Objective·To investigate the genetic etiology of a rare and complex case clinically suspected to be methylmalonic acidemia(MMA),but with negative whole exome sequencing(WES)results,using a multi-omics sequencing approach.Methods·DNA and RNA samples were extracted from the peripheral blood of the proband and both parents.Targeted MMA-related gene Panel sequencing and WES were first performed.Subsequently,RNA sequencing(RNA-seq)and whole genome sequencing(WGS)were conducted to comprehensively analyze the child's genetic variants,their origins and potential inheritance patterns.Results·No pathogenic variants associated with the patient's phenotype were identified through the MMA Panel or standard WES analysis.Extended analysis of WES suggested the possibility of uniparental disomy(UPD)of chromosome 4.WGS revealed a homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the metabolism of cobalamin associated A(MMAA)gene.The variant was located in the 5'untranslated region(5'UTR),specifically at the second base downstream of the splice donor site of exon 1(reference sequence:NM_172250).In genomic coordinates(hg19),the variant was located at base 146540561 on chromosome 4(chr4:146540561).Sanger sequencing confirmed that the mother was heterozygous for this variant,while the father did not carry it.RNA-seq showed no detectable expression of the MMAA gene on chromosome 4 in the patient.This was further confirmed by reverse transcription real time quantitative PCR,indicating nearly absent mRNA expression,suggesting that the non-coding splice-site variant affected transcriptional expression.Conclusion·A homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the MMAA gene—outside the coverage of WES—is likely the pathogenic cause in this case,presumably resulting from maternal UPD of chromosome 4.

Objective·To investigate the genetic etiology of a rare and complex case clinically suspected to be methylmalonic acidemia(MMA),but with negative whole exome sequencing(WES)results,using a multi-omics sequencing approach.Methods·DNA and RNA samples were extracted from the peripheral blood of the proband and both parents.Targeted MMA-related gene Panel sequencing and WES were first performed.Subsequently,RNA sequencing(RNA-seq)and whole genome sequencing(WGS)were conducted to comprehensively analyze the child's genetic variants,their origins and potential inheritance patterns.Results·No pathogenic variants associated with the patient's phenotype were identified through the MMA Panel or standard WES analysis.Extended analysis of WES suggested the possibility of uniparental disomy(UPD)of chromosome 4.WGS revealed a homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the metabolism of cobalamin associated A(MMAA)gene.The variant was located in the 5'untranslated region(5'UTR),specifically at the second base downstream of the splice donor site of exon 1(reference sequence:NM_172250).In genomic coordinates(hg19),the variant was located at base 146540561 on chromosome 4(chr4:146540561).Sanger sequencing confirmed that the mother was heterozygous for this variant,while the father did not carry it.RNA-seq showed no detectable expression of the MMAA gene on chromosome 4 in the patient.This was further confirmed by reverse transcription real time quantitative PCR,indicating nearly absent mRNA expression,suggesting that the non-coding splice-site variant affected transcriptional expression.Conclusion·A homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the MMAA gene—outside the coverage of WES—is likely the pathogenic cause in this case,presumably resulting from maternal UPD of chromosome 4.

Objective To find out the causes of food poisoning in a school banquet and identify the pathogenic bacteria, so as to provide evidence for the treatment of food poisoning. Methods A field epidemiological survey was conducted to search for cases, find suspicious poisoning meals and food, and collect cases and food samples for laboratory testing, to determine pathogenic pathogens and virulence genes. The pulsed field gel electrophoresis (PFGE) was applied to identify the homology of the pathogens. Results A total of 92 poisoning cases were found, and the incidence rate was 46.94%. The main clinical manifestations were diarrhea (93.48%), abdominal pain (86.96%), nausea (39.13%), vomiting (34.78%) and fever (17.39%). The median of onset latency was 17 hours. Vibrio parahaemolyticus was detected in samples from 3 patients (2 stools and 1 anal swab). The virulence gene trh was positive and the similarity coefficient of PFGE banding was 97.4%. Pathogenic bacteria were not detected in 10 food samples. The results of the case-control study showed that six types of food were suspicious (OR values were 15.75, 10.14, 8.44, 5.93, 5.56 and 4.71 respectively, P<0.05 and OR 95%CI>1), including couple lung slices and California bass with extremely high risk of exposure (OR values>10). Conclusion 　The food poisoning resulted from this enrollment banquet was caused by trh-positive Vibrio parahaemolyticus and no suspicious food was identified (the possibility of contamination by couples^' lung slices and California sea bass was high). It is suggested that the supervision and management of catering units and food safety publicity and education should be strengthened to reduce the occurrence of food-borne diseases from the source.

OBJECTIVETo assess the effectiveness of multiple cleaning and disinfection interventions in the homes and kindergartens, in reducing gastrointestinal and respiratory illnesses of children.

METHODSFrom October 2010 to September 2011, we performed a prospective, controlled study in China. 408 children under 5 years old were recruited and group randomized into intervention and control groups. Families and kindergartens in the intervention group were provided with antibacterial products for hand hygiene and surface cleaning or disinfection for one year. Each child's illness symptoms and sick leave were recorded every day.

RESULTSA total of 393 children completed the study, with similar baseline demographics in each of the 2 groups. Except for abdominal pain, the odds of symptoms (fever, cough and expectoration, runny nose and nasal congestion, diarrhea), illness (acute respiratory illness and gastrointestinal illness), and sick leave per person each month were significantly reduced by interventions. The rates of fever, diarrhea, acute respiratory illness, gastrointestinal illness and sick leave per person per year were significantly decreased as well.

CONCLUSIONNot only the acute respiratory and gastrointestinal illness but the sick leave rate in children were significantly reduced by multiple interventions.

Administration, Cutaneous ; Anti-Bacterial Agents ; administration & dosage ; Child, Preschool ; China ; epidemiology ; Disinfection ; Female ; Gastrointestinal Diseases ; epidemiology ; prevention & control ; Hand Hygiene ; Humans ; Incidence ; Male ; Prevalence ; Respiratory Tract Infections ; epidemiology ; prevention & control

To investigate the chemical constituents from the aerial parts of Lygodium japonicum. Methods:Various chromatographic techniques were employed for isolation and purification of the constituents. The structures were elucidated by chemical evidence and spectral methods. Results:A new stigmasterol glycoside,(24R)-stigmastan-3β,5α,6β-triol 3-O-β-D-glucopyranoside (1),together with three known phenolic glycosides:6-O-p-coumaroyl-D-glucopyranose (2),6-O-caffeoyl-D-glucopyranose (3),1-O-(E)-caffeoyl-β-D-gentiobiose (4) were obtained and identified by spectroscopic methods. Conclusion:All compounds were isolated from Lygodiaceae for the first time.

OBJECTIVETo study the expression of tenascin in normal and fibrotic rat liver.

METHODSLiver fibrosis induced in rat with CCl(4) were divided into three stages: the stage of hepatic injury (4 weeks), early stage of hepatic fibrosis (8 weeks) and later stage of hepatic fibrosis (12 weeks). Tenascin expression in liver tissue was observed by immunohistochemical method and in situ hybridization using digoxigenin-labelled DNA probe.

RESULTSIn normal rat liver there was a weak staining for tenascin. In both liver injury stage and early stage of hepatic fibrosis, both mRNA signal and immunostaining for tenascin were significantly increased as compared to that in normal liver. In later stage of hepatic fibrosis, mRNA signal and immunostaining for tenascin were decreased compared with that in early stage of hepatic fibrosis. The cellular source of tenascin in liver mainly restricted in mesenchymal cells.

CONCLUSIONSTenascin is a component of the extracellular matrix of liver tissue. Plays a role in early matrix organization during liver fibrogenesis.

Animals ; Carbon Tetrachloride ; Disease Models, Animal ; Extracellular Matrix Proteins ; biosynthesis ; genetics ; Image Processing, Computer-Assisted ; Immunohistochemistry ; In Situ Hybridization ; Liver Cirrhosis ; chemically induced ; metabolism ; Male ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Tenascin ; biosynthesis ; genetics

AIM: To observe the expression of undulin(Un) in liver tissue and to clarify the diagnostic significance of serum Un in experimental rat liver fibrosis. METHODS: Rat liver fibrosis was induced by carbon tetrachloride. The expression of Un in liver tissue was detected by immunohistochemical method. Serum Un levels was measured by enzyme immunoassay.RESULTS: Expression of Un increased in fibrotic liver than normal liver, and it was mainly distributed in portal tract stroma, central veins and fibrotic septa in fibrotic liver. Also, the level of serum Un was significantly higher in fibrotic liver than normal liver.CONCLUSION: These data suggest that Un should be a component of the hepatic extracellar matrix, and its expression could be increased greatly in fibrotic rat liver. Serum Un levels may be used as an indicator in liver fibrosis diagnosis.