1.Cloning and bioinformatics analysis of abscisic acid 8'-hydroxylase from Pseudostellariae Radix.
Jun LI ; Deng-Kai LONG ; Tao ZHOU ; Ling DING ; Wei ZHENG ; Wei-Ke JIANG
China Journal of Chinese Materia Medica 2016;41(13):2397-2403
Abscisic acid 8'-hydroxylase was one of key enzymes genes in the metabolism of abscisic acid (ABA). Seven menbers of abscisic acid 8'-hydroxylase were identified from Pseudostellaria heterophylla transcriptome sequencing results by using sequence homology. The expression profiles of these genes were analyzed by transcriptome data. The coding sequence of ABA8ox1 was cloned and analyzed by informational technology. The full-length cDNA of ABA8ox1 was 1 401 bp,with 480 encoded amino acids. The predicated isoelectric point (pI) and relative molecular mass (MW) were 8.55 and 53 kDa,respectively. Transmembrane structure analysis showed that there were 21 amino acids in-side and 445 amino acids out-side. High level of transcripts can detect in bark of root and fibrous root. Multi-alignment and phylogenetic analysis both show that ABA8ox1 had a high similarity with the CYP707As from other plants,especially with AtCYP707A1 and AtCYP707A3 in Arabidopsis thaliana. These results lay a foundation for molecular mechanism of tuberous root expanding and response to adversity stress.
2.Cloning and expression analysis on full length CDS of zeaxanthin epoxidase gene in Pseudostellaria heterophylla.
Wei ZHENG ; Tao ZHOU ; Jun LI ; Deng-Kai LONG ; Wei-Ke JIANG ; Ling DING
China Journal of Chinese Materia Medica 2017;42(4):669-674
Zeaxanthin epoxidase plays an important role in indirect pathway of plant abscisic acid biosynthesis. According to the data of Pseudostellaria heterophylla transcriptome, zeaxanthin epoxidase gene was isolated and named as PhZEP. The results of bioinformatics analysis showed that the coding sequence of PhZEP was 1 263 bp long and encoded 420 amino acids. The putative protein molecular weight was 47.34 kDa and its theoretical isoelectric point was 6.64. The characteristic structure domains were predicted, including binding site of lipoprotein and flavoprotein monooxyenase. A signal peptide was discovered at the N-terminal of amino acids. The Real-time PCR revealed that PhZEP had a higher expression level in leaves than other tissues of P.heterophylla. Highly expressed PhZEP was also observed at 10 d and 40 d tuberous root after flowering. PhZEP presented a different expression after treatment with ABA, fluridone and ABA +fluridone compared to the control. The expression of PhZEP in tuberous root after ABA treatment was close to that in control group, while PhZEP showed significant up-regulation in the fluridone treatment group. In this study, the PhZEP gene from P. heterophylla was cloned and this result has important significance for its functional identification. This research provides a basis for the further analysis on functional mechanism of ABA during development of P. heterophylla.
3.Cloning and expression analysis of four carotenoid cleavage dioxygenases genes from Pseudostellaria heterophylla.
Jun LI ; Deng-Kai LONG ; Jun LI ; Tao ZHOU ; Wei ZHENG ; Ling DING ; Wei-Ke JIANG
China Journal of Chinese Materia Medica 2016;41(13):2404-2410
To investigate the molecular mechanism of quality formation of Pseudostellaria heterophylla, the carotenoid cleavage dioxygenases (CCDs) genes were cloned from the transcriptome database of P. heterophylla, and analyzed them with bioinformatics analysis and expression analysis. The sequence length of four new gene were 1 617, 1 461, 1 746, 1 875 bp, and subsequently, named as PhCCD1,PhNCED2,PhNCED3 and PhCCD4 according to its genetic relationship with Arabidopsis thaliana. The sequence analysis showed that four new gene were all containing REP65 domains and binding sites of ferrous ion, such as histidine, glutamates and aspartates. Analysis phylogeny showed that PhNCED2 and PhNCED3 were the cluster of NCEDs, PhCCD1 and PhCCD4 were the cluster of CCDs. In addition, PhCCD1 and AtCDD1 of Arabidopsis thaliana, PhCCD4 and AtCCD4 of A. thaliana,PhNCED2, PhNCED3 and AtNCED3 of A. thaliana have high similarities. Analysis of real-time fluorescence quantitative showed that PhNCED2 and PhNCED3 were expressed mainly in underground part, the expression quantity of PhNCED2 reached the highest in fibrous root, PhNCED3 keeps higher in phloem and xylem, it may be the key enzymes of ABA biosynthesis genes. Moreover,PhCCD1 and PhCCD4 were expressed mainly in aerial part,the expression quantity of PhCCD1 reached the highest in leaf,PhCCD4 keeps higher in stem and leaf.It may be involved in the biosynthesis of carotenoids for P. heterophylla. The study obtained CDDs gene of P. heterophylla for the first time,this would lay the foundation of developing the response mechanism of P. heterophylla about external stress further,and then exploring the biological approach of quality formation in P. heterophylla.
4.Breeding and extension of Pseudostellaria heterophylla new variety "Shitai No.1" in Guizhou province.
Cheng-Hong XIAO ; Wei-Ke JIANG ; Tao ZHOU ; Ming-Wu LIAO ; Chang-Gui YANG ; En ZHANG
China Journal of Chinese Materia Medica 2016;41(13):2381-2385
Based on collections and researches of Pesudostellaria heterophylla germplasm resources from different areas of China, by using Shibing SB-4 provenance as materials, the new variety "Shitai No.1" was bred by mass selection, small plot variety comparative test, regional variety comparative test and field trial planting. Compared with "Qian taizishen No.1" and P. heterophylla land races. The disease and lodging resistance, root yield, polysaccharide content and the first grade rate of "Shitai No.1" have obvious advantages. In addition, it is relatively stable of yield in "Shitai No.1" in different places. It is demonstrated that "Shitai No.1" is a fine variety that adapt to the producing areas of P. heterophylla in Guizhou province, it is worthy to be promoted.
5.Pesudostellariae Radix industry status and development countermeasures.
China Journal of Chinese Materia Medica 2016;41(13):2377-2380
In recent years, with increasing role in traditional Chinese medicine industry of many provinces in china, Pseudostellariae Radix is one of the rapid development traditional Chinese medicine. Based on references and on-site investigation of main producing area, we clear the process of change from the small of the Panax ginseng to the Pseudostellariae Radix in aspects of history, and the change of producing area from Shandong, Jiangsu provinces to Fujian, Anhui and Guizhou provinces. And we clear how the Pseudostellariae Radix become a commonly used Chinese medicinal herbs from a small variety based on the market supply and demand and price changes, and it has gradually become a hot topic of the research. This paper summarized the status quo and the existing problems in the Pseudostellariae Radix industry. On this basis, we put forward countermeasures and suggestions for the development of the Pseudostellariae Radix industry.
6.Recent research on tofacitinib in the treatment of pediatric rheumatic diseases.
Shi-Hai ZHOU ; Ya-Qun XIONG ; Ya CHEN
Chinese Journal of Contemporary Pediatrics 2022;24(4):447-453
Tofacitinib is a Janus kinase inhibitor and can block the Janus kinase-signal transducer and activator of transcription signal transduction pathway and reduce the production and release of a variety of cytokines. It has great potential in the treatment of various rheumatic diseases with a rapid onset of action and can reduce corticosteroid dependence and related adverse events. The therapeutic effect of tofacitinib in adult patients has been confirmed, and it has been increasingly used in pediatric patients in recent years. This article reviews the clinical application of tofacitinib in the treatment of pediatric autoimmune diseases.
Adult
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Child
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Humans
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Janus Kinases/metabolism*
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Piperidines/therapeutic use*
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Protein Kinase Inhibitors/therapeutic use*
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Pyrimidines/therapeutic use*
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Rheumatic Diseases/drug therapy*
7. Correlation Between RhoA Gene in Wnt/PCP Signaling Pathway and Traditional Chinese Medicine Syndrome of AECOPD
Jun ZHANG ; Zheng-xing GE ; Xun ZHOU ; Yi YANG
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(16):73-77
Objective: To investigate the correlation between Ras homolog gene family member A (RhoA) gene in Wnt/PCP signaling pathway and acute exacerbation chronic obstructive pulmonary disease (AECOPD) traditional Chinese medicine(TCM)syndrome, attempting to provide an objective standard for the diagnosis of AECOPD TCM syndrome. Method: The 100 AECOPD patients were collected and divided into 5 groups:phlegm turbid obstructing lung syndrome,, phlegm-heat obstructing lung syndrome, syndrome of orifices confused by phlegm, deficiency of pulmonary and renal Qi, and edema due to yang deficiency, with 20 people in each group. 15 normal people were selected as a normal control group. All patients received fasting hemospasia, using a kit to extract blood total RibonucleicA(RNA) according to instructions. Real-time quantitative polymerase chain reaction (Real-time PCR) was adopted to detect the mRNA expression of RhoA gene in blood of patients with AECOPD TCM syndrome, and to explore the correlation. Result: There was no difference between phlegm-heat obstructing lung syndrome group and syndrome of orifices confused by phlegm group. The mRNA expression of RhoA gene in phlegm turbid obstructing lung syndrome group, phlegm-heat obstructing lung syndrome group, syndrome of orifices confused by phlegm group, deficiency of pulmonary and renal Qi group, and edema due to Yang deficiency group were significantly higher than that in normal group (P<0.01). The mRNA expression of RhoA gene in five TCM syndrome groups was as follows:phlegm turbid obstructing lung syndrome < phlegm-heat obstructing lung syndrome/syndrome of orifices confused by phlegm < deficiency of pulmonary renal Qi < edema due to Yang deficiency. There was a positive linear correlation between the relative expression of RhoA mRNA and above five AECOPD TCM syndrome groups. Conclusion: The significant difference in mRNA relative expression of RhoA gene in Wnt/PCP signaling pathway among the five AECOPD TCM syndrome groups may provide some objective diagnostic criteria for AECOPD TCM syndromes and reveal their disease severity.
8.Study on commodity specification and grade standard and quality evaluation of Dipsaci Radix.
Wang-Yin FENG ; Tao ZHOU ; Cheng-Hong XIAO ; Wei-Ke JIANG ; Zhi-Lai ZHAN ; Chang-Gui YANG ; Cheng-Gang ZHANG ; Yan LI
China Journal of Chinese Materia Medica 2019;44(14):2996-3001
Through the records of herbal and market investigations,the samples collected from different market and origin were analyzed,and the appearance character indexes were determined in order to revise the commodity specification and grade standard of Dipsaci Radix combined with production practice. This is also to analyze association of the appearance with quality different of intrinsic components. The investigation results indicated the root's long and thick was better,and atrovirens of fracture surface,there are most of the market is dominated by the ungraded goods. Through principal component analysis,variance analysis and cluster analysis,combining with the feasibility of actual operation and herbal record,the length,middle diameter and phloem color were filtrated and the specification was divided into 2 ranks: the selection and ungraded goods,and the grade was divided into 2 ranks for the selection goods: big,small selected goods. Moreover,there were no significant correlation between the appearance and the intrinsic quality index of Dipsaci Radix,the content of extractum were significantly positive correlation with the moisture,but had extremely negative correlation with the total ash. Multiple comparisons indicated that the content of the moisture,extractum and asperosaponin VI of the selected goods were higher than the ungraded goods,while the total ash content were lower,and they hasn't significant difference. The commodity specification and grade standard of Dipsaci Radix as a basis provide commodity specification and grade standard of communities and standardizing market trade order.
Cluster Analysis
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Dipsacaceae
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chemistry
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Drugs, Chinese Herbal
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standards
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Plant Roots
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chemistry
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Plants, Medicinal
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chemistry
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Principal Component Analysis
9.Changes of blood routine parameters in elderly apheresis platelet donors
Chang TAN ; Xin CUI ; Yuan ZHOU
Chinese Journal of Blood Transfusion 2022;35(11):1151-1154
【Objective】 To analyze the changes of blood routine and platelet related parameters of the elderly apheresis platelet donors since the first donation, so as to ensure the health and platelet quality of the elderly apheresis donors. 【Methods】 From 2019 to 2021, a total of 69 apheresis platelet donors in our center, aged 55-60 years old, involving 3400 occasions of donation, were enrolled in this study. The change trends of blood routine and platelet related parameters before and after the age of 55 were retrospectively analyzed. One way ANOVA was used for group comparison, and Pearson correlation analysis of platelet parameters was performed. 【Results】 The values of blood routine and platelet related parameters were higher in men than in women (P<0.05) when stratified by ages. As for the females in comparison of the parameters before 55 years old, RBC, WBC, Hb and Hct after 55 years old decreased significantly (P<0.05), while the platelet parameters of PDW, MPV and P-LCR elevated (P<0.05), and Plt remained unchanged (P>0.05). As for the males in comparison of the parameters before 55 years old, WBC decreased significantly after 55 years of age (P<0.05), while the platelet parameters of PLT, PDW, MPV and P-LCR elevated(P<0.05), and RBC, Hb and Hct remained with no statistical differences (P>0.05). The changes of PDW, MPV and P-LCR were consistent and showed a linear positive correlation. 【Conclusion】 The blood routine tests of the elderly apheresis platelet donors were all within the normal range and the proportion was relatively stable. Personalized blood donation schemes should be customized for the elderly blood donors and the health management of blood donors should be implemented.
10.Determination of militarine in Rhizoma Bletillae by HPLC.
Xun HE ; Aimin WANG ; Yongjun LI ; Yonglin WANG ; Shuyong ZHOU
China Journal of Chinese Materia Medica 2009;34(16):2076-2078
OBJECTIVETo develop an HPLC method for determination of militarine in Rhizoma Bletillae.
METHODAnalysis was carried out on a Symmetry C18 analytical column, and a C18 guard column eluted with mobile phase consisted of acetonitrile and water (37:63) militarine the detection wavelength was set at 244 nm.
RESULTThe contents of in 12 samples of Rhizoma Bletillae in market were in the ranges of 1.45%-3.32%.
CONCLUSIONThe method is simple, accurate and reproducible, and is suitable for the quality control of Rhizoma Bletillae.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Orchidaceae ; chemistry ; Succinates ; analysis