Endothelin (ET), recently isolated from culture supernatant of porcine aortic endothelial cells, is a novel potent vasoconstrictive substance. In this paper a highly specific rabbit antisera against endothelin was used to study the localization of ET in the isolated endothelial cells, from human umbilical veins and porcine arota, and that of ET in the cryostat cross sections of porcine aorta by using ABC immunohistochemical technique. The results demonstrate that a varied amounts of specific ET-like immunoreactive substance exist in the endothelial cytoplasm of human umbilical veins and porcine aorta which provides the mor- phological evidence for elucidating the endocrine function of vascular endothelial cells.

Brain natriuretic peptide(BNP) is a recently discovered novel neuropeptide of 26 amino acid residues, isolated from porcine brain, that is of similar potency to atrial natriuretic factor(ANF) in natriuretic, diuretic, hypotensive and smooth muscle relaxant activities. we used a highly selective antisera against BNP raised in rabbit to observe its distribution and localization in some brain areas and some other peripheral tissues by utili zing high sensitive avidin-biotin-peroxidase complex technique. Positive brain natriuretic peptide immunoreactive (BNPir) fibers and cell bodies were observed in the lateral hypothalamic area, caudate-putamen, hippocampus, amygdala, and supraoptic and paraventricular nuclei. A lot of BNPir granules were also found in rat atria. They were of similar localization to that of ANF. Most of the specific granules were accumulated in cytoplasm at both nuclear poles of atrial myocytes. The BNP immunoreactivity is less intense than that of the ANF. Some of scanty, diffuse and fine BNPir granules could also be observed in ventricular myocytes.The coexistence of both BNP and ANF in the brain and heart indicates that BNP may function as a neuropeptide and circulating hormone, and suggests the possibility that the physiological effects such as diuretic natriuretic, hypotensive and smooth muscle relaxant activities so far thought to be mediated by ANF may be regulated through a dual mechanism involving both BNP and ANF. In addition, some BNPir positive cells were also present in the anterior and intermediate lobes of rat pituitary gland. The significance of BNP in hypophysis would be elucidated in the further studies.

Objective: To study the influence of Centipede Acidic Protein(CAP) on the proliferation and collagen synthesis of cultured neonatal rat cardiac fibroblasts(CFb) induced by angiotensinⅡ(AngⅡ),and to explore the mechanisms of CAP on cardiac fibrosis.Methods: Neonatal rat cardiac fibroblasts were treated with AngⅡ to produce fibrosis model.The effects of CAP on proliferation of CFb were observed by MTT colorimetric assay,synthesis of collagen was observed by the hydroxyproline concentration.The NO contents were measured by Nitric acid reductase method.The c-myc expression was examined by semi-quantitative RT-PCR analysis.Results: Compared with that of control group,the proliferation,collagen synthesis and the levels of c-myc mRNA expression of CFb in the model group increased,while the NO contents decreased obviously(P

Precision medicine for cancer patients aims to adopt the most suitable treatment options during diagnosis and treatment of individuals.Detecting circulating tumor cell (CTC) or circulating tumor DNA (ctDNA) in plasma or serum could serve as liquid biopsy,which would be useful for numerous diagnostic applications.Liquid biopsies can help clinicians screen and detect cancer early,stratify patients to the most suitable treatment and real-time monitoring of treatment response and resistance mechanisms in the tumor,evaluate the risk for metastatic relapse,and estimate prognosis.We summarized the advantages and disadvantages of tissue and liquid biopsies.We also further compared and analyzed the advantages and limitations of detecting CTCs,ctDNAs,and exosomes.Furthermore,we reviewed the literature related with the application of serum or plasma CTCs,ctDNAs,and exosomes for diagnosis and prognosis of cancer.We also analyzed their opportunities and challenges as future biomarkers.In the future,liquid biopsies could be used to guide cancer treatment.They could also provide the ideal scheme to personalize treatment in precision medicine.

Matteuccia struthiopteris is a nature plant, which contains a lot of potential active components. In the present study, we investigated the effect of polysaccharides extracted from Matteuccia struthiopteris on lupus-like syndrome induced by Campylobacter jejuni CJ-S131 in BALB/c mice. Mice were randomly divided into normal, model control, SLE model (vehicle treated), Matteuccia struthiopteris polysaccharides treated (30 and 15 mg x kg(-1)) groups and prednisone 5 mg x kg(-1) treated groups. The effect of Matteuccia struthiopteris polysaccharides (Ms) on weight and organ index of BALB/c mice was detected. Autoantibodies and total IgG production were measured by enzyme linked immunosorbent assay. Proteinuria was measured and kidneys were examined by light microscopy. Compared with SLE model group, treatment with Matteuccia struthiopteris polysaccharides 30 and 15 mg x kg(-1) reduced weight loss and Matteuccia struthiopteris polysaccharides 15 mg x kg(-1) reduced spleen swelling (P < 0.05). The increased production of autoantibodies and total immunoglobulin G (IgG) were also significantly inhibited. Matteuccia struthiopteris polysaccharides protected kidney against glomerular injury in BALB/c mice with reduced immunoglobulin deposition and lowered proteinuria (P < 0.01). Matteuccia struthiopteris polysaccharides had a protective effect on lupus-like syndrome induced by CJ-S131 in BALB/c mice.

Laser scanning confocal microscope (LSCM) is currently the only equipment to observe fluorescence. However, this technique has disadvantages such as high cost and long test process. In this study, we developed a new system of laser-induced fluorescence (LIF) for microfluidic chip applied to detecting the expression of green fluorescent protein (GFP) in Bacillus subtilis. This novel system was comprised of laser device, optics unit, microfluidic chip, photomultiplier and computer treatment unit. The tests indicated that microfluidic chip could detect the expression of GFP as sensitively as LSCM in Bacillus subtilis. Moreover, this LIF detection system could instead of PCR to identify the positive clone in this special case. Nevertheless, the LIF system only was suitable to detect the fluorescent strength of GFP, and could not meet the request of some cases for example protein location. Therefore, this system will be applied in environmental detection with microbe, drug discovery and other cases.
Bacillus subtilis ; isolation & purification ; metabolism ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Microfluidic Analytical Techniques ; methods

Objective To study the mechanism of Wuzi Yanzong Wan (Five-Seed Progeny Pill) on improving heat stress resistance of sertoli cells in rats based on data from the whole genome microarray.Methods Sertoli cells in primary culture were randomly divided into Wuzi Yanzong Wan (WZYZW) group, normal group and model group.The cells in three groups were given WZYZW solution and equal volume of PBS respectively.After 24 hours, rats in WZYZW group and model group were placed in water bath at 43 ℃ for 15 minutes, while rats in normal group were put in water bath at 37 ℃ for 15 minutes, then total RNA of three groups were extracted respectively and data was analyzed with gene chip technology.Results Compared with the model group, there were totally 1,203 differently genes in WZYZW group based on Sertoli cell gene expression profiles, in which 697 were down regulated and 506 were up-regulated.There were totally 473 differently expressed genes in the normal group, including 168 up-regulated genes and 305 down regulated genes.Among these down-regulated gene expressions, RGD1305298, Trpt1, Dll3, Abi3 and Ppp1r12b were significant in the WZYZW group and normal group, while Dusp7 was significant among up-regulated gene expressions.The genes mentioned above were related to cell cycle, apoptosis, cell metabolism, cell adhesion and Notch signal pathway, regulation of actin cytoskeleton, MAPK signaling pathway and tight junction pathway, etc.Conclusion The mechanism of Wuzi Yanzong pill in improving heat stress resistance of sertoli cells may involve maintenance of tight junctions between sertoli cells, inhibition of sertoli cells apoptosis and promotion of cell differentiation and maturation.

Aptamers are DNA or RNA fragments that can specifically bind to target substances.Because of the excellent properties such as strong binding force,high specificity,small physical size,chemical synthesis and modification,good biocompatibility,and low immunogenicity aptamers show wide application propects in biomedical researches.Aptamers can also bind specifically to receptors on the surface of cell membranes,and mediate the endocytosis of nanoparticles into cells,making them ideal drug targeting ligands.Organic nanomaterials have excellent application value in nanodrug delivery system because of their good biocompatibility and degradability.In this paper,the recent research progress of aptamers and organic nanomaterials drug delivery systems was reviewed.

BACKGROUND: Diabetic wound healing remains a major challenge due to the impaired functionality of angiogenesis by persistent hyperglycemia. Mesenchymal stem cell exosomes are appropriate candidates for regulating the formation of angiogenesis in tissue repair and regeneration. Here, we explored the effects of exosomes derived from human amniotic mesenchymal stem cell (hAMSC-Exos) on the biological activities of human umbilical vein endothelial cells (HUVECs) treated with high glucose and on diabetic wound healing and investigate lncRNAs related to angiogenesis in hAMSC-Exos. METHODS: hAMSCs and hAMSC-Exos were isolated and identified by flow cytometry or western blot. A series of functional assays such as cell counting kit-8, scratching, transwell and tube formation assays were performed to evaluate the potential effect of hAMSC-Exos on high glucose-treated HUVECs. The effect of hAMSC-Exos on diabetic wound healing were tested by measuring wound closure rates and immunohistochemical staining of CD31. Subsequently, the lncRNAs profiles in hAMSC-Exos and hAMSCs were examined to screen the lncRNAs related to angiogenesis. RESULTS: The isolated hAMSC-Exos had a size range of 30–150 nm and were positive for CD9, CD63 and CD81. The hAMSC-Exos facilitate the functional properties of high glucose-treated HUVECs including the proliferation, migration and the angiogenic activities as well as wound closure and angiogenesis in diabetic wound. hAMSC-Exos were enriched lncRNAs that related to angiogenesis, including PANTR1, H19, OIP5-AS1 and NR2F1-AS1. CONCLUSION Our findings demonstrated hAMSC-Exos facilitate diabetic wound healing by angiogenesis and contain several exosomal lncRNAs related to angiogenesis, which may represent a promising strategy for diabetic wound healing.

Panax ginseng(PG)and Panax notoginseng(PN)are highly valuable Chinese medicines(CM).Although both CMs have similar active constituents,their clinical applications are clearly different.Over the past decade,RNA sequencing(RNA-seq)analysis has been employed to investigate the molecular mechanisms of extracts or monomers.However,owing to the limited number of samples in standard RNA-seq,few studies have systematically compared the effects of PG and PN spanning multiple conditions at the transcriptomic level.Here,we developed an approach that simultaneously profiles transcriptome changes for multiplexed samples using RNA-seq(TCM-seq),a high-throughput,low-cost workflow to molecularly evaluate CM perturbations.A species-mixing experiment was conducted to illustrate the accuracy of sample multiplexing in TCM-seq.Transcriptomes from repeated samples were used to verify the robustness of TCM-seq.We then focused on the primary active components,Panax notoginseng sa-ponins(PNS)and Panax ginseng saponins(PGS)extracted from PN and PG,respectively.We also char-acterized the transcriptome changes of 10 cell lines,treated with four different doses of PNS and PGS,using TCM-seq to compare the differences in their perturbing effects on genes,functional pathways,gene modules,and molecular networks.The results of transcriptional data analysis showed that the tran-scriptional patterns of various cell lines were significantly distinct.PGS exhibited a stronger regulatory effect on genes involved in cardiovascular disease,whereas PNS resulted in a greater coagulation effect on vascular endothelial cells.This study proposes a paradigm to comprehensively explore the differences in mechanisms of action between CMs based on transcriptome readouts.