1.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
2.Microsurgical resection of the recurrent craniopharyngiomas.
Xiang-en SHI ; Yong-li ZHANG ; Zhong-qing ZHOU ; Bo LIU
Chinese Journal of Surgery 2004;42(13):769-772
OBJECTIVETo study the effective method of microsurgical resection of the recurrent craniopharyngiomas.
METHODSMicrosurgical resection underwent in 40 cases with the recurrent craniopharyngiomas that accounted for 24 men and 16 women with mean age 35.1 year old. Visual deterioration was mainly complained in 25 patents, headache of symptoms in 9 patients, defect of visual field in 2 patients, amenorrhea in 2 patients, hyposexuality in one patient and diabetes insipidus in the one. Average history was 2.9 years. The superior sellar tumors on MR imaging grew in 19 cases, the superior-inferior sellar mass in 9 cases, intra-sellar in 5 cases and the tumors into the third ventricle in 7 cases. The huge calcification tumors were found in 5 cases, cystic tumors in 21 cases, and solid tumors 2 cases. Hydrocephalus presented in 12 cases. Evolution of tumor residuum was revealed in 31 cases after initial surgery and recurrent tumor after completing total removal of the tumor in 9 cases. The pterional approach was employed in 33 cases. The longitudinal fissure to the corpus callus approach in 2 cases was available for resection of the third ventricular tumor through the fornix column and septum pellucidum spaces.
RESULTSIn 33 cases with the pterional approaches, total removal of the tumors were completed in 22 cases, subtotal removal of tumors in 9 cases, and partial removal in 2 cases. In 5 cases with subfrontal approach, 4 patients the total removal of tumors obtained in 4 cases and one subtotal removal of tumor in one. Of 2 cases with the longitudinal fissure to the corpus callus approaches, one case was achieved with the total removal of tumor and the other with subtotal removal of tumor. The pituitary stalk was preserved in 8 cases, the pituitary stalk was severed in 9 cases and the pituitary stalk was not identified in 23 cases when surgery. 17 patients experienced diabetes insipidus and 12 patients had the hypothalamic hypofunction after surgery. One death occurred from the hypothalamic hypofunction 35 day after surgery. By follow-up from 3 months to 3 years, 22 patients returned normal life, 11 patients can carry out their self-life, and 6 patients need care.
CONCLUSIONSThe desirable removal of recurrent craniopharyngioma could be completed in the majority of patients although the reoperation of the tumors was performed very difficulty owing to the tumor adhesive to the surrounding hypothalamic structures.
Adolescent ; Adult ; Child ; Craniopharyngioma ; surgery ; Female ; Humans ; Male ; Microsurgery ; Middle Aged ; Neoplasm Recurrence, Local ; surgery ; Neurosurgical Procedures ; methods ; Pituitary Neoplasms ; surgery ; Reoperation ; Retrospective Studies ; Treatment Outcome
3.Changes of hypothalamus-pituitary hormones in patients after total removal of craniopharyngiomas.
Zhong-qing ZHOU ; Xiang-en SHI
Chinese Medical Journal 2004;117(3):357-360
BACKGROUNDThis paper aimed to elucidate the changes of hypothalamus-pituitary hormones in patients after total removal of craniopharyngiomas.
METHODSA total of 40 patients with craniopharyngiomas received surgery. The levels of triiodothyronine (T3), thyroxine (T4), thyrotropic hormone (TSH), antidiuretic hormone (ADH), and adrenocorticotropin (ACTH) were measured in the 40 patients before surgery and one week after surgery respectively.
RESULTSTwenty-eight patients (70%) had hypothyroidism before surgery, but 38 (95%) had hypothyroidism after surgery (P < 0.01). Twenty-three patients (57.5%) had diabetes insipidus (DI) before surgery and 38 (95%) had DI after surgery (P < 0.001). The pre- and post-operative levels of ADH were (2.49 +/- 0.30) pg/ml and (2.80 +/- 0.29) pg/ml respectively (P > 0.05), whereas those of ACTH were (23.97 +/- 2.69) pg/ml and (15.60 +/- 1.91) pg/ml respectively (P < 0.05).
CONCLUSIONSHormone deficits after total removal of craniopharyngioma appear to be the common complication of surgery. Hypothyroidism and diabetes insipidus are more frequent after surgery than before surgery. Thyroxine and glucocorticoids should be administered routinely after total removal of craniopharyngioma.
Adolescent ; Adult ; Child ; Craniopharyngioma ; surgery ; Female ; Humans ; Male ; Middle Aged ; Pituitary Hormones ; secretion ; Pituitary Neoplasms ; surgery ; Postoperative Complications ; Thyroid Hormones ; secretion
4.Effect of inducible nitric oxide synthase on intestinal microcirculation in endotoxic shock.
En-Yi SHI ; Xiao-Jing JIANG ; Han BAI ; Tian-Xiang GU ; Nakajima YOSHIKI
Acta Physiologica Sinica 2005;57(1):39-44
To investigate the changes of intestinal microcirculation in endotoxic shock and the effect of inducible nitric oxide synthase (iNOS) on intestinal microcirculation, endotoxic shock was induced by intravenous injection of lipopolysaccharide (LPS) in mice. Mean arterial pressure (MAP) was monitored throughout the experimental procedure. The velocity and flux of red blood cell (RBC) in villus tip arteriole and capillaries were measured by FITC-labeled erythrocytes and intravital microscopy. The effect of iNOS was determined by targeted disruption of mice iNOS-gene and administration of S-methylthiourea sulfate (SMT), a selective inhibitor of iNOS, before LPS injection. No significant differences in MAP, RBC velocity and flux at baseline were found among wild type mice, SMT pretreated mice and iNOS-gene knockout mice. LPS induced a dramatic fall of MAP in wild type mice. The decrease of MAP was significantly restored in iNOS-gene knockout mice and in wild type mice received SMT before LPS injection. The velocity and flux of RBC in villus tip arteriole and capillaries decreased markedly after LPS injection in wild type mice, while significantly higher velocity and flux of RBC were found in iNOS-gene knockout mice and SMT-pretreated mice both 60 and 120 min after LPS injection. The results demonstrate that iNOS plays an essential role in the intestinal microcirculation disturbance which occurs in endotoxic shock.
Animals
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Intestines
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blood supply
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Lipopolysaccharides
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Male
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Mice
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Mice, Knockout
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Microcirculation
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physiology
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Nitric Oxide Synthase Type II
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genetics
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physiology
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Shock, Septic
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chemically induced
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physiopathology
5.The correlation between Chlamydiae pneumonia and carotid atherosclerosis.
Zhi-wei TANG ; Xiang-en SHI ; Yong-kai CHANG ; Zhen WANG
Chinese Journal of Surgery 2013;51(5):421-425
OBJECTIVESTo study the correlation between Chlamydiae pneumonia and carotid atherosclerosis, and the correlation between the infection of Chlamydiae pneumonia and ischemic events.
METHODSThe study group consisted of 19 patients who underwent unilateral carotid endarterectomy surgery during the period from January 2010 to December 2011, and the atherosclerotic plaque specimens were harvested from these patients. The control group consisted of 10 patients who underwent extracranial-intracranial bypass surgery during the same period, and the normal external carotid artery specimens were got from these patients. The clinical data between the two groups had comparability. The presence of Chlamydiae pneumonia in atherosclerotic plaque and normal artery tissue were investigated by immunohistochemistry. The expression of Toll-like receptor 2 (TLR2), tumor necrosis factor-α (TNF-α) and vascular cell adhesion molecule-1 (VCAM-1) in the atherosclerotic plaque infected with Chlamydiae pneumonia were also detected. Data were analyzed using Fisher's exact test.
RESULTChlamydiae pneumonia was found in 9 of 19 atherosclerotic plaques, while no positive result was found in control group. The statistical analysis showed a significant difference (P = 0.011). Among the 19 patients in study group, 15 of them had ischemic events, and Chlamydiae pneumonia was found in 9 of these 15 patients; while the other 4 patients didn't have any ischemic events and no Chlamydiae pneumonia was found in them, but there was no statistical different between them (P = 0.087). Through immunohistochemistry, the expression of Chlamydiae pneumonia, TLR2, TNF-α and VCAM-1 were found in same area.
CONCLUSIONSThere is a correlation between Chlamydiae pneumonia and carotid atherosclerosis.And there might be a correlation between Chlamydiae pneumonia and cerebral ischemic events.
Carotid Artery Diseases ; metabolism ; microbiology ; Case-Control Studies ; Chlamydophila Infections ; complications ; Chlamydophila pneumoniae ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Plaque, Atherosclerotic ; microbiology ; Toll-Like Receptor 2 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism
6.Diagnosis and therapy of primary malignant tumors of the heart and pericardium.
Lei YU ; Tian-xiang GU ; En-yi SHI ; Zong-yi XIU ; Qin FANG
Chinese Journal of Oncology 2009;31(3):230-232
OBJECTIVETo summarize and analyze the experience in diagnosis and treatment of primary malignant tumors of the heart and pericardium.
METHODSThe clinical data of 24 patients with malignant tumors of the heart and pericardium treated in our hospital between Jun. 1980 and Mar. 2008 were retrospectively analyzed and the related literature was reviewed.
RESULTSAll 24 patients received surgical treatment. Radical resection in 10 cases and partial resection in 6 cases were performed, and the remaining 8 patients received only surgical exploration due to unresectable tumors. One patient died of cardiopulmonary failure in early postoperative stage and all the other patients died within 4 years after operation due to recurrence or metastases.
CONCLUSIONEchocardiography, CT, MRI, 3D-CT, CT of coronary artery and cardiac angiography are helpful for the diagnosis and selection of operation mode. Histological examination is necessary for the final diagnosis. Early diagnosis and surgical resection of the tumor as complete as possible, and combination with post-operative radiotherapy and/or chemotherapy may improve the survival of the patients.
Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Heart Neoplasms ; diagnosis ; surgery ; Humans ; Male ; Mesothelioma ; diagnosis ; surgery ; Middle Aged ; Myxoma ; diagnosis ; surgery ; Pericardium ; surgery ; Retrospective Studies ; Survival Rate ; Young Adult
7.Prognosis of different operative procedures of coronary artery bypass grafting in the senile patients.
Lei YU ; Tian-xiang GU ; En-yi SHI ; Qin FANG ; Chun WANG ; Yu-hai ZHANG
Chinese Journal of Surgery 2010;48(9):694-696
OBJECTIVETo investigate myocardial protection and prognosis of the different operative procedures of coronary artery bypass grafting (CABG) for the senile patients with coronary artery disease.
METHODSFrom July 2008 to June 2009, 125 cases (age ≥ 65 years old) underwent CABG were divided into 3 groups.Group A (70 cases) undertook off-pump CABG, group B (33 cases) undertook on-pump beating heart CABG. group C (22 cases) undertook on-pump CABG with cardiac arrest. Serum level of cardiac troponin I (cTnI) and creatine kinase MB isoenzyme (CK-MB) activity were examined right before operation and 0, 24, 72 and 168 h after operation. The complications during perioperative period and the mortality were investigated.
RESULTSNo significant difference was found in the right before operation and 168 h after operation of the serum level of cTnI and CK-MB in the 3 groups (P > 0.05). The serum level of cTnI and CK-MB activity of group A was the lowest and group B was lower than group C. Group A was the best concerning the complications during perioperative period. The mortality of group A was lower than group B but no significant difference was found in the mortality between group A and C. No significant difference was found in the complications during perioperative period and the mortality between group B and C.
CONCLUSIONOff-pump CABG could reduce myocardial injury and the complications during perioperative period in the senile patients.
Aged ; Aged, 80 and over ; Cardiopulmonary Bypass ; Coronary Artery Bypass ; methods ; Coronary Artery Bypass, Off-Pump ; Coronary Disease ; surgery ; Female ; Humans ; Male ; Prognosis
8.Effect and mechanism of panaxoside Rg1 on neovascularization in myocardial infarction rats.
Xiao-dong WANG ; Tian-xiang GU ; En-Yi SHI ; Chun-mao LU ; Chun WANG
Chinese journal of integrative medicine 2010;16(2):162-166
OBJECTIVETo investigate the effects and mechanisms of panaxoside Rg1 on the new vessel formation in acute myocardial infarction (AMI) rats.
METHODSThe AMI model of male Sprague-Dawley (SD) rats was established, and rats were randomly divided into the AMI model group, the treatment group of panaxoside Rg1, the placebo group and the treatment group of panaxoside Rg1 plus rapamycin. Cardiac creatases were determined with 1 mL blood drawn from vena caudalis of the rats 48 h after the model was successfully made. After 4 weeks, Evans blue was injected into the aorta roots of the rats, and then, red tetrazoline was dyed again and the myocardial infarction area was evaluated. The microvessel density (MVD) of infarction area was determined by the immunohistochemistry of CD31; enzyme-linked immunosorbent assay (ELISA) was used to detect the protein content of CD31 and hypoxia inducible factor-1alpha (HIF-1alpha) of the infarction area.
RESULTSThe MVD in the infarction area and the contents of CD31 and HIF-1alpha in the Rg1 treatment group were higher than those in the AMI model group significantly (P<0.05). The cardiac creatase and infarction area were lower in the Rg1 treatment group than those in the AMI model group significantly (P<0.05). The above effects, however, disappeared when rapamycin, the antagonist of mammalian target of rapamycin (mTOR), was administered simultaneously.
CONCLUSIONSPanaxoside Rg1 could increase the expression of HIF-1alpha and CD31 of myocardium and stimulate the angiogenesis. The above mentioned role of panaxoside Rg1 might be related to the excitation of mTOR receptor.
Animals ; Cell Count ; Collateral Circulation ; drug effects ; Drug Evaluation, Preclinical ; Ginsenosides ; administration & dosage ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Intracellular Signaling Peptides and Proteins ; metabolism ; physiology ; Male ; Microvessels ; pathology ; Myocardial Infarction ; drug therapy ; metabolism ; pathology ; Neovascularization, Physiologic ; drug effects ; Placebos ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; physiology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Sirolimus ; administration & dosage ; pharmacology ; TOR Serine-Threonine Kinases
9.Surgical treatment of vertebral artery-posterior inferior cerebellar artery dissecting aneurysms.
Zhi-Wei TANG ; Xiang-En SHI ; Yong-Li ZHANG ; Zhong-Qing ZHOU
Chinese Journal of Surgery 2010;48(23):1805-1810
OBJECTIVETo study an effective method for vertebral artery-posterior inferior cerebellar artery (VA-PICA) dissecting aneurysms.
METHODSFive patients with VA-PICA dissecting aneurysms were treated surgically between December 2007 and February 2010, who were all males, aged from 40 to 55 years. Four of them presented with subarachnoid hemorrhage (SAH), 1 of them suffered from an intermittent headache on occipital area about 4 months. All the aneurysms were trapped via a far-lateral approach after revascularization of the PICAs by occipital artery-posterior inferior cerebellar artery (OA-PICA) bypass.
RESULTSAmong the 5 patients, 1 of them got a good outcome without any neurological deficit after surgery, 3 of them had postoperative lower cranial nerve palsy, 1 of them complicated with bleeding in the operative field after postoperative antithrombotic treatment, and suffered from contralateral hemiplegia after second surgery of removing hematoma. Postoperative cerebral angiographies (received by 2 patients) and CT angiographies (received by 3 patients) all showed patent bypasses and no filling of the aneurysms. During the follow-up from 2 to 29 months, 4 patients got good outcomes, 1 patient still suffered from moderate hemiplegia.
CONCLUSIONSTrapping the aneurysms after revascularization of PICA by OA-PICA bypass should be an effective method to treat the VA-PICA dissecting aneurysms, but individualized strategies also need to be considered based on different cases.
Adult ; Cerebellum ; blood supply ; Follow-Up Studies ; Humans ; Intracranial Aneurysm ; surgery ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Vertebral Artery
10.Microsurgical treatment of ophthalmic segment aneurysms of internal carotid artery: 28 cases report.
Yong-li ZHANG ; Xiang-en SHI ; Yu-ming SUN ; Fang-jun LIU
Chinese Journal of Surgery 2010;48(12):911-914
OBJECTIVETo investigate the operative modalities and outcomes of 28 cases of ophthalmic segment aneurysms of internal carotid artery.
METHODSTwenty-eight cases of ophthalmic segment aneurysms of internal carotid artery were operated on from May 2004 to August 2009. Of all 28 cases, 20 were large or giant. Nineteen aneurysms were directly clipped or resected with internal carotid artery revascularization. Since 2006, high-flow extracranial-intracranial (EC-IC) bypass were available and performed in 9 patients of large or giant aneurysms and then the aneurysms were resected or trapped.
RESULTSSeventeen patients underwent digital subtraction angiography (DSA), computed tomography angiography (CTA) or magnetic resonance angiography (MRA) postoperatively. The images demonstrated that 5 grafts of bypass were in patency, and 2 were occluded. Only 1 aneurysm was partially clipped and the others disappeared on imaging. The 78% of these cases had good results (GOS 4-5). One patient died after EC-IC bypass due to neck hematoma.
CONCLUSIONSThe treatment of ophthalmic segment aneurysms of internal carotid artery, especially the large and giant ones, remains a challenge for neurovascular neurosurgeon. The accessory high-flow EC-IC bypass procedures and selection of suitable aneurysm clips are very important to improve the effectiveness of the operation.
Adolescent ; Adult ; Child ; Female ; Humans ; Intracranial Aneurysm ; surgery ; Male ; Microsurgery ; Middle Aged ; Ophthalmic Artery ; Retrospective Studies ; Treatment Outcome ; Vascular Surgical Procedures ; methods ; Young Adult