Objective To design and construct the vector of protein-rich tyrosine kinase 2 small RNA interference and investigate the effect of the recombinant plasmid on the proliferation of adult rat cardiac fibroblasts. Methods The pAVU6+27-PYK_2 siRNA expression vector was constructed by gene recombination, then transfected into the cultured adult rat cardiac fibroblasts by DOTAP method. PYK_2 mRNA and protein in cardiac fibroblasts were determined by RT-PCR and Western blotting respectively. Cell proliferation was tested by MTT and ~3H-TdR incorporation. Results The pAVU6+27-PYK_2 siRNA expression vector was successfully constructed. The recombinant plasmid can inhibit the proliferation of adult rat cardiac fibroblasts, as compared with control. Conclusion pAVU6+27-PYK_2 siRNA expression vector is efficient to suppress DNA synthesis of cardiac fibroblasts, which will be beneficial to further study of myocardial fibrosis.

Acute Coronary Syndrome ; blood ; Aged ; Cell Membrane ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Thromboplastin ; metabolism

Objective To study the inhibitory effect of panax notoginseng saponins (PNS) on 3-nitrotyrosine (3-NT) formation in brain induced by heme/NO2 -/H2O2 or ONOO - pathways in vitro. Methods According to the two major pathways of 3-NT formation in vivo, the models of protein nitration induced by heme/NaNO2/H2O2 or ONOO-system were established, respectively, in vitro. Bovine serum albumin (BSA)/rat plasma protein or rat brain homogenate protein were utilized as reactive substrates in both systems. Samples were divided into blank-control group, 3-NT group and PNS group (including low-, medium-and high-concentration subgroups). In 3-NT group, samples were exposed to heme/NaNO2/H2O2 or ONOO-system, respectively, at 37℃for 30 min, whereas in PNS group, samples were pre-incubated with PNS (at final concentrations of 50 mg/L, 100 mg/L, and 200 mg/L) at 37℃for 5 min before the nitrating system exposure. The 3-NT level in each group was detected by Western blot assy. Results Compared with the blank-control group, both heme/NaNO2/H2O2 and ONOO-system can induce significant 3-NT generation in BSA/rat plasma protein or rat brain homogenate protein (P<0.05). Compared with model group, PNS pre-treatment markedly inhibited 3-NT expression in BSA/rat plasma protein in a dose-dependent manner (P<0.05), the inhibitory effect of low intervention on the level of 3-NT in rat brain homogenate protein was not significant (P>0.05). Medium- and high-concentrations of PNS pre-treatment markedly inhibited 3-NT accumulation, with maximum effect at the concentration of 200 mg/L (P<0.05). Conclusion Medium- and high-concentrations of PNS can inhibit 3-NT formation in brain tissue mediated by either heme/NO2-/H2O2 or ONOO-pathways, implying that potential neuroprotective action against 3-NT involves pathological conditions, like trauma, stroke, and neurodegenerative diseases.

Objective: To investigate the clinical effect of combination therapy on ankylosing spondylitis. Methods: 160 Cases were divided into treatment group and control group randomly. The patients of treatment group were given Bushenqingdu Decoction and Hanbiwaiyongfang by external use. The patients of control group were given Wangbi Granule. The symptoms and signs were observed before and after treatment in both groups. Results: There was significant difference between treatment group and control group in total effective rate as well as morning stiffness and signs (P

Objective To explore the potential of breast cancer stem cells in differentiating into vascular endothelial cells and participating in the angiogenesis of breast tumor.Methods The expressions of mutant P53, CD31 ,VEGF and Her-2 amplification in tumor blood vessels were detected and the single cell suspension of breast cancer was prepared.CD44+/CD24-/low cells were selected and cultured in culture system.The expressions of CD31 and CD105 on endothelial cell surface were detected by EGM-2 endothelial cell culture medium.The ability of uptake of acetylated low-density lipoprotein was measured.The endothelial cells were cultured in vitro to observe their vascular structure.Results The expressions of CD31,VEGF and mutant P53 were found in paraffin sections of breast cancer tissue samples, and they were arranged along the vessel lumen or blood vessel sphere. Immunofluorescence showed that CD31 and DAPI signals were expressed intravascularly in breast cancer tissues. Four samples of Her-2 positive amplification and 9 cases of non-amplification were detected in the samples.The successful separation of breast cancer stem cells was carried out by immunomagnetic sorting.The percentage of CD44+ cells in the pre-sorting cells was (7.5±2.6)% and that of CD44+ cells was about (94.3±4.7)% after magnetic sorting (P<0.05).The ratio of CD24+ cells was (48.2±9.4)% before sorting,and that of CD24+ cells was (4.3±1.6)% after immunomagnetic bead sorting.The proportion of CD105+ cells in mammary gland cells was (4.5±0.9)% and the proportion of CD31+ cells was (6.2±1.3)%.After cultured for three generations,the percentages of CD105+ cells and CD31+ cells were (79.6±9.3)% and (84.1±10.7)%,respectively (P<0.05). DiL-Ac-LDL could be phagocytized by endothelial cells cultured in endothelial cell culture system.Vessel-like structure was found in the endothelial cell group while the control group did not have the tendency of vascular changes.Conclusion Breast cancer stem cell-derived endothelial cells differentiate into endothelial cells in vitro and have the ability of vascular formation,suggesting that breast cancer stem cells may be involved in the formation of tumor blood vessels.

Using the high efficient copper-adsorbing yeast strain Y17 as absorbing material, the major affect factors including pH, original concentration of Cu2+, cell biomass, adsorption time and temperature were examined, and then the absorbing sites of the Y17 was determined. The results showed that the solution pH was the most dominate factor which affected the biosorption of Cu2+, the other affecting factors were the ini- tial concentration of Cu2+, the cell biomass added, and adsorption time, respectively; the temperature had lit- tle effect on the rate of biosorption. The orthogonal experiment showed that the optimal absorption condition was as follow: the solution pH was 5.0, the absorption time was 40 min, the cell biomass of Y17 added was 5.0 g/L, and the concentration of Cu2+ was 8 mmol/L; the highest adsorbing rate was up to 82.7% at this condition. Based on the results of different pretreatments and the desorption of Cu2+, the cell wall of Y17 was identified as the main place occurring boisorption process, and the -NH2 group, -COOH group on the surface of the yeast cells played an important role on the boisorption process.

OBJECTIVETo discuss the results and significance of the detection of the CFTR gene mutation in azoospermia patients with congenital unilateral absence of the vas deferens (CUAVD).

METHODSWe collected peripheral blood samples from 6 azoospermia patients with CUAVD for detection of the CFTR gene mutations and single nucleotide polymorphisms. We analyzed the genome sequences of the CFTR gene in comparison with the website of the UCSC Genome Browser on Human Dec. 2013 Assembly.

RESULTSMissense mutation of c. 592G > C in exon 6 was found in 1 of the 6 azoospermia patients with CUAVD and splicing mutation of c. 1210-12T[5] was observed in the noncoding region before exon 10 in 2 of the patients, both with the V470 haplotype in exon 11.

CONCLUSIONMutations of the CFTR gene can be detected in azoospermia patients with CUAVD and the detection of the CFTR gene mutation is necessary for these patients.

Azoospermia ; genetics ; Cystic Fibrosis Transmembrane Conductance Regulator ; genetics ; Exons ; Humans ; Male ; Male Urogenital Diseases ; genetics ; Mutation, Missense ; genetics ; Vas Deferens ; abnormalities

Designing of natural product-like compounds using natural products as template structures is an important strategy for the discovery of new drugs. Gambogic acid (GA), which is a Garcinia natural product with a unique caged xanthone scaffold, inhibits potent antitumor activity both in vitro and in vivo. This review summarized the researches on the identification of the antitumor pharmacophore of GA, and the design, structural optimization and structure-activity relationship (SAR) of natural product-like caged xanthones based on it.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Biological Products ; chemical synthesis ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Screening Assays, Antitumor ; Garcinia ; chemistry ; Humans ; Molecular Structure ; Structure-Activity Relationship ; Xanthones ; chemical synthesis ; chemistry ; isolation & purification ; pharmacology

Wnt/β-catenin signaling pathway plays an important role in the proliferation, growth, invasion, and metastasis of human cancers. Moreover, β-catenin/T-cell factor 4 (TCF4) interaction regulates the transcription of the key oncogenes in Wnt/β-catenin signaling pathway. Therefore, β-catenin/TCF4 interaction would be a promising therapeutic target for the development of highly selective anticancer agents. At present, most ongoing small-molecule inhibitors targeting β-catenin/TCF4 interaction, including PKF222-815, iCRT3/5/14, LF3, and sanguinarine, have been developed in preclinical studies for human cancer therapeutics. In this review, we summarized the research advances of up-to date inhibitors targeting β-catenin/TCF4 interaction, including the molecular structure and cellular functions of β-catenin in canonical Wnt signaling pathway. This review holds a hopeful avenue for the development of novel and highly selective Wnt inhibitors targeting β-catenin/TCF4 interaction for future anticancer strategy.

OBJECTIVEIn our study, the function of the third-order branches of the mesentenc artery was measured by microvascular ring technique, which can be used to detect microvascular function in some disease related to microvascular dysfunction.

METHODSIsolated, fixed, standardized and then activated the third-order branches of rat mesenteric artery. Microvascular tone was measured by systolic and diastolic drags respectively, with the help of DMT tension apparatus and PowerLab data acquisition system.

RESULTSThe third-order branches of rat mesenteric artery showed excellent response to vasoactive drugs. The contraction effect of norepinephrine (NE) reached 19 in mN. When acetylcholine (Ach) or sodium nitroprusside (SNP) of 10(9)-10(5)mol/L was added, vascular tones showed gradient drop: 80% of maximal relaxation when adding ACh, while 95% of maximal relaxation when adding SNP.

CONCLUSIONThe third-order branches of the mesenteric artery function was successfully detected by using microvascular ring technique.

Acetylcholine ; pharmacology ; Animals ; In Vitro Techniques ; Male ; Mesenteric Arteries ; drug effects ; physiology ; Nitroprusside ; pharmacology ; Norepinephrine ; pharmacology ; Rats ; Vasoconstrictor Agents ; pharmacology ; Vasodilation ; physiology ; Vasodilator Agents ; pharmacology