Acute oral toxicity assessment and anti-hyperuricemic activity of Alocasia longiloba extracts on Sprague-Dawley rats

Hyperuricemia is defined as a metabolic abnormality that occurs when serum uric acid (UA) level is abnormally high in the body. We previously reported that A. longiloba possesses various important phytochemicals and in vitro xanthine oxidase activity. Despite A. longiloba ethnomedicinal benefits, its toxicity and anti-hyperuricemic effects have not been reported. The present study was carried out to ensure the safety and investigate the anti-hyperuricemic effects of A. longiloba fruit and petiole ethanolic extracts on rats. In the acute toxicity study, extracts were orally administered at a dose of 2000 mg/kg bodyweight and closely monitored for 2-week for any toxicity effects. The rats were then sacrificed and samples were collected and analyzed for hematological, biochemical, and histopathological parameters. The anti-hyperuricemic effect of A. longiloba fruit or petiole extract was investigated through determination of UA levels on potassium oxonate (PO)-induced hyperuricemic rats. Extracts or standard drug treatments were orally administrated 1-h after PO administration for 14-day. Animals were euthanized and samples were collected for further experiments. The toxicity results show, no significant changes were observed in behavioral, bodyweight changes in experimental groups compared to the control. Moreover, there were no significant changes in hematological, biochemical, and histological parameters between extracts treated and control group. In the anti-hyperuricemia study, the fruit and petiole extracts treatments significantly reduced the level of UA in serum compared to the hyperuricemic model group. This study demonstrated that the extracts of A. longiloba have anti-hyperuricemic activity and was found to be non-toxic to rats in acute toxicity test.     

益气活血中药复方对CVB3大鼠心肌细胞感染模型ATP6基因表达的影响

目的：研究益气活血中药复方对CVB3大鼠心肌细胞感染模型ATP6（ATP synthase F0 subunit 6）基因表达的作用机制。方法：本实验用新生2-3dWistar大鼠心肌细胞,建立CVB,病毒感染模型,通过改良的抑制性消减杂交技术（SSH）,克隆了受CVB,攻击的心肌细胞中被中药（益气活血中药复方）调控的基因。结果：ATP6基因,在中药组中高表达,而病毒组中表达减弱或抑制。结论：益气活血中药复方能影响受CVB,病毒攻击的宿主细胞ATP6基因的表达,有效保护心肌、阻断病程进度,从而实现治疗病毒性心肌炎的目的。     

Development and characterization of microsatellites of an important medicinal plant Orthosiphon stamineus (misai kucing)

Orthosiphon stamineus (Lamiaceae) is an important medicinal plant in Malaysia that has great potential in the herbal medicine industry. In this study, we developed and characterized ten microsatellites for O. stamineus. The microsatellite loci were checked for variability using 28 individuals of O. stamineus sampled from 10 nurseries. The number of alleles ranged from two to four, and the observed heterozygosity ranged from 0.143 to 0.963. Neighbor-joining analysis revealed two main genetic clusters corresponding to the two varieties (white and purple). The application of these microsatellites will facilitate the cultivar/variety identification and germplasm management in the species.     

Ex vivo expansion of primitive hematopoietic cells for cellular therapies: An overview

Sources of hematopoietic cells for bone marrow transplantation are limited by the supply of compatible donors, the possibility of viral infection, and autologous (patient) marrow that is depleted from prior chemo- or radiotherapy or has cancerous involvement. Anex vivo system to amplify hematopoietic progenitor cells could increase the number of patients eligible for autologous transplant, allow use of cord blood hematopoietic cells to repopulate an adult, reduce the amount of bone marrow and/or mobilized peripheral blood stem and progenitor cells required for transplantation, and reduce the time to white cell and platelet engraftment. The cloning of hematopoietic growth factors and the identification of appropriate conditions has enabled the development of successfulex vivo hematopoietic cell cultures. Purification systems based on the CD34 marker (which is expressed by the most primitive hematopoietic cells) have proven an essential tool for research and clinical applications. Present methods for hematopoietic cultures (HC) on stromal (i.e. accessory cells that support hematopoiesis) layers in flasks lack a well-controlled growth environment. Several bioreactor configurations have been investigated, and a first generation of reactors and cultures has reached the clinical trial stage. Our research suggests that perfusion conditions improve substantially the performance of hematopoietic reactors. We have designed and tested a perfusion bioreactor system which is suitable for the culture of non-adherent cells (without stromal cells) and readily scaleable for clinical therapies. Eliminating the stromal layer eliminates the need for a stromal cell donor, reduces culture time, and simplifies the culture system. In addition, we have compared the expansion characteristics of both mononuclear and CD34⁺ cells, since the latter are frequently assumed to give a superior performance for likely transplantation therapies.Abbreviations BFU0-E burst forming unit-erythroid - BM bone marrow - CB cord blood - CFU-C colony forming unit-culture - CFU-E colony forming unit-erythroid - CFU-F colony forming unit-fibroblast - CFU-GEMM colony forming unit-granulocyte, erythroid, macrophage, megakaryocyte - CFU-GM colony forming unit-granulocyte, macrophage - CFU-Mix colony forming unit-mixed (also known as CFU-GEMM) - CML chronic myeloid leukemia - CSF colony stimulating factor - DMSO dimethyl sulfoxide - ECM extracellular matrix - EPO erythropoietin - FL fetal liver - HC hematopoietic culture - LTBMC long-term bone marrow culture - LTC-IC long-term culture initiating cell - LTHC long-term hematopoietic culture - MNC mononuclear cells - PB peripheral blood     

Therapeutic effects of herbal extracts and constituents in animal models of psychiatric disorders

A search for novel pharmacotherapy from medicinal plants for psychiatric illnesses has progressed significantly in the past decade. This is reflected in the large number of herbal preparations for which psychotherapeutic potential has been evaluated in a variety of animal models. The objective of this review is to provide an overview of herbal extracts and constituents that have significant therapeutic effects in animal models of psychiatric illnesses. Eighty five individual herbs reviewed were classified as anxiolytic, antidepressant, neuroleptic, antidementia, or anti-substance abuse herbs. The full scientific name of each herb, herbal part used, active constituent, extract, dose range and route, animal model, possible mechanisms of action, and pertinent references are presented via synoptic tables. The herbal mixtures were also mentioned. A considerable number of herbal constituents whose behavioral effects and pharmacological actions have been well characterized may be good candidates for further investigations that may ultimately result in clinical use. The investigation of a large portion of the herbal extracts and herbal mixtures is in its infancy. Herbal remedies that have demonstrable psychotherapeutic activities have provided a potential to psychiatric pharmaceuticals and deserve increased attention in future studies.     

Herbal bioactivation: the good, the bad and the ugly

It has been well established that the formation of reactive metabolites of drugs is associated with drug toxicity. Similarly, there are accumulating data suggesting the role of the formation of reactive metabolites/intermediates through bioactivation in herbal toxicity and carcinogenicity. It has been hypothesized that the resultant reactive metabolites following herbal bioactivation covalently bind to cellular proteins and DNA, leading to toxicity via multiple mechanisms such as direct cytotoxicity, oncogene activation, and hypersensitivity reactions. This is exemplified by aristolochic acids present in Aristolochia spp, undergoing reduction of the nitro group by hepatic cytochrome P450 (CYP1A1/2) or peroxidases in extrahepatic tissues to reactive cyclic nitrenium ion. The latter was capable of reacting with DNA and proteins, resulting in activation of H-ras oncogene, gene mutation and finally carcinogenesis. Other examples are pulegone present in essential oils from many mint species; and teucrin A, a diterpenoid found in germander (Teuchrium chamaedrys) used as an adjuvant to slimming diets. Extensive pulegone metabolism generated p-cresol that was a glutathione depletory, and the furan ring of the diterpenoids in germander was oxidized by CYP3A4 to reactive epoxide which reacts with proteins such as CYP3A and epoxide hydrolase. On the other hand, some herbal/dietary constituents were shown to form reactive intermediates capable of irreversibly inhibiting various CYPs. The resultant metabolites lead to CYP inactivation by chemical modification of the heme, the apoprotein, or both as a result of covalent binding of modified heme to the apoprotein. Some examples include bergamottin, a furanocoumarin of grapefruit juice; capsaicin from chili peppers; glabridin, an isoflavan from licorice root; isothiocyanates found in all cruciferous vegetables; oleuropein rich in olive oil; dially sulfone found in garlic; and resveratrol, a constituent of red wine. CYPs have been known to metabolize more than 95% therapeutic drugs and activate a number of procarcinogens as well. Therefore, mechanism-based inhibition of CYPs may provide an explanation for some reported herb-drug interactions and chemopreventive activity of herbs. Due to the wide use and easy availability of herbal medicines, there is increasing concern about herbal toxicity. The safety and quality of herbal medicine should be ensured through greater research, pharmacovigilance, greater regulatory control and better communication between patients and health professionals.     

Herbal medicine Sho-saiko-to (TJ-9) increases expression matrix metalloproteinases (MMPs) with reduced expression of tissue inhibitor of metalloproteinases (TIMPs) in rat stellate cell

We have reported that Sho-saiko-to (TJ-9) prevents liver fibrosis in vivo. To gain further insights into the effect of TJ-9, the matrix metalloproteinases (MMPs)/tissue inhibitors of metalloproteinases (TIMPs) balance was examined. Hepatic stellate cells (HSCs) were isolated from male Wistar rats and cultured with TJ-9 (0-1000 microg/ml) on uncoated plastic dishes for 4 days. To elucidate the effects on the MMPs/TIMPs balance by TJ-9, quantitative analysis of type IV collagen-degrading activity, gelatin zymography and reverse zymography were carried out. Northern blot analysis was performed to determine the expression of MMP-2, 13 and TIMP-1 mRNAs. TJ-9 treatment resulted in dose-dependent upregulation of MMP-2, 13 mRNA and downregulation of TIMP-1 mRNA up to 500 microg/ml. Gelatin zymography, reverse zymography and quantitative analysis of type IV collagen-degrading activity confirmed that TJ-9 increased MMP-2 activity and prevented TIMP-1, 2 activities in a dose-dependent manner. SB203580 diminished the reduction of mRNA as well as the activity of TIMP-1 by TJ-9 and induction of mRNA as well as the activity of MMP-2. These results show that TJ-9 increased MMP-2, 13 activity with reduced TIMP-1, 2 activities on HSCs possibly via P38 pathway.     

Circulating tumor cell isolation,culture, and downstream molecular analysis

Circulating tumor cells (CTCs) are a major contributor of cancer metastases and hold a promising prognostic significance in cancer detection. Performing functional and molecular characterization of CTCs provides an in-depth knowledge about this lethal disease. Researchers are making efforts to design devices and develop assays for enumeration of CTCs with a high capture and detection efficiency from whole blood of cancer patients. The existing and on-going research on CTC isolation methods has revealed cell characteristics which are helpful in cancer monitoring and designing of targeted cancer treatments. In this review paper, a brief summary of existing CTC isolation methods is presented. We also discuss methods of detaching CTC from functionalized surfaces (functional assays/devices) and their further use for ex-vivo culturing that aid in studies regarding molecular properties that encourage metastatic seeding. In the clinical applications section, we discuss a number of cases that CTCs can play a key role for monitoring metastases, drug treatment response, and heterogeneity profiling regarding biomarkers and gene expression studies that bring treatment design further towards personalized medicine.