Facilitating the gut brain axis by probiotic bacteria to modulate neuroimmune response on lead exposed zebra fish models

Unravelling the efficacy of gut biome has a major impact on health. An unbalanced microbiome composition is linked to many common illnesses such as gut dysbiosis, mental deformities and immunological imbalance. An optimistic influence on the gut biome can be made by consumingprobiotics. This would stimulate neuroprotection and immunomodulation intended by heavy metals pollution. Lead is a major source of neurotoxin that can induce neural deformities. Lactobacillusspecies isolated from curd were characterized to confirm its specificity. Zebra fish was reared at standard conditions and preclinical assessment on the intensity of induced neurotoxin lead was performed. The embryo toxic assay, immunomodulation effects and animal behavioural models endorsed the consequence of neurotoxicity. Different concentrations of bacterial isolate with standard antidepressant was considered for analysing the vigour of toxicity and its influence on cognitive behaviour by novel tank diving method. The restrain in the animal behaviour was also conferred by all the test samples with a decreased bottom dwelling time which was authenticated with haematology and histopathological studies. The alterations in morphology of the lymphocytes were balanced by the treated test samples. This study paves a twofold potential of probiotic as neuroprotectant and immune modulator against heavy metal toxicity.

Study on the Heart Protective Effect of Panax quinquefolium Extract Based on Zebra Fish Model / 中国药房

OBJECTIVE：To study the heart protective effect of Panax quinquefolium extract of Zebra fish. METHODS ：The drug dosage of P. quinquefolium extract and the optimal modeling dose of terfenadine were determined by tolerance test ；24 hours post fertilization （24 hpf），Zebra fish were divided into control group （0.5％ dimethyl sulfoxide ），model group （2.5 μg/mL terfenadine），positive control group （2.5 μ g/mL terfenadine + 30 μ g/mL resveratrol） and different mass concentration P. quinquefolium extract groups （2.5 μg/mL terfenadine+5，10，25 μg/mL P. quinquefolium extract，by raw material ）. 48 h after incubation，the heart rate and sinus venous-bulbus arteriosus （SV-BA）interval of Zebra fish were used as indexes ，and the heart injury repair rate was calculated to investigate the heart protection of P. quinquefolium extract. Another 24 hpf Zebra fish were divided into blank control group ，model group ，positive control group ，20 batches of P. quinquefolium extract from different producing area groups （5 μg/mL，by raw material ），with same administration route and 48 h of incubation. The heart rate ，SV-BA interval of heart was determined ，heart injury repair rate and heart injury relative repair rate were calculated ；the heart protective effect of P. quinquefolium extract was further validated. RESULTS ：Compared with blank control group ，heart rate was decreased significantly（P＜0.01），while SV-BA interval was increased significantly in model group （P＜0.01）. Compared with model group，heart rate of Zebra fish was increased significantly in positive control group and 5，10，25 μg/mL P. quinquefolium extract groups（P＜0.01），while SV-BA interval of Zebra fish was shortened significantly in 5，10 μg/mL P. quinquefolium extract groups （P＜0.05 or P＜0.01）；heart injury repair rate of Zebra fish in 5 μg/mL P. quinquefolium extract group reached 73.77％. In validation test ，compared with model group ，heart rate was increased significantly （P＜0.01），while SV-BA interval of Zebra fish was decreased significantly in 20 batches of P. quinquefolium extract groups （P＜0.01）；heart injury repair rate ranged 70.45％-85.78％，and heart injury relative repair rate ranged 75.48％-98.12％. CONCLUSIONS ：P. quinquefolium extract has good heart protective effects.

Angiogenesis Activity of Morindae Officinalis Radix from Different Origins Based on Zebrafish Model / 中国实验方剂学杂志

Objective: To study the effect of Morindae Officinalis Radixe extract from different origins on the vascular growth of damaged internodes of zebra fish, so as to discuss regional differences of Morindae Officinalis Radix from different origins. Method: A zebra fish model of internode vascular injury induced by PTK787 was established to study the effect of Morindae Officinalis Radix extract from Guangdong, Guangxi and Fujian on the growth of internode vascular in zebra fish, and detect the number of internode angiogenesis in zebra fish treated with PTK787.On the basis of principal component analysis, relative angiogenesis rate was taken as the index, and the experimental data was analyzed by SPSS 17.0 statistical software. Result: The extract of Morindae Officinalis Radix had a repairing effect on the growth of damaged internode blood vessels of zebra fish, and the extracts of Morindae Officinalis Radix from Guangdong Gaoyao, and Guangdong Yunan, were better than those from other producing areas. The average number of internode angiogenesis was 9, the average length of angiogenesis was 448-504 μm, and the relative rate of angiogenesis was between 7.9% and 8.86%. The results of cluster analysis showed that Morindae Officinalis Radix of 12 different origins were classified into four categories, Guangdong Deqing, and Guangdong Gaoyao, Guangxi Cangwu and Guangxi Cenxi, and Guangdong Yunan, and Guangxi Cangwu could be better clustered. Conclusion: There are obvious regional differences in the pro-angiogenic effect of Morindae Officinalis Radix from different origins. Relevant studies provide basic data for the quality assessment of Morindae Officinalis Radix medicinal materials.

Iridoid glycoside extraction technology and anti-thrombotic activity of Osmanthus fragrans seeds / 中草药

Objective: To optimize the extraction technology of two major iridoid glucosides (specnuezhenide and nuezhenoside G13) from four kinds of Osmanthus fragrans (OF) seeds, and to evaluate the anti-thrombotic activity of OF seeds. Methods: The orthogonal-test experiment was employed to optimize the parameters including ethanol concentration, liquid-material ratio, and extraction time for three extraction methods (ultrasonic extraction, reflux extraction, and microwave extraction). The extraction yield, content, and total peak area of iridoid glucosides were selected for weighted analysis to determine the best extraction method and technology. Additionally, an anti-thrombotic zebra fish model was established for biological evaluation of OF seeds. Results: Microwave extraction was the best method for iridoid glucosides extraction with the optimal conditions of ethanol concentration 55%, material-liquid ratio 1∶10, and microwave time 15 min. HPLC analysis showed that there was no significant difference in chemical composition among the four kinds of OF seeds. In zebra fish biological screening model, OF seeds displayed a weak inhibitory effect on the growth of thrombus and exhibited a pericardial edema effect in high dose-treated group. Conclusion: In this paper, extraction technology of two iridoid glucosides from four different kinds of OF seeds and preliminary anti-thrombotic activity evaluation of OF seeds were investigated. These results can provide the reference for further development and utilization of the agricultural waste of OF seeds.

Applicability of Zebra Fish Thrombosis Model in Antithrombotic Activity Screening of Chinese Materia Medica / 中国中医药信息杂志

Objective To investigate the applicability of zebra fish thrombosis model in antithrombotic activity screening of Chinese materia medica.Methods The living zebra fish thrombosis model was induced by adrenaline hydrochloride. Zebra fish were randomly divided into blank control group, model group, positive medicine group and medication group. Each group was given the corresponding medicine or embryo culture water. O-anisidine staining solution was used to stain and calculate the staining intensity of erythrocytes in zebra fish heart, and quantitative analysis was carried out. The platelet aggregation of transgenic zebra fish was observed and under qualitative analysis. Results Compared with the model group, 100μg/mL salvianolic acid B, 300, 900μg/mL aqueous extract of Salvia miltiorrhiza, 45μg/mL 95% ethanol extract and 400, 1200μg/mL hypothalamus could significantly inhibited the formation of zebra fish thrombosis (P<0.01).ConclusionZebra fish thrombosis model has good applicability in antithrombotic activity screening of Chinese materia medica.

Application advances on zebra-fish model in study on Chinese materia medica / 中草药

Zebra fish, as a model organism, has been widely used in genetics and developmental biology. In recent years, researches on the zebra fish have expanded and extended the field of Chinese materia medica (CMM) since it is a useful model of choice for in vivo pharmacology dynamic screening. Through zebra fish, we are able to fully detect and identify the substances of activity and/or toxicity of CMM with the advantage of real-time dynamic monitoring, thus achieving high-throughput screening. In this article, we outline the features of the zebra fish model and summarize its progress in screening toxicity and active substances of CMM as well as metabolic study in vivo.

Research of zebrafish quality standardization / 中国实验动物学报

At present, zebrafish has played an increasingly important role in models for human development and diseases and several areas of life sciences.As a newly laboratory animal resource, standardization research has become the technical bottleneck to be solved and an inevitable trend.In this review, we summarized the research history and character-istics of zebrafish and the status of quality standardization.We also discussed the main problem facing by the standardiza-tion research of zebrafish as a newly laboratory animal.We hope that the data can provide useful reference for the develop-ment of zebrafish quality standardization research.

Acute effects of mercuric chloride on glycogen and protein content of Zebra fish, Danio rerio.

Presence of mercury and other heavy metals above permissible levels in water bodies across the globe is posing a serious threat to aquatic biota and public health. Occurrence of mercury above the permissible limits in the aquatic ecosystem of Hyderabad city is well established. In this context, we carried out static- renewal bioassays on the zebra fish, Danio rerio exposed to different concentrations of mercuric chloride, and the 96-h median lethal concentration (LC50) was found to be 0.077 mgl-1. Behavioral manifestations like loss of scales, hyper secretion of mucus, surfacing and darting movements, loss of balance, irregular swimming patterns were noticed in the fish exposed to 0.077 mgl-1. The present study also examined the toxic effects of mercuric chloride on vital biochemical constituent’s total glycogen and total protein. Significant decrease (p<0.001) in glycogen and protein content of fish exposed to 0.077 mgl-1

Bioactive molecules: current trends in discovery, synthesis, delivery and testing

Important bioactive molecules are molecules that are pharmacologically active derived from natural sources and through chemical synthesis. Over the years many of such molecules have been discovered through bioprospective endeavours. The discovery of taxol from the pacific yew tree bark that has the ability in stabilising cellular microtubules represents one of the hallmarks of success of such endeavours. In recent years, the discovery process has been aided by the rapid development of techniques and technologies in chemistry and biotechnology. The progress in advanced genetics and computational biology has also transformed the way hypotheses are formulated as well as the strategies for drug discovery. Of equal importance is the use of advanced drug delivery vehicles in enhancing the efficacy and bioavailability of bioactive molecules. The availability of suitable animal models for testing and validation is yet another major determinant in increasing the prospect for clinical trials of bioactive molecules.

Expression of pax5 gene in wildtype zebrafish embryos / 第二军医大学学报

Objective To study the role and related mechanisms of pax5 gene during the development of the embryos, especially in the early development of B cell and midbrain, and to observe the expression of pax5 gene in wild type zebra fish embryos. Methods Total RNA of Tue Un gen wild type zebra fish embryos was extracted to obtain cDNA of pax5 gene by RT-PCR with specific primers. The c DNA of pax5 gene and pCS2+ vector were double digested with EcoR I and Xba I, and then ligated by T4 DNA ligase. The recombinant vector was verified by double digestion, colony PCR screening and sequencing. The verified recombinant vector was then used to synthesize digoxin labeled anti-sense mRNA probe of pax5 gene using T3 RNA polymerase in vitro transcription system. The generated probes were used to detect pax5 gene expression in zebra fish embryos by whole-mount in situ hybridization. Results The pCS2+-pax5 recombinant plasmid and the probe of digoxin-labeled anti-sense mRNA of pax5 gene was successfully constructed. Whole-mount in situ hybridization showed that pax5 gene was expressed in the cerebellum and midbrain hindbrain boundary from 18-72 h post-fertilization (hpf). In the cochlea pax5 gene was expressed from 24-72 hpf and the expression was increased as time went by. From 18-48 hpf pax5 gene is found in the notochord. Conclusion Pax5 gene is highly expressed in the brain and notochord of zebra fish embryos. Pax5 gene expression is found in the cochlea of the Tue bingen wild type zebra fish' s embryos for the first time. It is suggested that pax5 may play an important role in the development of the nervous system in the early zebra fish embryos.

A light- and electron microscopic study of primordial germ cells in the zebra fish (Danio rerio)

In sexually reproducing organisms, primordial germ cells (PGCs) give rise to the cells of the germ line, the gametes. In many animals, PGCs are set apart from somatic cells early during embryogenesis. This study explores the origin of primordial germ cells (PGCs) of the zebra fish and examines their morphology during early development (1st day-15th day). PGCs were selectively stained by the alkaline phosphatase histochemical reaction and viewed by light and electron microscopy from the time they are first detectable in the yolk sac endoderm. PGCs occurred in the subendodermal space on the syncytial periblast; differing from the surrounding endodermal cells. Later the PGCs moved to between the blastoderm and yolk sac and transferred to the dorsal mesentery where they formed gonadal anlage with mesoderm cells. PGCs were easily distinguished from somatic cells by their morphology and low electron density of their nuclei. Under light microscopy, PCGs were rounded with a distinct cytoplasmic membrane.