Objective:To explore the effects of enriched environment combined with melatonin on learning and memory function and DNA oxidative damage in senescence accelerated mouse prone 8 (SAMP8) mice.Methods:Twenty-four 6-month-old SPF healthy male SAMP8 mice were randomly divided into model group, enriched environment group, melatonin group and enriched environment+ melatonin group, with 6 mice in each group. Six homologous SAMR1 mice of the same age were used as the control group. The mice in the enriched environment group and the enriched environment+ melatonin group were fed in the enriched environment. At the same time, the mice in the melatonin group and the enriched environment+ melatonin group were subcutaneously injected with melatonin (8 mg /(kg·d)) once a day for 28 d. The mice in the model group, the control group and the enriched environment group were subcutaneously injected with an equal volume of 0.9% sodium chloride solution once a day for 28 days. Aging score was used to evaluate the aging of mice. Morris water maze and Y maze tests were used to evaluate the learning and memory ability of mice. The cell morphology of hippocampus in mice was observed by hematoxylin-eosin staining, and the level of Aβ 1-42 protein in hippocampus of mice was detected by immunohistochemical staining. The levels of γ-H2A histone family member X(γ-H2AX) and 8-hydroxy-2 deoxyguanosine(8-OHdG) proteins in hippocampus of mice were detected by Western blot and Enzyme-linked immunosorbent assay. SPSS 25.0 statistical software was used to process the data. One-way analysis of variance was used for comparison among multiple groups, and LSD- t test was used for further pairwise comparison. Results:(1)There was a statistical difference in aging scores among the 5 groups of mice after intervention ( F=126.4, P<0.01). After intervention, the aging scores of mice in the enriched environment group, melatonin group, and enriched environment+ melatonin group were lower than that in the model group (all P<0.05), and the score of the enriched environment+ melatonin group was significantly lower than that in the enriched environment group ( P<0.05). (2)The time and group interaction, group main effect and time main effect of the escape latency among the 5 groups of mice were statistically significant ( F=11.2, 799.9, 121.8, all P<0.01). From day 2 to day 4, the escape latencies of mice in the enriched environment group, melatonin group and enriched environment+ melatonin group were significantly lower than that in the model group (all P<0.05). There was a statistically significant difference in the target quadrant residence time and cross-platform times among the 5 groups ( F=70.38, 48.83, both P<0.01). The target quadrant residence time and cross-platform times of mice in the enriched environment group, melatonin group, and enriched environment+ melatonin group were significantly higher than that in the model group (all P<0.05). (3) There were significant differences in the total number of alternations and correct rates among the 5 groups ( F=291.328, 113.482, both P<0.01). The total numbers of alternations and correct rates in melatonin group ((29.46±3.75)times, (53.16±3.47)%) and the enriched environment+ melatonin group((32.57±3.52)times, (58.60±4.13)%)were significantly higher than those in the model group ((18.62±3.96)times, (43.61±3.92) %)(all P<0.05). (4)The results of hematoxylin-eosin staining and immunohistochemistry staining showed that compared with the model group, the cell structure and morphology of the hippocampus of mice in enriched environment group, melatonin group, and enriched environment+ melatonin group were significantly improved, and the expression of Aβ 1-42 was significantly reduced (all P<0.05). (5) There were statistically significant differences in the levels of γ-H2AX and 8-OHdG proteins in the hippocampus of the 5 groups of mice ( F=78.09, 117.20, both P<0.01). The levels of γ-H2AX and 8-OHdG of mice in the enriched environment+ melatonin group ((1.37±0.26), (4.79±0.35)pg/μg) were significantly lower than those in the enriched environment group ((2.83±0.25), (7.23±0.41)pg/μg) and the melatonin group ((2.43±0.22), (6.69±0.28)pg/μg) (all P<0.05). Conclusion:Both enriched environment and melatonin can significantly improve the learning and memory function of SAMP8 mice, and the combined treatment effect is more significant.The mechanism may be related to the reduction of DNA oxidative damage in hippocampus.

Objective:To evaluate the diagnostic capabilities of PCR-flow Fluorescence Hybridization technology in prenatal genetic diagnosis of thalassemia.Methods:8 005 cases of prenatal genetic diagnosis of thalassemia in Guangdong Women and Children Hospital from September 2017 to December 2020 were retrospectively analyzed. All samples were diagnosed by traditional genetic methods include multiple Gap-PCR, PCR-RDB, MLPA and Sanger sequencing. Meanwhile, PCR-flow Fluorescence Hybridization technology was used as a verification platform for detecting common mutation sites of thalassemia. The results were analyzed to evaluate the diagnostic capabilities of PCR-flow Fluorescence Hybridization technology compared with traditional methods in prenatal genetic diagnosis of thalassemia.Results:By traditional methods, 1 939 cases (24.22%, 1 939/8 005) were normal and 6 066 cases (75.78%, 6 066/8 005) were diagnosed as thalassemia, including 4 513 cases of α-thalassemia, 1 475 cases of β-thalassemia, and 78 cases of αβ-thalassemia. By PCR-flow Fluorescence Hybridization technology, 7 845 samples were successfully diagnosed after initial interpretation by software. Compared with traditional methods, all the sensitivity, specificity and accuracy were 100%. The other 160 samples which failed in the initial interpretation can be successfully interpreted after review or manual interpretation.Conclusion:There were no differences between the two methods on the detecting of common mutation sites of thalassemia.

Objective: To investigate the blood lipid levels and prevalence of dyslipidemia in people with hypertension and diabetes in Henan province. Methods: From April 2016 to April 2017, multi-stage cluster sampling was adopted to investigate 71 285 local residents aged between 35 and 75 from 6 districts and counties in Henan province including Zhongmu county of Zhengzhou city, Huojia county of Xinxiang city, Hualong district of Puyang city, Qi county of Hebi city, Xigong district of Luoyang city, and Wugang city of Pingdingshan city. Blood samples were collected. According to the diagnostic criteria of hypertension and diabetes, the study population was divided into control group (n=29 427), hypertension group (n=21 965), diabetes group (n=8 009) and hypertension-diabetes group (n=11 884). Comparisons on blood lipid levels and dyslipidemia between 4 groups were performed. Results: The total cholesterol (TC) level of all subjects was 4.37 (3.78, 5.05) mmol/L. The triglyceride (TG) level was 1.27 (0.97, 1.80) mmol/L, the low-density lipoprotein cholesterol (LDL-C) level was 2.34 (1.88, 2.88) mmol/L and the high-density lipoprotein cholesterol (HDL-C) level was 1.31 (1.08, 1.59) mmol/L. Except for the TC level in women aged 65-75 years and LDL-C levels in women aged 55-64 and 65-75 years, there were significant differences in TC, TG, LDL-C and HDL-C levels between subjects of control group, hypertension group, diabetes group, and hypertension-diabetes group in different age ranges (including 35-44, 45-54, 55-64,and 65-75 years) and genders(all P<0.01).Except for the LDL-C and HDL-C in men aged 35-44 years and LDL-C in women aged 65-75 years, there were significant differences in the dyslipidemia rates of TC, TG, LDL-C and HDL-C between subjects of control group, hypertension group, diabetes group and hypertension-diabetes group in different age ranges and genders(P<0.01 or <0.05). After adjusting for age, gender, smoking, drinking, snoring, region, and body mass index, multivariate logistic regression analysis showed that hypertension (OR=1.221, 95%CI 1.113-1.339, P<0.01), diabetes (OR=1.636, 95%CI 1.461-1.833, P<0.01) and hypertension-diabetes (OR=1.832, 95%CI 1.658-2.023, P<0.01) were independent risk factors for TC abnormality. Hypertension (OR=1.566, 95%CI 1.478-1.659, P<0.01), diabetes (OR=2.182, 95%CI 2.031-2.342, P<0.01) and hypertension-diabetes (OR=2.655, 95%CI 2.492-2.829, P<0.01) were also independent risk factors for TG abnormality. Diabetes (OR=1.510, 95%CI 1.309-1.742, P<0.01) and hypertension-diabetes (OR=1.461, 95%CI 1.285-1.661, P<0.01) were independent risk factors for LDL-C abnormality. Diabetes (OR=1.261, 95%CI 1.180-1.346, P<0.01) and hypertension-diabetes (OR=1.195, 95%CI 1.126-1.268, P<0.01) were independent risk factors for HDL-C abnormality. Conclusion The prevalence of dyslipidemia in patients with hypertension and diabetes is high in Henan province, so adequate blood lipid education and control should be applied to people with risk factors as soon as possible.

Objective To analyze the value of dynamic enhanced multi-slice spiral computed tomography (MSCT) combined with two-dimensional (2D) curved reconstruction technique in the differentiation of benign and malignant intraductal papillary mucinous neoplasm (IPMN) of pancreas,and compare with magnetic resonance cholangiopancreatography(MRCP).Methods MSCT and MRCP data of a total of 50 patients with IPMNs confirmed by pathology after surgery was retrospectively reviewed.The benign and malignant IPMNs were differentiated based on the presence of mural nodules,main pancreatic duct (MPD) ≥ 10 mm,septum thickness ≥2 mm,calcification,surrounding vascular infiltration,enlarged peripancreatic lymph nodules,distant metastatic lesions and maximal branch duct type IPMN lesions ≥30 mm shown in the images.The sensitivity,specificity and accuracy were calculated and the receiver-operating-characteristics (ROC) analysis were drawn.Area under the curve (AUC) was calculated.Results Mural nodules in MSCT had a sensitivity,specificity,and accuracy of 77.1％ (27/35),80.0％ (12/15) and 78.0％ (39/50) for diagnosing malignant IPMN,respectively;which in MRCP were 77.1％ (27/35),86.7％ (13/15),and 80.0％ (40/50) in comparison.When MPD diameter ≥10 mm was used for diagnose malignancy,MSCT and MRCP had the sensitivity,specificity,and accuracy of 96.3％ (26/27),81.8％ (9/11),92.1％ (35/38),and 96.3％ (26/27),90.9％ (10/11),94.7％ (36/38),respectively.For thick septum ≥2 mm,MSCT and MRCP had the sensitivity,specificity,and accuracy of 4.8.6％ (17/35),93.3％ (14/15),62.0％(31/50),and 51.4％ (18/35),93.3％ (14/15),64.0％ (32/50),respectively.Out of 50 cases,calcifications were detected on MSCT in 6 patients,and 5 of them were pathologically diagnosed as malignant IPMN.MRCP failed to identify calcifications in any of these lesions.For MSCT,the AUC of MPD diameter ≥ 10 mm,mural nodules and thick septum ≥ 2 mm were 0.973 (P =0.000),0.825 (P =0.002) and 0.704(P =0.051),respectively.For MRCP,the AUC of the three factors above were 0.976(P =0.000),0.825(P =0.002),0.722 (P =0.034),respectively.For the predicting of IPMN malignancy,MSCT had an overall sensitivity,specificity,and accuracy of 94.3％ (33/35),73.3％ (11/15) and 88.0％ (44/50),respectively;in comparison,MRCP had values of 94.3％ (33/35),80.0％ (12/15) and 90.0％ (45/50),respectively.Conclusions Presence of mural nodules,MPD ≥10 mm and thick septum ≥2 mm on MSCT combined with 2D curved reconstruction or MRCP have a high value for predicting the malignancy of IPMN.The values of MSCT and MRCP were basically consistent in the differentiation of benign and malignant IPMN.MSCT can be used as the preferred examination for diagnosing IPMN in the primary hospitals without MR equipment.

Objective To investigate the CT features of early infection stage of thoracic and pulmonary paragonimiasis. Methods Medical records of 56 patients with thoracic and pulmonary paragonimiasis from January 2010 to June 2017 were collected, and the patients were diagnosed and treated at Yongjia County People's Hospital, and the results of laboratory examination and CT imaging features were analyzed retrospectively. Results The absolute value of eosinophils in peripheral blood of 56 patients was (5.61 ± 3.18) × 109/L, and the percentage of eosinophils was (35.90 ± 19.16)％, all of which increased to varying degrees. Forty-two patients had different degrees of pleural effusion and 52 cases with lung lesions. Lung lesions demonstrated one or several kinds of foci at the same time, randomly distributed in the lung field, mostly located in the sub-pleural lung tissue. There were 12 cases with pulmonary ground glass shadow, 4 cases with peribronchitis, 31 cases with pulmonary invasive lesions and 28 cases with pulmonary nodular/strip shadow. The size of most nodules were 0.5 - 1.0 cm, accompanied with halo sign. Conclusions The CT features of early infection stage of thoracic and pulmonary paragonimiasis are diverse. The size of 0.5 - 1.0 cm lung nodules with halo sign has certain characteristics in the diagnosis of paragonimiasis. Peribronchitis, infiltrative lesions, pleural effusion and increased peripheral blood eosinophil percentage can suggest diagnosis.

Female ; Humans ; Mutation ; Pregnancy ; Prenatal Diagnosis ; alpha-Globins ; genetics

Objective To devolope a method for extracting DNA from dried blood spots (DBS)and optimizing the operating procedure,which could be applied to clinical gene diagnosis of thalassemia.And the cross contamination of DBS punching and the storage stability of DBS were studied.Methods A total of 1 50 blood specimens were collected,and DBS were prepared.Circles (3 mm in diameter)were punched in the DBS,and eluted with lysis buffer.The eluting method and operating procedure were opti-mized.Genomic DNA extracted from the elution solution by magnetic beads,and were performed thalassemia gene test.Finally jud-ging whether the results of DBS and whole blood were consistent.Two methods of thalassemia gene test were used in DBS and the compatibility of DBS processing method was verified.Judging whether there was cross contamination of DBS punching by the thalassemia gene test results of blank hole which were punched in the blank filter paper between thalassemia positive DBS.The DBS storage stability in thalassemia gene test was verified by detecting the DBS which were dry stored at room temperature for 6 and 9 months.Results 5 circles (3 mm in diameter)DBS were vibrating eluted at 55 ℃ for 1 hour,the DNA concentration extracted from the elution solution was 10-20 ng/μL,which was dissolved in 50 μL solution,and the DNA quality was good.The thalassemia gene test results of DBS and whole blood were the same,and the DBS results of two thalassemia gene test methods were the same too. The cross contamination of DBS punching was not detected in thalassemia gene test.The DBS which were dry stored at room tem-perature for 6 and 9 months could be stably performed thalassemia gene test.Conclusion DBS could be used to perform thalassemia gene test,which is accurate,convenient and stable.It is an ideal way for specimen referral of thalassemia gene test.

OBJECTIVEDiagnosis and prenatal diagnosis to a family of hemoglobin variant with α-thalassemia.

METHODSWhole blood cell analysis, hemoglobin analysis by capillary zone electrophoresis (CZE), Gap-PCR, polymerase chain reaction-reverse dot blot (PCR-RDB) assay and DNA sequencing.

RESULTSHb Zurich Albisrieden with α°-thalassemia lead to severe anemia. The genotype of fetus is also Hb Zurich Albisrieden with α°-thalassemia.

CONCLUSIONAbnormal hemoglobin with α-thalassemia may lead to severe anemia, Prenatal diagnosis of thalassemia has the vital significance for eugenic birth.

Adult ; Base Sequence ; Child, Preschool ; Female ; Fetal Diseases ; blood ; diagnosis ; genetics ; Hemoglobins, Abnormal ; genetics ; metabolism ; Humans ; Male ; Molecular Sequence Data ; Pregnancy ; Prenatal Diagnosis ; Young Adult ; alpha-Thalassemia ; blood ; diagnosis ; embryology ; genetics

OBJECTIVETo employ single nucleotide polymorphisms (SNP) microarray to detect copy number variations (CNVs) for the diagnosis of disease and molecular classification.

METHODSFor a patient with split-hand/split-foot malformation, genome-wide copy number variants SNP microarray was applied. Tiny copy number variations were verified by real-time fluorescent quantitative PCR.

RESULTSThe results of SNP microarray has revealed that the patient has carried a 0.39 Mb duplication in 10q24.31-24.32 (102 955 122-103 348 688), which has encompassed genes including LBX1, BTRC and POLL. By real-time fluorescent quantitative PCR, duplicate area encompassing the pathogenic genes have been verified. The results for LBX1, BTRC, POLL genes were all consistent with the SNP microarray test. Moreover, a duplication was detected in exon 9 of FBXW4 gene which is in nearby.

CONCLUSIONSNP chips can efficiently identify tiny CNVs (< 1.0 Mb). In combination with real-time fluorescence quantitative PCR, this may provide valuable information for prenatal diagnosis.

Adult ; Asian Continental Ancestry Group ; genetics ; China ; Chromosome Duplication ; DNA Copy Number Variations ; DNA Polymerase beta ; genetics ; Homeodomain Proteins ; genetics ; Humans ; Limb Deformities, Congenital ; genetics ; Male ; Polymorphism, Single Nucleotide ; Transcription Factors ; genetics ; beta-Transducin Repeat-Containing Proteins ; genetics

To study the chemosensitivity and the mechanisms of recombinant adenovirus vector expressing ING4 and IL-24 (Ad-ING4-IL-24) on lung adenocarcinoma in vitro and in vivo, the expression of ING4 and IL-24 in A549 cells was detected by RT-PCR and Western blotting. The growth inhibition, apoptosis rate and apoptosis body were measured by MTT, flow cytometry and Hoechst staining respectively. For in vivo study, we first established the A549 tumor model by grafting A549 cells in athymic nude mice; and then injected Ad-ING4-IL-24 into the tumors. Two weeks after injection, we killed the mice, removed the tumors, weighted and calculated the ratios of tumor-suppression. We also detected the expressions of ING4, IL-24, bax, bcl-2, VEGF with immunohistochemistry. The results indicated that ING4 and IL-24 were proved successfully transcription and expression in A549 cells. More interestingly, the joint group inhibited the growth of A549 cells and induced apoptosis. The in vivo data showed that the joint group suppressed the tumor growth conspicuously through up-regulating the expression of bax, and down-regulating the expression of bcl-2, VEGF. The study proved that Ad-ING4-IL-24 significantly enhanced the chemosensitivity to anticancer drug DDP in lung adenocarcinoma, which may related with cell apoptosis and antiangiogenesis.
Adenocarcinoma ; drug therapy ; metabolism ; Adenoviridae ; genetics ; metabolism ; Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; Cell Cycle Proteins ; biosynthesis ; genetics ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Homeodomain Proteins ; biosynthesis ; genetics ; Humans ; Interleukins ; biosynthesis ; genetics ; Lung Neoplasms ; drug therapy ; metabolism ; Mice ; Mice, Nude ; Neoplasms, Experimental ; drug therapy ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Transfection ; Tumor Suppressor Proteins ; biosynthesis ; genetics