2.Enhancement of artemisinin biosynthesis in transgenic Artemisia annua L. by overexpressed HDR and ADS genes.
Ya-Xiong WANG ; Shi-Ping LONG ; Li-Xia ZENG ; Li-En XIANG ; Zhi LIN ; Min CHEN ; Zhi-Hua LIAO
Acta Pharmaceutica Sinica 2014;49(9):1346-1352
Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua
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genetics
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metabolism
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Artemisinins
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metabolism
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Biosynthetic Pathways
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Drugs, Chinese Herbal
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Mixed Function Oxygenases
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genetics
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Oxidoreductases
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genetics
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Plant Proteins
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genetics
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Plants, Genetically Modified
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genetics
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metabolism
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Plants, Medicinal
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genetics
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metabolism
3.Lyman NTCP model analysis of radiaton-induced liver disease in hypofractionated conformal radiotherapy for primary liver carcinoma
Zhi-Yong XU ; Shi-Xiong LIANG ; Ji ZHU ; Jian-Dong ZHAO ; Xiao-Dong ZHU ; Xiao-Long FU ; Guo-Liang JIANG ;
Chinese Journal of Radiation Oncology 2005;0(06):-
Objective To-identify the factors associated with radiation-induced liver disease (RILD) and to describe the probability of RILD using the Lyman normal tissue complication(NTCP) model for primary liver carcinoma(PLC) treated with hypofractionated conformal therapy (CRT).Methods A total of 109 PLC patients treated with hypofractionated CRT were prospectively followed according to the Child-Pugh classification for liver cirrhosis,93 patients in class A and 16 in class B.The mean dose of radi- ation to the isocenter was (53.5?5.5) Gy,fractions of (4.8?0.5) Gy,with interfraction interval of 48 hours and irradiation 3 times per week.Maximal likelihood analysis yielded the best estimates of parameters of the Lyman NTCP model for all patients;Child-Pugh A and Child-Pugh B patients,respectively.Results Of all the patients,17 developed RILD (17/109),8 in Child-Pugh A(8/93 ) and 9 in Child-Pugh B(9/ 16).By multivariate analysis,only the Child-Pugh Grade of liver cirrhosis was the independent factor (P= 0.000) associated with the developing of RILD.The best estimates of the NTCP parameters for all 109 pa- tients were n=1.1,m=0.35 and TD_(50) (1)=38.5 Gy.The n,m,TD_(50) (1) estimated from patients with Child-Pugh A was 1.1,0.28,40.5 Gy,respectively,compared with 0.7,0.43,23 Gy respectively,for patients with Child-Pugh B.Conclusions Primary liver cancer patients who possess Child-Pugh B cirrho- sis would present a significantly greater susceptibility to RILD after hypofractionated CRT than patients with Child-Pugh A cirrhosis.The predominant risk factor for developing RILD is the severity of hepatic cirrhosis in the liver of PLC patients.
4.Optimal Supply of Precursors for CoQ10 Production by Rhodopseudomonas Palustris
Shi-Yun JIANG ; Long-Jiang YU ; Xin XIONG ; Xiao-Lin SHEN ; Yan JIAN
Progress in Modern Biomedicine 2008;8(5):845-850
CoQ10 has been used not only as a medicine but also as food supplements because of its various physiological and biochemical activities. A full-factorial central composite design and response surface methodology were used for optimizing three precursors Solanesol, 4-hydroxybenzoic acid and methionine to maximize the production of CoQ10 by Rhodopseudomonas palustris J001. The optimization of the model predicted a maximum response 40.6 [(mg CoQ10)(g dried biomass)-1] CoQ10 production with 124.8 mg l-1 Solanesol, 267.7 mg l-14-hydroxybenzoic acid and 130.2 mg l-1 methionine, respectively. A new combination was prepared according to the result. The observed response was 40.5 ± 0.2 [(mg CoQ10)(g dried biomass)-1] and was 109.8%higher than in the control with no addition of the three precursors.
5.Effect of simulated microgravity on human monocytic cell proliferation and tissue factor mRNA expression.
Xia YU ; Lei ZHENG ; Shi-long XIONG ; Zhen CAI ; Qian WANG
Journal of Southern Medical University 2011;31(6):1020-1022
OBJECTIVETo investigate the effect of simulated microgravity on the proliferation of human monocytic cells THP-1 and the expression of tissue factor (TF) mRNA.
METHODSTHP-1 cells were cultured under a simulated microgravity environment using the rotating cell culture system (RCCS). The changes in the cell proliferation after microgravity culture were assessed by cell counting and cell cycle analysis with flow cytometry. RT-PCR was used to detect the changes in the expression of TF mRNA in THP-1 cells.
RESULTSCulture under simulated microgravity resulted in a significant decrease in the cell number of THP-1 cells in comparison with that of the control cells (P<0.01). After a 24-h culture under microgravity, the G0-Gl phase cells increased from the control level of (46.57∓1.64)% to (67.64∓2.71)% (P<0.05). The cells in both groups showed a low level of TF mRNA expression in the absence of LPS stimulation. A 4-h stimulation with LPS caused up-regulated expression of TF mRNA in both cells, but the microgravity group showed a significantly smaller increase in the expression (2.301∓0.179) than the control group (9.210∓1.328) (P<0.05).
CONCLUSIONMicrogravity can inhibit the proliferation of THP-1 cells and suppress the cellular expression of TF mRNA.
Cell Proliferation ; Cells, Cultured ; Humans ; Monocytes ; cytology ; metabolism ; RNA, Messenger ; genetics ; Thromboplastin ; genetics ; metabolism ; Weightlessness ; Weightlessness Simulation
6.Preliminary study on outcome assessment system of treatment of stroke.
Shi-long LAI ; Xin-feng GUO ; Wei-xiong LIANG
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(3):197-201
OBJECTIVEIn order to scientifically reflect the real efficacy of TCM treatment and to preliminarily establish a definitely valid and reliable assessment system of stroke treatment, with multi-dimensional outcome assessment indexes, including efficacy evaluation system of conventional western medicine, syndrome evaluation criteria in TCM and quality of life assessment system.
METHODSAn integrative approach of cross-sectional survey and prospective follow-up was adopted. Two hundred and forty-five case-episodes of stroke patients were assessed by determining such parameters as nerve functional deficit scale, grading of total status of living ability, activity of daily living (ADL), TCM stroke criteria of diagnosing-treatment, TCM syndrome related symptoms/signs, Health Survey Questionnaire (Short form 36, SF-36), and index of quality of life (QOL), and their construction and relation were analyzed by such methods as multivariate relation, partial relation, linear regression, factor analysis, Cronbach's alpha coefficients, and the responsiveness estimation.
RESULTSThe multi-dimensional assessment system of stroke treatment, containing 57 indexes with clarified constructions and classification, was created, and its validity and reliability confirmed after assessement. Various degrees of relationship were found between different TCM Syndromes and different domains of QOL.
CONCLUSIONThe assessment system of stroke treatment preliminarily created has satisfied reliability and validity. It could be expected to reflect the real efficacy of TCM treatment more inclusive and accurate. TCM Syndrome indexes are considered to be the factor related to both domains of mental and physical health, particularly with the former, therefore, to improve the TCM Syndrome would imply improvement of the mental health, physical health and QOL of the patients.
Aged ; Cerebral Infarction ; drug therapy ; Cross-Sectional Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Intracranial Hemorrhages ; drug therapy ; Male ; Middle Aged ; Outcome Assessment (Health Care) ; Phytotherapy ; Prospective Studies ; Quality of Life ; Stroke ; drug therapy ; Surveys and Questionnaires
7.Biomechancial issues in traditional Chinese medcine research
Journal of Medical Biomechanics 2013;28(3):E259-E262
Traditional Chinese medicine (TCM) has been formed and developed in medical practices over thousands of years, while biomechanics as a new research direction in TCM has been developing rapidly in recent years. Biomechanics in TCM encompasses abundant contents, such as hemodynamics in meridians, substance transport of Yin-Yang and the Five Elements, hemodynamics of pulse manifestation, heat conduction in tongue diagnosis, biomechanics of orthopaedics and traumatology, mechanics of acupuncture, massage and manipulation, TCM medical device, biomechnopharmacology and etc. Journal of Medical Biomechanics has published some papers about TCM biomechanics in the past years, and in this issue, a special column of TCM biomechanics in three research directions is organized, including massage and manipulation, fracture fixator by TCM treatment, and Tai Chi Quan, so as to arouse readers’ interest in TCM biomechanics.
8.Brain function connectivity of patients with non-fluent aphasia after subacute stroke based on functional near infrared spectroscopy
Maoqing CHEN ; Guohui JIANG ; Xiaoming WANG ; Zijuan SHI ; Tao XIONG ; Qingwen LONG ; Aimin HU ; Yujun LI
Chinese Journal of Neuromedicine 2023;22(11):1144-1150
Objective:To explore the neural mechanism of language dysfunction in patients with subacute stroke using functional near-infrared spectroscopy (fNIRS).Methods:Sixteen patients with non-fluent aphasia after subacute stroke (aphasia group), 16 patients with non-aphasia after stroke (non-aphasia group), and 16 healthy middle-aged and elderly subjects (control group) were enrolled into our study. The 6-min resting-state data of fNIRS were collected. Four language-related regions, Broca area, Wernicke area, dorso lateral prefrontal cortex (DLPFC), and supplementary motor area (SMA), were selected as regions of interest (ROIs), and the whole brain functional connection strength and functional connection strength in ROIs and between each two ROIs were analyzed by NirSpark software.Results:Compared with the control group (0.53±0.15) and non-aphasia group (0.47±0.12), the aphasia group had significantly decreased whole brain functional connection strength (0.29±0.14, P<0.05). Compared with the control group and non-aphasia group, the aphasia group had significantly decreased functional connection strength in the left Wernicke area, right Wernicke area, left Broca area, left SMA area, right SMA area and left DLPFC area ( P<0.05, FDR). Compared with the control group and non-aphasia group, the aphasia group had significantly decreased functional connection strength in the right Wernicke-left Wernicke area, right Wernicke-right Broca area, right Wernicke-left Broca area, right Wernicke-right DLPFC area, right Wernicke-left DLPFC area, right Wernicke-right SMA area, right Wernicke-left SMA area, left Wernicke-right Broca area, left Wernicke-left Broca area, left Wernicke-right DLPFC area, left Wernicke-left DLPFC, left Wernicke-right SMA area, left Wernicke-left SMA area, right Broca-left Broca area, right Broca-left DLPFC area, right Broca-right SMA area, right Broca-left SMA area, left Broca-right DLPFC area, left Broca-left DLPFC area, left Broca-right SMA area, left Broca-left SMA area, right DLPFC-left DLPFC area, right DLPFC-right SMA area, right DLPFC-left SMA area, left DLPFC-right SMA area, left DLPFC-left SMA area, and right SMA-left SMA area ( P<0.05, FDR). Conclusion:Abnormal functional connectivity strength of the whole brain and language-related key brain areas might be the neural mechanism of language dysfunction in patients with non-fluent aphasia after subacute stroke.
9.Prognostic factor of primary liver cancer treated by hypofractionated three-dimensional conformal radiotherapy.
Shi-xiong LIANG ; Guo-liang JIANG ; Xiao-dong ZHU ; Xiao-long FU ; Fu-xiang LI ; Qi-fang HUANG ; An-yu WANG ; Long CHEN ; Hai-jie LU
Chinese Journal of Oncology 2005;27(10):613-615
OBJECTIVETo evaluate the toxicity and efficacy of primary liver cancer (PLC) treated by hypofractionated three-dimensional conformal radiotherapy (3DCRT) and investigate the prognostic factors.
METHODSBetween April 1999 and August 2003, 128 PLC patients received hypofractionated 3DCRT. According to UICC/AJCC staging system, there were 83 T3 patients, 45 T4, with none of them having lymph node metastasis. The mean value of gross tumor volume (GTV) was (458.92 +/- 429.8) cm(3) (6.2-2097 cm(3)). Thirty-four patients had portal vein tumor thrombosis (PVTT). 108 patients had Child-Pugh Grade A liver cirrhosis and 20 Child-Pugh Grade B liver cirrhosis. All patients received a total dose of (53.6 +/- 6.6) Gy/4-8 Gy per fraction/3 fractions per week. Forty-eight of these patients received 3DCRT combined with transarterial chemoembolization (TACE).
RESULTSSeven patients died within 3 months after the treatment were dismissed from the series. The response rate (CR + PR) was 55% (67/121). The overall 1-, 2-, and 3-year survival rate was 65.0%, 43.3%, and 33.1%, respectively. T stage (P = 0.001), GTV (P = 0.0001), PVTT (P = 0.0001) and Child-Pugh Grade (P = 0.0001) had significant impact on the overall survival. However, only GTV and Child-Pugh Grade were independent significant prognostic factors by Cox-regression analysis, (P = 0.044 and P = 0.015).
CONCLUSIONT stage, GTV, PVTT and Child-Pugh Grade have significant impact on the overall survival in primary liver cancer patients treated by three-dimensional conformal radiotherapy. But only GTV and Child-Pugh Grade are independent prognostic factors.
Adult ; Aged ; Carcinoma, Hepatocellular ; radiotherapy ; Female ; Follow-Up Studies ; Humans ; Imaging, Three-Dimensional ; Liver Neoplasms ; radiotherapy ; Male ; Middle Aged ; Prognosis ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Conformal ; methods ; Regression Analysis ; Retrospective Studies
10.Quantitative research on the vertical force homogeneity of Yizhichan manipulation
Jie LÜ ; Jin-feng CAO ; Long-long MA ; Shi-xiong XU
Journal of Medical Biomechanics 2012;27(4):E456-E459
Objective To make a quantitative research on the vertical force homogeneity of Yizhichan manipulation and look for the quantitative index and reference value which can describe the force homogeneity of Yizhichan manipulation effectively. Methods After the force homogeneity was described mathematically, the vertical force signals from Yizhichan manipulation operated by 5 experts, 5 skilled doctors and 5 beginners were measured and analyzed, respectively. Results The period homogeneities and the waveform homogeneities were obtained. No significant differences were found in the period homogeneities among the three groups, but significant differences appeared in waveform homogeneities between the beginner and expert group or the beginner and skilled doctor group. Conclusions The waveform homogeneities are more suitable for evaluating the vertical force homogeneity of Yizhichan manipulation by different operators, and 0.927 can be used as a reference value to evaluate the vertical force homogeneity of the operator.