe craniocerebral trauma after tracheotomy. It is necessary to pay attention to predictive value of above risk factors,improve sensitivity and specificity of diagnosis and treatment. Antifungal agent should be selected rationally according to the results of drug sensitivity test. At the same time,early prophylactic or empirical antifungal treatment should be given in time for high risk patients with above factors.

An ultra-high performance liquid chromatography-tandem mass spectrometry ( UHPLC-MS/MS ) method was developed and validated for the simultaneous determination of 9 kinds of trace endocrine disrupting chemicals in biological samples using ultrasonic-assisted extraction followed by purification with gel permeation chromatography ( GPC) and silica gel columns. The sample extracts were purified by Bio beads S-X3 GPC columns with cyclohexane/ethyl acetate (1:1, V/V) as mobile phase, and the target compounds were eluted in the fraction of 12-28 mL retention volume. Electrospray ionization source operated in positive mode and atmospheric pressure chemical ionization source operated in negative mode were used for mass spectrometric detection. Data acquisition was carried out in multiple reaction monitoring mode. Recoveries were predominately within 65 . 2%-118 . 0%. Method quantification limits were 0 . 1-9 . 7 ng/g dw ( dry weight ) . This method was successfully applied to the analysis of the target endocrine disrupting chemicals in carps collected from the Pearl River. with the exception of carbanilide and triclocarban, the rest analytes were detected in fish tissue samples, with the concentrations varied within the range of 0. 1-22. 6 ng/g dw.

OBJECTIVE:To observe the efficacy and safety of mifepristone cycle therapy in the treatment of perimenopausal dysfunctional uterine bleeding. METHODS:Totally 116 patients with perimenopausal dysfunctional uterine bleeding were randomly divided into control group and observation group. Control group was given Mifepristone tablet 10 mg from 3 d after diagnostic cu-rettage on an empty stomach for continuous 90 d,once a day. Observation group was given Mifepristone tablet 10 mg from 3 d af-ter diagnostic curettage on an empty stomach,once a day,and it was stopped after continuous 5 d. Then it continued 5 d from the first day of menstruation,and lasted 3 menstrual cycles. The clinic data was observed,including clinical efficacy,estradiol(E2),fol-licle-stimulating hormone (FSH),luteinizing hormone (LH),progesterone (P),hemoglobin (Hb),endometrial thickness,men-strual conditions and recurrence in 2 groups before and after treatment were observed,and the incidence of adverse reactions were recorded. RESULTS:After treatment,E2,FSH,LH and P in 2 groups were significantly lower than before,and observation group was lower than control group,the differences were statistically significant(P<0.05). Hb in 2 groups was significantly higher than before,endometrial thickness was significantly lower than before,the differences were statistically significant(P<0.05);however, there was no significant difference between 2 groups(P>0.05). Total effective rate in observation group significantly higher than control group,normal proportion of menstruation were significantly better than control group,the differences were statistically sig-nificant(P<0.05). There was no significant difference in the incidence of a dverse reactions and dysfunctional uterine bleeding be-tween 2 groups(P>0.05). CONCLUSIONS:Mifepristone cycle therapy has significant efficacy in the treatment of perimenopausal dysfunctional uterine bleeding,can significantly improve the hormone level and the establishment of normal menstrual cycle and re-duce endometrial thickness,with good safety.

AIM:To observe the effect of plumbagin on the mRNA and protein expression of nicotinamide ade -nine dinucleotidephosphate oxidase 4 ( Nox4 ) , reactive oxygen species ( ROS ) level and protein expression of α-smooth muscle actin (α-SMA) in the HSC-LX2 cells stimulated with transforming growth factor β1 (TGF-β1) in vitro.METH-ODS:HSC-LX2 cells were cultured in vitro and divided into blank group, model group, high-, medium-and low-dose (2, 1.5 and 1 μmol/L) plumbagin groups .After incubated with each drug for 72 h, the mRNA expression of Nox4 was detec-ted by RT-PCR.ROS levels were tested by in situ loading probe method.The protein contents of Nox4 and α-SMA were measured by Western blot .RESULTS:Compared with model group , after treated with plumbagin for 72 h, the mRNA ex-pression of Nox4, ROS level and α-SMA protein were significantly decreased in high-and medium-dose plumbagin groups (P<0.01).CONCLUSION:Plumbagin inhibits the activation of HSC-LX2 cells via decreasing the expression of Nox4, thus decreasing ROS levels .

OBJECTIVETo survey the quality of life in children and adolescents with type 1 diabetes.

METHODSNinety-eight children and adolescents with type 1 diabetes who participated in Diabetes Summer Camp held in Chongqing, Wuhan and Cheng during 2012 April and December were recruited in the study. The American juvenile diabetes patients quality of life scale Diabetes Quality of Life for Youths was used to assess the quality of life and SPSS19.0 was used for statistical analysis.

RESULTSThe scale had satisfactory reliability and validity with a Cronbach's Alpha score of 0.942 and a validity score of 0.679. All three dimension of scales: scales of impact, scales of worries and scales of satisfaction were significantly correlated with self-health assessment (P<0.01). The scores of impact and worries accounted for >50% of total scores as the same for the self health assessment scores. The score of disease course, diet and blood glucose control were positive correlated with each other. Age and HbA1c were positively correlated with the scale of impact, while gender has negative correlation with satisfaction scale (P<0.05). The diabetes diet had significant effects on the quality of life.

CONCLUSIONThe quality of life in children and adolescents with type 1 diabetes is decreased, especially for those with longer disease course and female adolescents. The form of Diabetes Quality of Life for Youth used in the study has good reliability and validity, which can reflect the quality of life of Chinese diabetic children and adolescents.

Adolescent ; Child ; Diabetes Mellitus, Type 1 ; Female ; Humans ; Male ; Quality of Life ; Sickness Impact Profile ; Surveys and Questionnaires ; Young Adult