2.Cloning, prokaryotic expression, and functional identification of a sesquiterpene synthase gene (AsSS4) from Aquilaria sinensis.
Liang LIANG ; Qing-Mei GUO ; Zheng ZHANG ; Yan-Hong XU ; Xiao-Min HAN ; Juan LIU
Acta Pharmaceutica Sinica 2014;49(12):1724-1729
A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis
Alkyl and Aryl Transferases
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biosynthesis
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genetics
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Azulenes
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Cloning, Molecular
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DNA, Complementary
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Escherichia coli
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Open Reading Frames
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Polyisoprenyl Phosphates
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Recombinant Proteins
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biosynthesis
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Sesquiterpenes
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metabolism
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Sesquiterpenes, Guaiane
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Thymelaeaceae
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enzymology
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genetics
3.Effect of Shenfu Injection on Neuron Apoptosis Caused by Hypoxic-Ischemic Brain Damage in Neonatal Rats′ Hippocampus
jun, WANG ; li-juan, YANG ; yan-fang, HE ; qun-e, CHEN ; shao-mei, ZHANG
Journal of Applied Clinical Pediatrics 2006;0(21):-
Objective To study the effect of shenfu injection on the neuron apoptosis in hippocampal CA1 region of newborn with hypoxic-ischemic brain damage(HIBD).Methods The experiment included 2 parts.One was to measure the apoptosis rate by the flow cytometry,and the other was to investigate the expression of Bcl-2 and Bax of neurons in left hippocampal CA1 region.Models of postnatal 7-day Sprague-Dawley(SD)rats with HIBD were established,and were equally divided into 4 groups:sham operation(group S),control group(group C),shenfu injection pretreatment(group P),and shenfu injection treatment(group SF).The neuron apoptosis rate in hippocampal CA1 region in every group was measured at 2 h before and 2,12,24 h,3,7,14 and 28 d after hypoxic ischemic(HI) insult.The expressions of Bcl-2 and Bax were performed by immunohisochemistry.Results The apoptosis of neuron in hippocampal CA1 region in group P and group SF after HI insult was significantly less than that of group C(P
4.Association of HAb18G with clinicopathologic features and prognosis in non-small cell carcinoma of lung.
Xiao-yan XU ; Cheng ZHI ; Yu-mei LI ; Wen-juan QI ; Juan-juan MEI ; Zhi-min YAN ; Hong SHEN
Chinese Journal of Pathology 2012;41(3):151-155
OBJECTIVETo study the association between HAb18G expression, tumor parameters, metastatic potential and prognosis in non-small cell lung carcinoma (NSCLC).
METHODSImmunohistochemical study for HAb18G protein using SP methods was carried out in 144 cases of NSCLC. Nineteen cases of benign lung lesions and 41 cases of normal lung tissue were used as controls. The intensity (positive unit/PU) of HAb18G expression was assessed quantitatively by image analysis software. The results were correlated with tumor parameters, metastatic potential and follow-up data.
RESULTSThe intensity of HAb18G protein expression was significantly higher in NSCLC than that in controls (P = 0.000). In squamous cell carcinomas and adenocarcinomas, the expression of HAb18G protein in well-differentiated tumors was lower than that in moderately to poorly differentiated tumors (P = 0.001). Tumors of TNM stage IV had stronger expression than tumors of lower stages (P = 0.000). HAb18G PU was greater in tumors with lymph node metastasis than those without nodal metastasis (P = 0.045). The PU value of tumors with maximal diameter greater than 5 cm was higher than that of the smaller tumors (P = 0.000). It was also higher in male than in female patients (P = 0.046). There was no association between HAb18G protein expression and age of patients, history of smoking, tumor types and gross morphology (P > 0.05). The five-year survival rate in cases with low HAb18G protein expression was higher than that in cases with high expression (P = 0.006). Univariate analysis indicated that patients with high HAb18G protein expression carried a poor prognosis (P = 0.007). Multivariate analysis showed that expression of HAb18G protein was an independent prognostic factor in patients with NSCLC (P = 0.032, relative risk 3.962).
CONCLUSIONSHAb18G protein expression is associated with tumor progression and prognosis. It may represent a useful biomarker for prognostic evaluation.
Adenocarcinoma ; metabolism ; pathology ; Basigin ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Sex Factors ; Survival Rate ; Tumor Burden
5.Clinicopathological features and prognostic factors of 216 cases with primary gastrointestinal tract non-Hodgkin's lymphoma.
Wen-juan YIN ; Mei-juan WU ; Hai-yan YANG ; Xiu ZHU ; Wen-yong SUN
Chinese Journal of Hematology 2013;34(5):377-382
OBJECTIVETo investigate the clinicopathological features of primary gastrointestinal non-Hodgkin's lymphomas (PGI-NHL) and their prognostic values.
METHODSThe clinical and pathological data of 216 patients diagnosed as PGI-NHL from Zhejiang Cancer Hospital were analyzed retrospectively. χ² test, log-liner model analysis, COX proportional hazard regression analysis and Life-table survival analysis were used to analyze the survival status of the patients by SAS 8.2 software, and Log-rank test was performed to couple the overall survival rates with different prognostic factors.
RESULTSTotally, the age of onset was 8 to 89 years with the median age of 56.5 years. Male versus female was 1.27∶1(121∶95). The most frequently involved location was stomach (147 cases, 68.1%), followed by ileocecus (25 cases, 11.6%), large intestine (20 cases, 9.3%), small intestine (17 cases, 7.9%) and multiple GI involvement (5 cases, 2.3%). 182 cases were classified as B cell lymphomas, 22 cases as T cell lymphomas, and 12 cases not classified exactly due to insufficient data. The 3-year and 5-year survival rates of the patients were 69.4% and 53.3%, respectively. Univariate analysis revealed that age>60 years, ECOG≥2, high LDH level, stage Ⅲ-Ⅳ, IPI≥2, T cell type and intestinal involvement were predictors for poor prognosis. IPI≥2, T cell type and intestinal involvement were independent adverse predictors for prognosis by multiple COX proportional hazard regression analysis. Among different treatment groups, cases received chemotherapy combined with local radiotherapy gained the best survival status.
CONCLUSIONB-cell lymphoma was the main pathological type in PGI-NHL; IPI≥2, T-cell type and intestinal involvement are independent adverse predictors for prognosis; chemotherapy combined with local radiotherapy might be the choice of approach for advanced stage and aggressive PGI-HNL.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Gastrointestinal Neoplasms ; drug therapy ; pathology ; Humans ; Lymphoma, Non-Hodgkin ; drug therapy ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult
6.Cloning and expression analysis of cinnamate 4-hydroxylase (C4H) reductase gene from Aquilaria sinensis.
Liang LIANG ; Xiao-Min HAN ; Zheng ZHANG ; Qing-Mei GUO ; Yan-Hong XU ; Juan LIU ; Yong-Cui LIAO
China Journal of Chinese Materia Medica 2014;39(10):1767-1771
The study aimed to clone the open reading frame of cinnamate 4-hydroxylase (C4H) from Aquilaria sinensis and analyze the bioinformatics and expression of the gene. One unique sequence containing C4H domain was discovered in our previous reported wound transcriptome dataset of A. sinensis. The open reading frame of C4H was cloned by RT-PCR strategy with the template of mixed RNA extracted from A. sinensis stem which treated by different wound time. The bioinformatic analysis of this gene and its corresponding protein was performed. C4H expression profiles in responds to MeJA (methyl jasmonate) application were analyzed by real-time PCR. The length of C4H open reading frame (ORF) was 1 515 bp, encoding 514 amino acids. The GenBank accession number is KF134783. Inducible-experiments showed that the genes were induced by mechanical wound as well as MeJA induction, and reached the highest expression level at 8 h and 20 h, respectively. The full-length cDNA of C4H and its expression patterns will provide a foundation for further research on its function in the molecular mechanisms of aromatic compounds and flavonoids biosynthesis.
Amino Acid Sequence
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Cloning, Molecular
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Models, Molecular
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Molecular Sequence Data
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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 Proteins
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chemistry
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genetics
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metabolism
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Thymelaeaceae
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chemistry
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enzymology
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genetics
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Trans-Cinnamate 4-Monooxygenase
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chemistry
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genetics
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metabolism
7.Retrospective analysis of 71 cases of multiple myeloma.
Ping YANG ; Wen-Juan ZHANG ; Hong-Mei JING ; Xiao-Yan KE
Journal of Experimental Hematology 2009;17(6):1573-1576
The aim of this study was to investigate the efficacy of different chemotherapy regimens in patients with multiple myeloma (MM). The therapeutic effects of 71 MM patients receiving standard chemotherapy regimens were retrospectively analyzed and evaluated. The results showed that 44 out of 71 new-diagnosed MM patients gained remission in varying degree, total effective rate was 61.9%, in which the rate of complete remission (CR) plus nearly complete remission (nCR) was 21.1%. 21 MM patients received M2 regimen gained total effective rate of 57.1%, in which the CR plus nCR were found in 5 MM patients, and partial remission (PR) was observed in 7 MM patients. 8 MM patients received MP chemotherapy regimen gained total effective rate of 37.5%, in which the CR + nCR was not found, but the PR was observed in 3 MM patients. 30 MM patients received VAD regimen gained total effective rate of 63.3%, in which CR + nCR and PR were found in 6 and 13 MM patients respectively, 12 MM patients received combined bortezomib regimen gained total effective rate of 83.3%, in which CR + nCR and PR were found in 4 and 6 MM patients respectively. The median time of progression and the median time of survival in 72 MM patients were 22.1 and 29.5 months respectively. The 3 and 5 year survival rates in 72 MM patients were 41.2% and 20.6% respectively. In conclusion, the chemotherapy regimen for new-diagnosed MM patients should be selected according to their clinical features and subtypes, the bortezomib-combined regimen may be considered as a new and effective regimen for MM patients.
Adult
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Aged
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Aged, 80 and over
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Antineoplastic Combined Chemotherapy Protocols
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therapeutic use
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Boronic Acids
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administration & dosage
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therapeutic use
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Bortezomib
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Female
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Humans
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Male
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Middle Aged
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Multiple Myeloma
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drug therapy
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Pyrazines
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administration & dosage
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therapeutic use
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Retrospective Studies
8.Influence of H102 on the expression of amyloid protein and amyloid precursor protein in the hippocampus of APP695 transgenic mice.
Yan-Ling XU ; Juan ZHAO ; Rui-Yu MA ; Shu-Mei XU
Chinese Journal of Applied Physiology 2010;26(3):302-306
OBJECTIVETo observe the influence of H102 on the expression of amyloid protein and amyloid precursor protein in the hippocampus of APP695 transgenic mice.
METHODSThe 9-month-old APP695 transgenic mice were randomly divided into the model group and the H102 group; C57BL/6J mice were adopted as normal control group. The H102 group were injected with H102 in a dose of 3 microl/per mouse in lateral ventricle, once a day, for ten days; while the model group and the control group were injected with saline. The hippocampus and temporal cortex of the brain sections from transgenic mice and wild type female mice were subjected to immunohistochemistry and Congo red histological staining, and observed the difference of the protein expression under microscope. The expression of the APP protein was detected by Western blot.
RESULTSAbeta and APP immunohistochemistry showed density of positive cell in the CA1 region of hippocampus of control group were less than model group. H102 peptide reduced the area, and density of positive cells. Congo red staining showed there were lots of amyloid plagues in the brains of model mice but not in the brains of normal control. And the Western blot showed the content of the APP protein of the model group was much higher than the H102 group. H102 significantly decreased the amyloid plagues.
CONCLUSIONThe expression of APP, Abeta are increased in APP695 transgenic mice, and H102 can decrease the level of APP, Abeta in transgenic mice.
Amyloid beta-Protein Precursor ; metabolism ; Amyloidogenic Proteins ; metabolism ; Animals ; Brain ; metabolism ; Female ; Gene Expression ; Hippocampus ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic
9.Chemical constituents of Lepidium meyenii.
Wen-juan LIANG ; Hong-bo XU ; Cai-yan YANG ; Chang-an GENG ; Zhang XUE-MEI ; Ji-jun CHEN
China Journal of Chinese Materia Medica 2015;40(23):4531-4535
To study the chemical constituents of Lepidium meyenii, the air-dried rhizome of L. meyenii was extracted with 70% EtOH. The extract was condensed to a small amount of volume and extracted with petroleum ether, EtOAc and n-BuOH, successively. The compounds were isolated and purified by column chromatography, and identified based on spectral analyses (1H-NMR, 13C-NMR, HRESIMS). Eighteen compounds were isolated from L. meyenii, including 7 alkaloids and 4 fatty acids and 7 other compounds. They were characterized as (3-hydroxybenzyl) carbamic acid(1), phenylmethanamine(2), N-benzylformamide (3), N-benzylacetamide (4), pyridin-4-ylmethanamine(5), n-(4-methoxybenzyl) aniline(6), uracil(7), succininc acid(8), decanedioic acid(9), n-hexa- decanoic acid methyl ester(10), heptanoic acid(11), solerole(12), pyromucic acid methyl ester(13), 5-hydroxymethyl-2-furancar- boxadehyde(14), 5-(methoxymethyl)-1H-pyrrole-2-carbaldehyde(15), 1,7-dihydroxy-2,3, 4-trimethoxyxanthone (16), 1,7-di- hydroxy-3,4- dimethoxy-xanthone(17), (+)-pinoresinol(18). Meanwhile, compounds 1-18 were obtained from L. neyenii for the first time.
Lepidium
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chemistry
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Molecular Structure
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Plant Extracts
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chemistry
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Spectrometry, Mass, Electrospray Ionization
10.Inhibition of osthole for resorption of rats femur tissue in vitro.
Jian ZHOU ; Xue-mei REN ; Xiao-ni MA ; Yu-hai GAO ; Li-juan YAN ; Wen-gui SHI ; Ke-ming CHEN
China Journal of Orthopaedics and Traumatology 2015;28(9):832-837
OBJECTIVETo investigate osthole effect on femoral tissue resorption activity of rat in vitro.
METHODSSix SD rats weighted (80 ± 5) g were used to isolate and culture femoral tissue (diaphyses and metaphysis) in vitro. The cultured tissue were devided into control group, estradiol group and osthole group. The femoral tissue was treated with final concentration of 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol culture in vitro at 48 hours after cultured. Tartrate-resistant acid phosphatase (StrACP) activity, glucose and Lactic acid content, StrACP, MCSF (Macrophage colony stimulating factor) and CTSK (Cathepsin K) mRNA was detected by Real-Time RT-PCR were detected.
RESULTSConcetration of Alkaline phosphatase activity were 2226 and 2498 in 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol respectively. As compared with control group, the activity of StrACP of 1 x 10(-5) mol/L osthole and 1 x 10(-8) mol/L estradiol were inhibited at 6, 9, 12 days (P < 0.05); under treatment of in l x 10(-5) mol/L osthole, the content of Lactic acid were increased and the content of glucose were decreased at 3, 6, 9 days (P < 0.05); StrACP, MCSF and CTSK mRNA expression level were inhibited at 6, 9 days (P < 0.05).
CONCLUSIONOsthole can inhibit bone resorption and raise the level of nutrition metabolism of femurs tissue.
Acid Phosphatase ; metabolism ; Animals ; Bone Resorption ; prevention & control ; Coumarins ; pharmacology ; Estradiol ; pharmacology ; Femur ; drug effects ; Glucose ; analysis ; Lactic Acid ; analysis ; Male ; Rats ; Rats, Sprague-Dawley