Background and purpose:CpG methylation in promoter region is an essential mechanism for the dysfunction of the tumor suppression gene.Folate metabolism provides active methyl for organism methylation.Methionine synthase(MS) plays a vital role in the process of folate metabolism.This study aimed to explore the genetic polymorphism of MS, CHD5 methylation and their association with breast cancer morbidity.Methods:Fortyseven patients with primary breast cancer, 52 healthy subjects and 15 breast hyperplasia patients were enrolled in this experiment.The mRNA expression of CHD5 was determined by reverse transcription PCR(RT-PCR).Methylationspecific PCR(MSP) was used to detect the methylation status of CHD5.Polymorphisms in the MS gene were analyzed through the polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).Results:The mRNA expression of CHD5 in breast cancer tissues(0.27?0.19) and adjacent non-cancerous tissues(0.33?0.17) were both significantly lower than that in the breast hyperplasia tissues(0.67?0.14)(P

Objective:To investigate the methylation status of N-myc downstream regulated gene-1(NDRG-1) gene in breast cancer and the effects of methylation enzyme inhibitor 5-Aza-2'-deoxycytidine (5-Aza-CdR) on growth and expression of NDRG-1 mRNA in human breast cancer cell line T47D.Methods:Sensitive methylation-specific (MSP)-PCR was used to detect the methylation status in the promoter regions of NDRG-1 gene in 47 samples of breast cancer and tumor adjacent tissues and 15 cases of benign breast disease. The change in expression of the tumor suppressor gene NDRG-1 mRNA in cultured T47D cells was detected by RT-PCR before and after 5-Aza-CdR treatment. Cell proliferation was observed by MTT assay.Results:Hypermethylation frequencies of NDRG-1 gene promoter were 46.8% in breast cancer tissues and 21.3% in tumor adjacent tissues. No hypermethylation of NDRG-1 gene was observed in the tissues of breast benign disease. The growth of T47D cells was suppressed obviously after 5-Aza-CdR treatment compared with the control group. RT-PCR showed that compared with the control group, NDRG-1 mRNA expression was increased at different concentrations in 5-Aza-CdR treatment group. Conclusion:The promoter methylation status of NDRG-1 gene was significantly related with the occurrence of breast carcinoma. 5-Aza-CdR could effectively reverse the methylation of NDRG-1 gene and recover its expression, thereby inhibiting the growth of tumor cells.

Aim Toinvestigatetheinducementeffect of isatin on apoptosis of breast cancer cell line MCF-7 , andexploreitsdetailedmechanism.Methods MCF-7 cell lines were exposed to isatin at different concentra-tions(0,50,100,200 μmol·L-1 )for 48 h.Apop-totic features were demonstrated by nuclei staining with Hoechst 33258.Bcl-2,Bax and p53 mRNA were ana-lyzed by RT-PCR.Caspase-9 activation and mitochon-drial depolarization were assayed by flow cytometry. Bcl-2,Bax,p53 and cytochrome c proteins were ana-lyzedbyWesternblot.Results Isatininducesapopto-sis of MCF-7 cells.Furthermore,Bcl-2 expression was decreased and the ratio of Bcl-2 to Bax was significant-ly decreased by isatin.While,p53 expression relative-ly decreased.The mitochondrial transmembrane poten-tial was markedly reduced and the release of cyto-chrome c into the cytosol was increased after treatment with isatin.Simultaneously,caspase-9 was activated. Conclusions Isatinsignificantlyinducedtheapopto-sis of MCF-7 cells in vitro.These results strongly sug-gest that the p53 dependent mitochondrial pathway is involved in apoptosis.

Objective:To explore the effects of problem-solving based telephone and texting interventions for medication adherence of discharged patients with schizophrenia.Method:A total of 178 discharged patients with schizophrenia were randomly assigned to telephone intervention (TI)group (n =63),text-messaging intervention (TMI)group (n =61),and control group (n =54).All patients were routinely treated,and patients in TI and TMI groups were given problem-solving based interventions.Medication adherence and psychotic symptoms were as-sessed by pill counting and the Positive and Negative Symptom Scale (PANSS)at baseline and the first,third, sixth,ninth and twelfth month.Results:Overall,patients with TI had higher medication adherence than those with TMI (P <0.001),and patients with TMI had higher medication adherence than that in control group (P <0.001). PANSS scores in TI and TMI groups were not significantly different (P >0.05),but significantly lower than control group (P <0.01).There was higher rate of re-admission in control group in comparison with TI and TMI groups (9.3% vs.[0% and 1.6%],P <0.05).Conclusion:It suggests that problem-solving based telephone and texting interventions are effective for improving medication adherence in patients with schizophrenia after their discharge from hospital,so as to reducing psychotic symptoms and lowering re-hospitalization risk.In comparison with TMI, TI has a better effect in improving medication adherence.

Objective:To investigate the mRNA expression of the WIF-1 gene and the methylation of its promoter in breast can-cer, and to determine the correlation between the epigenetic aberrant WIF-1 DNA methylation and the clinicopathological significance of WIF-1 in breast cancer . Methods:RT-PCR and sensitive methylation-specific-PCR (MSP) were used to detect WIF-1 mRNA ex-pression and the methylation of the WIF-1 promoter in 30 breast cancer samples as well as in tumor-adjacent tissue samples and 9 be-nign breast tissues. Results:The WIF-1 mRNA expression in 30 breast cancer samples significantly decreased compared with those of the other two groups. In addition, WIF-1 methylation was more frequent in breast-tumor tissues compared with those in tumor-free tis-sues. Meanwhile, WIF-1 mRNA expression in breast cancer tissues involved the abnormal methylation of its promoter. Clinicopatholog-ical correlation analysis showed that the abnormal methylation of the WIF-1 gene promoter was not associated with age, TNM stage, histotype, or lymph node metastasis. Conclusion:WIF-1 mRNA expression loss due to abnormal methylation may be a crucial factor in breast cancer development and can thus be used in the prognosis and progression of the disease.

Objective To investigate the effects of simulated microgravity by rotary cell culture system (RCCS) on expression profiles of miRNA in mouse fibroblasts L929 cell line.Methods L929 cells were cultured in vitro and divided into simulated microgravity (SMG) group and normal gravity (NG) group according to the simple random method.Samples of two groups were collected on 7th day of culture and the total RNAs were extracted,labeled,and hybridized in sequence.Feature Extraction Software was used to collect the array images and get raw data,which were analyzed by Genespring Software.Differentially expressed miRNAs were identified and then validated by qRT-PCR.Target genes of differentially expressed miRNAs were predicted by the databases of Targetscan and microRNAorg,and the intersections of databases were identified as potential regulatory target genes.Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis were applied to determine the roles of these target genes.Relevant biological functions and/or signaling pathways of the regulated genes especially related with wound healing process were categorized based on their enrichments.Then integration predictions of the miRNA and mRNA expression profiles had been proposed to refine the functional miRNA-mRNA relationships.The miRNA-mRNA functional network and miRNA-mRNA control network were constructed.Results Four miRNA genes were up-regulated significantly including mmumiR-669j,-122-5p,-30a-3p,-6516-3p,among which mmu-miR-669j was up-regulated at 52.84 folds with the greatest significance (P ＜ 0.05).Seventeen miRNA genes were down-regulated significantly including mmu-miR-21a-3p,-miR-28a-5p,-218-5p,-210-3p,-miR-19a-3p,-miR-31-3p,and-miR-19b-3p,among which mmu-miR-28a-5p was down-regulated at 15.47 folds with the greatest significance (P ＜ 0.05).The qRT-PCR showed a high concordance with the microarray results (P ＜ 0.05).Target gene prediction and functional enrichment analysis suggested that a variety of biological processes and signaling pathways involved in wound repair were significantly enriched (P ＜ 0.05).Function network and regulation network of miRNA-mRNA covered all the differentially expressed miRNAs,which suggested that miR-21 a-3p and predicted target gene Smad3 might play an important role in wound healing under microgravity.Conclusions Simulated microgravity by RCCS can significantly affect the expression of stress-related miRNAs in mouse fibroblasts L929.The miRNA target gene prediction and functional enrichment analysis based on gene chip technology may provide theoretical basis for illustrating the mechanism and management of weightlessness stress injury.

Objective To investigate the status of depression in female infertility patients and its influencing factors in reproductive medicine center. Methods Totally 147 cases of female infertility patients from the reproductive medicine center of a tertiary hospital in Beijing were collected as the research object by convenient sampling method from July to September 2015. The questionnaire survey was used to investigate the patients′general information, disease related data and depression mood. Results In the study, the average depression score of 147 participants was (0.45±0.12). Among them, there were 32 cases (21.8%) of mild depression, 15 cases ( 10. 2%) of moderate depression and 4 cases ( 2. 7%) of severe depression. Multiple regression analysis showed that patients with depression were affected by academic qualifications, age and whether there was a history of transplantation. Conclusions The outpatients of the reproductive medicine center have a more severe depression mood and will be affected by the patient′s education background, age, and whether there is a history of transplantation. It is recommended to consider the patient′s education background, age, and whether there is a history of transplantation in patients′health education.

A male child aged two years and six months was admitted to the Affiliated Women and Children′s Hospital of Qingdao University in June 2023 due to wide eye distance, stubby fingers, koilosternia, cryptorchidism, and short tongue frenum. After clinical data collection, whole exome sequencing (WES) was conducted and bio-informatics analysis was performed to search for possible mutation sites on the patient. Family lineage verifications were conducted through Sanger sequencing. WES results showed that the patient carried c.2432dupT frameshift hemizygote variation of the FGD1 gene on the X chromosome (Xq11.22). Sanger sequencing confirmed that the mother was a carrier of the c.2432dupT heterozygous variant but not the father. According to the sequence interpretation guidelines of the American College of Medical Genetics and Genomics (ACMG), this variant is pathogenic (ACMG: PVS+2PP). The patient was diagnosed with Aarskog-Scott syndrome(ASS), which is a rare X-linked disorder characterized by facial, skeletal and genital anomalies. This case study has enriched the variant spectrum of the FGD1 gene and provided guidance for clinical screening and diagnosis of ASS.

Objective:To analyze food intolerance status in children in Qingdao by detecting the serum levels of food-specific IgG (sIgG).Methods:In this cross-sectional study, a total of 4 249 children aged 0 to 14 years (all were permanent residents of Qingdao City) admitted to Women and Children′s Hospital Affiliated to Qingdao University from May 2017 to December 2020 for suspected food intolerance were selected as the study objects with the whole sampling method. According to the age, the objects were divided into 4 groups: 0-<1 year group (440 cases), 1-<3 years group (1 761 cases), 3-<6 years group (1 193 cases), and ≥6 years group (855 cases). Positive condition of serum sIgG antibodies of 14 kinds of food in the children were detected by enzyme-linked immunosorbent assay. Chi-square test was used to compare the positive rate of the antibodies among different foods, gender and age groups.Results:The total positive rate of food sIgG antibody in 4 249 children was 95.32% (4 050/4 249), the highest positive rate was found in eggs (81.50%) and the lowest positive rate was found in pork (1.15%). The positive rates of sIgG antibody in milk (54.98% vs 49.69%, χ2=11.627), crab (5.59% vs 3.71%, χ2=8.049) and shrimp (4.62% vs 2.75%, χ2=9.784) in boys were significantly higher than those in girls, and the positive rates of sIgG antibody in tomato (49.19% vs 45.54%, χ2=5.510), cod (8.53% vs 5.96%, χ2=10.512) and beef (2.58% vs 1.70%, χ2=3.959) in girls were significantly higher than those in boys (all P<0.05). The total positive rate of sIgG antibody in 14 foods was the lowest in 0-<1 year group (89.09%), and it was the highest in 3-<6 years group (96.98%) ( χ2=63.950, P<0.001). The highest positive rate in 0-<1 year group was found in tomato (56.36%), and it was eggs (85.29%, 88.94%, 85.50%) in all the other 3 groups. The positive rates of corn and beef decreased with age ( χ2=44.098, 20.106, P<0.001), while those of cod and mushroom increased with age ( χ2=32.315, 40.338, P<0.001). The positive rate of wheat (57.13%, χ2=42.273), tomato (57.01%, χ2=209.862), soybean (24.99%, χ2=92.580), crab (6.81%, χ2=33.201), shrimp (6.25%, χ2=47.863) were all the highest in 1-<3 years group among the 4 groups (all P<0.001), and the positive rate of chicken was the highest in 3-<6 years group (7.88%, χ2=29.875; P<0.001). Conclusions:Children in Qingdao have a high level of food intolerance, and the highest positive rate is for eggs. Milk, crab and shrimp should be focused on for boys, while tomatoes, cod and beef shoud be paid more attention to for girls. Children of different ages have different kinds of food intolerance, and their diets should be adjusted reasonably according to the characteristics.

To analyze the positivity rate and titer of antinuclear antibody (ANA), as well as nuclear pattern and target antigen of ANA in healthy pregnant women during early pregnancy in Qingdao area. A prospective cohort study design was used to include a total of 9 528 healthy pregnant women registered at the Women and Children′s Hospital Affiliated to Qingdao University from March 2023 to June 2024.Indirect immunofluorescence assay (IIF) was used to detect ANA, its titer and cell staining pattern. Fifteen specific antibodies were tested using the magnetic bar code immunofluorescent luminescence method. Logistic regression model was used to analyze the risk factors of pregnancy with autoimmune disease(AID). The results showed that among 9 528 pregnant women in early pregnancy, 1 346 cases (14.1%) were positive of ANA, including 1 011 cases with a titer of 1∶100 (10.6%), 236 cases (2.5%) with a titer of 1∶320, and 99 cases (1.0%) were detected at a titer >1∶320. Among the 1 346 ANA-positive pregnant women, nuclear granular type accounted for the highest proportion (483 cases, 35.9%), followed by speckled type (347 cases, 25.8%) and cytoplasmic type (176 cases, 13.1%).Then, pregnant women with ANA titers ≥1∶100 were detected 15 specific antibodies.Anti-SSA was tested in 121 cases accounted for the majority, followed by 110 cases with anti-Ro-52, 56 cases with anti-SSB, 51 cases with anti-mitochondrial M2 subtype antibodies and 37 cases with anti-centromere B. In conclusion,in healthy pregnant women in Qingdao area, ANA positivity rate was 14.1%, and the titer of ANA was mainly at 1∶100.The predominant nuclear patterns were nuclear granular and speckled types.The specific autoantibodies were mainly anti-SSA antibodies and anti-Ro-52 antibodies.The detection of ANA and specific autoantibodies is of great significance for early prediction, diagnosis, and intervention of autoimmune diseases during pregnancy.