Research on Anti-bacterial Effects of 5 Kinds of Chinese Herb Extracts on Ameromonas hydrophila in vitro

[Objective] The aim was to study the anti-bacterial effects of 5 kinds of Chinese herb extracts on Ameromonas hydrophila in vitro. [Method] in vitro anti-bacterial effects of 5 kinds of Chinese herb extracts like Galla Chinensis,Syzygium aromaticum,Salvia miltiorrhiza,Punica granatum L. and Terminalia chebula Retz on Ameromonas hydrophila were studied; furthermore,cure rates of the Chinese herb extracts with better anti-bacterial effects were determined to find out the optimal drug dosage. [Result] Under the same experimental conditions,Galla Chinensis,Punica granatum and Terminalia chebula Retz had relatively strong anti-bacterial effects on Ameromonas hydrophila,among them the anti-bacterial effect of Galla Chinensis was significantly higher than those of the others （P0.05）. The optimal treatment dose of Galla Chinensis treating bacterial septicemia caused by Aeromonas hydrophila was that they were treated with medicated bath for 40 min by 0.5 mg/ml Galla Chinensis extract,and the cure rate was 100%. [Conclusion] The research provides a scientific drug basis for the control and prevention of outbreak bacterial diseases of fish.

Effects of Stylosanthes scabra Forage Supplementation on in Vitro Gas Production and Fiber Degradation of Eragrostis Grass Hay

Natural pastures constitute a major component of ruminant livestock feed, and are the most cost-effective feed resource available for smallholder subsistence farmers. However, this feed resource does not meet animal nutritional requirement due to deficiency in nitrogen, energy and minerals. In addition, at maturity lignification is the major concern since it reduces digestibility and contributes to methane emission. Thus, the objective of this study was to evaluate the effect of supplementing low-quality Eragrostis grass hay with five (9281, 11,252, 11,255, 11,595 and 11,604) selected Stylosanthes scabra accessions on in vitro ruminal fermentation and neutral detergent fiber degradation. Therefore, in vitro study was conducted on grass hay, accessions and the mixture of grass hay with each accession included at two (15%, 30%) levels. The substrates (grass hay, accessions and the mixtures) were incubated in separate serum bottles for 72 h. Neutral detergent fiber (NDF) of the accessions ranged from 300 to 350 g/kg DM with crude protein (CP) value ranging from 177.5 to 184.1 g/kg DM. Eragrostis grass hay had NDF value of 813 g/kg DM, with CP value of 34.3 g/kg DM. Grass hay fermented slowly, it took 30 h for grass hay to produce gas volume above 50 mL, while Stylosanthes scabra accessions took 12 h. Supplementing grass hay with accessions significantly improved fermentation. However, it was observed that 15% inclusion took 30 h to produce gas volume above 50 mL, whereas at 30% inclusions it took 24 h for accession 9281, 11,595 and 11,604. Accession 11,604 improve grass fermentation by almost three times the value of grass hay in 2 h. Grass hay supplemented with accession 11,604 at 30% had a positive associative effect and significantly improved NDF degradability. In conclusion, accession 11,604 may be fed strategically as forage supplement to low-quality forage for ruminants.

In vitro prebiotic effects of seaweed polysaccharides

Although prebiotic activities of alginate and agar oligosaccharides isolated from seaweeds have been reported, it remains unknown whether seaweed polysaccharides have prebiotic activity. In this study, we isolated polysaccharides from four species of seaweeds, such as Grateloupia fi licina(GFP), Eucheuma spinosum(ESP), Ulva pertusa(UPP), and A scophyllum nodosum(ANP), and characterized their structures and prebiotic ef fects in vitro. The results showed that these polysaccharides were dif ferent in total sugar and sulfate contents as well as monosaccharide composition. GFP and ESP significantly promoted bifi dobacterium proliferation and 0.1% ESP and 0.4% GFP resulted in the highest proliferation rates of beneficial bacteria, whereas UPP and ANP inhibited the growth of beneficial bacteria at all tested concentrations(0.1%–0.5%). The different behaviors of the four seaweed-originated polysaccharides might be refl ected by differences in monosaccharide composition and structure. Therefore, polysaccharides isolated from GFP and ESP could be utilized as prebiotics. However, more studies must be carried out in vivo.

Toxicity Evaluation of in vitro Cultures of Freshwater Cyanobacterium Microcystis aeruginosa:Ⅰ.Hepatotoxic and Histopathological Effects in Rats

Laboratory cultures of freshwater cyanobacterium (blue-green alga) Microcystis aeruginosa PCC 7806 was cvaluated for its hepatotoxic effects in rats. The lyophilized cell extract injected intraperitoneally at 1 and 2 LD50 (15.8 and 31.6 mg/kg, respectively) produced significant increase in liver-specific enzymes viz. plasma alkaline phosphatase,γ-glutamyl transferase, lactate dehydrogenase with a concomitant decrease in hepatic glutamic pyruvic transaminase. A corresponding increase in liver body weight index and histopathological changes in liver (degeneration of hepatocytes, congestion and hemorrhage etc.) are indicative of a dose and time dependent hepatotoxic nature of the algal extract

Effects of natural-cerebrolysin-containing serum on neurotoxicity and synaptogenesis in amyloid-beta 1-40-induced Alzheimer's disease in vitro models

BACKGROUND： Neuronal loss, synapse mutilation, and increasing malnourished axons are pathologically related to Alzheimer＇s disease. Microtubule-associated protein 2 （MAP2） is of importance for neuronal, axonal, and dendritic generation, extension, and stabilization, as well as for the regulation of synaptic plasticity. OBJECTIVE： To investigate the antagonistic effects of natural-cerebrolysin-containing serum on beta amyloid protein 1-40 （Aβ1-40）-induced neurotoxicity from the standpoints of cell proliferation, synaptogenesis, and cytoskeleton formation （MAP2 expression）. DESIGN, TIME AND SETTING： A paralleled, controlled, neural cell, and molecular biology experiment was performed at the Institute of Integrated Chinese and Western Medicine, Shenzhen Hospital, Southern Medical University between February 2006 and April 2008. MATERIALS： PC12 cells, derived from the rat central nervous system, were purchased from Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China. A β1-40 was provided by Sigma, USA. Natural-cerebrolysin was provided by Shenzhen Institute of Integrated Chinese and Western Medicine, China. The natural-cerebrolysin was predominantly composed of Renshen （Radix Ginseng）, Tianma （Rhizoma Gastrodiae）, and Yixingye （Ginkgo Leaf） in a proportion of 1：2：2. Following conventional water extraction technology, an extract （1：20） was prepared. Each gram of extract equaled 20 grams of crude drug. In a total of 12 adult male New Zealand rabbits, six underwent intragastric administration of natural-cerebrolysin extract for 1 month to prepare natural-cerebrolysin-containing serum, and the remaining six rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： An AIzheimer＇s disease in vitro model was induced in PC12 cells using Aβ1-40. The cells were incubated with varying doses of natural-cerebrolysin-containing serum （2.5%, 5%, and 10%）. Normal blank serum-treated PC12 cells served as a blank control group. MAIN OUTCOME MEASURES： Through the use of inverted phase contrast microscope, cell morphology and neurite growth were observed, neurite length was measured, and the percentage of neurite-positive cells was calculated. Cell proliferation rate was determined by MTT assay, and MAP 2 expression was detected by fluorescent immunocytochemistry. RESULTS： Following Aβ1-40 treatments, some PC12 cells were apoptotic/dying, and only a few short neurites were observed. Following interventions with natural-cerebrolysin-containing serum, the PC12 cells proliferated, there was an increased number of neurites, and neurite length was enhanced. After middle- and high-dose natural-cerebrolysin treatments, the percentage of neurite-positive cells, as well as the average length of neurites, was significantly greater than the normal blank serum-treated PC12 cells （P 〈 0.05 or P 〈 0.01）. Compared with the blank control group, MAP2 expression in the Aβ1-40-treated PC12 cells was significantly inhibited, and the cell proliferation rate was significantly decreased （P 〈 0.01）. Following incubations with natural-cerebrolysin-containing serum, MAP2 expression and cell proliferation rate in the PC12 cells were significantly increased in a dose-dependent manner, compared with treatments with blank control serum （P 〈 0.05 or P 〈 0.01 ）. CONCLUSION： Natural-cerebrolysin exhibited antagonistic effects on neurotoxicity in Aβ1-40 induced Alzheimer＇s disease in vitro models. These effects were likely related to cell proliferation and the upregulation of intracellular MAP2 expression.

Antioxidants, anti-proliferative, anti-inflammatory, anti-diabetic and anti-microbial effects of isolated compounds from Swertia corymbosa (Grieb.) Wight ex C.B. Clark – An in vitro approach

The present study,antioxidant,enzyme inhibitory,anti-inflammatory and antimicrobial activity of isolated compounds such as Decussatin(1),Gentiacaulein(2),Swertianin(3),1,8-dihydroxy-2,6-dimethoxy xanthone(methylswertianin)(4)8-hydroxy-1,2,4,6-tetramethoxyxanthone(5)and 1,2-dihydroxy-6-methoxyxanthone-8-O--d-xylopyranosyl(6)were investigated using an in vitro model.Results of antioxidant studies revealed that the compound 6 possessed an efficient 2,2-diphenyl-1-picryl-hydrazyl(DPPH•)(IC5007.19±4.56μmol/mL),2,2-azinobis(3-ethylbenzothiozoline-6-sulfonic acid)(ABTS•^+)(42.62±0.25 mmol/L TE/g),superoxide(57.89±3.45μmol/mL),nitric oxide(18.45±1.23μmol/mL)and hydroxyl(12.13±2.76μmol/mL)radical scavenging activities,ferric reducing antioxidant power(14.76±0.10 molar Fe(II)/g),metal chelating(213.85±27.18 mg EDTA/g)ability.Compounds 6 and 3 exhibited significant anti-proliferative activity.Compound 6 displayed strongest antibacterial activity against Streptococcus pneumoniae and Escherichia coli with MIC value of 3.90μg/mL and 21.21±0.25 and 20.27±0.11 mm zone of inhibition at 25μg/mL concentration respectively.In the membrane stabilization and protein denaturation test 3 was the most potent with an IC50 value of 12.57,18.75μmol/mL respectively.

THE SPECIFIC PHOTODYNAMIC EFFECTS OF MONOCLONAL ANTIBODIES CONJUGATED WITH HEMATOPORPHYRIN DERIVATIVE ON GASTRIC CANCER IN VITRO AND IN VIVO

Murine monoclonal antibody (MoAb) BB4.3, raised against the human gastric cancer cell line BGC823, was puriffied with Protein A-Sepharose CL-4B affinity chromatography and identified as IgG2a. It was then conjugated with a hematoporphyrin derivative (HPD) by using carbodiimide. The qualitative analysis of this conjugate showed that the amount of free HPD was negligible and there were no IgG aggregates among the conjugates. The conjugate retained both the antibody and photochemical activity of HPD.In vitro, the phototoxic effect of this HPD-BB4.3 conjugate on target cells was about 15 times higher than that of free HPD. The quality of selective photocytotoxicity was proven by the greater cytotoxi-city this conjugate showed than that of corresponding normal mouse IgG (NIgG) conjugated with HPD. It showed less cytotoxicity to colon cancer cell line B-80 (negative reaction to MoAb BB4.3) than to BGC825. Moreover, its cytotoxicity to BGC823 cells could be blocked specifically by excess BB4.3 antibody, but not by another MoAb 3G9, which combines with BGC823 at different binding sites from MoAb BB4.3.Nude mice inoculated with 2 × 10- BGC823 cells were given HPD-BB4.3, HPD, HPD-NIgG, HPD plus BB4.3 and PBS, respectively then exposed to light. Four out of six animals treated with the HPD-BB4.3 conjugate remained tumor-free for a long period. Although two developed tumors, there was a significant difference between the HPD-BB4.3-treated group and all the control groups in tumor induction time, tumor growth rate, and survival time (p<0.001). The HPD-BB4.3 conjugate inhibited the growth of established tumors by more than 40% in comparison with control groups (p<0.05).

Preclinical Verification of Modulated Electro-Hyperthermia —Part I. In Vitro Research

Modulated electro-hyperthermia (mEHT) targets tissue’s natural electric and thermal heterogeneities to heat the cancer cells selectively. The applied 13.56 MHz radiofrequency (RF) is a carrier of the low-frequency modulation. The high-frequency part was chosen to select the malignant lesion using the specialties of the tumor: the higher conductivity and dielectric constant of the tumor than its host. The electric field selects the tumor, and the low-frequency amplitude modulation polarizes and excites the transmembrane proteins of the malignant cells. The dominant absorption of the energy by the microscopic clusters of the membrane rafts acts like nanoparticle heating. Exciting the membrane produces various apoptotic signals. The processes were modeled using silico and phantom experiments, which proved the concept. The preclinical verification was made in vitro and in vivo, and in the end, clinical proofs validated the method. Our objective is to follow all the development steps from the laboratory to the clinics in a trilogy of articles. This present is the first part, which deals with in silico, phantom, and in vitro research.

STUDY ON EFFECTS OF Yb^(3+) AND ITS COMPLEXES ON CELLS IN VITRO

This paper reports the effects of the Yh^(3+) and its four complexes on cultured cells.The results indicate that the 5 mmol YbCl_3 has evident toxicity on cultured cells,and after it is coordinated with ligands,its toxicity is decreased.These.four complexes neither destroy the growth nor inhibit the adliesion of diploid cell to the surface of the flasket,but they inhibit the adhesion of cancer cells,and result in cell death.The complex of Yb~?(3+) with ligand(1) can destroy the cancer cells in division phase.forming aberrant mitotic cells,but the other three complexes of Yb^(3+) with ligands can destroy the cancer cells in interphase.These effects depend on the dose of the complex.

EFFECTS OF BLEOMYCIN A5 COMBINED WITH CALMODU-LIN INHIBITOR ON THE PROLIFERATION OF S-180 CELLS IN VITRO

The effects of bleomycin A5 (BLM A5) alone and combined with calmodulin inhibitor N-(4-aminobutyl)-5-chloro-2-naphthalene sulfonamide (W-13) on the proliferation on S-180 cells in vitro were studied. IC50 of BLM used alone for the cells was about 2.63 μg/ml, but it was reduced to 1/3.8 and 1/9.5 of 2.63 μg/ml when plus W-13 1, 5 μg/ml respectively. The results indicated that nontoxic doses of W-13 enhanced the hinibition of cell proliferation under the condition of BLM 0.5 - 2.5 μg/ ml. In colony forming test, the survival fraction of S-180 cells treated with BLM plus W-13 was decreased to 1/87 - 240 of that of the cells treated with BLM alone. The results suggest that W-13 can enhance antitumor activity of BLM in vitro and may be used as an synergist of BLM A5 in vivo.

IN VITRO ANTI-HEPATOMA EFFECTS OF MONOCYTES AND KUPFFER CELLS ISOLATED FROM HEPATOMA PATIENTS AFTER TREATMENT WITH BIOLOGICAL IMMUNE STIMULANTS

Monocytes (MC), lymphocytes (LC) and Kupffer cells (KC) were isolated respectively from blood and surgical liver samples of patients suffering from he-patocellular carcinoma (HCC). 13 patients were given BCG, mixed bacterium vaccine (MBV) and human white blood cell interferon (IFN), the other 3 patients were not treated with any biological immune stimulants (BIS) and served as controls. The cytosta-tic and cytotoxic effects of MC and KC on human hepatoma SMMC-7721 (TC) were assayed in vitro and the numbers of T total (Tt), T helper (Th) and T suppressor (Ts) cells were counted using CD monoclonal antibody immunofluorescence. The results were as follows: (1) On the 7th day after the first administration of BIS, the cytostatic and cytotoxic effects of MC on TC showed obvious increase over pre-administration. The activity of BIS was 1 ?5 times as high as that in the controls. (2) After 3 administrations, the cytostatic effect of MC on TC increased to the normal level (84%), while the controls remained as before (45%). (3) On the 7th day after first administration, cytostatic and cytotoxic effects of KC on TC were 0.5 and 1 times higher respectively than those of the controls. (4) The numbers of Tt and Th of patients given BIS increased continuously; on the contrary Ts decreased in number. These results indicate that combined use of BCG, MBV and IFN can actively enhance the immune anti-hepatoma function of patients suffering from HCC.

Effects of Buyang Huanwu decoction and Astragalus mongholicus on platelet activating factor receptor activity in rabbits in vitro

BACKGROUND: The pharmacological action of traditional Chinese medicine compound is the comprehensive effect of the various ingredients, and the interactions of various ingredients are closely correlated with the final effect. In order to reveal the compatibility mechanism of BHD's prescription in treating and preventing ischemic cerebrovascular disease, we needed explore the effect and relation of ingredients in the prescription. OBJECTIVE: To observe the effect of Buyang Huanwu decoction (BHD) and Astragalus mongholicus on the activity of platelet activating factor receptor (PAFR) in the platelet of rabbits in vitro, and investigate the mechanism of Astragalus mongholicus. DESIGN: A decomposed recipes study. SETTING: Guangzhou University of Traditional Chinese Medicine. MATERIALS: Five New Zealand rabbits, weighing 2-3 kg, both sexes, were used. BHD was composed of Sheng Huang Qi 120 g, Dang Gui Wei 6 g, Chi Shao 4.5 g, Chuan Xiong 3 g, Di Long 3 g, Tao Ren 3 g, Hong Hua 3 g. The prescription for activating blood circulation consisted of Dang Gui Wei 6 g, Chi Shao 4.5 g, Chuan Xiong 3 g, Di Long 3 g, Tao Ren 3 g and Hong Hua 3 g. The prescription for invigorating qi consisted of 120 g Sheng Huang Qi. The prepared herbal pieces were purchased from the traditional Chinese medicine Dispensary of Foshan Second People's Hospital, and appraised by Professor Xu from Science of Chinese Materia Medica College, Guangzhou University of Traditional Chinese Medicine. 3H-PAF was supplied by Amersham Co., Ltd. (specific activity: 6. 475 TBq/mmol; batch number: 200402); PAF standard by Biomol Co., Ltd. (batch number: P1318V). METHODS: The experiments were carried out in the Laboratory of Nuclear Medicine, Guangzhou University of Traditional Chinese Medicine from September to December 2004. ① Injections of BHD, prescriptions for activating blood circulation and invigorating qi were prepared by the decoction and alcohol sedimentation technique. Rabbit common carotid artery blood (40 mL) was drawn via intubation to prepare platelet suspension of (0.8-1.0)×1010 L-1. ② Determination of 3H-PAF and washed PAFR binding: The general combination tube (T) contained washed platelet-rich plasma (WPRP) 380 μL + 3H-PAF (0.35 nmol/L)10 μL+distilled water 5 μL; The nonspecific binding tube (P) contained WPRP 380 μL+3H-PAF(0.35 nmol/L)10 μL+cold PAF (1 μmol/L) 5 μL; The sample tube (Y) contained WPRP 380 μL+3H-PAF(0.35 nmol/L)10 μL+experimental medicine (injection of BHD, prescriptions for activating blood circulation or invigorating qi) 5 μL. The test was conducted for three times for each sample in the same way as mentioned above. The samples were shaken on the oscillator for 30 s, then bathed at 25 ℃ for 40 minutes, and the reaction was terminated with cold Tris buffer containing 0.1% BSA, multichannel cell detachment separator was used for vacuum suction to filter the separated free 3H-PAF, and the filter paper was washed with cold Tris buffer for four times, then dried in the baking oven (80 ℃) for 1 hour, and placed in xylol liquid scintillator, and the radioactivity was determined automatically by the liquid scintillation detector. The mean of the three parallel tubes was calculated. The specific binging inhibition rate was calculated: SBIR=[(T-Y)/(T-P)]×100%]. ③ Univariate analysis of variance was conducted. And for comparison of each paired groups, the q test was adopted. MAIN OUTCOME MEASURES: Effect of BHD whole prescription, prescriptions for activating blood circulation and invigorating qi on the specific binding inhibition rate of 3H-PAF and PAFR. RESULTS: BHD, prescriptions for activating blood circulation and invigorating qi were all able to inhibit the specific binding of 3H-PAF to PAFR, the specific blinding inhibition rates were (45.90±7.50)%, (97.90±1.84)% and (26.75±2.48)%, respectively, and there were significant differences between every two groups (P < 0.01). CONCLUSION: Single Astragalus mongholicus (120 g) can inhibit the specific blinding of PAFR in the platelet of the rabbit with 3H-PAF, but the combination of Astragalus mongholicus with the drugs for activating blood circulation in BHD can significantly decrease the inhibiting action of the latter on PAFR activity of the platelet, reflecting the combined mechanism of 'removing blood stasis without injuring the vital qi' in BHD.

Synergistic effects of brain-derived neurotrophic factor and retinoic acid on inducing the differentiation of bone marrow stromal cells into neuron-like cells in adult rats in vitro

BACKGROUND： Under induction of retinoic acid （RA）, bone marrow stromal cells （BMSCs） can differentiate into nerve cells or neuron-like cells, which do not survive for a long time, so those are restricted to an application. Other neurotrophic factors can also differentiate into neuronal cells through inducing BMSCs; especially, brain-derived neurotrophic factor （BDNF） can delay natural death of neurons and play a key role in survival and growth of neurons. The combination of them is beneficial for differentiation of BMSCs. OBJECTIVE： To investigate the effects of BDNF combining with RA on inducing differentiation of BMSCs to nerve cells of adult rats and compare the results between common medium group and single BDNF group. DESIGN： Randomized controlled animal study SETTING： Department of Neurology, Affiliated Hospital of Xuzhou Medical College MATERIALS： The experiment was carried out in the Clinical Neurological Laboratory of Xuzhou Medical College from September 2003 to April 2005. A total of 24 SD rats, of either gender, 2 months old, weighing 130-150 g, were provided by Experimental Animal Center of Xuzhou Medical College [certification： SYXK （su） 2002-0038]. Materials and reagents： low-glucose DMEM medium, bovine serum, BDNF, RA, trypsin, separating medium of lymphocyte, monoclonal antibody of mouse-anti-nestin, neuro-specific enolase, glial fibrillary acidic protein （GFAP） antibody, SABC kit, and diaminobenzidine （DAB） color agent. All these mentioned above were mainly provided by SIGMA Company, GIBCO Company and Boshide Company. METHODS： Bone marrow of SD rats was selected for density gradient centrifugation. BMSCs were undertaken primary culture and subculture; and then, those cells were induced respectively in various mediums in total of 3 groups, including control group （primary culture）, BDNF group （20 μg/L BDNF） and BDNF＋RA group （20 μg/L BDNF plus 20 μg/L RA）. On the 3^rd and the 7^th days after induction, BMSCs were stained immunocytochemically with nestin （sign of nerve stem cells）, neuron-specific enolase （NSE, sign of diagnosing neurons） and GFAP （diagnosing astrocyte）, and evaluated cellular property. MAIN OUTCOME MEASURES ： Induction and differentiation in vitro of BMSCs in 3 groups RESULTS： （1） Induction and differentiation of BMSCs： Seven days after induction, cells having 2 or more apophyses were observed. Soma shaped like angle or erose form, which were similar to neurons and glial cells having strong refraction. （2） Results of immunocytochemical detection： Three days after induction, rate of positive cells in BDNF＋RA group was higher than that in BDNF group and control group [（86.15±4.58）%, （65.43±4.23）%, （4.18±1.09）%, P 〈 0.01]. Seven days after induction, rate of positive cells was lower in BDNF group and BDNF＋RA group than that in both groups at 3 days after induction [（31.12±3.18）%, （29.35±2.69）%, P 〈 0.01]; however, amounts of positive cells of NSE and GFAP were higher than those at 3 days after induction （P 〈 0.01）; meanwhile, the amount in BDNF＋RA group was remarkably higher than that in BDNF group （P 〈 0.01）. CONCLUSION： Combination of BDNF and RA can cooperate differentiation of BMSCs into neurons and astrocyte, and the effect is superior to single usage of BDNF.

The Effects of Dietary Restriction and Aging on in Vivo and in Vitro Binding of Aflatoxin B_1 to Cellular DNA

Laboratory animals maintained on a reduced calorie but nutritionally adequate diet have extended life spans and lowered incidences of spontaneous and chemically induced cancers compared to ad libitum- fed counterparts. Many of the effects of dietary restriction on laboratory animals have been suggested to be related to a deceleration of the aging process. The inhibition of age-related changes in xenobiotic metabolizing enzyme activities by dietary restriction has previously been reported. Alterations of these enzyme activities may cause changes in metabolic activation of carcinogens and, therefore, carcinogen-DNA binding. DNA-repair capability has also been reported to be enhanced in diet-restricted rats. Using AFB1 as a model carcinogen, we have studied in vivo and in vitro hepatic AFB1 -DNA binding, demonstrating that dietary restriction (60% of ad libitum consumption) may decrease the metabolic activation of AFB1, and subsequently reduce AFB 1-DNA binding. Our preliminary results obtained from the AFB 1-DNA binding experiments in isolated hepatocytes suggest that the observed age-dependent reduction in AFB 1-DNA binding which may be attributed to a loss of metabolic activating capability was delayed in the diet-restricted rats.

Effects of GW5074 in the process of imDCs inducing differentiation of naive CD4^+ T cells into Treg cells in vitro

Objective To establish a stable and efficient method of culturing imDCs in vitro,and to explore the effect of GW5074,which blocks ERK1 /2 signal pathway in the process of immature dentritic cells ( imDCs) on inducing differentiation of the naive allogeneic CD4 + T

In Vitro Probiotic Effect of Pseudostellaria heterophylla Fibrous Roots on Feed Bacillus subtilis and Preliminary Study on Its Solid Fermentation

Traditional Chinese medicine Pseudostellaria heterophylla fibrous roots have been widely studied and applied in non-antibiotic clinical breeding of livestock and poultry. In vitro probiotic effect of P. heterophylla fibrous roots extract on feed probiotics Bacillus subtilis BD-K010 was studied, and the feasibility of solid fermentation of P. heterophylla fibrous roots was preliminarily evaluated. For in vitro probiotic effect, the increased concentration of P. heterophylla fibrous roots extract dependently increased the total biomass of B. subtilis BD-K010;1.0% P. heterophylla fibrous roots extract received the best effect, and the final p H of 1.0% experimental group was closer to neutral. Meantime, B. subtilis BD-K010 with optimum concentration of P. heterophylla fibrous roots showed significantly higher in vitro antibacterial effect than the control group(P<0.01),and the antibacterial effects on Escherichia coli and Staphylococcus aureus were improved by 51.99% and 63.16%, but it was ineffective to Salmonella. For solid fermentation, the profile of substrate complex and appendage flocculent structure on substrate surface at the end of fermenta-tion in experimental groups added with B. subtilis BD-K010 and cellulase plus BD-K010 were more complex;the live bacteria number, polysaccharide content and saponin content at fermentation end-point in two experimental groups were extremely higher than those in the control group(P<0.01).P. heterophylla fibrous roots extract had good in vitro probiotic effect on B. subtilis BD-K010 and promoted its antibacterial effect, and it is feasible to use probiotics for solid fermentation of P. heterophylla fibrous roots and to improve effective components. It is of great significance to further de-velop and utilize P. heterophylla fibrous roots resources in modern animal husbandry.

Effects of Mirazid~ and myrrh volatile oil on adult Fasciola gigantica under laboratory conditions

Objective:To evaluate the effects of Mirazid and myrrh volatile oil on adult Fasciola gigantica(F.gigantica) under laboratory conditions.Methods:The effects of oleoresin extract of myrrh(Mirazid) and myrrh volatile oil on the surface morphology of adult F.gigantica following treatment in vitro had been determined by scanning electron microscopy.The results were compared with those observed in the fluke tegument following incubation in triclabendazole sulphoxide(TCBZ-SO),active form.(Fasinex,Ciba-Geigy).Results:Observations of the efficacy of Mirazid oleoresin extract and myrrh volatile oil indicated that both products showed dosedependent anthelmintic efficacy.The anterior half of the fluke was consistently more severely affected than the posterior half.The surface changes induced by Mirazid oleoresin extract were less severe than those observed after exposure to either myrrh volatile oil or TCBZ-SO.Flukes showed swelling after these treatments,but its level and blebbing were much greater with myrrh volatile oil;in which patches of tegumental sloughing were observed in the apical cone and the posterior mid-body region of flukes.This was not observed after treatment with Mirazid oleoresin extract.Conclusions:The comparatively more disruption,observed in myrrh volatile oil exposed specimens,compared to that exposed to Mirazid oleoresin extract might suggest that the anthelmintic activity of Mirazid oleo resin extract was attributed to its content of volatile oil.So,increasing the concentration of myrrh volatile oil in Mirazid might possibly help to developing its anthelmintic activity._______________________________________________

Radiation Induced Bystander Effect: From in Vitro Studies to Clinical Application

In the past 20 years, the classic paradigm in radiobiology recognizing DNA as the main target for the action of radiation has changed. The new paradigm assumes that both targeted and non-targeted effects of radiation determine the final outcome of irradiation. Radiotherapy is one of the main modality treatments of neoplastic diseases with intent to cure, or sometimes to palliate only, thus radiation-induced non-targeted effect, commonly referred to as the radiation-induced bystander effect (RIBE) may have a share in cancer treatment. RIBE is mediated by molecular signaling from radiation targeted cells to their non-irradiated neighbors, and comprises such phenomena as bystander effect, genomic instability, adaptive response and abscopal effect. Whereas first three phenomena may appear both in vitro and in vivo, an abscopal effect is closely related to partial body irradiation and is a systemic effect mediated by immunologic system which synergizes with radiotherapy. From the clinical point of view abscopal effect is particularly interesting due to both its possible valuable contribution to the treatment of metastases, and the potential harmful effects as induction of genetic instability and carcinogenesis. This review summarized the main results of investigations of non-targeted effects coming from in vitro monolayer cultures, 3-dimentional models of tissues, preclinical studies on rodents and clinically observed beneficial abscopal effects with particular emphasis on participation of immunotherapy in the creation of abscopal effects.

The Inhibitory Effect of Extracts From Fructus lycii and Rhizoma polygonati on in vitro DNA Breakage by Alternariol

Alternariol caused DNA single-strand breakage. Conversion of the closed circular double-stranded supercoiled DNA (pBR 322) to the nicked circular form and linear form was used to investigate the effect of extracts of some Chinese medical herbs on DNA nicking induced by alternariol. Some substances in the extracts of Rhizoma polygonati (RP) and Fructus lycii (FL) were shown to protect DNA from the attack by alternariol.Some substance in the RP may bind to plasmid DNA, and this binding reduces the electrophoretic mobility of DNA. These results indicate that substances from FL and RP may be used as DNA protectors. It is possible that they play an important role in preventing cancer.