Bioactive chemical constituents, in vitro anti-proliferative activity and in vivo toxicity of the extract of Curcuma singularis Gagnep rhizomes.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcuma singularis Gagnep is a Vietnamese medicinal plant which has been commonly used as a medicinal remedy in traditional and folk medicines for improving health as well as for treating some diseases, like rheumatoid arthritis, kidney failure. However, pharmacological effects, including anti-cancer activity and the safety of this plant has not been fully investigated. AIM OF THE STUDY: This study aimed to investigate the in vitro anti-growth activity of an extract derived from Curcuma singularis rhizome extract (CSE) against cell lines as well as determine its phytochemical composition. The other goal of our study was to assess the safety of CSE in rats. MATERIALS AND METHODS: The main constituents in the extract were identified and quantitatively analyzed. The in vitro cytotoxicity of CSE was evaluated in several cancer and normal cell lines. The apoptotic activity of CSE and the expression of the apoptosis-related genes were investigated in AGS cells to clarify the underlying molecular mechanisms. The in vivo toxicity of CSE was assessed via acute and subacute oral studies on Sprague-Dawley rats, respectively according to the guidelines 425 and 407 of the Organization for Economic Cooperation and Development (OECD). The drug-related toxicity signs, mortality, body and organ weights were recoreded during the experimental period. In addition, the selected hematological and biochemical parameters, and histological alterations were determined at the end of the subacute toxicity test. RESULTS: Germacrone, ar-turmerone, and curcumol were three sesquiterpene components found in the extract. CSE showed cytotoxic effects in different cancer cells, but had minimal effects on normal cells. Apoptosis in AGS cells was caused by CSE in a concentration-dependent pattern through increase of Bax/Bcl-2 ratio, and release of cytochrome c, which leads to activation of caspase-3/-7, caspase-9, as well as cleavage of PARP. In the acute toxicity test, no signs of toxicity and no mortality were recorded in rats at both doses of 1000 and 5000 mg/kg. In the subacute toxicity study, CSE showed no drug-related adverse effects on water and food consumption, body and organ weights. CSE at a dose of 1000 mg/kg slightly increased WBC and platelet values in female rats, while it increased WBC values in male rats in all tested doses. The decrease of total cholesterol and triglyceride levels were found in female rats treated CSE at doses of 250 or 500 mg/kg. In addition, the increase of serum ALT and AST levels in rats treated at the dose of 1000 mg/kg were noted. No significant changes in histopathological structures of kidneys, spleen, heart and lungs, except liver tissue with minor modifications was found. CONCLUSIONS: Our findings indicated that CSE exhibited in vitro anti-proliferative effects on AGS cells by mainly activating the caspase-dependent mitochondrial apoptotic pathway. CSE also showed in vivo toxicity signals at the dose of 1000 mg/kg with proven minor hepatic injuries, which should be avoided the high dose for prolonged use. Curcuma singularis rhizomes may be used as a chemotherapeutic agent for the treatment of gastric cancer with in vitro anti-cancer investigation and in vivo biological safety evaluation.

Allogeneic umbilical cord-derived mesenchymal stem cell transplantation for treating chronic obstructive pulmonary disease: a pilot clinical study.

INTRODUCTION: Chronic obstructive pulmonary disease (COPD) is the third leading cause of death worldwide. COPD results from chronic inflammation of the lungs. Current treatments, including physical and chemical therapies, provide limited results. Stem cells, particularly mesenchymal stem cells (MSCs), are used to treat COPD. Here, we evaluated the safety and efficacy of umbilical cord-derived (UC)-MSCs for treating COPD. METHODS: Twenty patients were enrolled, 9 at stage C and 11 at stage D per the Global Initiative for Obstructive Lung Disease (GOLD) classification. Patients were infused with 106 cells/kg of expanded allogeneic UC-MSCs. All patients were followed for 6 months after the first infusion. The treatment end-point included a comprehensive safety evaluation, pulmonary function testing (PFT), and quality-of-life indicators including questionnaires, the 6-min walk test (6MWT), and systemic inflammation assessments. All patients completed the full infusion and 6-month follow-up. RESULTS: No infusion-related toxicities, deaths, or severe adverse events occurred that were deemed related to UC-MSC administration. The UC-MSC-transplanted patients showed a significantly reduced Modified Medical Research Council score, COPD assessment test, and number of exacerbations. However, the forced expiratory volume in 1 s, C-reactive protein, and 6MWT values were nonsignificantly reduced after treatment (1, 3, and 6 months) compared with those before the treatment. CONCLUSION: Systemic UC-MSC administration appears to be safe in patients with moderate-to-severe COPD, can significantly improve their quality of life, and provides a basis for subsequent cell therapy investigations. TRIAL REGISTRATION: ISRCTN, ISRCTN70443938. Registered 06 July 2019.

Downregulation of Kinesin spindle protein inhibits proliferation, induces apoptosis and increases chemosensitivity in hepatocellular carcinoma cells.

BACKGROUND: Kinesin spindle protein (KSP) plays a critical role in mitosis. Inhibition of KSP function leads to cell cycle arrest at mitosis and ultimately to cell death. The aim of this study was to suppress KSP expression by specific small-interfering RNA (siRNA) in Hep3B cells and evaluate its anti-tumor activity. METHODS: Three siRNA targeting KSP (KSP-siRNA #1-3) and one mismatched-siRNA (Cont-siRNA) were transfected into cells. Subsequently, KSP mRNA and protein levels, cell proliferation, and apoptosis were examined in both Hep3B cells and THLE-3 cells. In addition, the chemosensitivity of KSP-siRNA-treated Hep3B cells with doxorubicin was also investigated using cell proliferation and clonogenic survival assays. RESULTS: The expression of endogenous KSP at both mRNA and protein levels in Hep3B cells was higher than in THLE-3 cells. In Hep3B cells, KSP-siRNA #2 showed a further downregulation of KSP as compared to KSP-siRNA #1 or KSP-siRNA #3. It also exhibited greater suppression of cell proliferation and induction of apoptosis than KSP-siRNA #1 or KSP-siRNA #3; this could be explained by the significant downregulation of cyclin D1, Bcl-2, and survivin. In contrast, KSP-siRNAs had no or lower effects on KSP expression, cell proliferation and apoptosis in THLE-3 cells. We also noticed that KSP-siRNA transfection could increase chemosensitivity to doxorubicin in Hep3B cells, even at low doses compared to control. CONCLUSION: Reducing the expression level of KSP, combined with drug treatment, yields promising results for eradicating hepatocellular carcinoma (HCC) cells in vitro. This study opens a new direction for liver cancer treatment.

Differentiation of umbilical cord lining membrane-derived mesenchymal stem cells into endothelial-like cells.

BACKGROUND: Stem cell therapy for the treatment of vascular-related diseases through functional revascularization is one of the most important research areas in tissue engineering. The aim of this study was to investigate the in vitro differentiation of umbilical CL-MSC into endothelial lineage cells. METHODS: In this study, isolated cells were characterized for expression of MSC-specific markers and osteogenic and adipogenic differentiation. They were induced to differentiate into endothelial-like cells and then examined for expression of the endothelial-specific markers, karyotype, and functional behavior of cells. RESULTS: Isolated cells expressed MSC-specific markers and differentiated into adipocytes and osteoblasts. After endothelial differentiation, they expressed CD31, vWF, VE-cadherin, VEGFR1, and VEGFR2 at both mRNA and protein level, but their morphological changes were not apparent when compared with those of undifferentiated cells. There were no significant changes in karyotype of differentiated cells. Furthermore, angiogenesis assay and LDL uptake assay showed that differentiated cells were able to form the capillary-like structures and uptake LDL, respectively. CONCLUSION: The results indicated that umbilical CL-MSC could differentiate into functional endothelial-like cells. Also, they are suitable for basic and clinical studies to cure several vascular-related diseases.

An ultrasensitive protein array based on electrochemical enzyme immunoassay.

An ultrasensitive electrochemical enzyme immunoassay (EEIA) for the detection of proteins on an 8x8 array is described. The assay is based on wired enzyme technology. Briefly, capture antibody was covalently immobilized on a self-assembled 11-mercaptoundecanoic acid (MUA) monolayer coated gold electrode. After incubating with a target protein (antigen), the gold electrode was treated sequentially in biotinylated detection antibody solution and in avidin-horseradish peroxidase conjugate (A-HRP) solution. A cationic redox polymer (electrochemical mediator) overcoating was applied to the gold electrode through layer-by-layer electrostatic self-assembly. The formation of a bilayer brought the HRP in electrical contact with the underlying electrode, making the bilayer an electrocatalyst for the reduction of hydrogen peroxide where the redox polymer acts as an artificial mediator. Consequently, the concentration of protein could be quantified amperometrically. This electrochemical immunoassay combined the specificity of the immunological reaction with the sensitivity of the electrochemical detection. The applicability of the system in protein detection was demonstrated with a snake toxin, beta-bungarotoxin, a neurotoxin from the venom of the snake Bungarus multicinctues. Under optimized experimental conditions, the assay allowed the detection of beta-bungarotoxin in the range of 20 pg/mL to 1.5 ng/mL with a detection limit of 10 pg/mL (20 fg). A higher detection limit of 25 pg/mL was obtained in serum.

Oral vaccine produced from transgenic plants

An important feature of immune system in general and gastrointestinal immune system in particular is the mobility of member cells. In DNA recombination technique, bacteria like E. coli are often used to make recombination protein including vaccine . The advantage of the bacteria is manipulating easily with their gene. There are two major methods to transfer gene in plants. One is based on Agrobacterium tumefactions, a bacterium that naturally live in the soil, which is able to transfer a number of DNA including one of its plasmids to plant cells to integrate into the plant's genome at cell nucleus. The other is transformation by Agrobacterium which is more common used to make vaccine transgenic plants

Kidney donor exchange - an efficacious solution to the lack of transplant organs

Currently world wide, the majority of kidney transplantations is realized with living donor, not using cadaver. In the past, in majority of cases, the donor and the recipient are familiar relatives, assuring immune compatibility. But at present basing on the new advances in antirejection measures, this technique can be applied in any pairs without blood relations. The detection of these pairs is very difficult but could be realized completely. The cases of exchange between uncomfortable pairs could give satisfied results

The value of code conversion of human genome in medicine

The Human Genome Project is an international cooperative project that launched in 1990 and completely finished in April 14, 2003. The discovery of pathogenic genes is promising that light up the pathogenesis of many diseases in human. In the future, the understanding of these genes is helpful for patients and physicians, but the first advantages are in diagnosis and prognosis. It’s hoped to bring about new more effective therapies tailored for each patient according to his genome in the next 5 years

Immune support from mother to infant

The child needs time to develop his own immune system and to have specific immune responds. Thus the child needs his mother's support to have immediate protection against bacteria from mother's gastrointestinal tract that infected the baby and have longer protection during his development of immune system. This immune supporting of mother for her baby was conducted thru placenta and breast milk. The immune supporting thru breast milk was directly and indirectly by indirect transmitting of resistance for baby thru breast milk, direct stimulating of baby immune system, and preventing some immune diseases