Isoxsuprine-induced alterations of pulmonary pressure-volume relationships in premature rabbits

Isoxsuprine hydrochloride was injected intramuscularly into 28-day-old rabbit fetuses (term is 31 days) three hours prior to delivery by cesarean section. When the drug-treated newborn rabbits were killed after 30 minutes of air breathing, their lungs retained more air on deflation, i.e., were more stable, than those of control animals that received saline. These findings are consistent with an increase alveolar surface active material in the drug-treated group, and indicate that isoxsuprine may enhance the release of surface active material into the alveolus that normally occurs at birth.     

Tonglian Decoction(通莲汤) Arrests the Cell Cycle in S-phase by Targeting the Nuclear Factor-Kappa B Signal Pathway in Esophageal Carcinoma Eca109 Cells

Objective:To investigate the anti-tumor activity and molecular mechanism of Tonglian Decoction(通莲汤,TLD) on esophageal carcinoma Eca109 cells.Methods:Eca109 cells were treated with TLD and its separated formulae,including the clearing-heat and detoxification formula(Q),activating-blood and promoting-qi formula(H) and nourishing-yin and blood formula(Z).Cell proliferation was measured using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay,cell morphology was observed using a microscope,the cell cycle was measured using flow cytometry and the activity of the nuclear factor-kappa B(NF-κB) signal pathway was detected by Western blot.Results:The half maximal inhibitory concentrations of TLD,Q and H were 386,771 and 729 mg/L,respectively.TLD,Q and H significantly inhibited cell proliferation,with 69.43%,60.84%and 61.90%of treated cells in the G1 phase of the cell cycle.The percentage of cells in S phase increased significantly after treatment with TLD,Q,and H compared with the control group(P0.05),and TLD showed the strongest effect.Z had no influence on the cell cycle compared with the control group(P0.05).Western blot detection indicated slight differences in the inhibition of the NF- k B pathway by the different formulae.TLD formula strongly inhibited IKKβ,NF-κB,interleukin-6 and tumor necrosis factor-α expression compared with the control group.Conclusions:TLD inhibited Eca109 cell proliferation by arresting cells in S phase.The possible mechanism might be related to inhibiting the NF- κB transduction cascade.The combination of the herbs found in the three separate formulae,H,Q and Z,work synergistically in TLD to produce the inhibitory effects of TLD treatment on Eca109 proliferation.     

CD109 deficiency induces osteopenia with an osteoporosis‐like phenotype in vivo

Osteoporosis is a global public health problem that is increasing along with an aging population. A major determinant of osteoporosis is high bone turnover, which results from osteoclast activation. CD109 is a glycosylphosphatidylinositol‐anchored glycoprotein, a deficiency that leads to a psoriasis‐like skin inflammation in mice. Although the expression of CD109 has been reported in mouse pre‐osteoclast cells, its function in osteoclasts in vivo remains largely unknown. To investigate the physiological role of CD109 in bone metabolism, we analyzed bones from wild‐type and CD109‐deficient adult mice. Micro‐computed tomography analysis of the femur (thigh bone) showed that bone volume was lower in CD109‐deficient mice than in wild‐type mice. Bone histomorphometric analysis showed not only a reduction in bone volume but also an increase in bone turnover in CD109‐deficient mice as compared with wild‐type mice. Additionally, we measured serum levels of several markers of bone turnover and found a significant increase in the N‐terminal telopeptide of type I collagen, a bone resorption marker, as well as alkaline phosphatase, a bone formation marker, in CD109‐deficient mice. These results indicate that CD109 deficiency induces a high‐turnover, osteoporosis‐like phenotype, which suggests that CD109 plays a role in bone metabolism in vivo.     

High-level expression of CD109 is frequently detected in lung squamous cell carcinomas

CD109 is a glycosylphosphatidylinositol (GPI)-anchored cell surface protein, which is a member of the alpha2-macroglobulin/C3, C4, C5 family of thioester-containing proteins. It has been reported that CD109 is expressed in a subset of hematopoietic cells, endothelial cells and several kinds of human tumors. Herein it is reported that the CD109 protein is preferentially expressed in lung squamous cell carcinomas compared with other types of lung carcinoma including adenocarcinomas, large cell carcinomas and small cell carcinomas. Immunohistochemical staining of surgically resected lung specimens using an anti-CD109 antibody detected CD109 expression in basal cells of bronchial and bronchiolar epithelia and myoepithelial cells of bronchial secretary glands, but not in bronchial and bronchiolar apical epithelial cells and alveolar epithelial cells. Furthermore, the CD109 immunoreactivity was observed in squamous cell carcinomas at a high frequency compared with other types of lung carcinoma. Although the detailed function of CD109 protein is unclear, these results suggest that CD109 expression may play a role in the development of lung squamous cell carcinoma.     

Genomic structure and tissue-specific expression of human and mouse genes encoding homologues of the major bovine seminal plasma proteins

Sperm capacitation is a maturation event that takes place in the female reproductive tract and is essential for fertilization. A family of phospholipid-binding proteins present in bovine seminal plasma (BSP proteins) binds the sperm membrane at ejaculation and promotes bovine sperm capacitation. Homologues of these proteins have also been isolated from boar, ram, goat, bison and stallion seminal fluid, suggesting that BSP proteins and their homologues are conserved among mammals. However, there have been no reports on BSP-homologous proteins in mice and humans to date. A search of the mouse and human genomes, using the nucleic acid sequences of BSP proteins, revealed the presence of three BSP-like sequences in the mouse genome, named mouse BSP Homologue 1 (mBSPH1), mBSPH2 and mBSPH3, and one sequence in the human genome (hBSPH1). Mouse epididymal expressed sequence tags corresponding to partial sequences of mBSPH1 and mBSPH2 were identified. The entire complementary DNA (cDNA) sequences of mBSPH1 and mBSPH2 from mouse epididymis and hBSPH1 from human epididymis were obtained by 5'-/3'-rapid amplification of cDNA ends (RACE) and encode predicted proteins containing two tandemly repeated fibronectin type II domains, which is the signature of the BSP family of proteins. Using RT-PCR, it was revealed that mBSPH1, mBSPH2 and hBSPH1 mRNA are expressed only in the epididymis. Expression of mBSPH3 was not detected in any tissue and probably represents a pseudogene. This work shows, for the first time, that BSP homologues are expressed in mouse and human and may be involved in sperm capacitation in these species.     

Nicotinic acid receptor subtypes and their ligands

Half a century ago, nicotinic acid (niacin) was introduced into the clinic as the first orally available drug to treat high cholesterol levels and to improve the balance between (V)low density lipoproteins (LDL) and high density lipoproteins (HDL). Remarkably, its putative mechanism of action has only been recently elucidated, particularly because of the cloning of a G protein-coupled receptor (HM74A or GPR109A). This receptor responds to both nicotinic acid and the ketone body beta-hydroxybutyrate, the latter thought to be the more probable endogenous ligand for HM74A. In this review, we will discuss the pharmacology and medicinal chemistry of this receptor subtype and a related one (HM74 or GPR109B). Although still in its infancy, the ligand repertoire is developing, and a number of compound classes have now been described, among which are both full and partial agonists. Antagonists, however, are still lacking, thus compromising thorough pharmacological studies. Mutagenesis experiments have provided clues regarding the ligand binding site; in particular, an arginine residue in transmembrane domain 3 of the receptor seems to recognize the acidic moiety present in nicotinic acid and related substances. HM74A has also been linked to one of the major side effects of nicotinic acid, that is, flushing, since this receptor subtype also occurs in skin immune cells. It is not known yet whether HM74 is also present on these cells. Since nicotinic acid is one of the few available medicines that raise HDL ("good cholesterol") levels, HM74A and HM74 appear promising targets for future pharmacotherapy.     

木犀草苷对人食管鳞状癌Eca109细胞生长的抑制作用及其机制

目的 研究木犀草苷对人食管鳞状癌Eca109细胞生长的抑制作用及其机制。方法 Eca109细胞经不同浓度木犀草苷处理后,MTT法检测Eca109细胞增殖;倒置显微镜观察Eca109细胞形态的变化;流式细胞术检测Eca109细胞周期变化及细胞凋亡情况;RT-PCR检测Eca109细胞cyclin D1、survivin和c-myc基因表达。结果 MTT实验表明,木犀草苷80、120、160、200、240 μmol/L对Eca109细胞均有抑制作用,且与浓度相关。木犀草苷可引起Eca109细胞形态改变、体积缩小,并与周围细胞脱离,浓度为240 μmol/L时使细胞呈出芽状,有的细胞伸出多个伪足样突起;浓度为160、240 μmol/L时,可将细胞阻滞于G₂/M期,诱导细胞凋亡;浓度为240 μmol/L并处理Eca109细胞48 h后,使Eca109细胞cyclin D1、survivin和c-myc基因表达低于对照组（P＜0.05）。结论 木犀草苷对食管鳞状癌Eca109细胞的生长有显著抑制作用,其通过改变细胞周期、诱导细胞凋亡及抑制相关基因的表达发挥抗肿瘤作用。     

Profiling of sperm proteins and association of sperm PDC-109 with bull fertility

Abstract

The composition of sperm proteins influences the fertilizing ability of sperm and hence the present study was conducted (i) to profile sperm proteins expression patterns in bulls of differing fertility index and (ii) to identify and relate the abundant sperm proteins with bull fertility. The semen samples were collected from Holstein-Friesian bulls (n?=?12) varying in conception rate (CR) (high/low). The frozen semen straws (three ejaculates, from each bull) were used to study (a) sperm kinetic parameters, (b) plasmalemma integrity, (c) mitochondrial membrane potential, and (d) chromatin distribution. Three bulls were randomly selected from each group (n?=?3) and the neat sperm pellets were subjected to percoll purification, followed by protein isolation using 0.1% Triton X100. The sperm kinetic parameters, plasmalemma integrity, mitochondrial membrane potential, and the chromatin distribution did not differ significantly between groups. The number of acidic (pI; 3.1–5.6, 37%) and basic (pI; 7.9–10.0, 27%) proteins and their pattern of expression varied significantly (p?<?0.05) between high and low fertile bulls. The abundant sperm protein spots in 2D-gel electrophoresis (2DE) were identified as seminal plasma protein PDC-109 (i.e., protein with N-terminus aspartic acid, D and carboxy terminus cystine, having 109 amino acids) and its isoform and spermadhesin-1 (SPADH1). The western blot analysis confirmed the presence of PDC-109 isoform proteins at 15.4?kDa (pI 5.3 and 5.5). The seminal plasma protein PDC-109 was abundant in the low fertile when compared to the high fertile group (p?<?0.05). This study suggests that the imbalance in acidic and basic sperm proteins may influence sperm fertility and sperm PDC-109 levels above a certain threshold affects bull fertility.     

香加皮宝藿苷-I对人食管癌细胞Eca-109侵袭力的影响

目的探索宝藿苷-I对人食管癌细胞Eca-109侵袭力及TFPI-2基因表达的影响。方法以12.5、25、50μg/ml浓度的宝藿苷-I处理Eca-109细胞48h,分别采用Transwell小室法检测细胞的侵袭能力、RT-PCR法检测细胞TFPI-2mRNA表达水平以及Westernblot法检测TFPI-2蛋白表达水平等。结果 12.5、25、50μg/ml宝藿苷-I均可抑制人食管癌细胞Eca-109的侵袭力并能上调TFPI-2mRNA及蛋白的表达,25、50μg/ml组与对照组相比均有显著差异（P〈0.05）。结论宝藿苷-I能抑制人食管癌细胞Eca-109的侵袭力,可能与上调TFPI-2基因的表达有关。