1.Verification of Differential Expression Genes after CacyBP/SIP Nuclear Translocation in Colon Carcinoma Cell Line
Anping WANG ; Yingying ZHAO ; Xin LIU ; Huihong ZHAI
Tianjin Medical Journal 2014;(5):410-413
Objective To verify the genes screened by the polymerase chain reaction (PCR) chip of cell cycle. Methods The colon cancer cells SW480 were randomized into two groups, the test group (with gastrin stimulation) and con-trol group (without gastrin stimulation). The method of Western blot was used to detect the expression of calcylin binding pro-tein/Siah-1 interacting protein (Cacybp/SIP) before and after gastrin stimulation. The differential expression genes, cyclin de-pendent kinase 8 (CDK8) and cyclin dependent kinase subunit (CKS2), were verified by using real-time quantitative PCR (qRT-PCR). Results It was found that before the stimulation, CacyBP/SIP was located and expressed in cytoplasm, and then in both cytoplasm and nucleus after gastrin stimulation. The qRT-PCR results of CDK8 and CKS2 genes were consis-tent with those of microarray detection. The expressions of CDK8 and CKS2 were up-regulated (P < 0.05). Conclusion The stimulation of human gastrin can lead to the nuclear translocation of CacyBP/SIP. The results of microarray are reliable, and the differentially expressed genes screened through gene chip deserve further study.
2.Protection effect of amentoflavone in Selaginella tamariscina against TNF-alpha-induced vascular injury of endothelial cells.
Xiaoke ZHENG ; Caixia LIU ; Yingying ZHAI ; Lingling LI ; Xiaolan WANG ; Weisheng FENG
Acta Pharmaceutica Sinica 2013;48(9):1503-9
This study is to observe the protection effect of amentoflavone (AMT) in Selaginella tamariscina against TNF-alpha-induced vascular inflammation injury of endothelial cells. On the basis of TNF-alpha induced human umbilical vein endothelial cell, observe the influence of AMT on endothelial active factor, the contents of SOD and MDA, the protein expression of vascular endothelial adhesion molecules and inflammatory factor; study the effect of its common related signal pathways such as NF-kappaB; research the effect of AMT against TNF-a induced human umbilical vein endothelial cell injury by means of MTT, ELISA, Western blotting and the cell immunofluorescence. The results showed that AMT could increase the content of NO and decrease the levels of VCAM-1, E-selectin, IL-6, IL-8 and ET-1; enhance the activity of SOD, reduce the content of MDA; downregulate the protein expressions of VCAM-1, E-selectin, NF-kappaBp65 and up-regulate IkappaBalpha, attenuate the NF-kappaBp65 transfer to cell nucleus. AMT has the effect of protect vascular endothelial and maybe via the signal pathway of NF-kappaB to down-regulate the inflammation factor and oxidative damage factor of downstream.
3.Noninvasive prenatal screen of trisomy-21 using maternal plasma fetal free RNA allelic ratio
Yajuan XU ; Shanshan ZHAI ; Xiaohua LUO ; Yingying ZHANG ; Limin RAN ; Lidan REN
Chinese Journal of Obstetrics and Gynecology 2015;(8):568-575
Objective Through the detections of the heterozygote frequencies tests of fetal specific genes PLAC4 and COL6A2 mRNA alleles in plasma of pregnant women, to explore its possibility of application in the noninvasive prenatal screenings of trisomy-21. Methods A toltal of 500 cases (males and females 250 cases respectively)of Han ethnic groups with Henan Provice of China who were subject to the physical checkup clinic of the Third Affiliated Hospital,Zhengzhou University from June to December, 2013 were selected as the healthy physical checkup group, and such techniques as DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) were adopted to the determinations of the heterozygote frequencies of the single nucleotide polymorphism(SNP)of the PLAC4 and COL6A2 genes in the maternal peripheral blood in the healthy physical checkup group, and the differential comparisons of the determination results of the SNP heterozygote frequencies and the corresponding heterozygote frequencies in the National Center for Biotechnology Information (NCBI) database;30 cases of healthy pregnant women who spontaneously underwent pregnancy checkups at the maternity clinic were randomly selected as the healthy pregnancy group, and real-time fluorescence quantitative reverse transcription-PCR technique was adopted for determining the expression levels of PLAC4 and COL6A2 mRNA in the peripheral blood of pregnant women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks;40 cases of the same phase were selected for acting as the specimens for the karyotype analyses of the amniotic fluid cells, among which 20 cases were trisomy-21, and the 20 cases of the negative control group, and reverse transcription-multiplex ligation dependent probe amplification (RT-MLPA) technique was adopted for screening the fetal trisomy-21. Results (1) The allele heterozygote frequencies of the SNP of the healthy physical checkup group:determinations of the genotypes and hybrid rates of the 10 SNP sites of the PLAC4 and COL6A2 genes indicated that those with higher heterozygote frequencies were respectively rs7717, rs559, rs1044598, rs59066201 and rs1042917, with population coverage of 98%. Among them, the allele hybrid rates of rs59066201 were never seen in the NCBI database;in the respective comparisons of the allele hybrid rates of rs8130833, rs9977003 and rs7844 with the hybrid rates of the NCBI database, the variations had statistical significance (P<0.05). (2) The expression levels of PLAC4 and COL6A2 mRNA of the different pregnancy weeks of the healthy pregnancy group: the levels of PLAC4 mRNA in the peripheral blood of women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks of pregnancy were respectively 7.22 ± 1.05, 8.02±1.41,9.51±1.69,11.33±2.11 and 13.31±2.58, with their expression levels rising along with the increase of the pregnancy weeks; among them, the comparison of pregnancy 8 weeks and pregnancy 10 weeks, the variations had no statistical significance (P>0.05);in the mutual comparisons among the expression levels of the various pregnancy weeks, the variations had statistical significance (P<0.05). The expression levels of COL6A2 mRNA in 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks were respectively 8.95 ± 1.28, 11.19 ± 1.36,15.00 ± 1.58,16.87 ± 1.72 and 18.96 ± 2.79, with their expression levels rising along with the increase of the pregnancy weeks, and in the mutual comparisons between the expression levels of the various pregnancy weeks, the variations all had statistical significance (P<0.05). (3) Prenatal screenings of trisomy-21 in the validation group of the trisome:a total of 5 sites of rs7717, rs559, rs1044598, rs59066201 and rs1042917 were selected from the allele heterozygote frequencies of SNP sites were selected from the subjects of the healthy physical checkup group, and 10 cases of trisomy-21 specimens and 10 cases of negative CTR specimens were accurately determined, with the sensitivity reached 80%(17/20), and the specificity reached 90%(18/20). One case of the trisomy-21 and two negative cases were both homozygotes, and among the trisomy-21 specimens of two cases, only one SNP was a heterozygote, and it was impossible to conduct screenings on these 5 cases, with the screening accuracy reaching 100%(35/35). Conclusions Fetal specific genes PLAC4 and COL6A2 mRNA are expressed in the peripheral blood of pregnant women in different gestational age;its expression level increases with the increase of gestational age. Among them, five SNP including rs7717, rs559, rs1044598, rs59066201 and rs1042917 show highest heterogeneity rate, which is different from the corresponding heterogeneity rate in NCBI database. RT-MLPA technology is a rapid, effective, noninvasive and low cost method of prenatal screening 21 trisomy.
4.Correlation between birth weight and cesarean section rate of term singleton live-birth newborns in recent 10 years
Chunfeng WU ; Yunfeng WANG ; Aiyang LI ; Huixia YANG ; Yingying QI ; Yanan ZHAI
Chinese Journal of Perinatal Medicine 2011;14(9):530-533
Objective To investigate the trend in birth weight of term infants delivered in Miyun Hospital in recent ten years, and its relationship with cesarean section rate, to provide evidence for health care during pregnancy and lowering the cesarean section rate. MethodsSingleton term live-birth newborns in Beijing Miyun Hospital from January 1, 2000 to December 31, 2009 were admitted into this study.Trends of birth weight, microsomia rate and cesarean section rate were analyzed. Results(1) There were 14 716 singleton term live-birth newborns delivered in the tenyear period, among which 7642 males and 7074 females with the proportion of male and female of 1.08 : 1. One thousand seven hundred and seventy-three (12.0%) microsomia were delivered, and the number of low birth weight infants was 180 (1.2%). (2) The birth weight of term infants was from 1200 g to 5850 g and the average birth weight was (3422±447) g. The average birth weight of each year increased, and the differences among each year showed statistical significance (F= 15. 337,P<0.01). (3) The percentage of microsomia of each year increased, and the differences among each year also showed statistical significance (x2= 125. 285, P<0. 01).(4) The cesarean section rate increased from 49.9% in 2000 to 57.1% in 2009 with statistical difference (x2 =180. 883, P<0.01).(5) The cesarean section rate increased with the average birth weight of the singleton live-birth term infants in the 10 years from 52.2 % in those <2500 g to 85.4 % in those 4500 g~, and the differences had statistical significance (x2 =518. 519, P<0. 01). ConclusionsCesarean section rate had close correlation with birth weight in term birth infants.It is suggested that prenatal care should be strengthened and nutrition guidance during pregnancy should be paid more attention.
5.Epidemiological surveillance of Henipavirus in Chongqing
Dan PENG ; Zhilei ZENG ; Dan ZHU ; Xiao CHEN ; Jianping YU ; Mingming XU ; Yingying ZHANG ; Qunling ZHAN ; Hong ZHAI ; Peng XIE
Chinese Journal of Microbiology and Immunology 2008;28(11):1050-1054
Objective To establish nucleic acid testing techniques for detecting Nipah virus (NiV) and Hendra virus (HeV), and to test the NiV and HeV in peripheral blood collected from domestic pigs, cows and goats in Chongqing. Methods Peripheral blood samples of 580 domestic pigs, 250 cows, 180 goats were collected from Chongqing since June 2007 to June 2008. The lymphocytes were separated by density gradient centrifugation and total RNA was extracted using Trizol method for detection of NiV and HeV with one-step real-time RT-PCR. Sequence identification and analysis were performed for positive PCR prod-ucts. Virus isolation and culture were adopted for positive samples, and epidemiologic reports were submit-ted. Results Nucleic acid detections searching for NiV and HeV were successfully performed in animal blood samples collected from Chongqing. "Takeoff points" were not found in fluorescence amplification curves of all samples. Curves kept the same slope, and assays were judged as negative. Conclusion Until now, Neither NiV or HeV infection has been found in domestic animals blood samples collected from Chongqing, which suggest a lower possibility of outbreaks of Nipah disease and Hendra disease in Chongqing in the near future.
6.Impact of prescribed adaptive statistical iterative reconstruction-V on image quality and radiation dosage of lung pure ground glass nodules: A phantom study
Ying LI ; Na'na AI ; Yingying ZHANG ; Yucun FU ; Jiong JIANG ; Dandan NIU ; Yanhui ZHAI ; Shouqiang JIA
Chinese Journal of Medical Imaging Technology 2018;34(5):775-778
Objective To investigate the impact of prescribed adaptive statistical iterative reconstruction V (Pre-ASiR-V) on imaging quality and radiation dosage of pure ground glass nodules (pGGN) in chest phantom,in order to obtain the optimal level of Pre-ASiR-V.Methods CT scanning for a chest phantom containing 4 artificial pGGNs was performed with Revolution CT,and the Pre-ASiR-V level was set as 0,20%,40%,60%,80% and 100% group,respectively.The mean noise,effective dose (ED) and the subjective scores of pGGN imaging were recorded and compared.Results The mean noise of groups (Pre-ASiR-V 0,20%,40%,60%,80% and 100%) was (17.93±2.20)HU,(17.30±3.68)HU,(18.20± 3.44)HU,(18.80±0.20)HU,(19.87±2.56)HU and (15.90±4.56)HU,respectively (F=0.568,P=0.723).ED of these groups was 7.40 mSv,5.16 mSv,3.36 mSv,1.97 mSv,0.97 mSv and 0.33 mSv,respectively.Compared with imaging of PreASiR-V 0,the reduction percentage of ED was 30.27%,54.59%,73.38%,86.89% and 95.54%,respectively.The subjective score of the image quality evaluated by the 2 observers had high agreement (Kappa=0.778,P=0.003),and all the scores were greater than 3.The subjective score of Pre-ASiR-V 80% and 100% group was slightly lower than those in other groups.Conclusion Different Pre-ASiR-V level slightly impacts the noise of high-resolution CT images of chest phantom,while reduces radiation dosage significantly.Pre-ASiR-V level of 60% is the optimal protocol.
7.Research progress of anti-tumor mechanism of effective components of angelica dahurica
Shengnan CUI ; Zhihui ZHOU ; Wei LI ; Yingying BAI ; Dongzhi ZHAI ; Xu CHAO ; Changhu DONG ; Hong LI
International Journal of Traditional Chinese Medicine 2018;40(4):381-384
Angelica dahurica is diaphoretic, commonly used in the treatment of cold, wet, and itchy rubella nasosinusitis. Studies have shown that the effective components of Angelica dahurica can be used in the treatment of malignant tumors. This paper summarizes the related literature home and abroad recently, and regards the effective components of Angelica dahurica as a major role in treatment of tumor by inhibiting tumor cell proliferation, promoting apoptosis, inhibiting tumor metastasis, inhibiting platelet aggregation, enhancing immunity, and anti-tumor drug resistance. Meanwhile, the paper finds the shortcomings of the present researches, and hopes to provide reference for the future experiment and clinical research.
8.Practice and thinking of pediatrics TBL teaching plan writing in medical simulation teaching mode
Guoyu ZHONG ; Shaoling YANG ; Shuling LAN ; Yingying ZHAI ; Dehui CHEN ; Songqing XU ; Huimin DENG
Chinese Journal of Medical Education Research 2019;18(3):231-235
The clinical practice teaching reform of pediatrics based on TBL combined with medical simulation teaching model was carried out. TBL teaching plan writing under the model of medical simulation teaching was researched. The principles, requirements, contents and procedures of compiling teaching plans were formed. Emphasis should be placed on medical simulation teaching, team cooperation, humanistic quality education and humanistic skills training. The purposes of the reform were to achieve standardized operation skills, improve clinical ability, ensure medical safety, and improve the quality of medical purposes, but still the reform need further improvement in the teaching practice.
9.Diagnosis of a Primary Pleomorphic Liposarcoma in a Tree Shrew(Tupaia belangeri subsp. yaoshanensis)
Zhuxin LI ; Liang LIANG ; Yingying CAO ; Shanshan ZHAI ; Yinhan DAI ; Xia HE ; Junyu TAO ; Jing LENG ; Haibo TANG
Laboratory Animal and Comparative Medicine 2023;43(6):647-653
A large spontaneous mass on the dorsal abdomen near the hip joint was found in an aged female Tupaia belangeri subsp. yaoshanensis after 5 years of routine feeding. The tumor in the diseased tree shrew was huge, with an intact surface and no ulceration; however, it caused inconvenience in movement while maintaining a good mental state. After inhalation anesthesia with isoflurane (2%-4%), the tree shrew was euthanized after the tree shrew entered deep anesthesia. Anatomical dissection of the tumor, the tumor boundary was unclear and infiltrated into surrounding tissues. HE staining showed that small focal pleomorphism and large areas of adipocytes were seen in the tumor tissue. The pleomorphic sarcoma area was mainly composed of atypical epithelioid cells with easily visible nuclear divisions. The size of the adipocytes was significantly different, and more pleomorphic adipocytes were seen. The cell volume was large, the nucleus was deeply stained and deformed, the edge was impressed, and the cytoplasm was seen with multiple vesicular lipid droplets. Immunohistochemical results showed that the tumor cells were positive for Vimentin, the small focal polymorphic adipocyte nucleus was positive for S-100, and Ki-67 exhibited a higher proportion of positivity. Combined with HE staining and immunohistochemical results, the spontaneous tumor in this tree shrew was comprehensively diagnosed as pleomorphic liposarcoma.
10.Diagnosis of Trichoepithelioma in a Tree Shrew and Observation of Cell Biological Characteristics
Shanshan ZHAI ; Liang LIANG ; Yingying CAO ; Zhuxin LI ; Qing WANG ; Junyu TAO ; Chenxia YUN ; Jing LENG ; Haibo TANG
Laboratory Animal and Comparative Medicine 2023;43(4):440-445
ObjectiveTo observe the primary tumor of tree shrews and to provide a basis for studying the pathogenesis and prevention of trichoepithelioma. MethodsA tumor was discovered in the chest and abdomen of a tree shrew during natural cultivation. The tree shrew was anesthetized, and the tumor was surgically removed. Hematoxylin and eosin (HE) staining and immunohistochemical staining were performed on the tumor tissue after paraffin section, and the tumor cells were isolated and cultured by passage. The isolated tumor cells were subcutaneously injected into healthy tree shrews and nude mice. The tumorigenesis of tumor cells in vivo was observed once a day, with nude mice continuously observed for 2 months and tree shrews observed for more than 6 months. ResultsHE staining showed that the basal cells in the dermis were arranged as a whole, like a string of petals, forming nests and stripe-like structures with clear boundaries. The observation results after magnification revealed that the tumor cells were arranged in a pallisade-like and basal pattern, with deep nuclear staining and minimal cytoplasmic. Immunohistochemical staining showed the high expression of CK protein and low proportion expression of ki-67 protein in tumor cells, as well as the high expression of vimentin and low expressions of Bcl2 and CD10 in tumor cell mesenchyme. The isolated tumor cells grew well in DMEM medium containing 10% fetal bovine serum and could be cultured by passage, but no tumor formation was observed in healthy tree shrews and nude mice inoculated with tumor cells. ConclusionCombined with the location of the tumor, overall morphology, HE staining, and immunohistochemical results, the thoracoabdominal mass of the tree shrew was diagnosed as a trichoepithelioma.