Correlations between content of linarin in Chrysanthemum indicum and climatic factors in habitats / 中国中药杂志

Chrysanthemi Indici Flos(CIF), the capitulum of Chrysanthemum indicum, is widely used in proprietary Chinese medicine and daily chemical products. At present, CIF is mainly produced from wild resources and rarely cultivated. This study aims to reveal the correlations between linarin content in CIF and climatic factors in different habitats, and provide a theoretical basis for suitable zoning and rational production of medicinal materials. The content of linarin in CIF was determined by HPLC. Grey relational analysis and Pearson correlation analysis were carried out for linarin content with climatic factors. The results showed that the content of linarin in CIF was significantly different among different habitats. The grey relational degrees of climatic factors with linarin content was in an order of average annual precipitation>annual average sunshine hours>annual average temperature>longitude>annual frost-free period>latitude>altitude. Longitude, annual average temperature and average annual precipitation had significantly positive correlations with the content of linarin in CIF, whereas latitude and altitude showed negative correlations with it. The annual frost-free period and annual average sunshine hours had no significant correlation with the content of linarin in CIF. The content of linarin in CIF varied significantly in different habitats. High longitude, low latitude, low altitude, high annual average temperature and high annual average precipitation could be used as indicators for the habitats of high-quality Ch. indicum. This study provides a reference for selecting suitable producing areas of Ch. indicum and establishing artificial cultivation system.

Application of ITS2 secondary structure phylogenetic information in DNA barcode identification of Chrysanthemum indicum and its related plants / 中国中药杂志

By exploring additional phylogenetic information hidden in ITS2 secondary structure,the possibility of identifying Chrysanthemum indicum and its related species with DNA barcode of ITS2 nucleic acid sequence and its structure information were discussed.The genomic DNA was extracted from 12 samples. The ITS2 fragments were amplified by PCR and sequenced bidirectionally to obtain ITS2 sequence information. 28 sequences of related species for Ch. indicum were downloaded from Gen Bank. Until all 40 ITS2 sequences were aligned,ITS2 secondary structure prediction and structure comparison were finished. Then ITS2 secondary structure information was coded. After comparing ITS2 structure information and nucleic acid information,MP phylogenetic trees were built. The results showed that the secondary structures of ITS2 shared the same structure model--a four-fingered hand. They not only have the common characteristics of ITS2 secondary structures in plants,but also have many other conservative sequences,and their overall conservativeness is high. Among all species used in this study,their ITS2 secondary structures had obvious difference. In addition,the number of mutation sites in the joint matrix compared with the nucleic acid sequences increased by nearly 90%,which greatly enriched the number of mutation sites. This method of information analysis distinguished Ch. indicum from its related species. At the same time,the support rate of the branches of evolutionary trees and the identification rate of species were significantly improved. Although there was no distinction between Ch. zawadskii and Ch. morifolium,it effectively distinguished the three species,namely,Ch. hypargyrum,Ch.oreastrum,and Ch. dichrum. Therefore,the authors suggest that the ITS2 sequence combined with its structural data information should be applied to the identification of Ch. indicum and its related species,and be widely applied to DNA barcode research.

Identification of Chrysanthemum indicum and its adulterants based on ITS2 barcode / 中国中药杂志

DNA barcode technology was used to establish a rapid identification method of Chrysanthemum indicum based on ITS2 sequences. The total DNA was extracted from 22 collected samples,and the ITS2 sequence was amplified by PCR and sequenced,and the information of ITS2 sequence was obtained. Another 14 items of the same family or the same genus were downloaded from Gen Bank.We aligned all 36 sequences,calculated the intraspecific and interspecific distances,and constructed Neighbor Joining( NJ) phylogenetic tree,using MEGA 7. 0. The difference of the secondary structure between the ITS2 sequences was compared. The results showed that the genetic distance of Ch. indicum and Ch. morifolium was overlapped,but the maximum intraspecific distance was far less than the minimum interspecific distance between and among Ch. indicum and other species,with an obvious barcoding gap. The NJ tree showed that Ch. indicum and Ch. morifolium shared a clade,and most of Ch. morifolium with some Ch. indicum were shared a subclade,while Inula lineariifolia,Sinosenecio oldhamianus and Senecio scandens belonged to one clade separately. ITS2 secondary structures for I. lineariifolia,S. oldhamianus and S. scandens were significantly different enough to identify completely but Ch. indicum and Ch. morifolium shared two secondary structures of A and B. It was proved that Ch. indicum was one of the evolutionary sources of Ch.morifolium. Therefore ITS2 sequence as DNA barcode can identify Ch. indicum and its adulterants accurately and quickly. The study provides an important basis for Ch. indicum for the identification of germplasm resources and the safety of clinical medication.

Identification of Chrysanthemum indicum in different geographical populations and Ch. morifolium based on DNA barcodes of psbA-trnH,matK and trnL / 中国中药杂志

DNA barcode technology was used to establish a rapid identification method of Chrysanthemum indicum and Ch. morifolium based on psbA-trn H,mat K and trn L sequences. The total DNA was extracted from 21 samples collected,and the psbA-trn H,mat K,trn L sequences were amplified by PCR and sequenced. The information of these sequences were obtained. We aligned all 63 sequences,calculated the intraspecific and interspecific distances,analysed the SNPs distribution of psbA-trn H+mat K+trn L combination sequences and constructed the Neighbor-joining( NJ) Tree,using MEGA 7. 0. The results showed that the genetic distances of Ch. indicum,Ch. indicum( Juhuanao)and Ch. morifolium were overlapped. The SNPs analysis of psbA-trn H+mat K+trn L combination sequences showed that there were 19 nucleotide polymorphism loci( SNPs) and nine parsim-informative sites in the combination sequences. In addition,Ch. indicum showed more obvious sequence polymorphism than those of Ch. indicum( Juhuanao) and Ch. morifolium. The psbA-trn H sequences showed obvious length variation.The NJ Tree showed that Ch. morifolium numbered C2-C5 were clustered into a single subbranch with a bootstrap value of 62%,and Ch.morifolium could be distinguished from Ch. indicum and Ch. indicum( Juhuanao). Moreover,Ch. indicum numbered Z9 and Z10 collected from Gansu province were singly clustered into one branch with a bootstrap value of 77%. It was also found that the changes of psbA-trn H and trn L sequences information of Ch. indicum samples from the northwest were obviously related to the geography and environment. Moreover,Ch.indicum and Ch. indicum( Juhuanao) had obvious differentiation,were also regarded as the evolutionary sources of Ch. morifolium. Therefore,psbA-trn H+mat K+trn L combination sequences as DNA barcode can identify Ch. indicum and Ch. morifolium accurately and rapidly,which provides an important basis for germplasm resources identification and species identification.

Histone deacetylase inhibitor, 2-propylpentanoic acid, increases the chemosensitivity and radiosensitivity of human glioma cell lines in vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Treatment for malignant glioma generally consists of cytoreductive surgery followed by radiotherapy and chemotherapy. In this study, we intended to investigate the effects of 2-propylpentanoic acid (VPA), a histone deacetylase inhibitor, on chemosensitivity and radiosensitivity in human glioma cell lines.</p><p><b>METHODS</b>Human glioma cell lines, T98-G, and SF295, were treated with temozolomide (TMZ) or irradiation (IR), with or without VPA (1.0 mmol/L). Then, cytotoxicity and clonogenic survival assay was performed. Cell cycle stage, apoptosis, and autophagy were also detected using flow cytometry and dansyl monocadaverin (MDC) incorporation assay. One-way analysis of variance (ANOVA) and t-test were used to analyze the differences among variant groups.</p><p><b>RESULTS</b>Mild cytotoxicity of VPA was revealed in both cell lines, T98-G and SF295, with the 50% inhibiting concentration (IC50) value of (3.85 ± 0.58) mmol/L and (2.15 ± 0.38) mmol/L, respectively; while the IC50 value of TMZ was (0.20 ± 0.09) mmol/L for T98-G and (0.08 ± 0.02) mmol/L for SF295. Moreover, if combined with VPA (1.0 mmol/L) for 96 hours, the sensitivity of glioma cells to TMZ was significant increased (P < 0.05). The surviving fractions at 2 Gy (SF2) of T98-G and SF295 cells exposed to IR alone were 0.52 and 0.58. However, when VPA was combined with IR, the SF2 of T98-G and SF295 dropped to 0.39 (P = 0.047) and 0.49 (P = 0.049), respectively. Treatment with VPA plus TMZ or IR also resulted in a significant decrease in the proportion of cells in the G2 phase and increased apoptotic rates as well as autophagy in T98-G and SF295 cell lines (P < 0.01).</p><p><b>CONCLUSION</b>VPA may enhance the activities of TMZ and IR on glioma cells possibly through cell cycle block and promote autophagy, and thus could be a potential sensitizer of glioma treatment.</p>

Correlation of angiotensin-converting enzyme 2 gene polymorphisms to essential hypertension and ischemic stroke / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the relationship between angiotensin-converting enzyme 2 (ACE2) gene polymorphisms and the risk factor for essential hypertension (EH) with concurrent ischemic stroke in southern Chinese population.</p><p><b>METHODS</b>The G9570A polymorphism in ACE2 gene were detected in 139 patients with EH and stroke using polymerase chain reaction-restriction fragment length polymorphism. Detailed clinical and biochemistrical data of the patients, including the pulse pressure, high sensitivity C-reactive protein (hsCRP), intima-media thickness (IMT), high-density lipoprotein cholesterol (HDL-C) and uric acid levels, were collected to study the relationship between ACE2 gene and the risk factor of EH and stroke.</p><p><b>RESULTS</b>The levels of hsCRP (OR=1.022), uric acid (OR=1.224), IMT and pulse pressure was positively correlated to the incidence of EH and stroke. The pulse pressure, hsCRP, IMT, and HDL-C levels in male stroke patients carrying A allele was significantly higher than those in patients carrying G allele (P<0.05). In female stroke patients, the pulse pressure, hsCRP, IMT, and HDL-C levels were also significantly different with regard to the genotype of ACE2 gene (P<0.05).</p><p><b>CONCLUSIONS</b>The patients with EH and ischemic stroke carrying the A/AA allele of ACE2 gene have higher risks than those carrying other allele, and can be also more vulnerable to stroke recurrence.</p>

Relationship of angiotensin-converting enzyme 2 gene polymorphism with the prognosis of hypertensive stroke patients / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the relationship between angiotensin-converting enzyme 2 (ACE2) gene G9570A polymorphisms and the clinical outcome of stroke patients with essential hypertension (EH) in South China Han population.</p><p><b>METHOD</b>The ACE2 gene polymorphisms were detected in 141 stroke patients with EH and 156 patients with EH using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The genetic marker was tested for its association with the baseline measurements and clinical outcomes of the patients over a median follow-up period of 22 months. As the ACE2 gene is X-linked, analyses were performed for male and female patients separately.</p><p><b>RESULTS</b>The A allele frequency in the stroke patients was significantly different from that in the EH patients, and the AA allele frequency in the female patients was significantly different between the two groups (P<0.01). Kaplan-Meier model analysis showed that ACE2 gene polymorphism was not associated with the the patients' prognosis (P>0.05). Multivariate Cox's proportional hazard regression model identified age (RR=1.057, 95%CI: 1.020, 1.095), blood glucose (RR=1.575, 95%CI: 1.178, 2.104), hypertriglyceridemia (RR=1.947, 95%CI: 1.503, 2.780), blood creatinine (RR=1.034, 95%CI: 1.001, 1.068), and blood uric acid (RR=1.056, 95%CI: 1.002, 1.097) as the risk factors associated with the mortality.</p><p><b>CONCLUSION</b>Stroke occurs more likely in hypertensive patients carrying the A/AA allele than those carrying other alleles. The ACE2 gene G9570A polymorphisms may be associated with the occurrence of stroke in EH patients in South China, but may not have a strong correlation to the prognosis.</p>

Glioblastoma stem cells resistant to temozolomide-induced autophagy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Recent studies have demonstrated the existence of a small fraction of cells with features of primitive neural progenitor cells and tumor-initiating function in brain tumors. These cells might represent primary therapeutic target for complete eradication of the tumors. This study aimed to determine the resistant phenotype of glioblastoma stem cells (GSCs) to temozolomide (TMZ) and to explore the possible molecular mechanisms underlying TMZ resistance.</p><p><b>METHODS</b>Freshly resected glioblastoma specimen was collected and magnetic isolation of GSCs was carried out using the Miltenyi Biotec CD133 Cell Isolation kit. The cytotoxic effect of TMZ on CD133(+) and CD133(-) glioblastoma cells was determined by using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Autophagy-related proteins (Beclin-1, LC3 and Atg5) and cleaved caspase-3 (p17) were analyzed by Western blotting. Immunofluorescent staining was used to detect Atg5, glial fibrillary acidic protein (GFAP) and CD133 expression in glioblastoma cells. Statistical analysis was carried out using SPSS 10.0 software. For all tests, the level of statistical significance was set at P < 0.05.</p><p><b>RESULTS</b>CD133(+) glioblastoma cells exhibited neurosphere-like growth in vitro and high expression of CD133 stem cell marker. The growth-inhibiting rate in CD133(-) glioblastoma cells treated with 5 or 50 micromol/L TMZ was significantly higher than that in CD133(+) glioblastoma cells ((14.36 +/- 3.75)% vs (2.54 +/- 1.36)% or (25.95 +/- 5.25)% vs (2.72 +/- 1.84)%, respectively, P < 0.05). Atg5, LC3-II and Beclin-1 levels were significantly lower in CD133(+) glioblastoma cells than those in autologous CD133(-) cells after TMZ treatment (P < 0.05). Caspase-3 was mildly activated only in CD133(-) glioblastoma cells after exposure to TMZ (P < 0.05). Immunofluorescent staining revealed elevated expression of Atg5 in GFAP(+) cells following TMZ treatment.</p><p><b>CONCLUSIONS</b>The GSCs display strong capability of tumor's resistance to TMZ. This resistance is probably contributed by the CD133(+) cells with down-regulation of autophagy-related proteins. Future treatment should target this small population of cancer stem cells in tumors to improve survival of patients.</p>

Relationship Between the Angiotensin-Converting Enzyme 2 Genotype and Effect of Irbesartan on Left Ventricular Structure and Function in Hypertensive Patients / 中华高血压杂志

Background Experimental data have shown tnat polymorpnisms in tne angiotensm-converting en- zyme 2(ACE2)gene are related to echocardiographically determined parameters of left ventricular mass,structure or function in the general population whether ACE2 genotype influences the effect of angi0tensin Ⅱ receptor blocker which improve left ventricular remodeling and function is unknown.Objective To investigate the association be- tween ACE2 gene G9570A polymorphism and the effect of irbesartan on left ventricular structure and function in hy- pertensive patients.Methods Two hundred and five male patients and 190 female patients who were preliminaryly diagnosised with mild and moderate essential hypertension were treated with irbesartan for 48 weeks with initial dose of 150 mg/d and titrated to 300 mg/d to reach the targed BP.Gene polymorphisms of ACE2 G9570A were detected by PCR-RFLP methods.The association between changes in the SBP,DBP,parameters of left ventricular struc- ture and function and genotypes of the ACE2 gene locus were analyzed.Results Irbesartan reducted in blood pres- sure in all patients(P