2.Reversal of drug resistance in human ovarian cancer cells by wild-type PTEN gene and its mechanisms
Hui-Juan WU ; Hai-Tao WU ; Dan-Hui WENG ; Hui XING ; Yun-Ping LU ; Ding MA ;
Chinese Journal of Obstetrics and Gynecology 2000;0(09):-
Objective To examine expression of PTEN gene in ovarian cancer cisplatin-sensitive cell line OV2008 cells and cisplatin-resistant cell line C13K cells,and evaluate the effect of wild-type PTEN gene on reversing cisplatin-resistance of C13K cells and underlying mechanisms.Methods The expression of PTEN mRNA and protein in OV2008 and C13K cells were detected by semi-quantitative RT-PCR and western blot.Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine 2000.The expression of PTEN mRNA was monitored by RT- PCR and the expression of PTEN,protein kinase B(AKT),phospho-AKT(p-AKT)protein were analyzed by western blot in PTEN transfected and untransfected C13K cells.Proliferation and chemosensitivity of cells to cisplatin were measured by methyl thiazolyl tetrazolium(MTT),and cell apoptosis was detected by flow cytometry after treatment with cisplatin.Results(1)The expression of PTEN mRNA and protein(1.02 ?0.05,1.02?0.07)in OV2008 cells were significantly higher than those in C13K cells,which were 0.45 ?0.03 and 0.55?0.03 respectively(P
3.Survey on the knowledge of epilepsy to patients and their family members.
Ding DING ; Guo-Xing ZHU ; Jin-Yang WANG ; Gang ZUO ; Jia YANG ; Jun-Chao LU ; Xian-Tao LI ; Zhen HONG
Chinese Journal of Epidemiology 2004;25(12):1059-1062
OBJECTIVETo study the epilepsy patients and their family members on their knowledge of the disease.
METHODSA 34-point questionnaire with 34 questions related to epilepsy knowledge was used for the survey on 170 pairs of epilepsy patients and their family members in Huashan hospital. Characters of the disease on the subjects were recorded.
RESULTSThe mean scores of the epilepsy knowledge of the patients and their family members were 16.5 +/- 8.2 and 16.1 +/- 8.5, respectively. The scores were quite low with no statistical difference between patients and their family members. The rate of correct answer in the urban subjects was obviously higher than those subjects living in the rural areas. All the subjects lacked the knowledge on the "cause of disease" when comparing with items as "diagnosis" and "treatment". Multivariate analysis showed that rural residents (P = 0.0001, OR = 52.963) and low education level (P = 0.0294, OR = 2.266) related to low epilepsy knowledge score among epilepsy patients. However, for family members, the factor related to low score was only living in the rural area (P = 0.0001, OR = 37.229).
CONCLUSIONEducation on the epilepsy knowledge should be strengthened, especially in the rural areas.
Adolescent ; Adult ; Epilepsy ; psychology ; Family Health ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Middle Aged ; Rural Health ; Surveys and Questionnaires
4.High-level production of neutral endoglucanase 1 in Pichia pastoris.
Shao-Jun DING ; Mei-Jing SONG ; Hong-Jun YANG ; Zeng-Tao XING ; Rui ZHOU ; Jie CAO
Chinese Journal of Biotechnology 2006;22(1):71-76
The gene (eg1) encoding for novel endoglucanase 1 was cloned previously from Chinese straw mushroom Volvariella volvacea. EG1 has high thermal stability and optimal pH at neutral and shows great potential in textile and paper industry applications. To improve the expression level of EG1 in Pichia pastoris, the increasing copy number of clone, and its high cell density fermentation in 3.2L fermenter for its high-level expression were investigated in this work. By electro-transformation of pPICZalphaB-egl into GS115EG11 integrated with single copy of eg1 gene, A resistant transformant with 3.8 times higher level expression than GS115EG11 was screened from YPDSZ plate containing 2000 microg/mL of Zeocin. The effect of initial cell density, pH and methanol on its expression and biomass accumulation was evaluated in shaking culture. Optimal EG1 production was observed when initial cell density OD600 was 5.0. EG1 production and biomass accumulation did not seem to vary when cells were induced at different pH values. Both of EG1 and cell density were found to increase with higher methanol concentrations, reaching 62.48 IU/mL and 31.7 (OD600) respectively after 120 h induction with 2.0% (V/V) methanol compared to 30.24 IU/mL and 17.79 (OD60) with 0.25% methanol induction. EG1 expression was further increased by 6.4 times higher than shaking culture after 95.5 hours induction with methanol in fed-batch fermentation, so totally 34 times higher than that for GS115EG11 was achieved by screening of high Zeocin resistant clone and high cell density fermentation. The production of EG1 with 543.36IU/mL CMC activity and 8.80mg/mL protein expression was obtained in Pichia pastoris.
Cellulase
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biosynthesis
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genetics
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Cloning, Molecular
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Culture Media
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Electroporation
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Escherichia coli
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genetics
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metabolism
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Fermentation
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Fungal Proteins
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biosynthesis
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genetics
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Pichia
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genetics
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metabolism
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Volvariella
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enzymology
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genetics
5.Effect of magnesium-free on glucocorticoid receptor expression in primary cultured cortical neurons of fetal rats in vitro.
Tao BO ; Lu YI ; Tuan-Mei WANG ; Jian LI ; Xing-Fang LI ; Ding-An MAO
Chinese Journal of Contemporary Pediatrics 2010;12(3):211-214
OBJECTIVETo study the changes of glucocorticoid receptor (GR) expression in embryonic rat cortical neurons exposed to transient Mg(2+)-free treatment.
METHODSSix days after rat cortical neuronal cultures, two groups were created based on the medium to which were transiently exposed. The control group was exposed to a physiological solution (PS), and the Mg(2+)-free group was exposed to the same medium as the control group except for the removal of magnesium. The expression of GR mRNA and protein was determined by real-time PCR and immunocytochemistry staining 1, 7 and 12 days after transient Mg(2+)-free treatment.
RESULTSCompared to the control group, the Mg(2+)-free group displayed the significantly less accumulated optical density (AOD) of GR immunoreactivity 12 days after transient Mg(2+)-free treatment (p<0.05). On the contrary, GR mRNA expression increased significantly 1 and 7 days after transient Mg(2+)-free treatment in the Mg(2+)-free group (p<0.05).
CONCLUSIONSGR expression is modified following Mg-free-induced injury in cultured developing neurons in rats.
Animals ; Cell Survival ; Cells, Cultured ; Cerebral Cortex ; metabolism ; Fetus ; metabolism ; Hypothalamo-Hypophyseal System ; physiology ; Magnesium ; physiology ; Neurons ; metabolism ; Pituitary-Adrenal System ; physiology ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Receptors, Glucocorticoid ; analysis ; genetics
6.Expression of CD40 and COX-2 and Its Relationship with Angiogenesis in Esophageal Squamous Cell Carcinoma
Liying XUE ; Xianghong ZHANG ; Xuemin LI ; Yuehong LI ; Tao DING ; Jianmin MI ; Junling WANG ; Xia YAN ; Shi WEN ; Xin XING ; Jinfeng CUI
Chinese Journal of Clinical Oncology 2010;37(1):36-39
Objective:To explore the putative role of CD40 and COX-2 expression in human esophageal squamous cell carcinoma(ESCC)and to analyze their possible relationship with angiogenesis in ESCC.Methods:The expression of CD40 and COX-2 was detected in 79 ESCC and 28 normal esophageal epithelial tissue samples with immunohistochemical staining.The microvessel density by CD34 was determined and the clinicopathological significance of CD40 and COX-2 expression in ESCC was analyzed.In addition,the expression of CD40 and COX-2 in esophageal squamous cell carcinoma cell line Eca109 and primary cultured normal esophageal epithelial cells in vivo was comparatively studied with immunocytochemical staining andWestern blot.Results:Compared with that in normal epithelial tissues,the expression of CD40 and COX-2 in ESCC was significantly higher(54.43%vs 10.71%:69.62%vs 17.86%:respectively,P<0.05).The positive expression rate of CD40 in ESCC cases with lymph node metastasis was significantly higher than that in those without lymph node metastasis(70.37%vs 46.1 5%.P<0.05).No correlation was found between CD40 expression and patient age,sex,tumor location,size and differentiation of tumors.No clinicopathological significanca of COX-2 expression in ESCC was found.There was a positive correlation between CD40 expression and COX-2 expression in esophageal squamous cell carcinoma(P<0.05,φ=0.446).The mean MVD value in ESCC was significantly higher than that in normal esophageal tissue(25.02±5.52 vs 12.09±4.55,P<0.05).MVD value in ESCC was closely correlated with lymph node metastasis.The mean MVD value in ESCC cases with positive CD40 and COX-2 expression was higher than that in those with negative CD40 and COX-2expression(26.37±6.02 vs 22.58±5.25.P<0.05).Western blot results showed that CD40 and COX-2 expression in Eca109 was higher than that in cultured normal esophageal epithelial cells (P<0.05).Conclusion:The expression of CD40 is involved in the carcinogenesis and progression of esophageal squamous cell carcinoma.CD40 may play an important role in the angiogenesis of ESCC through promoting COX-2 expression.
7.Effects of neonatal recurrent seizures on glucocorticoid receptor expression in the rat brain.
Tuan-Mei WANG ; Tao BO ; Man-Zhi WANG ; Xiao-Hua ZHU ; Jian LI ; Xing-Fang LI ; Ding-An MAO
Chinese Journal of Contemporary Pediatrics 2010;12(1):47-50
OBJECTIVETo investigate the effets of flurothyl-induced neonatal recurrent seizures on glucocorticoid receptor (GR) expression in the rat brain.
METHODSForty-eight seven-day-old Sprague-Dawley rats were randomly divided into two groups: control and seizure. Seizures were induced by inhalant flurothyl daily for six consecutive days. Brains were sampled on postnatal days 13, 15 and 19. The expression of GR protein in the cerebral cortex was detected by Western blot and immunohistochemical method.
RESULTSThe expression of GR in the cerebral cortical plasma protein was significantly lower in the seizure group than in the control group on postnatal day 15. The expression of GR protein in the cerebral cortical nuclear protein decreased significantly in the seizure group compared with that in the control group on postnatal days 15 and 19 (p<0.05). Compared to the control group, the accumulated optical density (AOD) of GR immunoreactivity (IR) decreased significantly in the parietal cortex on postnatal day 13 (p<0.05), the AOD of GR IR decreased significantly in the parietal cortex and the temporal cortex on postnatal day 15 (p<0.05), and the AOD of GR IR decreased significantly in the parietal cortex, temporal cortex and the frontal cortex in the seizure group on postnatal day 19 (p<0.05).
CONCLUSIONSRecurrent seizures in neonatal rats result in abnormal GR expression in the cerebral cortex which might play an important role in short-term brain injury induced by early recurrent seizures.
Animals ; Blotting, Western ; Cerebral Cortex ; chemistry ; Female ; Hypothalamo-Hypophyseal System ; physiology ; Immunohistochemistry ; Male ; Pituitary-Adrenal System ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Glucocorticoid ; analysis ; physiology ; Recurrence ; Seizures ; metabolism
8.Expression of serum FSTL-1 in bone metastasis of prostate cancer and its clinical implication.
Tao DING ; Xiao-Zhou HE ; Xian-Lin XU ; Hai-Yan XU ; Cui-Xing ZHOU ; Yu-Ji WANG
National Journal of Andrology 2014;20(12):1090-1092
OBJECTIVETo investigate the expression of follistatin-like protein 1 (FSTL-1) in bone metastasis of prostate cancer (BMPC), the correlation of serum FSTL-1 with the chronic inflammatory factor interleukin-6 (IL-6) and bone morphogenetic protein 6 (BMP6) , and the clinical application value of serum FSTL-1 in BMPC.
METHODSUsing ELISA, we measured the expression levels of serum FSTL-1, IL-6, and BMP6 in 35 patients with BMPC and another 30 with benign prostatic hyperplasia (BPH) and performed correlation analysis on the data obtained.
RESULTSCompared with the BPH controls, the BMPC patients showed a significantly decreased expression of serum FSTL-1 ([34.45 ± 12.35] μg/L vs [20.23 ± 8.69] μg/L, P < 0.01) and increased levels of IL-6 ([11.21 ± 8.62] μg/L vs [23.56 ± 20.12] μg/L, P < 0.05) and BMP6 ([293.50 ± 39.72] μg/L vs [428.30 ± 178.40] μg/L, P < 0.05). There was a significant negative correlation between the level of serum FSTL-1 and those of IL-6 and BMP6 in the BMPC patients, with correlation coefficients of -0.971 and -0.972, respectively (P < 0.05).
CONCLUSIONThe expression of serum FSTL-1 decreases in patients with bone metastasis of prostate cancer, and it is correlated with the levels of inflammatory factor and cell transformation factor. This finding offers a novel biological marker for the development and progression of prostate cancer as well as a new biological target factor for its intervention.
Aged ; Biomarkers, Tumor ; blood ; Bone Morphogenetic Protein 6 ; blood ; Bone Neoplasms ; blood ; secondary ; Disease Progression ; Follistatin-Related Proteins ; blood ; Humans ; Interleukin-6 ; blood ; Male ; Prostatic Hyperplasia ; blood ; Prostatic Neoplasms ; blood ; pathology
9.Data security and privacy protection of wireless medical sensor network based on hybrid encryption
Xing-Tao DING ; Bo-Cheng ZHONG ; Shu-Wen ZHU ; Bo HUANG
Chinese Medical Equipment Journal 2017;38(12):11-15
Objective To propose a hybrid encryption algorithm involving in elliptic curve cryptography (ECC) and advanced encryption standard (AES) to solve the problems of wiretapping and tampering of medical data in wireless channel transmission.Methods The sender used AES session key for medical data encryption and ECC public key for AES session key,and applied SHA-1 to obtaining the information summary of medical data and ECC public key to gaining information summary signature.The recipient employed ECC private key to verify the signature and then to decrypt AES session key,and finally decrypt medical data with AES session key.Results The proposed security solution gained advantages over the conventional ECC algorithm and hybrid cryptographic algorithm of RSA and AES in encryption and decryption time,safety strength and energy consumption,so that the confidentiality and integrity of medical data could be ensured during transmission.Conclusion The hybrid encryption scheme can meet the requirement of wireless medical sensor network in low time complexity,and realizes the protection of the patient data and privacy.
10.Introduction of pertussis surveillance system and evaluation of its operational effect in Tianjin.
Hai-Tao HUANG ; Zhi-Gang GAO ; Ya-Xing DING ; Jing SUN ; Yong-Cheng LI ; Ying ZHANG
Chinese Journal of Contemporary Pediatrics 2019;21(3):218-222
The pertussis surveillance system has been established since 2009 in Tianjin, and continuously improved over the past 10 years. This system determines the definition and classification of pertussis, establishes simple and feasible sampling methods and laboratory detection methods in clinical practice, standardizes the report management of pertussis cases and the treatment of epidemic situations. After the implementation of the surveillance system, the number of reported pertussis cases increased from 26 in 2009 to 802 in 2017, the number of diagnosed cases increased from 19 in 2009 to 662 in 2017, the reported incidence rate of pertussis increased from 0.16/100 000 in 2009 to 4.28/100 000 in 2017, and the number of medical institutions of reporting perutssis cases increased from 2 in 2009 to 53 in 2017. The specimen collection rate of the reported cases reached up to 93.66%. These results show that the sensitivity of pertussis surveillance has been improved and show that the data from the surveillance system may reflect more precisely the epidemical characteristics of perutssis in Tianjin.
Humans
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Incidence
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Infant
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Pertussis Vaccine
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Whooping Cough