OBJECTIVESince glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.

METHODSAccording to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio <1.00) and control group with 65 children (G6PD/6GPD ratio≥1.00). The primer design and PCR amplification conditions were optimized, and RT-PCR sequencing was used to analyze the complete coding sequence and verify the genomic DNA sequence in the two groups.

RESULTSIn the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).

CONCLUSIONSRT-PCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.

Adolescent ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Infant ; Male ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Allergic diseases such as allergic asthma, allergic der-matitis, allergic rhinitis, are polygenic diseases, involving the interaction between the environment, genes and immunity. In the past few decades, the incidence rate of allergic diseases in-creased predominantly and influenced the quality of people's lives seriously, so looking for new targets for the prevention and treat-ment of allergic diseases and drugs with less adverse reaction be-comes a hot topic for researchers. MicroRNAs(miRNAs)are a class of endogenous non-coding small RNAs that mediate nega-tively posttranscriptional regulation of gene expression by targe-ting specific mRNA sequences to inhibit the translation of mR-NAs. They are widely involved in the biological processes of cell differentiation, immune response and tumor development. The study shows that miRNAs can control the occurrence and devel-opment of allergic diseases. Studying the regulatory role of miR-NAs in allergic diseases has important implications for exploring the immunopathological mechanisms and discovering new thera-peutic targets of drugs.

Objective:To discuss the type, treatment and results of different therapies of biliary fistula after orthotopic liver tansplantation(OLT).Methods:Data of 24 patients who developed biliary fistula after OLT in the First Affiliated Hospital of Xi′an Jiaotong University from January 2000 to March 2019 were retrospectively analyzed. Patients with biliary fistula were classified into 4 types according to presence or absence of stricture. All patients were treated by endoscopic retrograde cholangiopancreatography (ERCP) or interventional therapy, including endoscopic nasobiliary drainage (ENBD), endoscopic retrograde biliary drainage (ERBD) or percuteneous transhepatic cholangial drainage (PTCD). Main outcome measurements were the onset time of biliary fistula, the site of biliary fistula, the complications of ERCP or PTCD, the time of removing abdominal or biliary drainage tube, and the onset of new biliary stricture.Results:Biliary fistula was found in (46.5±36.6) days (6-122 days) after OLT. The numbers of patients in four types of biliary fistula were 6, 14, 2 and 2, respectively. Biliary fistula was cured in 22 patients, with clinical cure rate of 91.7%. All patients underwent ERCP first, and the technical success rate and clinical cure rate were 87.5% (21/24) and 85.7% (18/21), respectively. The clinical cure rates of ERCP forⅠ-Ⅳ biliary fistula were 6/6, 84.6%(11/13), 1/2, and 0, respectively. The clinical cure rates of ENBD and ERBD were 8/10 and 6/8, respectively. Five cases in whom ERCP failed, underwent PTCD, with technical success and clinical cure rates of 4/5 and 3/4 respectively. Eight patients(33.3%)developed cholangitis after treatment, and the incidence rate seemed higher in type Ⅱ biliary fistula than that in type Ⅰ [35.7% (5/14) VS 16.7% (1/6)]. Incidence of cholangitis was higher in patients with non-anastomotic stricture than those with anastomotic stricture [83.3%(5/6) VS 16.7%(3/18)].Conclusion:The first line treatment for biliary fistula after OLT is ERCP, followed by PTCD. The best procedures of biliary fistula typeⅠ-Ⅳ were ENBD, ENBD combined with ERBD, ENBD and PTCD, respectively.

Bronchial asthma is a kind of respiratory disease which affects people 's life quality seriously. Many factors in-volved in the occurrence and development of such disease, of which the aberrant expression of E-cad plays a critical role in it. Research found that E-cad is an important cell adhesion molecu-lar, and its main function is to maintain the structural integrity of cells and participate in the improvement of airway remodeling as well as restoration of immune function. Further study showed that the role of mucosal barrier of airway epithelial cells in bronchial asthma patients was often damaged. Moreover, the protein ex-pression of E-cad decreased significantly in mucosal molecular, which suggested that the abnormal expression of E-cad was in-volved in the development of bronchial asthma. A review on the relations between the abnormal expression of E-cad protein and bronchial asthma has been discussed in this paper, also it in-cludes the discussion about the mechanisms of E-cad’ s disorder-induced bronchial asthma as well as explores the strategies of bronchial asthma treatment, which may provide references for the follow-up research and clinical treatment.

Objective To observe the effect oftert-Butylhydroquinone (tBHQ) on the expression of nuclear factor erythroid 2-related factor 2 (Nrf2),heme oxygenase (HO)-1 and phosphatidylinositol 3-kinase (PI3K) in high glucose cultured retinal Müller cells;and to investigate the anti-oxidative stress and anti-apoptotic effects oftBHQ.Methods Retinal Müller cells were divided into normal glucose group (5.5 mmol/L,N group),high glucose group (45 mmol/L,HG group) and tBHQ intervention group (HG+tBHQ group).After retinal Müller cells were cultured with high glucose for 48 hours,the pretreatment with tBHQ (20 μmol/L) induced the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and HO-1.The Müller cells were identified by immunofluorescence staining.The expressions of Nrf2,HO-1,PI3K,B-cell lymphoma-2 (Bcl-2) and Bax were detected by Western blot and real-time fluorescence quantitative PCR.Flow cytometry was used to detect the apoptosis of retinal Müller cells in rats.Results Müller cytoplasm and nucleus GS showed strong positive,large cell body,abundant cytoplasm,uniform green fluorescence;nuclear DAPI staining round or oval,clear boundary.The expression of Nrf2 protein (t=4.114,P=0.006),HO-1 protein (t=9.275,P=0.000),Nrf2 mRNA (t=7.292,P=0.000) and HO-1 mRNA (t=15.014,P=0.000) in the HG group were higher than those in the N group.The expressions of Nrf2 protein (t=7.847,P=0.000),HO-1 protein (t=7.947,P=0.000),PI3K protein (t=5.397,P=0.002),Bcl-2 protein (t=6.825,P=0.000),Nrf2 mRNA (t=18.046,P=0.000),HO-1 mRNA (t=39.458,P=0.000),PI3K mRNA (t=4.979,P=0.003) and Bcl-2 mRNA (t=9.535,P=0.000) in the HG+tBHQ group were significantly higher than those in the HG group.The protein and mRNA expressions of Bax protein in the HG+tBHQ group were significantly lower than those in the HG group (t=14.998,16.520;P=0.000,0.000).Flow cytometry showed that the apoptosis rate of Müiller cells in the HG group was significantly higher than that in the N group (t=39.905,P=0.000).The apoptosis rate of Müller cells in the HG+tBHQ group was significantly lower than that in the HG group (t=21.083,P=0.000).Conclusion tBHQ can inhibit the apoptosis of retinal Müller cells by up-regulating the expression ofNrf2,HO-1 and PI3K.

Objective To investigate the relationship between adiponectin gene polymorphisms and premature myocardial infarction (PMI).Methods A total of 306 PMI patients of Beijing Anzhen Hospital were identified from March 2013 to January 2015 using the case-control method,412 patients with normal coronary angiography results served as the control group.17 single nucleotide polymorphisms (SNPs) in adiponectin gene were selected and genotyped using mass spectrometry.The Chi-squared test was used to compare allele and genotype frequencies between PMI and control groups.The plasma adiponectin concentrations were measured using enzyme-linked immunoassay,and the relationship was analyzed between adiponectin level and SNPs.Results There was no significant difference in allele frequencies and in genotype frequencies between PMI patients and controls (P ＞0.05).Multivariate logistic regression models were used to estimate the correlation in additive,dominant and recessive genetic models,respectively.Analysis showed that rs2241766T ＞ G was significant associated with PMI risk in a addition model in Chinese Han population(OR =1.446,95％ C1 1.006-2.079,P =0.035),and rs12629945G ＞ A was significant association with PMI risk in a dominant model in Chinese Han population(OR =1.609,95％ CI 1.006-2.573,P =0.041).Adiponectin levels in PMI group were significantly lower than in control group ((863.3 ± 112.8)μg/L vs.(910.4 ± 117.1) μg/L,P =0.042).Adiponectin levels were significantly lower in patients with the homozygous mutation genotype (GG) compared to patients with the heterozygote mutant and wild type genotypes (GT + TT) in adiponectin gene rs2241766 T ＞ G polymorphisms ((859.4 ± 98.1) μg/L vs.(908.9 ± 113.1) μg/L,P =0.032).Conclusion The rs2241766T ＞ G and rs12629945G ＞ A polymorphisms in the adiponectin gene were associated with PMI in Chinese Han population.Rs2241766T ＞ G polymorphisms is linked with lower plasma adiponectin levels,and increased risk of PMI.

China Kadoorie Biobank (CKB) conducted the baseline survey from June 2004 to July 2008 in five rural areas in Zhejiang, Hunan, Gansu, Sichuan and Henan Provinces and five urban areas in Heilongjiang, Shandong, Jiangsu, Guangxi and Hainan Provinces (Autonomous Region). A total of 512 891 adults aged 30-79 years were recruited in the baseline survey, i.e. questionnaire survey, physical examination and blood sample collection. The first resurvey of 19 786 people was conducted from June to October in 2008, and the second resurvey of 25 239 people was conducted from August, 2013 to September, 2014. In 2016, with the support of the "Precision Medicine Research" Key Project, National Key Research and Development Program of China, CKB started a large natural population cohort demonstration study on the basis of the previous work. The third resurvey was conducted from August, 2020 to December, 2021 among 25087 people, including questionnaire survey (with additional aging related items), physical examination and biological sample collection (blood, urine, saliva, stool). By June, 2022, CKB had conducted the follow up in cohort population for an average 15 years, resulting an observation of 7.7 million person-years. which documented 74 000 deaths, 371 000 health insurance events (2.795 million episodes in total), 11 000 active follow-up events (12 000 episodes in total), 100 000 morbidity monitoring events (147 000 episodes). CKB Biobank has stored 1 292 000 blood samples, 150 000 urine samples, 780 000 DNA samples, 25 000 saliva samples, and 20 000 stool samples. CKB project team has developed the technical specifications for long-term follow-up, sample database construction and management, database and data sharing platform construction and management, which have been compiled and published as the Technical Specifications for Large Population Cohort Research. In addition, the group standards of field survey of large natural population cohort, long term follow up, biobank construction, data process and data security have been developed. Meanwhile, high-quality scientific research have been conducted consecutively based on the CKB cohort data, the research of the relationship between healthy lifestyle and major chronic diseases have provided specific evidence in Chinese population.

Objective VATER/VACTERL-like association is associated with adverse pregnancy outcomes.Genetic evidence of this disorder is sporadic.In this study,we aimed to provide genetic insights to improve the diagnosis of VACTERL. Methods We have described a Chinese family in which four members were affected by renal defects or agenesis,anal atresia,and anovaginal fistula,which is consistent with the diagnosis of a VACTERL-like association.Pedigree and genetic analyses were conducted using genome and exome sequencing. Results Segregation analysis revealed the presence of a recessive X-linked microdeletion in two living affected individuals,harboring a 196-380 kb microdeletion on Xq27.1,which was identified by familial exome sequencing.Genome sequencing was performed on the affected male,confirming a-196 kb microdeletion in Xq27.1,which included a 28%loss of the CDR-1 gene.Four family members were included in the co-segregation analysis,and only VACTERL-like cases with microdeletions were reported in X27.1. Conclusion These results suggest that the 196-380 kb microdeletion in Xq27.1 could be a possible cause of the VATER/VACTERL-like association.However,further genetic and functional analyses are required to confirm or rule out genetic background as the definitive cause of the VACTERL association.

De-identification of personal health information is essential in order not to require written patient informed consent. Previous de-identification methods were proposed using natural language processing technology in order to remove the identifiers in clinical narrative text, although these methods only focused on narrative text written in English. In this study, we propose a regular expression-based de-identification method used to address bilingual clinical records written in Korean and English. To develop and validate regular expression rules, we obtained training and validation datasets composed of 6,039 clinical notes of 20 types and 5,000 notes of 33 types, respectively. Fifteen regular expression rules were constructed using the development dataset and those rules achieved 99.87% precision and 96.25% recall for the validation dataset. Our de-identification method successfully removed the identifiers in diverse types of bilingual clinical narrative texts. This method will thus assist physicians to more easily perform retrospective research.
Algorithms ; *Data Anonymization ; *Electronic Health Records ; *Health Records, Personal ; Humans ; Multilingualism ; Natural Language Processing ; Research Design

OBJECTIVE To examine the synergistic inhibiory effect of combination of mammalian target of sirolimus (Rapamycin) (mTOR) inhibitor everolimus and AKT inhibitor MK-2206 on hepatocar-cinoma cell proliferation. METHODS HepG2 and BEL-7402 cells were treated with sirolimus and evero-limus alone for 0, 1, 3, 6, 12 and 24 h or in combination with insulin-like growth factor 1 receptor (IGF-1R) inhibitor NVP-AEW541 or AKT inhibitor MK2206 for 24 h. p70S6K and AKT kinase activityies were detected by Western blotting. Plate clone formation assay and CCK8 assay were used to detect the growth and proliferation of hepatocarcinoma cells treated with everolimus and MK2206 alone or in combi-nation. RESULTS Sirolimus and everolimus inhibited p70S6K activity while causing feedback activa-tion of AKT kinase activity at different time points (P<0.01). NVP-AEW541 and MK-2206 could inhibit AKT kinase feedback activation by everolimus (P<0.05). Colony formation of hepatocarcinoma cells treated with everolimus and MK-2206 in combination was significantly inhibited compared with everolimus or MK-2206 alone (P<0.01). Everolimus and MK-2206 in combination inhibited the proliferation rate of two types of hepatocarcinoma cancer cells by more than 45％ compared with everolimus used alone (P<0.01). CONCLUSION The resistance of sirolimus and its derivatives in hepatocellular carcinoma cells may be achieved throngh the feedback-activated PI3K/AKT pathway, and the combination therapy can synergistically inhibit the growth and proliferation of hepatocarcinoma cells.