Background and purpose:Abnormal DNA methylation is closely associated with the onset and progression of tumors.This study aimed to investigate the expression of intermediate filament family orphan 1(IFFO1),a methylation-driven gene(MDG)in pancreatic adenocarcinoma(PAAD),along with its effects on the invasion and metastasis of PAAD cells,as well as its potential as a diagnostic and prognostic biomarker.Methods:mRNA expression data(TCGA-PAAD-mRNA),DNA methylation data(TCGA-PAAD-meth,GSE53051,PACA-AU)of PAAD and adjacent normal tissues,as well as DNA methylation data of healthy individuals'blood(GSE69270),were obtained from the The Cancer Genome Atlas(TCGA),International Cancer Genome Consortium(ICGC)and Gene Expression Omnibus(GEO)databases.By performing differential expression analysis combined with differential methylation analysis,we screened for MDG in PAAD.In the TCGA database,Pearson correlation tests were employed to verify the relationship between IFFO1 promoter methylation level and its expression level.Additionally,Kaplan-Meier survival analysis was conducted to evaluate the relationship among IFFO1 promoter methylation level,expression level,and the prognosis of PAAD.Pathological sections of cancer tissues and corresponding adjacent tissues from 27 PAAD patients were obtained from Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine.All samples involved in this study were approved by the human ethics committee of Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine(ethics number:hospital ethics review[2017]No.53).Immunohistochemistry staining(IHC)was utilized to detect the expression of IFFO1 in cancer tissues and corresponding adjacent tissues from 27 PAAD patients.Based on the median expression level of IFFO1,patients in the TCGA database were classified into high-expression and low-expression groups.Subsequently,differential analysis,gene ontology(GO)enrichment analysis and gene set enrichment analysis(GSEA)were performed.Western blot and real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)were employed to assess the expression variations of IFFO1 between the normal pancreatic ductal epithelial cell line H6C7 and the PAAD cell lines MIA PaCa2,BxPC-3,AsPC-1 and Capan-2.The impact of IFFO1 overexpression on the migration and invasion capacities of PAAD cell lines AsPC-1 and Capan-2 was evaluated using scratch and invasion assays.Additionally,receiver operating characteristic(ROC)curves and Kaplan-Meier survival analysis were utilized to assess the diagnostic and prognostic significance of IFFO1 methylation levels in the TCGA pan-cancer cohort.Results:Through the cross-screening of five datasets,41 MDG in PAAD were identified.Among these,IFFO1 was found to be the gene most closely associated with the prognosis of PAAD[hazard ratio(HR)=0.28,P＜0.001].IFFO1 exhibited high methylation and low expression levels in PAAD.Moreover,a significant negative correlation was observed between the methylation level of its promoter and its expression level(r=-0.55,P＜0.001).IHC results indicated that IFFO1 expression was significantly lower in PAAD tissues than in adjacent non-tumor tissues(P＜0.05).TCGA survival analysis demonstrated that patients with high methylation or low expression of IFFO1 had poorer overall survival(P＜0.05).Both GO and GSEA analyses indicated that the pathway"Negative regulation of cell migration"was enriched in patients with high IFFO1 expression.Western blot and RTFQ-PCR results demonstrated that IFFO1 expression in normal pancreatic ductal epithelial cells H6C7 was significantly higher compared with PAAD cell lines MIA PaCa2,BxPC-3,AsPC-1,and Capan-2.Overexpression of IFFO1 significantly inhibited the migration and invasion of the PAAD cell lines AsPC-1 and Capan-2.Additionally,pan-cancer analysis revealed that IFFO1 exhibited abnormal promoter methylation and low expression across various cancer types,with its methylation levels demonstrating significant diagnostic and prognostic prediction value among different tumors.Conclusion:Promoter hypermethylation results in decreased expression of IFFO1 in PAAD.IFFO1 may suppress the invasion and migration abilities of PAAD cells.Furthermore,IFFO1 methylation holds great promise as a novel biomarker for the diagnosis and prognosis of PAAD.

Objective:To evaluate the safety of mass vaccination of inactivated 2019-nCoV vaccine in population aged ≥3 years in Guizhou Province.Methods:The open-label study was conducted in eligible volunteers in Yanhe County of Guizhou from June 2021 to July in 2022. In the study, two doses of the inactivated 2019-nCoV vaccine were given at (0, 28) days according to the immunization schedule. The information about adverse reaction (AR) within 30 minutes and during 0-28 days after vaccination were collected, and the incidence of AR was analyzed by age, doses, and health status.Results:The overall incidence of AR was 1.51% (294/19 458), all ARs, mainly pain at injection site, occurred within 7 days after the vaccination, the AR grade was 1-2. The incidence of AR was 1.01% in age group 3- years (58/5 721), 2.44% in age group 18- years (220/9 017), and 0.34% in age group ≥60 years (16/4 720). The differences were significant ( P<0.001). The incidence of AR after the first dose vaccination (1.20%, 233/19 458) was significantly higher than that after the second dose (0.37%, 61/16 368), the difference was significant ( P<0.001). In the elderly aged ≥60 years, the incidence of AR was 0.36% (9/2 520) in healthy group and 0.32% (7/2 200) in group with underlying medical conditions, the differences were not significant ( P=0.818). Conclusion:The domestic inactivated 2019-nCoV vaccine showed good safety in mass vaccination in population aged ≥3 years, including the elderly in both healthy group and group with underlying medical conditions.

Objective:To explore clinical evaluation and genetic analysis of patients with idiopathic hypogonadotropic hypogonadism (IHH).Methods:The clinical data and phenotypes of 22 patients with IHH diagnosed and treated in our department were reviewed and analyzed. Whole-exome sequencing(WES)and Sanger method were used for variant analysis and verification.Results:Among the 22 cases of IHH probands, 12 cases of Kalman syndrome (KS) and 10 cases of IHH (nIHH) with normal sense of smell. On physical examination, males showed short penis, small testicles, small or inconspicuous laryngeal knots, and a sharp voice. Mammary gland development, mammary gland dysplasia, primary amenorrhea, etc. in women. Sex hormone examination: Follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), estradiol (E2) levels are reduced or at the lower limit of normal. There were nine missense variants of CHD7 gene in 8 patients. Based on the American College of Medical Genetics and Genomics guidelines, the c. 307T>A(p.Ser103Thr), c.3143G>A(p.Gly1048Glu), c.6956G>T(p.Arg2319Leu) and c. 3145A>T(p.Ser1049Cys) variants of CHD7 gene were predicted to be likely pathogenic (PS1+ PP1+ PM2, PM2+ PM6+ PP2+ PP3, PM2+ PM5+ PM6+ PP2+ PP3 and PM2+ PM6+ PP2+ PP3), the remaining 14 cases of IHH patients had negative genetic screening. Conclusion:CHD7 gene variants may be related to IHH disease.

Objective: To identify pathogenic variants in 5 sporadic patients and two Chinese pedigrees affected with 17-hydroxylase deficiency (17-OHD). Methods: Peripheral blood samples were collected with informed consent. Variants of CYP17A1 gene were screened by PCR and Sanger sequencing. Suspected mutations were validated in other members of the pedigrees. Results: Gene sequencing has identified a homozygous c. 985_987delTACinsAA (Y329Kfs) mutation in exon 6 of the CYP17A1 gene in 4 patients and the sister of case 3. Case 1 was found to harbor compound heterozygous mutations c. 1459_1467del9 (p.D487_F489del) and c. 1244-3C>A. The parents and brother of cases 2 and 5 were heterozygous carriers of a c. 985_987delTACinsAA(Y329Kfs) mutation. Conclusion Mutations of the CYP17A1 gene probably underlie the pathogenesis of 17-OHD, for which c. 985_987delTACinsAA(Y329Kfs) is the most common. The c. 1244-3C>A is a novel mutation. Above results have facilitated genetic counseling for the affected families.

OBJECTIVE: To explore the genotype-phenotype correlation among 18 patients with 21-hydroxylase deficiency (21-OHD). METHODS: PCR-Sanger sequencing was used to analyze the 10 exons and flanking regions of the CYP21A2 gene among the 18 patients and 20 healthy controls. RESULTS: Seventeen patients had variants of the CYP21A2 gene. Eight patients (44.4%, 8/18) carried homozygous variants including p.Ile 173Asn (62.5%, 5/8), p.Pro31Leu (25.0%, 2/8), and IVS2-13A/C>G (12.5%, 1/8), respectively. Six patients (33.3%, 6/18) carried compound heterozygous variant, among which IVS2-13 A>G+p.Ile 173Asn were most common (50.0%). 94.4% (34/36) of the variant were pathogenic, with the most common variants being p.Ile173Asn (41.7%), IVS2-13A/C>G (19.4%), and p.Ile173Asn (7.5%). No variant was identified among the 20 healthy controls. CONCLUSION The majority of 21-OHD patients carried CYP21A2 gene variants in homozygous or compound heterozygous forms, among which the p.Ile173Asn was the most common one. There is a strong correlation between the genotypes and clinical phenotypes.
Adrenal Hyperplasia, Congenital ; genetics ; Genotype ; Humans ; Mutation ; Phenotype ; Steroid 21-Hydroxylase ; genetics

OBJECTIVE: To identify pathogenic variants in 5 sporadic patients and two Chinese pedigrees affected with 17-hydroxylase deficiency (17-OHD). METHODS: Peripheral blood samples were collected with informed consent. Variants of CYP17A1 gene were screened by PCR and Sanger sequencing. Suspected mutations were validated in other members of the pedigrees. RESULTS: Gene sequencing has identified a homozygous c.985_987delTACinsAA (Y329Kfs) mutation in exon 6 of the CYP17A1 gene in 4 patients and the sister of case 3. Case 1 was found to harbor compound heterozygous mutations c.1459_1467del9 (p.D487_F489del) and c.1244-3C>A. The parents and brother of cases 2 and 5 were heterozygous carriers of a c.985_987delTACinsAA(Y329Kfs) mutation. CONCLUSION Mutations of the CYP17A1 gene probably underlie the pathogenesis of 17-OHD, for which c.985_987delTACinsAA(Y329Kfs) is the most common. The c.1244-3C>A is a novel mutation. Above results have facilitated genetic counseling for the affected families.
Adrenal Hyperplasia, Congenital ; genetics ; Exons ; Female ; Humans ; Male ; Mutation ; Pedigree ; Steroid 17-alpha-Hydroxylase ; genetics

Objective To investigate the effect and its underlying mechanism of Linagliptin on mild cognitive impairment (MCI) in elderly type 2 diabetes mellitus (T2DM) patients.Methods Montreal Cognitive Assessment(MoCA)scale was used to prospectively screen T2DM patients for MCI in our hospital from December 2016 to June 2017,and a total of 98 elderly T2DM patients with MCI were recruited.They were randomly divided into the linagliptin group(Linagliptin + metformin,n=50)and the non-linagliptin group(gliclazide + metformin,n =48).Serum fasting plasma glucose (FPG),glycosylated hemoglobin(HbAlc),blood lipids and amyloid β-protein 1-42 (Aβ1-42) levels were determined,and MoCA score and homeostasis model assessment of insulin resistance(HOMA-IR)were calculated,and were compared between the two groups before and after 24 weeks of treatment.Results In the linagliptin group,serum FPG,HbA1c,HOMA-IR,Aβ1-42 levels were significantly decreased and MoCA score was increased after 24 weeks of treatment as compared with pre-treatment [(7.29± 1.00) mmol/L vs.(9.16 ± 1.60) mmol/L,(7.19 ± 0.99) ％ vs.(9.36 ± 1.07) ％,(3.05 ± 1.12) vs.(4.05±1.30),(0.463±0.093)g/L vs.(0.528±0.110)g/L,(24.48± 1.18) vs.(23.22± 1.37),all P＜0.05].In the non-linagliptin group as control,FPG and HbA1c levels were decreased after 24 weeks of treatment as compared with pre-treatment[(7.23±1.09)mmol/L vs.(9.20± 1.75) mmol/L,(7.23±1.03)％ vs.(9.69± 1.18)％,both P ＜ 0.05],while there was no significant difference in HOMA IR,Aβ1-42 level and MoCA score[(3.95 ± 1.00) vs.(4.19± 1.13),(0.517± 0.113)g/L vs.(0.526±0.119)g/L,(23.21±1.18) vs.(23.00±1.32),all P＞0.05].It is worth to pay close attention to the key discovery of this paper that HOMA-IR and Aβ1-42 levels were significantly lower and MoCA score was significantly higher in the linagliptin group than in the non-linagliptin group after 24 weeks of treatment(all P＜0.05).Conclusions Linagliptin as one of DPP-4 enzyme inhibitors can improve the cognitive function in elderly patients with T2DM,which might be relevant to reducing serum Aβ level and improving HOMA-IR.DPP-4 enzyme inhibitor may be a good option for treatment of mild cognitive dysfunction in T2DM patients in the future.

Objective To analyze gene mutations and the etiology of six fetuses with osteogenesis imperfecta detected by prenatal ultrasonography.Methods From March 2016 to May 2017,six gravidas of singleton pregnancy and their fetuses that were diagnosed with osteogenesis imperfecta by prenatal ultrasonography were enrolled in this study.Gravida 1 came to the Center of Prenatal Diagnosis of the First Affiliated Hospital of Zhengzhou University for prenatal diagnosis,while the other five were referred to that center after termination to identify genetic defects with their fetal tissues.Next-generation sequencing technology was carried out for exome sequencing in the genomes of six fetuses.Suspected mutations were confirmed by polymerase chain reaction and Sanger sequencing.Two hundred unrelated healthy individuals were analyzed with Sanger sequencing for validation of novel mutations.Results Fetus 1 carried a heterozygous mutation in collagen,type Ⅰ,alpha-1 (COL1A1) gene,c.724G＞C(p.Gly242Arg),which was found in the mother and brother but not in the father.Fetus 2 carried a known heterozygous mutation in COL1A 1 gene,c.2461G＞A(p.Gly821Ser),which was found in the mother but not in the father.Four heterozygous mutations,c.2282G＞A(p.Gly761Asp),c.1002+5G＞A in COL1A1 gene,c.1774G＞A(p.Gly592Ser) and c.3277G＞T(p.Gly1093Cys) in collagen,type Ⅰ,alpha-2 (COL1A2) gene,were respectively carried by fetuses 3 to 6,but not by their parents.Mutations of c.724G＞C(p.Gly242Arg),c.2282G＞A (p.Gly761Asp) and c.1002+5G＞A in COL1A1 gene and c.3277G＞T (p.Gly1093Cys) in COL1A2 gene were four novel mutations,which were not found in the 200 unrelated healthy individuals.The mother of fetus 1 who was highly suspected with osteogenesis imperfecta selected to continue the pregnancy because the family members had mild symptoms.After delivery,cord blood was collected for genetic test and the result was consistent with that of prenatal genetic diagnosis.Fetus 1 had no fractures during a six-month follow-up after birth.Conclusions Mutations in the COL1A1 and COL1A2 genes may be the etiology of osteogenesis imperfecta in these six fetuses.Results of this study could enrich the data on COL1A1 and COL1A2 mutations relating to osteogenesis imperfecta,and provide a basis for genetic counseling.

Objective To analyze the situation of table tennis competitions in emergency public health events, find the possible weak links in the emergency rescue and care work, put forward the hospital in response to emergency public health emergency preparedness plan, health emergency drills and emergency rescue personnel training and so on targeted reference views. Methods A total of four public health emergencies such as traffic accident, mass food poisoning, fire and bird flu were selected as the theme of the competition to simulate the emergency rescue process after the emergency,the examination of each team is divided into pre-hospital disposal and hospital disposal links. Results The score of Crash in Batches was 225.13 ± 8.38, followed by Fire accident 222.78 ± 16.57, Avian Influenza 210.32 ± 11.19, Group Food poisoning 204.80 ± 6.77, the difference was statistically significant (F=8.706,P=0.000). The scores of pre-hospital treatment in all four subjects were109.04±4.80, 108.62±10.12, 98.24±6.95, 101.14± 8.85,lower than those in hospital, which were 116.09±4.79, 114.16±7.61, 106.56±5.31, 109.18±4.24, the difference was statistically significant (t=-6.130--2.338,P＜0.01 or 0.05). Conclusions On the different topics exposed in the desktop exercise competition and the differences between the pre-hospital treatment and the hospital disposal, the nursing managers should strengthen the training and practice of public health emergencies with different themes, and pay attention to the pre-hospital emergency public health emergencies training and rehearsal,the emergency response plans for public health emergencies were revised and improved through the health emergency drills.

Objective To evaluate the efficacy of superficial temporal artery(STA)pressure-guided selective cerebral perfusion(SCP)during deep hypothermic circulatory arrest(DHCA)in patients undergoing aortic arch surgery.Methods Ninety-six patients of both sexes,aged 35-64 yr,with body mass index of 19-23kg/m2,of American Society of Anesthesiologists physical status Ⅲ or Ⅳ,undergoing aortic arch surgery,were divided into STA pressure group(group A)and clinical experience group(group B)using a random number table,with 48 patients in each group.In group A,STA catheterization was performed after tracheal intubation,and arterial pressure was monitored.SCP flow was adjusted to maintain the target value of STA pressure between 30 and 40mmHg during DHCA in group A.SCP flow rate was set at 5-10ml·kg-1·min-1 according to clinical experience in group B.The volume of fluid perfused during SCP,emergence time,extubation time and duration of intensive care unit stay were recorded.Neurological function was evaluated during length of hospitalization after surgery,and the development of permanent and transient neurological dysfunction and mortality in hospital were recorded.Results Compared with group B,the volume of fluid perfused during SCP was significantly decreased,the emergence time,extubation time and duration of intensive care unit stay were shortened,the incidence of permanent and transient neurological dysfunction(2% and 4%,respectively)was decreased(P < 0.05),and no significant change was found in the mortality rate in hospital in group A(P>0.05).Conclusion Maintaining STA pressure at 30-40mmHg is a reliable method for guiding SCP during DHCA in patients undergoing aortic arch surgery.