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ISSN 1674-2850
CN 11-9150/N5
 
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March 15,2016
Volume 9,Issue 5
Pages -
Subject Area:Chinese and Western Integrative Medicine,Biomedical Engineering
 
Title: Status quo and countermeasures about the internationalization of traditional Chinese medicine preparations
Authors: ZHANG Kai, ZHANG Yongtai, FENG Nianping
PP: 522-528
Abstract: In this paper, we make a thorough discussion on the main challenges faced by traditional Chinese medicine preparations internationalization through combing and reviewing the situation of international trade and foreign registration of traditional Chinese medicine preparations. We put forward measures and suggestions including perfecting the quality standards of traditional Chinese medicine preparations, upgrading traditional Chinese medicine dosage forms, promoting the use of new traditional Chinese medicine pharmaceutical excipients, conducting applied fundamental research on Chinese medicine preparations, and setting up policies for dealing with trade barriers. We also elaborate that traditional Chinese medicine preparation has to constantly improve its quality, set up quality criterions and technical standards both in line with the characteristics of traditional Chinese medicine and international norms for going abroad and becoming the international drug.
Keywords: pharmaceutics; internationalization of traditional Chinese medicine preparations; countermeasures; suggestions
 
Title: Preparation and characterization of calcium phosphate microspheres by ion exchange method
Authors: XIA Xiaoyu, GU Lihong, YANG Hua, SONG Xueying, GU Wei
PP: 518-521
Abstract: Calcium phosphate plays an important role in various fields, and it has a variety of preparation methods. The main purpose of this paper is to use the ion exchange method to prepare calcium phosphate by using calcium carbonate, and to confirm the process by various means. Firstly, the calcium carbonate microspheres were synthesized by using starch as the template, and then were converted to calcium phosphate microspheres by ion exchange reaction at room temperature and standard pressure. The calcium phosphate microspheres were characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and laser nano particle size analyzer. The particle size of the particles was obviously changed, and the results of the FT-IR spectrum and XRD map revealed the changes before and after, the characteristic peak of calcium carbonate disappeared, and the characteristic peak of calcium phosphate displayed. All the results showed that the calcium carbonate could be converted into calcium phosphate by ion exchange reaction.
Keywords: pharmaceutics; biomimetic preparation; calcium carbonate; calcium phosphate; template; ion exchange reaction
 
Title: Construction of K5 polysaccharide-gossypol nanocarrier for drug delivery
Authors: SHI Yu, PENG Huanhuan, TIAN Huihui, WANG Rui, XU Xiaoyu, CHEN Jinghua
PP: 512-517
Abstract: Gossypol is a chemotherapeutic agent for prostate cancer therapy, but its poor solubility, lack of selectivity and high side-effects limit the application. In this paper, a K5 polysaccharede-gossypol conjugate (KGP) based on boronate ester, which is pH sensitive, was prepared as a drug delivery system. The self-assembly, in vitro drug release behaviors, and anticancer efficacy were evaluated. The results showed that KGP could self-assemble into nanoparticles in water medium with the average size at 221.6 nm. In vitro drug release assay indicated the drug release rate could be adjusted by the variation of pH values. The nanoparticles exhibited faster drug release rate at acidic pH=5.0 than that at pH=7.4. The cytotoxicity assay in vitro illustrated that KGP could significantly inhibit the proliferation of prostate cancerous cells as compared with that of normal cells, showing favaroble selectivity. Moreover, fluorescein isothiocyanate (FITC) labeled KGP had rapid cellular internalization of PC-3 cells as compared with COS7 cells. In general, it suggested that KGP could be a promising nanocarrier for therapy of prostate cancer.
Keywords: pharmaceutics; K5 polysaccharide; gossypol; boronate ester; prostate cancer
 
Title: DOX-MNPs suppress gain-of-fuction of ras proto-oncogene in Caenorhabditis elegans and their toxicity
Authors: WANG Dong, ZHI Dejuan, YU Lan, DUAN Ziyun, ZHAO Longhe, ZHANG Qiqi, LI Hongyu
PP: 505-511
Abstract: Objective: Using Caenorhabditis elegans as a model organism to evaluate the potential antitumor activity and toxicity of doxorubicin-Fe3O4 magnetic nanoparticles (DOX-MNPs). Methods: The structure and its magnetic properties of DOX-MNPs were described. The influences of doxorubicin (DOX), Fe3O4 magnetic nanoparticles (MNPs) and DOX-MNPs on the multivulva phenotype, pharyngeal pumping rate and body length of ras (gain of function, gf ) mutant were evaluated. Results: The DOX-MNPs were spherical and the grain size distribution were homogeneous with an average particle diameter of (10.5±0.5) nm, showing excellent superparamagnetism and magnetotactic efficiency. DOX-MNPs with low concentrations (5 mg/L) could significantly suppress the multivulva phenotype of MT2124, while higher concentration (20 mg/L) of DOX showed a similar effect. DOX significantly decreased the pharyngeal pumping rate of worms. DOX-MNPs had little impact on pharyngeal pumping rate. Moreover, DOX-MNPs had lower influence on the body length of the MT2124 compared with DOX. Conclusion: DOX-MNPs with excellent magnetic properties could effectively inhibit the excessive activation of ras gene at low concentrations and showed the lower toxicity. The results indicated that when DOX was conjugated to MNPs, DOX-MNPs were more efficient and of lower toxicity than single DOX.
Keywords: pharmacodynamics; doxorubicin-magnetic nanoparticles; Caenorhabditis elegans; antitumor; toxicity
 
Title: Genetic diversity of Ralstonia solanacearum strains isolated from Pogostemon cablin (Blanco) Benth.
Authors: DENG Zhicheng, HE Hong, JIN Hua, ZHANG Yuyao, LI Zhuan
PP: 498-504
Abstract: Objective: To study on the genetic diversity and differentiation of Ralstonia solanacearum strains isolated from Pogostemon cablin (Blanco) Benth. Methods: Differences in the colonial morphology of R. solanacearum strains were observed on triphenyl tetrazolium chloride (TTC) medium. Further, genetic diversity among the 11 strains was analyzed by amplified 16S ribosomal DNA restriction analysis (16S-ARDRA) and BOX-A1R-based repetitive extragenic palindromic-PCR (BOX-PCR). Results: When observed on TTC medium, the strains could be divided into four groups according to their colonial morphology. Based on 16S-ARDRA patterns, the strains could be grouped into four 16S-ARDRA types. Type A was the main type, which contained 8 strains. At the 48% similarity level, similar results were obtained by using BOX-PCR fingerprint technique. While at 84% similarity level, type A could be further divided into three sub-groups by BOX-PCR fingerprint. Conclusion: A significant level of genetic diversity exists among R. solanacearum strains isolated from P. cablin (Blanco) Benth. Among the strains, BOX-PCR analysis shows higher discriminatory power than 16S-ARDRA technique in the determination of genetic diversity.
Keywords: medicinal botany; Pogostemon cablin (Blanco) Benth.; Ralstonia solanacearum; genetic diversity; 16S-ARDRA; BOX-PCR
 
Title: Screening of antioxidant activity of endophytic fungi from Rhodiola angusta
Authors: CUI Jinlong, GUO Tingting, WANG Mengliang, WANG Junhong, REN Zhenxing
PP: 491-497
Abstract: To evaluate the antioxidant capacities of endophytic fungi from Rhodiola angusta systematically, scavenging activities to1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, hydroxyl free radical, ferrous ions, superoxide anion free radical and nitrite free radical were determined to fungal fermentation. The results showed that 84%, 74%, 58%, 4% and 30% of isolates that had the scavenging rate more than 50%. Ra-R-32 was a promising isolate with scavenging rate more than 50% in each evaluation method. The represent research indicated that endophytic fungi from R. angusta had remarkable antioxidant capacities, which could be potential resource of exploring novel chemicals.
Keywords: medicinal botany; Rhodiola angusta; endophytic fungi; antioxidant; scavenging rate
 
Title: Effect of pachymaran on activation and migration of lymphocytes in GALT of immunosuppression mice
Authors: DENG Xiangliang, HUANG Rongrong, LUO Xia, ZHOU Lian
PP: 484-490
Abstract: Objective: To investigate the effect of pachymaran on lymphocytes activation and migration in gut associated lymphoid tissue (GALT) of immunosuppression mice induced by cyclophosphamide. Methods: 60 BALB/c mice were randomly divided into control, model and pachymaran high, medium and low dose groups. Mice in pachymaran groups were ig administered with pachymaran 400, 200, 100 mg/kg, once per day for 10 d, while those in control group and model group were ig administered with the same dosages of saline solutions. On the sixth day, mice in model group and pachymaran groups were ip administered with cyclophosphamide (100 mg/kg), and those in control group were ip administered with the same volume of saline solutions. The second day after the last administration, lymphocytes suspension from mesenteric lymph nodes (MLN) and Peyer’s patches (PPs) was prepared. The expression levels of CD69 and CCR7 molecules on lymphocytes surface were detected by flow cytometry. The morphologies of PPs were observed under microscope after hematoxylin-eosin (HE) staining. Results: Compared with control group, CD69 molecule expression levels were decreased on T and B cells of MLN and on T cells of PPs in model group, while that was increased on B cells of PPs (P<0.05). Compared with model group, CD69 molecule expression levels were recovery on B cells of MLN and PPs in pachymaran groups (P<0.05). CCR7 molecule expression levels in pachymaran groups were higher than those in model group, and it was significant different in pachymaran medium and low dose group compared with model group (P<0.05). HE staining results showed that the area of PPs was smaller in model group than that in control group, but it was significant recovered in pachymaran groups. Conclusion: Pachymaran can regulate the activation level of B cells and promote the migration of lymphocytes in GALT.
Keywords: Chinese and Western integrative medicine; pachymaran; gut associated lymphoid tissue; CCR7; CD69
 
Title: Diffuse fluorescence tomography based on radiative transfer equation for small-animal imaging
Authors: ZHANG Yan, LI Jiao, ZHAO Huijuan, ZHANG Limin, GAO Feng
PP: 476-483
Abstract: Diffuse florescence tomography (DFT) as a high-sensitivity optical molecular imaging tool, can be applied to in vivo visualize interior cellular and molecular events for small-animal disease model through quantitatively recovering biodistributions of specific molecular probes. In DFT, the radiative transfer equation (RTE) and its approximation P1, such as the diffuse equation (DE), have been used as the forward models. The RTE-based DFT methodology is more suitable for biological tissue having void-like regions and the near-source area as in the situations of small animal imaging. We present a RTE-based scheme for the steady state DFT, which combines the discrete solid angle method and the finite difference method to obtain numerical solutions of the 2D steady RTE, with the natural boundary condition and collimating light source model. The approach is validated using the forward data from the Monte Carlo simulation for its better performances in the spatial resolution and reconstruction fidelity compared to the DE-based scheme.
Keywords: biomedical engineering; diffuse fluorescence tomography; radiative transfer equation; discrete solid angle; natural boundary condition; small-animal imaging
 
Title: Interaction between RhoA/ROCK pathway and ERK pathway in cultured hypertrophy cardiomyocyts
Authors: LI Le, ZHANG Ke, TAO Houquan, SHANG Hao, ZHANG Cailing
PP: 470-475
Abstract: Objective: To investigate whether the interaction between RhoA (ras homologue A)/ROCK pathway and extra cellular signal regulated protein kinase (ERK) pathway in the cultured hypertrophy cardiomyocyts. Methods: The hearts of neonatal rats were isolated and cultured in vitro,then the cardiomyocyts were divided into four groups which stimulated by same dosage isoprenaline respectively: control group, RhoA group, ERK blocking drug (PD98059) group, RhoA+ERK blocking drug (PD98059) group. The expression of ROCK, ERK1/2 proteins, the content of phosphorylated ERK1/2 (p-ERK1/2) and ROCK proteins (p-ROCK) were detected by Western blotting,and the expression of ROCK mRNA in the cultured hypertrophy cardiomyocyts was detected by reverse transcription PCR (RT-PCR). Results: Compared with the control group, the contents of p-ROCK and p-ERK1/2 proteins in RhoA group were increased (P<0.05), while those in ERK blocking drug group were increased (P<0.05). There was no difference between control group and RhoA+ERK blocking drug group. Compared with RhoA group, the contents of p-ROCK and p-ERK1/2 proteins in ERK blocking agent group and RhoA+ERK blocking drug group were decreased (P<0.05). There was no difference in the expression of ROCK and ERK1/2 proteins among different groups. There were obvious differences in expression of ROCK mRNA among different groups. Conclusion: RhoA may directly activate ROCK proteins and ERK1/2 pathway, and the ERK1/2 pathway may be located in the downstream of the RhoA/ROCK pathway. The activation of ROCK proteins may partially depend on ERK1/2 pathway stimulation.
Keywords: pharmacology; cardiac hypertrophy cell; RhoA; ERK
 
Title: Preliminary study of effects of Pulsatilla saponin A on non-small cell lung cancer radiosensitivity
Authors: PENG Chaojun, JIAO Yang
PP: 464-469
Abstract: Objective: To investigate the potential application of Pulsatilla saponin A (PsA) as a radiosensitizer of human non-small cell lung cancer. Methods: MTT viability assay, clonogenic assay, flow cytometry (FCM) analysis, immunoflourescence staining, and single cell gel eletrophoresis assay were utilized to determine the viability, cell cycle distribution, and DNA damage of PsA treated H1975 cells and control cells, with or without X-ray exposure. Results: Compared with the effects on normal human bronchial epithelial cell line, PsA obviously suppressed the cell viability of non-small cell lung cancer cell line H1975. PsA significantly impaired the clonogenic ability of H1975 cells after exposed to X-ray irradiation, caused and enhanced the G2/M phase cell population of H1975 cells after irradiation. In addition, PsA could increase the X-ray induced DNA double strand breaks of H1975 cells. Conclusion: PsA could sensitize H1975 cells to X-ray irradiation, probably via its regulation on irradiation induced DNA damage, which indicates the possibility of PsA as a promising radiosensitizer for cancer radiotherapy.
Keywords: radioactive medicine; radiosensitivity; Pulsatilla saponin A; non-small cell lung cancer; ionizing irradiation; DNA double strand break
 
Title: Regulation of Kindlin-2 on tumor cell invasion and metastasis via inhibiting microRNA-200 family
Authors: QI Lihua, YU Yu, ZHANG Hongquan
PP: 456-463
Abstract: To investigate the molecular mechanism of integrin-interacting protein Kindlin-2 regulating tumor cell invasion and metastasis, by analysis of microRNA (miRNA) microarrays, we find that over-expression of Kindlin-2 obviously down-regulates the levels of miRNA-200 family (miR-200a/200b/200c/141). Meanwhile, results of mRNA microarrays showed that Kindlin-2 could regulate some gene expression of tumor related signaling pathway, including Wnt signaling genes, Hedgehog signaling genes and epithelial-mesenchymal transition (EMT) related genes. Particularly, Kindlin-2 promotes the expression of ZEB1, a target gene of miRNA-200. More importantly, we find that over-expression of miRNA-200 can partly rescue the promotion of Kindlin-2 on tumor cell invasion. Thus, this research will reveal some new molecular mechanism of regulation of Kindlin-2 on tumor cell invasion by analysis of gene microarray.
Keywords: medical cell biology; Kindlin-2; microRNA-200; cell invasion
 
Title: Effect of Chidamide on proliferation and apoptosis of luminal A breast cancer cell line MCF-7
Authors: SHI Xiuqing, MA Fei, LI Huihui, WANG Haijuan, ZHANG Xueyan, LIN Chen, QIAN Haili, XU Binghe
PP: 449-455
Abstract: Objective: To investigate effect of histone deacetylase inhibitor (HDACi) Chidamide on proliferation and apoptosis of luminal A breast cancer MCF-7 cells in vitro. Methods: Human breast cancer MCF-7 cell line was treated with different concentrations of Chidamide (2, 5, 10, 20, 50, 100 μmol/L), and blank group was established as control. Cell growth was assessed by MTS. Morphological changes of MCF-7 cells were observed under inverse microscopy, and apoptosis, cell cycles as well as stem cell proportion were analyzed by flow cytometry assay. Western blotting was used to detect expression of p53 protein between experimental and control groups. Results: Chidamide was able to inhibit proliferation of MCF-7 in vitro in positive dosage dependent manner (P<0.05). After being treated with Chidamide, cell morphology appeared great changes. Flow cytometry assay showed obvious apoptosis compared with control group with the increase of drug concentration and treatment time (P<0.05), but no alternation of cell cycles and stem cell proportion were detected. The expression of p53 protein increased obviously after treatment compared with control group. Conclusion: Chidamide could significantly inhibit proliferation of MCF-7 cells, and induce its apoptosis via upregulation expression of p53 protein.
Keywords: oncology; histone deacetylase inhibitor; Chidamide; breast cancer; apoptosis
 
Title: Construction of Streptococcus mutans dexA gene deletion strain by PCR ligation mutagenesis
Authors: YANG Yan, HONG Xiao, HU Tao, YANG Yingming
PP: 443-448
Abstract: Dextranase (DexA) synthesized by dexA gene of Streptococcus mutans has an important influence on the metabolism of extracellular polysaccharides and the structure of biofilm, via hydrolysis of α-1,6glycosides bond inside water soluble glucan (WSG). In the purpose of studying the further function of dexA gene, this paper was to construct the dexA gene knockout mutants of S. mutans by PCR ligation mutagenesis. In this study, the dexA gene deletion strain of S. mutans (SmudexA) was constructed with PCR ligation mutagenesis. The successful construction of the mutation was verified by PCR, gel electrophoresis. Fluorescence quantitative real-time PCR (RT-PCR) showed dexA gene expression deficiency and nucleotide sequence analysis confirmed dexA gene was knocked out. We have successfully constructed and authenticated the mutant strains of S. mutans for dexA gene deletion which lay foundation for the further subsequent exploration.
Keywords: stomatology; Streptococcus mutans; dexA gene; PCR ligation mutagenesis; mutant construction
 
Title: α-melanocyte-stimulating hormone inhibits pathological retinal neovascularization in a mouse model of oxygen-induced retinopathy
Authors: YANG Qianhui, ZHANG Yan, BO Qiyu, CAO Yunshan, YANG Wei, LI Xiaorong
PP: 435-442
Abstract: Objective: To study the effects of intravitreal injection of α-melanocyte-stimulating hormone (α-MSH) at different concentrations on pathological retinal neovascularization (RNV) in a mouse of oxygen-induced retinopathy (OIR). Methods: Healthy C57BL/6J pups of hygiene grade were randomly divided into OIR+α-MSH-1, OIR+α-MSH-5, OIR+α-MSH-10, OIR and normal control groups at postnatal day 7 (P7). The α-MSH intervention groups and OIR group were exposed to high oxygen [(75±2)%] for 5 d, then maintained under normal air condition for another 5 d; whereas the normal control group was raised under normoxia for 10 d. Retro-orbital injection of high molecular weight fluorescein isothiocyanate-dextran (FITC-dextran) was performed on P17 mice. The retina whole mounts were prepared to observe retinal vessels and quantify the relative area of vessel obliteration. The mouse eyeballs were subjected to paraffin sections and hematoxylin-eosin (HE) staining, and the average number of pre-retinal nuclei per section was quantified. Results: FITC-dextran-labeled retinal whole mounts showed that the relative vessel obliteration area was of statistical significance among the groups (F=19.64, P<0.05). No vessel obliteration was observed in normal control retinas, whereas the relative avascular area in the OIR group was (23.01±3.39)%, the difference between the two groups was of statistical significance (P<0.001). The relative avascular area in OIR+α-MSH-1, OIR+α-MSH-5, and OIR+α-MSH-10 groups were (18.14±7.20)%, (15.64±7.07)%, and (7.62±6.52)%, respectively. The relative avascular area in the OIR+α-MSH-10 group was significantly lower than that in the OIR group (P<0.01). HE staining of the paraffin sections showed that the average number of pre-retinal nuclei per section was 13.59±4.28 in the OIR group, and 1.48±1.27 in the normal controls, there is statistical significance between the two groups (P<0.001). The average numbers of pre-retinal nuclei per section in the α-MSH-treated groups were 6.35±2.34 for OIR+α-MSH-1 group, 4.96±1.79 for OIR+α-MSH-5 group, and 1.03±1.25 for OIR+α-MSH-10 group, and these numbers were all significantly less than that in the OIR group (P<0.001) and different compared with each group (F=147.87, P<0.05). Conclusion: Intravitreal injection of α-MSH dose-dependently reduces the relative area of vessel obliteration and the average number of pre-retinal nuclei in the retinas of OIR mouse model, thereby inhibiting RNV under the pathological condition.
Keywords: ophthalmology; oxygen-induced retinopathy; retinal neovascularization; α-melanocyte-stimulating hormone; pathology
 
Title: Physiological cyclic stretch induced proliferation of human bladder smooth muscle cells via PI3K-SGK1-Kv1.3 pathway
Authors: TIAN Ye, YUE Xuan, YANG Tongxin, CHEN Lin, LUO Deyi, WANG Kunjie
PP: 428-434
Abstract: Objective: To explore the different modes of stress induced proliferation of human bladder smooth muscle cells (HBSMCs) and to investigate the mechanism involved in this process. Methods: HBSMCs were seeded in a silicone membrane and subjected to cyclic stretch of 2.5%, 5%, 10% and 15% equibiaxial strain at frequencies of 0.05, 0.10, 0.20, 0.50, 1.00 Hz, respectively. HBSMCs without stess were employed as the control group. Bromodeoxyuridine (BrdU) assays were used to detect the proliferative activity of each group, and hereby to determine the optimal stress mode. To further determine the mechanism of the cell proliferation process triggered by physiological cyclic stretch, the expression of PI3K, SGK1, Akt, Kv1.3 was investigated at the transcriptional and translational levels (molecular and protein levels) by real-time PCR (RT-PCR) and Western blotting analysis, respectively. Results: Optimal physiological stretch was established as 5% strain at a frequency of 0.10 Hz, whereby HBSMCs revealed a statistical significant increase in proliferative activity compared with the other groups, including the control group, which served as the control (P<0.05). The expressions of PI3K, SGK1 and Kv1.3 (P<0.05), however, without Akt (P<0.05), were upregulated by cyclic stretch as compared with the control group. When separately treated with inhibitors of SGK1 and Kv1.3, increased stress induced proliferation was largely eliminated. Conclusion: Cyclic stretch induces the proliferation of HBSMCs and the PI3K-SGK1-Kv1.3 pathway is involved in this process, either fully or at least partially, rather than its related pathway, PI3K-Akt.
Keywords: urology; tissue engineering; smooth muscle cells; cell proliferation; mechanical stress; signal transduction