At present,birth defects and genetic diseases has become a major public health problem affecting children's health and quality of births.Genetic diagnostic testing of DNA or RNA by molecular biological methods is an important means of birth defects and genetic disease detection.This review will elaborate the applications and prospects of mainstream technology of genetic diagnosis in the fields of birth defects and genetic diseases detection.

Objective To investigate the expression of SOX4 and C/EBPα mRNA in chronic myeloid leukemia (CML) and their clinical significances. Methods Bone marrow samples from 68 cases of CML including 57 newly diagnosed patients and 11 patients treated with imatinib were collected, and peripheral blood mononuclear cells from 30 healthy people were collected as healthy control. The expression of SOX4 and C/EBPαmRNA and protein levels were detected by RT-PCR and Western blot, respectively. The relations between the expression of SOX4 and C/EBPα and the influences of imatinib on SOX4 and C/EBPα were analyzed. Results The expression level of SOX4 mRNA was increased in newly diagnosed CML patients compared with that of normal control group (6.545 5±1.495 2 vs. 0.059 6±0.018 8, t=3.139, P=0.002 3), but the expression level of C/EBPαmRNA was significantly decreased (0.238 8±0.033 8 vs. 0.810 5±0.056 2, t=9.240, P<0.000 1). The expression levels of SOX4 and C/EBPαmRNA had no significant correlation with age, gender, white blood cell count (WBC) and bcr-abl of newly diagnosed patients (all P>0.05). The expression level of SOX4 mRNA in 5 patients treated with imatinib was decreased (0.120 6 ±0.044 9 vs. 0.557 9±0.144 8, t=2.885, P=0.020 4), and the expression level of C/EBPαmRNA was increased (0.330 3±0.042 4 vs. 0.150 5±0.046 5, t=2.855, P=0.021 3). The expression level of SOX4 mRNA in 6 patients who developed blast phase during the treatment of imatinib was increased (0.469 9±0.123 0 vs. 0.050 2±0.036 6, t=2.370, P=0.039 3), and the expression level of C/EBPα mRNA was decreased (0.197 9 ±0.064 7 vs. 0.378 7±0.042 9, t=2.327, P=0.042 3). The expression of SOX4 mRNA was negatively correlated with C/EBPα mRNA (t=-0.554 6, P=0.002 8). Conclusions In newly diagnosed CML, the expression level of SOX4 is increased, C/EBPα is decreased compared with that of healthy control, and both have negative correlation. In the patients in blast phase after imatinib treatment, SOX4 gene is up-regulated, and C/EBPα is down-regulated. C/EBPα-SOX4 axis may play a role in the occurrence and development of CML. SOX4 may be a new molecular target for the treatment of CML.

Objective To explore the role of tumor inhibition of enhancer binding protein C/EBPα in the leukemic mice. Methods BALB/c nude mice were randomly divided into three groups. Three kinds of cells including pEGFP-C/EBPα-K562 cells, pEGFP-K562 cells and K562 cells as the control were injected into mice separately through the subcutaneous and tail vein, and subcutaneous tumors and leukemic models were formed. The changes of tumors were observed and the apoptosis of cells was detected by TUNEL; The capacity of proliferation of leukemia cells was observed in the bone marrow and the peripheral blood by Wright-Giemsa staining. The expression of genes of related to proliferation was detected by RT-PCR. Results The quality and the max diameter of tumors in the pEGFP-C/EBPα-K562 group were smaller than that of pEGFP-K562 group and K562 control group [(2.4±0.1) g vs (5.1±0.3) g and (5.7±0.4) g, both P ＜0.05; (11+2)mm vs (19+3) mm and (23+3) mm, both P ＜0.05]. More apoptosis cells were found in the pEGFP-C/EBPα-K562 group leukemic cells were found in the peripheral blood of leukemic models, and the proliferation of leukemic cells in the pEGFP-C/EBPo-K562 group were lower than that of other groups, accompany by the conspicuous cell differentiation. p53 was significantly elevated by RT-PCR, while down-regulated of c-myc.Conclusion Enhancer binding protein C/EBPα promote the apoptosis of cells and inhibit the proliferation of leukemia cells in leukemia mice, and further induce the cell differentiation. The inhibition of enhancer binding protein C/EBPα in the leukemia may have effect through the regulation of related genes.

Objective To investigate the effects of circadian clock gene Period2(Per2)on the proliferation,differentiation and apoptosis of K562 cells and its probable molecular mechanism.Methods The Per2 expression plasmid pcDNA3.1-Per2 and empty control plasmid were respectively transfected into K562 cells with cationic liposome,and the resistant cells stably expressing Per2 gene were obtained by G418 selection.Their morphological changes were observed under light microscope following Wright-Giemsa staining.Trypan blue excluding staining and MTT assay were employed to evaluate cell proliferation.Flow cytometry was performed to analyze cell cycle distribution and cell apoptosis,and electron microscopy was used to detect cell apoptosis.Meanwhile,the expressions of proliferation and apoptosis associated proteins,such as P53,Cyclin B1 and C-Myc,were respectively detected by RT-PCR and Western blot analysis at mRNA and protein level.Results The K562/Per2 cell line stably expressing Per2 gene was screened out.As compared with either the empty plasmid transfected group(K562/empty)or the untreated group(K562/untreated),K562/Per2 cells was smaller in volume and showed no obvious cellular differentiation.Circadian clock gene Per2 could significantly inhibit both growth and proliferation of K562 cells.The percentage of K562 cells in G2/M phase increased [K562/Per2 group(36.1?5.5)%,K562/empty group(12.5?2.9)%,untreated group(9.7?2.3)%,P

Objective:To explore the diagnosis, treatment and prognostic of pediatric intracranial atypical teratoid/rhabdoid tumor(AT/RT).Methods:A total of 15 pediatric patients with intracranial AT/RT were treated between January 2012 and June 2019 at the First Affiliated Hospital of Zhengzhou University.The clinical data were retrospectively analyzed.Overall survival (OS) rate and progression free survival (PFS) rate were calculated by adopting Kaplan- Meier method.The differences between the 2 groups were tested by performing Log- rank method, and the prognostic factors were analyzed by COX regression. Results:There were 12 males and 3 females, with the median age of 5.5 years (ranging from 8 months to 17.1 years). All patients underwent surgical resection.Gross-total resection (GTR) was achieved in 10 cases and subtotal resection (STR) was carried out in 5 patients.The conducted treatments were as follows: surgery+ radiotherapy+ chemotherapy+ intrathecal injection in 6 cases, surgery+ chemotherapy+ intrathecal injection in 4 cases, surgery+ radiotherapy in 2 cases, and surgery alone in 3 cases.Until January 2020, the median survival time of all the 15 patients was 18 months (ranged 1-27 months), and the survival rate was 33.3%.The 1-year OS rate and PFS rate for all 15 cases were 71.5% and 49.7%, respectively.The 2-year OS rate and PFS rate were 17.9% and 0, respectively. Log- rank analyses revealed that the 1-year OS rates of children less than 3 years old and those older than 3 years were 87.5% and 57.1%, respectively ( χ2=6.057, P=0.014). The 1-year OS rates of children with GTR and those with STR were 90.0% and 40.0%, respectively ( χ2=6.057, P=0.014). The 1-year OS rates of children with tumor dissemination and those without tumor dissemination were 100.0% and 33.3%, respectively( χ2=9.865, P=0.002). The 1-year OS rates of children in the standard-risk group and those in the high-risk group were 88.9% and 41.7%, respectively ( χ2=5.111, P=0.024). COX regression analyses proved that age, the extent of tumor resection, tumor dissemination and risk stratification are independent risk factors for prognosis [hazard radio( HR)=3.411, 3.795, 5.245, 3.397; P=0.025, 0.011, 0.001, 0.017]. Conclusions:Pediatric intracranial AT/RT is rare.The preliminary diagnosis and prognosis are difficult and poor, respectively.The complete resection of tumors with maximal safety remains the primary treatment.Age, the extent of tumor resection, tumor dissemination and risk stratification are independent prognostic factors for AT/RT children.

Objective:To investigate the utilization of health management services and its influencing factors among new urban population.Methods:It is a cross-sectional study. From July 2020 to March 2021, a stratified random sampling method was used to extract 1978 new urban population in Jining city, and an anonymous self-administered questionnaire survey was conducted using a self-made questionnaire ′Residents Health Questionnaire′. The survey included general demographic characteristics, personal behavior lifestyle and medical care status. The χ2 test and binary logistic regression were used to analyze the factors influencing the utilization of health management services by new urban population. Results:The overall utilization of health management services in the new urban population was 47.22%. There were significant differences in utilization of health management services among new urban population with different gender, age, education level, occupation and monthly income. Binary logistic regression analysis showed that female ( OR=1.354, 95% CI: 1.094-1.676), people aged over 60 years ( OR=1.873, 95% CI: 1.413-2.483), people with a mean monthly income over 3 000 yuan ( OR=1.498, 95% CI: 1.123-1.997), people engaged in light manual labor ( OR=1.596, 95% CI: 1.003-2.539), people who exercise regularly( OR=2.400, 95% CI: 2.028-2.841) and people having social basic medical insurance ( OR=2.633, 95% CI: 2.042-3.394) had better utilization of health management projects. People who sat more than 3 hours a day ( OR=0.630, 95% CI: 0.532-0.745) had lower utilization of health management care. Conclusion:The utilization of health management projects in the new urban population is low. Gender, age, monthly income, physical exercise, sedentary time, daily labor intensity and social basic medical insurance status are the main influencing factors.

Objective:To explore the clinical efficiency evaluation and prognostic factors of aspiration guided by neuronavigation in the treatment of pediatric brain abscess (PBA).Methods:A total of 47 patients with PBA were treated with aspiration guided by neuronavigation between January 2013 and January 2019 at the First Affiliated Hospital of Zhengzhou University.All clinical data were retrospectively analyzed.According to Glasgow Outcome Scale on discharge, all children were divided into 2 groups, namely good prognosis group and poor prognosis group.Prognostic factors were analyzed by using univariate analysis and binary Logistic regression multivariate analysis. Results:Among the 47 children, 38 children (80.9%) were assigned to the good prognosis group, and 9 children (19.1%) were assigned to the poor prognosis group.Univariate analysis proved that abscess volume>4 cm( χ2=5.650, P=0.017), multiple or multilocular abscess ( χ2=3.258, P=0.027), and abscess located in functional areas ( χ2=6.187, P=0.013) were correlated with poor prognosis.Multivariate analysis revealed that abscess volume>4 cm( OR=5.913, 95% CI: 2.241-25.917, P=0.023) and abscess located in functional areas ( OR=10.519, 95% CI: 3.918-62.513, P<0.001) were independent risk factors for poor prognosis. Conclusion:The treatment of PBA with aspiration guided by neuronavigation is safe, effective and minimal invasive, and the clinical efficiency is satisfactory.Abscess volume>4 cm and abscess located in deepbrain/functional areas are independent risk factors for poor prognosis.

Objective:To conduct periodic revalidation of the 15 items and 43 terms autoverification rules of blood analysis after 1 year of application, analyze the application suitability and make the rules improved.Methods:Track the results of 528 010 blood analysis samples of our hospital from August 1, 2019 to January 31, 2020, and analyze the pass rate and interception rate of autoverification; 600 specimens in total were selected randomly for microscope examination, including 300 specimens which touched autoverification rules (1 012 items of autoverification rules) and were intercepted by autoverification and 300 specimens which untouched autoverification rules and were released by autoverification. The abnormal characteristics and unacceptable Delta check of the specimens also need to be concerned at the same time.The false negative rate and false positive rate, true negative rate, true positive rate and pass correct rate of autoverification were verified and compared with the rate of the second phase verification when the autoverification rule was established. The false negative rate, false positive rate, true negative rate and true positive rate of the Delta check rule which 54 716 specimens touched were calculated and compared with the second phase verification rate when the autoverification rule was established.The results of microscopic examination were used as the gold standard for the calculation of the rates, and P<0.05 was considered as a significant difference. The false positive and true positive of 1 012 autoverification rules were analyzed item by item.The false positive and true positive of 108 specimens which touched blast cell autoverification rule were analyzed terms by terms. The mean TAT and median TAT of 528 010 specimens and 193 750 outpatient specimens were calculated respectively, and the report percentages of 528 010 samples that TAT<30, 30-60　and>60 min were calculated respectively. Analyze and evaluate the application suitability of autoverification rules to juge whether they meet the needs of doctors and laboratory. The design process and the rules and application process of autoverification were optimized and improved.Results:The autoverification pass rate was 63.06% (332 971/528 010), the interception rate was 36.94% (195 039/528 010). The false negative rate was 1.00% (1/600), the false positive rate was 12.67% (76/600), the true negative rate was 49% (294/600), the true positive rate was 37.33% (224/600), and the correct rate was 98% (294/300). The pass rate, true negative rate, true positive rate and correct rate of the periodic reverification group were higher than the second phase verification group, the false negative rate and false positive rate were lower than that the second phase verification group. The false negative rate and true positive rate of the Delta check of periodic verification group were lower than that the second phase verification group, the false positive rate and true negative rate were higher than the second phase verification group, there were significant differences in the comparition results. The mean TAT of 528 010 specimens was25 min, and the median TAT was 22 min. The mean TAT of 193 750 outpatient specimens was 23 min, and the median TAT was 20 min. The report percentages of 528 010 samples that TAT<30 min, 30 min-60 min and>60 min were 83.30% (439 819/528 010), 8.00% (42 250/528 010) and 8.70% (45 941/528 010), respectively.Conclusion:The results of periodic revalidation of autoverification after 1 years application show that the 15 items and 43 terms autoverification rules of blood analysis could meet requirements about the accuracy and efficiency of the laboratory, and have a good suitability for application.

Objective:To explore the markers predicting the efficacy of anti-programmed death receptor-1 (PD-1)/PD-1 ligand (PD-L1) immunotherapy for non-small cell lung cancer (NSCLC) and analyze their relationships with the tumor immune microenvironment.Methods:(1) Gene expression profile data and relevant clinical data of 55 NSCLC patients receiving anti-PD-1/PD-L1 monotherapy from two independent clinical cohorts were downloaded from the GEO database. Genes associated with progression-free survival (PFS) were screened using univariate Cox regression analysis. (2) Twenty-six pathological tissue specimens of NSCLC patients who received anti-PD-1/PD-L1 monotherapy in the Cancer Hospital of Chinese Academy of Medical Sciences (CICAMS) from April 2016 to August 2019 prior to the screening of genes were selected to verify the predictive value of the screened genes using immunohistochemistry. (3) The relationship between efficacy predictive markers and PD-L1 and infiltrating immune cells in the tumor immune microenvironment was analyzed using NSCLC gene expression profile downloaded from the TCGA database.Results:Univariate Cox regression analysis revealed that only T-Cell Surface Glycoprotein CD3 Gamma Chain (CD3G) was a risk predictive gene for PFS in NSCLC patients in both clinical cohorts of the GEO database (GSE93157: P = 0.049; GSE136961: P = 0.034). Further Kaplan-Meier survival curves showed that PFS was significantly prolonged in the high CD3G expression group ( P = 0.012). The results of survival analysis in the CICAMS clinical cohort also demonstrated that NSCLC patients with high CD3G expression had a better prognosis after receiving anti-PD-1/PD-L1 monotherapy ( P = 0.001) compared with those with low CD3G expression. Univariate and multivariate Cox regression analyses also showed that CD3G was an independent predictor of PFS (univariate: P = 0.002; multivariate: P = 0.001). The tumor immune microenvironment analysis showed that CD3G was positively correlated with PD-L1 expression (lung adenocarcinoma: r = 0.44, P < 0.001; lung squamous cell carcinoma: r = 0.37, P < 0.001) and the infiltration level of CD8 + T cells (lung adenocarcinoma: r = 0.13, P = 0.002; lung squamous cell carcinoma: r = 0.70, P < 0.001). CD3G was also positively correlated with the expression of CD8 + T cell chemokines CCL5, CXCL9, CXCL10 and CXCL11. Conclusion:Patients with NSCLC with high CD3G expression have greater clinical benefits after anti-PD-1/PD-L1 monotherapy compared with those with low CD3G expression. CD3G can be used as a potential marker to predict the efficacy of immunotherapy for NSCLC.