2.Effects of Nurr1 down-regulation on the expression of tyrosine hydroxylase and neurite extension in dopaminergic cells..
Yun-Cheng WU ; You-Qing CAI ; Yong-Bo ZHAO ; Jian FEI
Acta Physiologica Sinica 2006;58(4):351-358
In the experiment, we designed and synthesized two siRNAs based on the sequence of nuclear receptor-related factor 1 (Nurr1) mRNA. They were separately subcloned into the plasmid of pSilenCircle (pSC) containing U6 promoter. The pSC-Nurr1 vectors (pSC-N1 and pSC-N2) specific to Nurr1 gene and the negative control vector of short-hairpin RNA (shRNA) eukaryotic expression vector were constructed. We cultured the dopaminergic cell line MN9D and the verified vectors were transfected with LipofectamineTM 2000 in vitro. The positive cell clones transfected with pSC were obtained after being screened with 500 mug/ml G418. After that, the silencing effects of Nurr1 and TH mRNA or protein were detected by real time RT-PCR and Western blot. The neurite extension of MN9D cells was observed and photographed by inverted microscope. The results showed that Nurr1 mRNA expression in MN9D cells was specifically down-regulated by the vectors of pSC-N1 and pSC-N2, and the silencing effects were 62.3% and 45.6%, respectively. The dopaminergic phenotype of TH mRNA was also suppressed significantly and the silencing effects were 76.3% and 62.6%, respectively. Meanwhile, the expressions of Nurr1 and TH proteins were also significantly suppressed, and the silencing effects of Nurr1 and TH protein were 57.4%, 72.0% and 79.1%, 70.1% respectively. The negative control and liposome groups had no effect on the two genes. In conclusion, Nurr1 shRNA expressing vectors can inhibit the expressions of Nurr1 and TH mRNA or protein in MN9D cells, and Nurr1 might play a role in neurite extension of MN9D cells. Nurr1 shRNA expressing vector may provide a novel applicable strategy for the study on the function of the genes associated with Parkinson disease and the development of dopaminergic neuron.
Cell Line
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Dopaminergic Neurons
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cytology
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metabolism
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Down-Regulation
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Fetus
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Humans
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Mesencephalon
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cytology
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Neurites
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physiology
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Nuclear Receptor Subfamily 4, Group A, Member 2
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genetics
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metabolism
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RNA, Messenger
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genetics
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metabolism
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RNA, Small Interfering
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genetics
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Transfection
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Tyrosine 3-Monooxygenase
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genetics
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metabolism
3.Peripheral NK and NKT cell in pediatric subjects with chronic HBV infection.
Bin YANG ; Li-Ming CHEN ; Wen-Hua YE ; Hong-Fei ZHANG ; Yun CHENG
Chinese Journal of Experimental and Clinical Virology 2009;23(1):59-61
OBJECTIVETo investigate the characteristic of NK cells and NKT cells in HBV infected pediatric subjects for evaluation of their clinical implication.
METHODSFresh peripheral blood samples were obtained from 42 HBV-infected pediatric cases and 15 healthy counterparts. NK cells and NKT cells were analyzed by flow cytometry assay. The clinical data such as serum ALT level and HBV viral load was simultaneously recorded from each HBV carrier.
RESULTSHBV-infected children had an obviously increasing percentage of NK cells 12.071% +/- 7.100%, there were significant differences between the children with chronic B hepatitis and the healthy children (P <0.05). As far as percentage of NKT 3.048% +/- 1.937% was concerned, there were not differences. Furthermore the association was not found between serum HBV viral load level and the NK lymphocyte.
CONCLUSIONOur data may provide valuable information of NK and NKT lymphocyte for evaluation of disease progression of HBV infected children NKI cells.
Adolescent ; Cells, Cultured ; Child ; Female ; Hepatitis B virus ; immunology ; physiology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Killer Cells, Natural ; immunology ; Male ; Natural Killer T-Cells ; immunology
4.Benefit of network education to college students' knowledge about sexual and reproductive health in Ningbo city.
Guo-yao WANG ; Yun-xin JI ; Hui-qing DING ; Zhong-bao GUI ; Xiao-ming LIANG ; Jian-fei FU ; Yue CHENG
National Journal of Andrology 2015;21(12):1077-1081
OBJECTIVETo investigate how network education can improve college students' knowledge on sexual and reproductive health in Ningbo city.
METHODSFrom December 2012 to June 2013, we conducted a questionnaire investigation among college students in Ningbo city about the effects of network education on their knowledge about sexual psychology, sexual physiology, sexual ethics, and reproductive health.
RESULTSA total of 7 362 college students accomplished the investigation, of whom 2 483 (42.1% males and 57.9% females) received network education, while the other 4 879 (24.1% males and 75.9% females) did not. Approximately 47.1% of the male and 28.0% of the female students acquired sexual and reproductive knowledge via network education. Reproductive health-related network education significantly enriched the students' knowledge about the reproductive system and sex, pubertal development, sexual physiology, conception and embryonic development, methods of contraception, sexual psychology, sexually transmitted diseases and their prevention, pregnancy care and eugenics, and environment- and occupation-related reproductive health (P < 0.01). It also remarkably improved their cognitive attitude towards reproductive health knowledge (P < 0.01). Those who received reproductive health-related network education showed a significantly higher rate of masturbation (P < 0.01) but markedly later time of the first masturbation (P < 0.01) than those who did not.
CONCLUSIONNetwork education can enhance the effect of reproductive health education among college students and improve their sexual experience and health.
China ; Contraception ; Female ; Health Education ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Masturbation ; Pregnancy ; Reproduction ; Reproductive Health ; Sexual Behavior ; physiology ; psychology ; Sexually Transmitted Diseases ; Students ; Surveys and Questionnaires ; Universities
5.A chromatographic fingerprinting-based approach to evaluating the stability of production process of traditional Chinese medicine.
Chu-Chu HU ; Yun-Fei LI ; Yi-Yu CHENG
China Journal of Chinese Materia Medica 2006;31(14):1151-1162
OBJECTIVETo study a new method for the evaluation of the stability of production process of Traditional Chinese Medicines (TCM) based on chromatographic fingerprinting.
METHODTaking the extraction process of Red Ginseng for example, the fingerprints of extracts produced with fluctuating processing factors, i.e. ratio of solvent to liquid, solvent concentration, extraction durations and extraction times, were acquired. Meanwhile, the similarity of fingerprinting based on Euclidean distance was calculated and each fingerprint was fitwith cubic spline.
RESULTSimilarities of fingerprints of the extracts responded to the fluctuation of processing factors, and it showed that taking similarity as index, quantitative relation between chromatographic fingerprints and processing factors could be established.
CONCLUSIONThe method proposed in the paper was able to describe the quality of products resulting from fluctuating conditions in extracting process, and therefore, can be developed as a practicable strategy to evaluate the stability Traditional Chinese Medicines in production process.
Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; isolation & purification ; Evaluation Studies as Topic ; Ginsenosides ; isolation & purification ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Solvents ; Technology, Pharmaceutical ; methods
6.Practical experience and lessons learnt on COVID-19 prevention and control in Huangpu District, Shanghai
Hua-dong ZHUANG ; Fei SU ; Yun-hui LU ; Jing WANG ; Wen-yu ZHOU ; CHENG CHENG
Shanghai Journal of Preventive Medicine 2020;32(5):397-
Corona virus disease 2019 (COVID-19) quickly spread from Wuhan, Hubei Province to other provinces in China since December 2019.Huangpu District of Shanghai was one of the areas where COVID-19 was found at an earlier time in people coming from Wuhan.By summarizing and reviewing the experience and lessons learnt in Huangpu District, the article explores the working mode for public health interventions to prevent and control COVID-19, providing reference for other cities in this regard.
7.Grades evaluation of Cortex Moutan decoction pieces based on quality constant of Chinese medicine
Cheng-Kai XIA ; Yun-Fei HU ; Si-Yu CHEN ; Cheng-Wu FANG
Chinese Traditional Patent Medicine 2018;40(3):647-653
AIM To determine the quality constant of Chinese medicine and to establish a method of grades evaluation of Cortex Moutan decoction pieces.METHODS Cortex Moutan decoction pieces' morphological indexes should be observed and measured,the contents of phenol and paeoniflorin were measured by HPLC.Cortex Moutan decoction pieces was graded according to the quality constant of Chinese medicine which was calculated.RESULTS Cortex Moutan decoction pieces could be divided into three grades by the quality constant of Chinese medicine,the cutoff point of the firsts was greater than or equal to 1.56;the second was greater than or equal to 1.12 but less than 1.56;while the third was less than 1.12.CONCLUSION This method combines the apparent parameters of Cortex Moutan decoction pieces with its intrinsic quality,which can devide grades of it objectively and provides a reference for its classification.
8.Cloning and induced expression analysis of 4-hydroxy-3-methyl-but-2-enyl diphosphate reductase gene (smHDR) of Salvia miltiorrhiza.
Qi-Qing CHENG ; Yun-fei HE ; Geng LI ; Chao JIANG ; Yuan YUAN ; Wei GAO ; Lu-Qi HUANG
Acta Pharmaceutica Sinica 2013;48(2):236-242
This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.
Amino Acid Sequence
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Cloning, Molecular
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DNA, Complementary
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genetics
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Diterpenes, Abietane
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biosynthesis
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metabolism
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Gene Expression Regulation, Plant
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Open Reading Frames
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Oxidoreductases
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chemistry
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genetics
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metabolism
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Phylogeny
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Plant Roots
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genetics
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metabolism
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Plants, Medicinal
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genetics
;
metabolism
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Protein Structure, Secondary
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RNA, Messenger
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metabolism
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Salvia miltiorrhiza
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genetics
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metabolism
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Sequence Alignment
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Silver Nitrate
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pharmacology
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Synthetic Biology
9.The status of glycemic control in type 2 diabetes mellitus: a cross-sectional survey in Jiangsu province
Yan BI ; Aziguli MAIHEMAITI ; Dalong ZHU ; Shiwei CUI ; Wei LI ; Fei WANG ; Jinluo CHENG ; Yan ZHU ; Ning XU ; Yun HU ; Shanmei SHEN
Chinese Journal of Endocrinology and Metabolism 2011;27(7):575-579
Objective To determine the status of glycemic control in type 2 diabetic patients in Jiangsu province. Methods Survey was conducted among outpatients with type 2 diabetes mellitus from 56 diabetes centers including 23 tertiary hospitals, 18 secondary hospitals and 15 primary hospitals, covering 13 cities in Jiangsu province. Data were obtained by patient interview and by reviewing medical records of the most recent laboratory assessment and clinical examinations. Blood samples were collected during patient′s visits for assessment of HbA1C. Results The mean HbA1Cof 2 966 patients was 7.18%±1.58%. Mean age of patients was(56.4±11.2) years, mean duration of diabetes was(6.3±5.7) years, and mean body mass index was(24.5±3.3) kg/m2. The proportion of good control was 40.2% defined as HbA1C<6.5%. 42.2% of the patients in the tertiary hospitals, 36.5% of the cases in the secondary hospitals, and 36.2% of cases in the primary hospitals achieved that goal with HbA1C<6.5%, while age, body mass index, and duration of diabetes revealed no difference among these 3 groups of patients. Conclusions The status of glycemic control in Jiangsu province has been improved as compared with the result from national survey performed 3 years ago. Patients from tertiary hospitals are better controlled than those from secondary and primary hospitals. Structural interventions to improve diabetes care in secondary and primary hospitals are needed.
10.Investigation of ulinastatin on protection of organ functions in patients with severe disease
Fei WU ; Xiao-Yun YU ; Yong LEI ; Jian-Ming ZHU ; Yan GAO ; Wen ZHU ; Ai-Rong LI ; Xiao-zhen WAN ; Mei-cheng LE
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(05):-
Objective To study the mechanism and protection of ulinastatin on organ functions in patients with severe disease.Methods Sixty patients in the intensive care unit(ICU)from October 2005 to July 2007 were randomly divided into a control group and an ulinastatin treatment group(each 30 cases).The patients in the control group received the conventional therapy,and the cases in the other treatment group accepted ulinastatin and conventional therapy.According to the disease situations,ulinastatin was administered 200-400 kU once,2-4 times a day,sequentially for 5-7 days.On the day of admission and 3, 5,and 7 days after admission in ICU respectively,blood samples were obtained for measuring alanine aminotransferase(ALT),aspartate aminotransferase(AST),creatinine(Cr),blood urea nitrogen(BUN), activated partial thromboplastin time(APTT),fibrinogen(FIB)and oxygenation index(PaO_2/FiO_2); whether breathing machine or hematodialysis was used and the end results were recorded.Results The rate of usage of breathing machine(23.3%),the incidences of hepatosis(3.3%)and renal dysfunction(10.0%) and fatality(3.3%)in ulinastatin treatment group were obviously lower than those of the control group (63.3%,23.3%,46.7%,10.0%,P0.05).Only one patient received bematodialysis in control group.Conclusion Ulinastatin can protect liver,renal and lung functions markedly and lower the incidence of multiple organ dysfunction syndrome and mortality in patients with severe disease.