1.Effects of Budesonide on pulmonary vascular development and the expressions of vascular endothelial growth factor, nucleotide binding oligomerization domain-like receptor protein 3 in newborn rats with bronchopulmonary dysplasia caused by intrauterine infection
Guofeng LAN ; Yijin WANG ; Yunfang LI ; Qinghua WEI ; Fenglang SHI ; Qiliang CUI ; Hussnain MIRZA ; Xuekai SHI
Chinese Journal of Applied Clinical Pediatrics 2021;36(2):128-133
Objective:To investigate the effect of Budesonide (BUD) on pulmonary vascular development and the expression of vascular endothelial growth factor (VEGF) and nucleotide binding oligomerization domain-like receptor protein 3 (NLRP3) in newborn rats with bronchopulmonary dysplasia (BPD) caused by intrauterine infection.Methods:The 15-day-pregnant SD rats were divided into control group and infection group [intraperitoneal injection of 0.35 mg/(kg·d) lipopolysaccharide], and the newborn rats born by the above groups were divided into 3 groups: BUD group (0.5 mg of BUD suspension), normal control group (NC group, equal amount of 9 g/L saline), BPD group (equal amount of 9 g/L saline), with 40 rats in each group, all of them were inhaled twice a day for 14 days.Ten newborn rats were selected at birth, on the 3 rd, 7 th and 14 th day after administration.Pulmonary histopathological changes and radial alveolar counts (RAC) were observed after HE staining, and the thickness of alveolar respiratory membrane was measured; the platelet-endothelial cell adhesion molecule (PECAM-1/CD 31) in lung tissue was detected by immunohistochemistry, and the density of pulmonary microvessels was calculated; the expressions of VEGF, NLRP3 and Caspase-1 were detected by Western blot; and the levels of serum interleukin( IL)-1β and IL-18 were measured by enzyme-linked immunosorbent assay. Results:With the increase of day-old, the lung tissue of newborn rats in NC group was gradually developed and matured, the structure of alveoli was clear, the size was uniform, the count was significantly increased, and no obvious pathological changes were observed.In BPD group, the lung tissue structure was disordered, the alveoli were different in size and few in count, and inflammatory cells were exuded from the alveoli or the alveoli space.Compared with BPD group, the pathological changes of lung tissue in BUD group were significantly reduced.On the 3 rd, 7 th and 14 th day after administration, compared with NC group, the RAC, average integral optical density of CD 31 positive cells, density of pulmonary microvessel and level of VEGF protein in lung tissue of BPD group and BUD group were lower, and the differences were statistically significant (all P<0.05); while the thickness of respiratory membrane, level of NLRP3, Caspase-1 proteins in lung tissue and serum levels of IL-1β, IL-18 were significantly higher, and the differences were statistically significant(all P<0.05). Compared with BPD group, the RAC, average integral optical density of CD 31 positive cells, density of pulmonary microvessel and level of VEGF protein in lung tissue of BPD group and BUD group were significantly higher, and the differences were statistically significant (all P<0.05); while the thickness of respiratory membrane, level of NLRP3, Caspase-1 proteins in lung tissue and serum levels of IL-1β, IL-18 were significantly lower, and the differences were statistically significant (all P<0.05). Conclusions:The occurrence and development of pathological changes of BPD newborn rats caused by intrauterine infection can affect the development of pulmonary vessels through the inflammatory response of lung tissue.BUD can alleviate pathological changes in lung tissues of BPD newborn rats by reducing inflammatory reaction and up-regulating VEGF expression, promoting pulmonary vascular remodeling, and increasing pulmonary microvascular density.
2.Genetic Polymorphisms of STR Loci D18S53, D18S59 and D18S488 in Fetus in Tianjin
Xiaozhou LI ; Jing LIU ; Yunfang SHI ; Rui JU ; Yan LI ; Ying ZHANG ; Tianfu YUE
Tianjin Medical Journal 2014;(2):105-108
Objective To investigate the genetic polymorphisms of 3 short tandem repeat (STR) loci D18S53, D18S59 and D18S488 on chromosome 18 in fetus of Tianjin Han population, and to provide basic data in the use of 3 STR lo-ci in the prenatal diagnosis of Edward syndrome (ES). Methods A total of 64 villus samples and 374 amniotic fluid sam-ples were collected from gravida in Tianjin Han population. QF-PCR and ABI PRISM 377 sequence were used in this study. The frequencies of the genotypes were tested with H-W equilibrium. Genetic analysis was performed to conclude some data of population genetics such as the frequency of the alleles, the heterozygosity of observation (Ho), the polymorphism informa-tion content (PIC), the probability of discrimination power (DP), and the probability of exclusion (PE). Results The 15, 13 and 15 alleles of D18S53, D18S59 and D18S488 were observed respectively. The frequencies of the genotypes were in good agreement with H-W equilibrium. The Ho of 3 STR loci were 0.797, 0.847 and 0.792. The PIC was 0.81, 0.75 and 0.73. The DP was 0.944, 0.901 and 0.881. The PE was 0.593, 0.689 and 0.585. Conclusion D18S53, D18S59 and D18S488 STR lo-ci were the favorable genetic markers of chromosome 18, which can be used in prenatal genetic diagnosis of ES.
3.Regulating effects of Neiguan(PC6)and Gongsun(SP4)on hypothalamic-pituitary-adrenal axis in rats with functional dyspepsia
Bowen XING ; Simin QIN ; Lifen ZHAN ; Yunfang XIE ; Pingxiang WU ; Yu JIANG ; Haibing SHI ; Ke HE ; Weiai LIU ; Yanping YANG
Journal of Acupuncture and Tuina Science 2023;21(4):247-253
Objective:To investigate the effects and the possible mechanisms of Neiguan(PC6)and Gongsun(SP4)on the hypothalamic-pituitary-adrenal(HPA)axis in rats with functional dyspepsia(FD),thus to provide a theoretical basis for the clinical application of the Eight Confluent Points.Methods:Forty specific-pathogen-free Sprague-Dawley rats were divided into a blank group,a model group,an electroacupuncture(EA)group,and a Western medicine group by the random number table method,with 10 rats in each group.Rats in the blank group did not receive modeling or intervention.Rats in the other three groups were subjected to the FD with mood disorder model using the compound etiology modeling method.After the successful modeling,rats in the model group did not receive any interventions,rats in the Western medicine group received deanxit and mosaprid intervention,and those in the EA group received EA intervention on the ipsilateral Neiguan(PC6)and Gongsun(SP4)for 21 d.The sugar-water consumption rate was measured before the experiment and before and after interventions to assess the emotional status.The gastric emptying rate was measured after interventions to assess the gastrointestinal dynamics.The expression levels of hypothalamic corticotropin-releasing hormone(CRH),pituitary adrenocorticotropic hormone(ACTH),and adrenal corticosterone(CORT)were measured by enzyme-linked immunosorbent assay.Results:Compared with the blank group,the sugar-water consumption rate and the gastric emptying rate were decreased(P<0.01),and the hypothalamic CRH,pituitary ACTH,and adrenal CORT expression levels were increased(P<0.01)in the model group.Compared with the model group,the sugar-water consumption rate and the gastric emptying rate were significantly increased(P<0.01),while the expression levels of hypothalamic CRH,pituitary ACTH,and adrenal CORT were significantly decreased(P<0.01)in the EA group and the Western medicine group.The differences between the EA group and the Western medicine group were not statistically significant(P>0.05).Conclusion:The Eight Confluent Points Neiguan(PC6)and Gongsun(SP4)can improve the mood and gastrointestinal dynamics in FD rats,which may be achieved by down-regulating the hypothalamic CRH,pituitary ACTH,and adrenal CORT,as well as by correcting the HPA axis hyperfunction.
4.Construction of evaluation index system of Hunan Medical Science and Technology Award
Jie GUO ; Jingcheng SHI ; Yunfang WANG ; Xiaolei ZHONG
Chinese Journal of Medical Science Research Management 2020;33(2):91-95
Objective:To establish an evaluation index system for Hunan Medical Science and Technology Award.Methods:Based on the literature review and expert demonstration, combined with the previous review experiences of Hunan Medical Science and Technology Award, a review system framework was initially developed. Delphi method was used for expert consultation, outcome analysis were also adopted for the modification and reconstruction of this index system.Results:The standard evaluating system was set up with four first-level indicators, fifteen second-level indicators and their scoring standards.Conclusions:The evaluation index system established has the characteristics of scientific, systematic, pertinent, practical, simple and operable, also provides a reference and basis for scientific, objective and accurate evaluation of Medical Science and Technology Awards.
5.Application study of the evaluation index system of Hunan Medical Science and Technology Award
Jie GUO ; Yunfang WANG ; Jingcheng SHI ; Jianhua ZHU
Chinese Journal of Medical Science Research Management 2020;33(5):340-345
Objective:To test the practicality and effectiveness of Evaluation system of Hunan Medical Science and Technology Awards by empirical data.Methods:The empirical data of this paper is based on the 48 award-winning projects of the 2018 Hunan Medical Science and Technology Award. The actual evaluation process was simulated and peer experts were invited to grade the researching samples by index system to collect measured data. Use AMOS and SPSS as tools to test the reliability, validity and the correlation of the Evaluation system.Results:The internal consistency reliability of Cronbach α is 0.820, which shows this evaluation system with good reliability; sample data of confirmatory factor analysis matches the theoretical model, which shows this evaluation system with good validity; The results of analysis of variance showed that the differences among the three winning groups were statistically significant( P<0.05); The correlation coefficient of spearman rank correlation analysis is 0.335( P<0.05), which shows the positive correlation between the theoretical results and the actual evaluation results. Conclusions:The evaluation system of Hunan Medical Science and Technology Awards performs well and possesses strong applicability and practicality. The higher the score of the project measured by this indicator, the higher the quality of the project, and the higher the order of rewards that can actually be obtained. It could be used as an objective reference for the evaluation of Hunan Medical Science and Technology Award.
6.Wnt3a is Important in The Differentiation From Neural Stem Cell Into Dopaminergic Neuron In vitro
Shu HAN ; Wei SHI ; Yanhua LI ; Hailei YAO ; Xiaoyan XIE ; Lin CHEN ; Shuangshuang SHI ; Cixian BAI ; Xue NAN ; Fang YAN ; Yunfang WANG ; Xuetao PEI
Progress in Biochemistry and Biophysics 2006;0(10):-
Wnt signaling is implicated in the control of cell growth and differentiation during neural stem cell(CNS) development.Wnt3a, one of wnt gene family members, has effect on regeneration neurospheres and differentiation into neurons.Wnt3a inhibits regeneration of neurospheres, and promotes its differentiation. In vitro neurosphere was cultured in a serum-free defined medium DMEM/F12 supplemented with bFGF and EGF. Dissociated cells were plated onto poly-d-lysine-coated coverslips and propagated in medium containing recombined Wnt3a-adenovirus. Plenty of Nurr1 were detected by RT-PCR after 3 days. Wnt3a combined AA would improve NSC differentiation into dopaminergic (DA) neuron. The quantity of DA neuron is obviously more than the AA alone group's. Moreover, the expression of TH mRNA is 1.86 fold in Wnt3a combined AA group. Induced cells were immunostained for TH and DAT. The proportion of TH-positive was (37.42 ? 2.54) % (P
7.Genetic polymorphisms of HPRTB, DXS6803 and DXS6809 STR loci in Tianjin Han female population
Na ZHANG ; Yunfang SHI ; Xiaozhou LI ; Yan LI ; Duan JU ; Li XIN ; Jingyi YAO ; Xiaoyuan XIE ; Dianqin LIU ; Xiaohui YANG ; Tianfu YUE ; Ying ZHANG
Tianjin Medical Journal 2015;(1):12-16
Objective To investigate genetic polymorphisms of HPRTB, DXS6803 and DXS6809 STR loci in Tianjin Han female population, and to provide experimental data in the prenatal diagnosis of aneuploidies accurately and rapidly. Methods A total of 150 blood samples were collected in Tianjin Han population. QF-PCR and capillary electrophoresis were used in this study. The relevant data were analyzed by ABI Prism GeneMapper v3.0 software. Two homozygotes were se?lected from each locus for sequencing. The frequencies of the genotypes were checked using Chi-square test to verify Hardy-Weinberg Equilibrium. Data of genetic polymorphisms were calculated by PowerStatsV12 software. Results A total of 150 samples were successfully amplified in 24 hours. The 10, 6 and 10 alleles and 22, 12 and 29 genotypes were found respec?tively in HPRTB, DXS6803 and DXS6809 loci. The most common alleles were 14, 13 and 14. The higher frequencies of gen?otypes were 14-14, 12-13 and 13-14. No significant deviations from the Hardy-Weinberg equilibrium were observed in these three STR loci (χ2=10.554, 5.783 and 15.355, respectively, P>0.05). Values of He were 0.748, 0.649 and 0.806 for these three STR loci respectively. Values of Ho were 0.607, 0.700 and 0.713 respectively. Values of PIC were 0.706, 0.599 and 0.775 respectively. Values of PD were 0.894, 0.814 and 0.931 respectively. And values of PE were 0.299, 0.428 and 0.449 respectively. Conclusion HPRTB, DXS6803 and DXS6809 STR loci were highly polymorphic, which are favorable genetic markers on chromosome X and can be used in rapid prenatal genetic diagnosis.
8.Clinical application and evaluation of health economics for non-invasive prenatal testing of fetuses in Tianjin.
Ruiyu MA ; Xiaozhou LI ; Song XU ; Yunfang SHI ; Duan JU ; Yan LI ; Fanrong MENG ; Xiuyan WANG ; Xinxin DU ; Naiwei XU ; Ying ZHANG
Chinese Journal of Medical Genetics 2023;40(2):135-142
OBJECTIVE:
To assess the clinical efficacy and health economic value of non-invasive prenatal testing (NIPT) for the prenatal screening of common fetal chromosomal aneuploidies.
METHODS:
10 612 pregnant women from October 2017 to December 2019 presented at the antenatal screening clinic of the General Hospital of Tianjin Medical University were selected as the study subjects. Results of NIPT and invasive prenatal diagnosis and follow-up outcome for the 10 612 pregnant women were retrospectively analyzed and compared. Meanwhile, NIPT data for two periods were analyzed for assessing the health economic value of NIPT as the second- or first-tier screening strategy for the prenatal diagnosis of fetal trisomies 21, 18 and 13.
RESULTS:
The NIPT was successful in 10 528 (99.72%) subjects, with the sensitivity for fetal trisomies 21, 18 and 13 being 100%, 92.86% and 100%, and the positive predictive value (PPV) being 89.74%, 61.90% and 44.44%, respectively. The PPV of NIPT for sex chromosome aneuploidies was 34.21%. Except for one false negative case of trisomy 18, the negative predictive value for trisomy 21, trisomy 13 and other chromosomal abnormalities were 100%. For pregnant women with high risk by serological screening, advanced maternal age or abnormal ultrasound soft markers, NIPT has yielded a significantly increased high risk ratio. There was no statistical difference in the PPV of NIPT among pregnant women from each subgroup. NIPT would have higher health economic value as a second-tier screening until 2019, while compared to 2015 ~ 2017, its incremental cost-effectiveness ratio as a first-tier screening had declined clearly.
CONCLUSION
The screening efficacy of NIPT for trisomies 21, 18 and 13 for a mixed population is significantly better than conventional serological screening, but it is relatively low for sex chromosomal abnormalities. NIPT can also be recommended for populations with relatively high risks along with detailed pre- and post-test genetic counselling. From the perspective of health economics, except for open neural tube defects, it is possible for NIPT to replace the conventional serological screening in the future as its cost continues to decrease.
Pregnancy
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Female
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Humans
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Trisomy/genetics*
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Retrospective Studies
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Prenatal Diagnosis/methods*
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Down Syndrome/genetics*
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Aneuploidy
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Chromosome Aberrations
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Trisomy 18 Syndrome/genetics*
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Sex Chromosome Aberrations
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Fetus
9.Genetic analysis of a fetus with mosaicism Y chromosome aberration.
Fanrong MENG ; Duan JU ; Xiuyan WANG ; Yunfang SHI ; Meng YANG ; Xiaozhou LI
Chinese Journal of Medical Genetics 2023;40(11):1414-1419
OBJECTIVE:
To carry out prenatal diagnosis for a fetus with mosaicism Yq deletion.
METHODS:
A fetus with high risk of sex chromosomes indicated by non-invasive prenatal testing (NIPT) at Tianjin Medical University General Hospital in July 2021 was selected as the study subject. Prenatal diagnosis of the fetus was performed with combined G-banded chromosomal karyotyping, fluorescence in situ hybridization (FISH), copy number variation sequencing (CNV-seq), real-time fluorescence PCR (QF-PCR), and ultrasound examination.
RESULTS:
Analysis of the amniocytes at 23 gestational weeks had yielded a 45,X karyotype. However, FISH had shown signals of Y chromosome. Re-examination by cordocentesis had shown a mosaicism of 46,X,+mar[33]/45,X[17]. FISH showed that 69% of the cells had contained Y chromosome signals. The result of CNV-seq was seq[19]del(Y)(q11.1q12)(mos) chrY: g.13200001_ 28820000del (mosaicism rate = 64%), which suggested mosaicism for a Yq deletion, which encompassed the azoospermia factor (AZF) region. Deletion of the AZF region was verified by QF-PCR. The fetal karyotype was ultimately determined as mos46,X,del(Y)(q11.1)[33]/45,X[17]. Although ultrasound examination had shown no abnormality in the fetus, the couple had opted to terminate the pregnancy, and the induced fetus had a normal male appearance.
CONCLUSION
The combined use of multiple techniques is beneficial for accurate and rapid prenatal diagnosis. For fetuses with mosaicism chromosomal abnormalities, it may be difficult to accurately predict the postnatal phenotype. It is therefore necessary to further explore their genotype-phenotype correlation in order to provide better guidance upon genetic counseling.
Female
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Pregnancy
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Male
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Humans
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Mosaicism
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DNA Copy Number Variations
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In Situ Hybridization, Fluorescence
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Y Chromosome
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Fetus
10.Effect of herbal formula GAPT on cholinergic system in mice with scopolamine?induced memory impairment
Yunfang DONG ; Huili FENG ; Fang CHEN ; Ning SHENG ; Ying REN ; Jingduo YANG ; Jing SHI ; Jinzhou TIAN ; Pengwen WANG
Acta Laboratorium Animalis Scientia Sinica 2018;26(1):107-113
Objective To investigate the effect of a Chinese medicine formula GAPT, a combination containing Chinese herbs ginseng,Acorus tatarinowii, Polygala and tuber curcuma, on the behavior and cholinergic system in mice with scopolamine-induced memory impairment, and to explore the neuroprotective mechanism of GAPT. Methods ICR mice were randomly divided into normal control group(solution of 0.5% carboxy methyl cellulose, CMC), model group (0.5% CMC),positive control group(donepezil, 0.92 mg/(kg·d)), GAPT high, medium and low dose groups(20 mg/(kg·d),10 mg/(kg·d),5 mg/(kg·d)),18 mice in each group, were given intragastric gavage once a day for 30 days. After the last administration,the control group and model group received intraperitoneal injection of normal saline,and the other groups intraperitoneal injection of scopolamine(3 mg/(kg·d)), dissolved in 0.9% normal saline. Morris water maze test was performed to assess the learning and memmory ability. Then the mice were killed and the Ach content and AchE and ChAT activity in the cortex and hippocampus were detected. Results GAPT increased the swimming dis-tance,swimming time and the residence time in target quadrant of the model mice,increased the content of Ach and the ac-tivity of ChAT,and decreased the activity of AchE in the brain of the model group. Conclusions GAPT can improve the learning and memory ability of mice induced by scopolamine,and its mechanism may be related to cholinergic system.