Objective To establish an improved model of exercise-induced glycometabolism in type II diabetic rats,and to provide a theoretical reference for the establishment of exercise prescription for type II diabetes.Methods Forty-five 8-week old SPF male Wistar rats were used in this study.Of which 32 were fed with high-fat diet for 7 weeks,and intraperitoneal injection of 30 mg/kg STZ was given to establish the rat model of type II diabetes.The normal rats and successful model rats were divided into four groups:The normal control group (C group),normal exercise group (CE group),diabetic group (DM group) and diabetic exercise group (DME group).The exercise group was assigned by the Ploug training protocol,6 days/week,60 min/day,for a total of 8 weeks.After the high fat diet fed for 7 weeks,blood sample was taken from the tail vein,FBG and serum insulin were detected after baseline and 8 weeks exercise,and blood sample was collected from the tail vein to determine the FBG.Serum insulin (FINS) was detected by orbital blood sampling at the end of 8 weeks of exercise,and HOMA-IR was calculated.Results 1.After 7 weeks of high fat diet,compared with the groups C and CE,the levels of FBG,FINS and HOMA-IR were significantly higher in the DM and DME groups.2.After 8 weeks of exercise intervention,compared with the groups C and CE,FINS was significantly lower in the groups DM and DME,but the FBG and HOMA-IR were higher.Compared with the DM group,the level of FINS was significantly higher in the DME group,and the levels of FBG and HOMA-IR were significantly lower.The body weights of DM and DME groups were significantly lower than those of the groups Cand CE,the body weight had no significant difference between the DME and DM groups,and similar result was between the groups CE and C.Conclusions 1.The rat model of type II diabetes is successfully established with high fat diet for 7 weeks plus STZ injection(30 mg/mL).2.Aerobic exercise 60 min/day for a total of 8 weeks can improve the glycometabolism in type 2 diabetic rats,to be an ideal animal model for study of the mechanism of prevention and amelioration of type II diabetes.

Objective To investigate the value of prenatal diagnosis in identifying the etiology and predicting the prognosis of fetal pleural effusion (FPE).Methods Forty-two cases of FPE were recruited in this study from January 2012 to September 2016.Ultrasound scan and genetic tests were performed on all fetuses.Seven fetuses with severe FPE were given pleurocentesis.Pregnancy outcomes of all the fetuses were followed up.Results FPE was commonly accompanied with other abnormalities,such as ascites,hydrops,hydramnion,hygroma colli,abnormal posturing,joint contractures,arrhythmia and micromandible.Chromosomal abnormality was detected in 11 fetuses (26.2％),of which ten were further confirmed by karyotype analysis,including six with 45,X,three trisomy 21 and one trisomy 18,and one was detected with a 9.83 Mb uniparental disomy (UPD) located at 12q24.21q24.31 by gene chip.One fetus was diagnosed with--SEA/--SEA thalassemia.All of the 12 families decided to terminate the pregnancies after genetic counseling.Among the other 30 fetuses,seven with severe FPE and normal karyotype underwent pleurocentesis.Five of the seven cases were with favorable outcomes,one with progressive hydrops was aborted and one neonate with severe hydrops died after birth.Spontaneous regression of FPE with good outcome was found in two cases.Parents of the other 21 fetuses chose to terminate the pregnancies.Conclusions Prenatal diagnosis is important to identify the etiology and predict the outcome of FPE.Chromosomal abnormality is a relatively common cause of FPE,and 45,X and trisomy 21 are the most common abnormalities.Intrauterine intervention is beneficial for FPE without chromosomal or other definite genetic abnormalities.Genetic test may be of great value for pregnant counseling.

OBJECTIVE: To assess the value of combined copy number variation sequencing (CNV-seq) and chromosomal karyotyping for the diagnosis of amniocytic mosaicisms, in addition with a literature review. METHODS: Forty cases of amniocytic mosaicisms detected at the Genetic and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University from January 2018 to December 2021, in addition with 245 mosaicisms retrieved from 11 recent literature were evaluated in terms of detection rate, consistency rate, and pregnancy outcomes. RESULTS: The detection rate of amniocytic mosaicisms was 0.46% (40/8 621) in our center. And its consistency rate with chromosomal karyotyping was 75.0% (30/40). After genetic counseling, 30 (75.0%) couples had opted to terminate the pregnancy, 5 (12.5%) had decided to continue with the pregnancy, 3 (7.5%) fetuses were born alive, and 2 cases (5.0%) were lost in touch. By contrast, 245 cases (0.39%) of mosaicisms were identified among 63 577 amniotic samples, with a consistency rate of 62.8% (103/164) with other techniques. Among these, 114 cases (55.1%) were terminated, 75 (36.2%) were born alive, and 18 (8.7%) were lost during the follow up. CONCLUSION Combined CNV-seq and chromosomal karyotyping has a high value for the detection of amniotic mosaicisms.
Pregnancy ; Female ; Humans ; Mosaicism ; Chromosome Disorders/genetics* ; DNA Copy Number Variations ; Chromosome Aberrations ; Karyotyping ; Prenatal Diagnosis/methods*

Objective:To explore the clinical significance of the combined application of prenatal cellular and molecular genetics in the diagnosis of fetal sex chromosome mosaicism.Methods:A retrospective analysis was conducted on 14 034 pregnant women aged 20-46 years (mean age 27±3 years) who came to the Genetic Counseling Clinic of the First Affiliated Hospital of Zhengzhou University from May 1st 2017 to January 31th 2020 for G-banding karyotype analysis of fetal amniotic fluid chromosomes. They were 17-32 weeks pregnant and had no consanguineous marriage. The patients diagnosed as sex chromosome mosaicism were screened out and their prenatal diagnostic indications were analyzed. The results of whole-genome copy number variation sequencing (CNV-seq)/single nucleotide polymorphism-array(SNP-array)/fluorescence in situ hybridization(FISH) were comopared, combined with ultrasound findings, and the pregnancy outcomes of all pregnant women were followed up by access to our hospital′s electronic medical record system or telephone.Results:A total of 46 cases of sex chromosome mosaicism were found. There were 43 cases with two types of karyotype mosaicism, accounting for 93.48%, and 3 cases with three types of karyotype mosaicism, accounting for 6.52%. Comparison of karyotype and CNV-seq/SNP-array/FISH results showed that 4 cases had consistent results, 9 cases had consistent results but different proportion, and 10 cases had inconsistent results. Combined with the results of the cytogenetic/molecular genetic analysis and/or ultrasound findings, pregnant women will decide to continue or terminate the pregnancy.Conclusion:The combination of prenatal cytogenetic and molecular genetic methods is helpful for rapid diagnosis of fetal sex chromosome mosaicism, providing scientific basis for pregnant women′s pregnancy selection.

Objective:Cytogenetic and molecular genetic analysis was performed on two consecutive antenatal abnormal fetuses and their parents in a family to clarify the copy number variation(CNV) and its mechanism.Methods:The karyotypes of two fetuses and their parents were analyzed by conventional karyotyping techniques, and CNVs of two fetuses and their mother were analyzed by low-coverage whole-genome copy number variation sequencing (CNV-seq) techniques.Results:The amniotic fluid karyotype results of fetus 1 and 2 were 46, XN, der(4)t(4;10)(q35;p13). The mother′s peripheral blood karyotype result was 46, XX, t(4;10)(q35;p13), and the father′s karyotype was normal. The CNV-seq results of fetus 1 and 2 were seq[hg19]6q22.31(122740000-125440000)X1; 10p15.3p13(120000-17260000)X3, suggesting that there was a heterozygous deletion of about 2 700 000 bp in fetal 6q22.31 and a duplication of about 17 140 000 bp in fetal 10p15.3p13. The CNV-seq result of their mother was seq[hg19]6q22.31(122740000-125440000)X1, suggesting that there was a heterozygous deletion of about 2 700 000 bp in 6q22.31. The pregnant woman and her family chose to terminate the pregnancy after genetic consulting.Conclusion:The combined application of karyotyping and CNV-Seq is significantly beneficial to detecting microdeletions or microduplications of fetal chromosomes and effectively preventing the birth of defective children.

Objective To investigate the value of local myocardial blood flow and myocardial function parameters in monitoring the dynamic changes of radiation induced heart disease (RIHD) using 13NNH3 PET gated myocardial perfusion imaging(GMPI).Methods Six healthy male Beagle dogs underwent 13N-NH3 PET GMPI 1 week before irradiation and 3,6 and 12 months after irradiation in the anterior wall of the left ventricle with a single dose of 20 Gy.Global myocardial function parameters including left ventricular ejection fraction (LVEF),end-diastolic volume (EDV),end-systolic volume (ESV),and regional myocardial function parameters including wall motion (WM),wall thickening (WT),end-diastolic perfusion (EDP),end-systolic perfusion (ESP) before and after irradiation were compared by repeated measures analysis of variance and paired t test.Results There were no significant changes between EDV,ESV and LVEF at baseline and those at 3 months after irradiation.EDV at 6 months after irradiation still had no change,compared with baseline value and EDV at 3 months after irradiation,but ESV was increased and LVEF was decreased.Twelve months after irradiation,ESV was further expanded,LVEF was further reduced,and EDV began to increase (F values:20.974-177.846,all P＜0.05).Compared with the baseline,WM,WT,EDP and ESP were increased in 10％(2/20),20％(4/20),10％(2/20) and 15％(3/20) of myocardial segments at 3 months after irradiation (t values:14.446-672.315,all P＜0.05);those parameters were decreased in 15％(3/20),20％(4/20),15％(3/20) and 25％(5/20) of myocardial segments at 6 months after irradiation (t values:18.171-723.156,all P＜0.05),and were decreased in 35％(7/20),45％(9/20),40％(8/20) and 60％ (12/20) of myocardial segments at 12 months after irradiation (t values:14.783-711.259,all P＜0.05).Conclusions 13N-NH3 PET GMPI could be used to detect RIHD early and monitor the dynamic development of RIHD.Compared with the global left ventricular function parameters,regional myocardial function parameters (WM,WT,EDP and ESP) are more sensitive,which may be served as the early monitoring indicators for RIHD.

Objective:To explore the clinical application value and accuracy of cell-free fetal DNA (cff-DNA) technique in prenatal screening.Methods:The results of quantitative fluorescent PCR (QF-PCR) and karyotype of amniotic fluid cells were analyzed retrospectively in 2 398 monocyesis pregnant women who had been amniocentesis at the First Affiliated Hospital of Zhengzhou University from May 2013 to December 2019, and the results of 359 cases who had been examined by single-nucleotide polymorphism array (SNP array).Results:Cff-DNA test of 2, 398 cases indicated 987 cases of trisomy 21, 351 cases of trisomy 18, 135 cases of trisomy 13, 566 cases of sex chromosome abnormality, and 359 cases of other chromosome abnormality. Chromosome karyotype analysis detected 826 cases of trisomy 21, 213 cases of trisomy 18, 17 cases of trisomy 13, 221 cases of sex chromosome abnormality, and 26 cases of other chromosome abnormality. The detection rate were 83.69% (826/987), 60.68% (213/351), 12.59% (17/135), 39.04% (221/566) and 7.24% (26/359), respectively. QF-PCR detected 1 046 cases of trisomy and 188 cases of sex chromosomes abnormality, and the detection rate was 99.05% (1 046/1 056) and 85.07% (188/221), respectively. Compared with the abnormal number detected by chromosome karyotype analysis, 10 cases of trisomeric chimerism and 24 cases of sex chromosome were missed by QF-PCR. Among the 359 other chromosomal abnormalities detected by SNP array, 64 cases were consistent with the results of cff-DNA, and the detection rate was 17.83% (64/359), which was 10.59% higher than the karyotype result.Conclusions:Karyotype analysis is the gold standard for diagnosing chromosomal abnormalities. QF-PCR could diagnose common chromosome aneuploidy rapidly and accurately, and it could be used as an auxiliary detection technique for karyotype analysis. The incidence of sex chromosome chimerism is high, so missed diagnosis should be warned. SNP array could be given priority to verify chromosome microdeletion or microduplication detected by cff-DNA.

OBJECTIVE: To assess the value of low-depth whole-genome copy number variation sequencing (CNV-seq) for the analysis of chromosomal copy number variations among fetuses with echogenic bowel (EB). METHODS: A total of 163 fetuses were included in this study. Amniotic fluid (162 cases) or chorionic villi (1 case) were collected and subjected to CNV-seq for the analysis of CNVs. RESULTS: Thirteen (8.0%) pathogenic CNVs were detected, including 9 (5.5%) aneuploidies and 4 (2.4%) CNVs. The detection rate of the isolated EB group and combined EB group were 1.7% (1/58) and 11.4% (12/105), respectively. There was a significant difference between the two groups (P < 0.05). A Xp22.1 duplication was detected in both groups, and the fetuses were predicted as female DMD carriers and born healthy. Nine cases of aneuploidies and 2 (likely) pathogenic CNVs were identified in the combined EB group, all of them have warranted induced labor. CONCLUSION The prevalence of chromosomal aneuploidies and pathogenic CNVs in fetuses with combined EB was much higher than isolated EB, and most of them may warrant termination of pregnancy. Compared with isolated EB, more attention should be paid to combined EB, for which prenatal diagnosis and genetic counseling should be carried out in time.
Amniotic Fluid ; Aneuploidy ; Chromosome Aberrations ; DNA Copy Number Variations ; Echogenic Bowel ; Female ; Humans ; Pregnancy ; Prenatal Diagnosis ; Technology

OBJECTIVE: To assess the diagnostic value of copy number variation sequencing (CNV-seq) in the genetic etiology of fetuses with nasal bone dysplasia (NBD). METHODS: A total of 217 fetuses discovered with NBD from December 2017 to December 2020 were divided into the isolated NBD group and NBD combined with other anomalies group, for which copy number variations (CNVs) were analyzed. RESULTS: A total of 40 fetal abnormalities were detected in 217 cases, with an overall abnormal rate of 18.4%. These included 31 cases with aneuploidies (14.3%, 31/217) and 9 cases with genomic CNVs (4.1%, 9/217). Five cases of trisomy 21 (3.5%, 5/144) and two CNVs cases with unknown clinical significance (1.4%, 2/144) were detected in the isolated group. As for the combined NBD group, 26 aneuploidies (35.6%, 26/73), including 19 cases with trisomy 21, 6 cases with trisomy 18, 1 case with trisomy 13, 5 cases with pathogenic CNVs (6.8%, 5/73), and 2 cases with CNVs of unknown clinical significance (2.7%, 2/73) were detected. A significant difference was detected between the two groups (P < 0.01). CONCLUSION The detection rate of CNV-seq is high for chromosomal aneuploidies and pathogenic CNVs in fetuses with NBD, particularly in those combined with other ultrasonic abnormalities.
Aneuploidy ; Bone Diseases, Developmental ; Chromosome Aberrations ; DNA Copy Number Variations ; Down Syndrome/genetics* ; Female ; Fetus/abnormalities* ; Humans ; Pregnancy ; Prenatal Diagnosis ; Trisomy

AIM: To study the inhibitory and scavenging effects of ambroxol combined with levofloxacin on the bacterial biofilm of Klebsiella pneumoniae, and to provide a new strategy to treat and antagonize the formation of the biofilm. METHODS: We collected Klebsiella pneumoniae of different resistance and divided them into sensitive group (wild bacteria group), ESBLs group and CRKP group with 15 strains in each group and performed biological semi-quantitative detection of its biofilm by crystalline violet staining method. After selecting 3 strains with similar membrane yields from each group, we determined the minimum inhibitory concentration (MIC) of ambroxol and levofloxacin against Klebsiella pneumoniae by micro broth dilution method.In addition, we determined the effects of ambroxol in different concentrations on the MIC of levfloxacin by the micrdilution checkerboard techniques and calculated the partial inhibitory concentration index (FIC) to determine the joint effect and select the best synergistic concentration. Finally, the effects of ambroxol and levofloxacin in different concentrations on the inhibition formation test and removal test of Klebsiella pneumoniae biofilm were observed by crystal violet method combined with confocal laser scanning microscopy. RESULTS: We found that all three groups of bacterial biofilms became mature on the 5th day, and the sensitive group was easier to form and produce biofilm more than the ESBLs and CRKP groups (F=3.725, P=0.032). It was showed that the geometric average of levofloxacin MIC value in the three groups decreased significantly. And the selected strains all showed a synergistic effect on the two-drug combination.In the biofilm test, as the concentration of ambroxol increased, its inhibition rate reached more than 75%, but its biofilm removal rate did not reach 70%. CONCLUSION: Our study support that ambroxol combined with levofloxacin can inhibit the bacterial biofilm of Klebsiella pneumoniae early, and its optimal synergistic concentration is 0.49 mg/mL (ambroxol) + 4 μg/mL (levofloxacin).