2.Correlation of macrophage-related cytokines and silent information regulator 1 and forkhead box protein O3 levels in peripheral blood mononuclear cells in patients with active pulmonary tuberculosis
Lang XIAO ; Hui-min LIU ; Qing-liang HU
China Tropical Medicine 2022;22(12):1179-
Abstract: Objective To explore the correlation between the levels of silent information regulator 1 (SIRT1) and forkhead box protein O3 (FOXO3) in peripheral blood mononuclear cells of patients with active pulmonary tuberculosis (APTB) and macrophage-related cytokines-inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1). Methods A total of 64 APTB patients who were treated in Yubei Hospital, the First Affiliated Hospital of Chongqing Medical University from January 2020 to December 2021 were gathered as the APTB group, 59 people with latent tuberculosis infection (LTBI) were gathered as the LTBI group, and 62 healthy people were gathered as the control group. Quantitative real-time PCR (qPCR) method was performed to measure the levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells. The enzyme-linked immunosorbent assay (ELISA) was performed to measure serum iNOS and Arg-1 levels; ROC curve was used to analyze the value of SIRT1 mRNA and FOXO3 mRNA levels in the differential diagnosis of LTBI and APTB; Pearson correlation was performed to analyze the correlation of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients with serum iNOS and Arg-1 levels. Results The levels of SIRT1 mRNA, FOXO3 mRNA and serum iNOS in peripheral blood mononuclear cells decreased in control group, LTBI group and APTB group, and the level of serum Arg-1 increased in turn (P<0.05). The AUCs of SIRT1 mRNA and FOXO3 mRNA in differential diagnosis of LTBI and APTB were 0.876 and 0.887, respectively, the sensitivity was 71.2% and 76.3%, and the specificity was 96.9% and 90.6% respectively. The levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients were positively correlated (r=0.500, P<0.05), and they were positively correlated with serum iNOS and negatively correlated with serum Arg-1 (P<0.05). The SIRT1 mRNA, FOXO3 mRNA and serum iNOS in peripheral blood mononuclear cells of APTB patients after 6 months of treatment were higher than those before treatment, and serum Arg-1 was lower than before treatment (P<0.05). Conclusions The levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients are low, and they are positively correlated with macrophage-related cytokine iNOS and negatively correlated with Arg-1.
3.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
4.Changed accumulation of active ingredient in different localities and growth period of Hemsleya zhejiangensis (Cucurbitaceae).
Wang-Wei YANG ; Zu-Pei LEI ; Wei-Min WANG ; Wei-qing LIANG ; Wei-Qing ZHOU ; Xiao-Feng JIN
China Journal of Chinese Materia Medica 2014;39(16):3123-3127
In this paper, the content of moisture, ethanol-soluble extractives, total saponins and polysaccharide of different tuber samples of Hemsleya zhejiangensis, from different localities, years and seasons, were detected based upon Chinese Pharmacopoeia 2010 version. The samples of roots, stems and leaves in summer were detected as well. The results are mainly as follows. (1)With tuber quality increasing, the content of total saponins increased and then decreased. The individual quality of tubers getting 594.06 g, the content of total saponins reached the peak. (2) The content of active ingredients in different localities was significantly different, and the population of Wuyanling had the maximum content of total saponins and polysaccharide. (3) The content of active ingredients revealed stability between the years 2012 and 2013, but the content of polysaccharide was significantly different. The content in 2012 was higher than that of 2013. (4) The content of active ingredients reached the peak in autumn, which was the best harvest season. (5) Among different component content detection of nutritional organs, tubers had the maximum content of ethanol-soluble extractives, total saponins and polysaccharide. Leaves also contained higher content of ethanol-soluble extractives and total saponins than roots and stems. All of these provide theoretical basis for plant, harvest and production of H. zhejiangensis, which is an endemic, rare, and endangered medicinal plants.
China
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Cucurbitaceae
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chemistry
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growth & development
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metabolism
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Drugs, Chinese Herbal
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analysis
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metabolism
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Plant Leaves
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chemistry
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growth & development
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metabolism
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Plant Roots
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chemistry
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growth & development
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metabolism
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Plant Tubers
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chemistry
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growth & development
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metabolism
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Plants, Medicinal
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chemistry
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growth & development
;
metabolism
5.Efficacy of water knife needle release combined with bone peptide injection for heel pain.
Wen-zhi CHEN ; Qing-liang SHEN ; Hui-min WANG ; Han-qing WU
Journal of Southern Medical University 2010;30(8):1953-1955
OBJECTIVETo evaluate the efficacy of water knife needle release combined with bone peptide injection in the management of heel pain.
METHODSThirty-five patients with unilateral heel pain were treated with water knife needle release and bone peptide injection under local anesthesia. The deep tissue with the tenderness was released in the operation, and the result was evaluated 1 week after the surgery to decide whether to conduct another surgery. No more than 3 treatment sessions were administered. The efficacy was evaluated according to nimodipine method by the principles of Chinese clinical drug guidance, and the complications of the surgery were observed.
RESULTSSix months after the surgery, 28 cases had excellent results, 3 had good outcomes, 2 showed improvement, and 2 failed to respond favorably, with a rate of good and excellent result of 94.2%. No adverse side effect was recorded in the follow up of the patients.
CONCLUSIONWater needle knife release combined with bone peptide injection can produce a good result in the treatment of heel pain.
Adult ; Aged ; Female ; Foot Diseases ; therapy ; Heel ; Humans ; Injections ; Male ; Middle Aged ; Needles
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
;
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.Expression of mCD_(14) on Alveolar Macrophage of Neonates with Respiratory Distress Syndrome Treated by Me-chanicalventilation
li-min, KANG ; lei, CAO ; cui-qing, LIU ; xi-qun, JIA ; qiu-jin, LIANG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To investigate the mechanism of mCD14 expression on AM in the pathogenes of neonatal respiratory distress syndromes( NRDS). Methods The expression of mCD14 on AM was analyzed with flow cytometry. Enzyme - linked immunosorbent assay was performed for detecting the concentration of IL- 1? and IL-8.Results The percentage of mCD14 positive AM in experimental group [(54.772 ?17 .341)%] was higher than that in control group [(14.023? 10. 713)% ](t= -7.739 P
8.Simultaneous quantitation of artemisinin, arteannuin B, artemisic acid, and scopoletin in mice plasma by HPLC-MS.
Qing MIN ; Wei LU ; Man-Yuan WANG ; Dong ZHANG ; Tian-Yan ZHOU ; Liang LI
China Journal of Chinese Materia Medica 2014;39(17):3306-3310
The objective of this study is to develop a sensitive and reliable high-performance liquid chromatography mass spectrometry (LC-MS) method for simultaneous determination of artemisinin, arteannuin B, artemisic acid, and scopoletin, and study the pharmacokinetics of the four constituents in mouse serum after oral administration of the four components to mice. The analytical column used was Agilent Zorbax SB-C18 (2.1 mm x 150 mm, 5 mm). The mobile phase was acetonitrile: 0.5% acetic acid (60: 40) and the flow rate was 0.3 mL x min(-1). The temperature of the column was 40.0 degrees C. In this condition, we established an analysis method to simultaneously determine the four components. A sensitive and specific liquid chromatography-mass spectrometric (LC-MS) method was developed and validated for the determination of artemisin in derivatives in mice plasma. The method we established has a linear range of 5-3 000 μg x L(-1) with a good sensitivity and specificity for all of the four components. This method is simple, rapid, accurate and suitable for the determination of the content of the four compounds.
Animals
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Artemisinins
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blood
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pharmacokinetics
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Chromatography, High Pressure Liquid
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instrumentation
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methods
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Dose-Response Relationship, Drug
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Male
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Mice
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Reproducibility of Results
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Scopoletin
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blood
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pharmacokinetics
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Spectrometry, Mass, Electrospray Ionization
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methods
9.Mutating Escherichia coli by atmospheric and room temperature plasmas for succinic acid production from xylose.
Qing WAN ; Weijia CAO ; Changqing ZHANG ; Rongming LIU ; Liya LIANG ; Kequan CHEN ; Jiangfeng MA ; Min JIANG
Chinese Journal of Biotechnology 2013;29(11):1692-1695
Escherichia coli AFP111 is a spontaneous mutant with mutations in the glucose specific phosphotransferase system (ptsG) in NZN111 (delta pflAB deltaldhA). In AFP111, conversion of xylose to succinic acid generates 1.67 molecule of ATP per xylose. However, the strain needs 2.67 molecule ATP for xylose metabolism. Therefore, AFP111 cannot use xylose due to insufficient ATP under anaerobic condition. Through an atmospheric and room temperature plasma (ARTP) jet, we got a mutant strain named DC111 that could use xylose under anaerobic condition in M9 medium to produce succinic acid. After 72 h, DC111 consumed 10.52 g/L xylose to produce 6.46 g/L succinic acid, and the yield was 0.78 mol/mol. Furthermore, the reaction catalyzed by the ATP-generating PEP-carboxykinase (PCK) was enhanced. The specific activity of PCK was 19.33-fold higher in DC111 than that in AFP111, which made the strain have enough ATP to converse xylose to succinic acid.
Atmosphere
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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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Industrial Microbiology
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Metabolic Engineering
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Mutation
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Plasma Gases
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pharmacology
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Succinic Acid
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metabolism
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Temperature
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Xylose
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metabolism
10.Approach to the patients with transient neonatal diabetes mellitus
Qing GU ; Zhe ZHANG ; Min CHEN ; Yin CHEN ; Hua LIANG ; Chengjiang LI
Chinese Journal of Endocrinology and Metabolism 2016;32(6):510-513
[Summary]_ Transient neonatal diabetes mellitus is a kind of rare special types of diabetes. It should be distinguished from type 1 diabetes. Genetic analysis can be used to define the subtype of neonatal diabetes mellitus, which helps us to select the most appropriate treatment and to predict the disease recurrence. Sulfonylureas is able to improve insulin secretion in most patients with transient neonatal diabetes mellitus and provide effective glycemic control. A case of transient neonatal diabetes mellitus is reported in order to call attention to the diagnosis and treatment of this disease.