Xuefu zhuyu decoction containing serum in vitro induced expressions of desmin and alpha-actin: an experimental research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe whether Xuefu Zhuyu Decoction (XZD) could induce the differentiation of mesenchymal stem cells (MSCs) into cardiac myoid cells, thus seeking for safe and effective inducers.</p><p><b>METHODS</b>The serum pharmacological method was used to induce. XZD containing serum was prepared. MSCs were isolated and cultured. The serum cytotoxicity was detected by MTT. The third generation of favorably grown cells was selected in this experiment. Cells were divided into three groups, i.e., the vehicle control group, the XZD containing serum induced group, and the 5-azacytidine induced group. Expressions of Desmin and alpha-actin were detected by immunocytochemical staining method.</p><p><b>RESULTS</b>Before induction protein expressions of Desmin and alpha-actin were negative, and few was weakly positive. There was no statistical difference in the weak positive expression rate among the 3 groups (P > 0.05). After induction protein expressions of Desmin and alpha-actin were negative, and few was weakly positive in the vehicle control group. Protein expressions of Desmin and alpha-actin were positive in the XZC containing serum induced group and the 5-azacytidine induced group. There was statistical difference in the positive expression rate when compared with the vehicle control group (P > 0.05).</p><p><b>CONCLUSIONS</b>XZD played a role in in vitro inducing differentiation MSCs to cardiac myoid cells. It might participate in expressions of Desmin and alpha-actin.</p>

Activity of heme oxygenase-1 affects expression levels of hypoxia inducible factor-1 gene in vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>One effect of solid tumors is severe hypoxia of local tissues. Heme oxygenase-1 (HO-1) is highly expressed in a variety of human tumor tissues; its induction and activity are closely related to growth of solid tumors. Hypoxia inducible factor-1 (HIF-1) is a transcription factor that regulates hypoxia signal transduction and plays a central role in tumor hypoxia regulation. However, whether and how changes in HO-1 activity affect HIF-1 gene expression has not been reported previously.</p><p><b>METHODS</b>Hypoxia-inducible models were established using gastric cancer cell lines (SGC-7901) in a hypoxia incubator. Cells were placed in four groups: Group A, transfected by plasmid harboring HO-1 shRNA; Group B, transfected with scrambled shRNA vector; Group C, treated with hemin; and Group D, exposed to hypoxia only. Expressions of HO-1 and HIF-1 mRNAs were quantified by reverse transcription-polymerase chain reaction. Expressions of HO-1 and HIF-1 proteins were determined by immunohistochemistry and Western blotting.</p><p><b>RESULTS</b>mRNA and protein levels of HO-1 and HIF-1 in the control group were significantly higher than in Group A (P < 0.01), but lower than in Group C (P < 0.01). Chromatin immunoprecipitation analysis showed that HIF-1 was identified as the direct HO-1 target gene.</p><p><b>CONCLUSION</b>While affected by HIF-1, HO-1 up-regulation promotes the expression of HIF-1 and the down-regulation of HO-1 suppresses the expression of HIF-1 gene.</p>

Expression and localization of annexin A7 in the rat lithium-pilocarpine model of acquired epilepsy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Annexin A7 (synexin, ANXA7) is a member of annexins, which plays an essential role in the regulation of calcium homeostasis. Considerable evidence shows that the pathogenetic mechanism of acquired epilepsy (AE) has been related to the imbalance of calcium homeostasis. The aim of this study was to investigate ANXA7 expression and cellular localization in the cortex and hippocampus in the rat lithium-pilocarpine model of AE.</p><p><b>METHODS</b>Totally 81 adult healthy male Wistar rats were randomly divided into control group (n = 9) and experimental group (n = 72), the experimental group contained eight subgroups according to sacrifice time (n = 9) (6-hour, 24-hour, 48-hour, 72-hour, 7-day, 15-day, 1-month, and 2-month). In the experimental group, rats were intraperitoneally injected by lithium-pilocarpine to induce AE model. We examined the expression and localization of ANXA7 via immunohistochemistry, double-label immunofluorescence with the use of neuron specific enolase (NSE) antibody, glial fibrillary acidic protein (GFAP) antibody and propidium iodide (PI), respectively. The data of optical density value were analyzed by analysis of variance.</p><p><b>RESULTS</b>ANXA7 expression increased significantly in the experimental groups especially in the acute period (6 hours, 24 hours, and 48 hours after the onset of seizure) using immunohistochemistry. Double-label immunofluorescence and confocal microscopy disclosed that ANXA7 localized in the neurons but not in astrocytes and did not localize in the nucleus, which were performed with anti-NSE, anti-GFAP and PI respectively.</p><p><b>CONCLUSION</b>ANXA7 may play a potential role in the pathogenetic mechanisms of the rat lithium-pilocarpine model of AE.</p>

Studies on in vitro capacitation of goat spermatozoa by heparin treatment / 生物工程学报

Systematical studies are lacking on the influencing factors and mechanisms of the heparin enhanced sperm capacitation, although many studies have shown that heparin enhanced sperm capacitation. The effect of heparin concentration and exposure time, incubation temperature and co-culture with oviductal epithelial cells or cumulus cells on goat sperm capacitation were investigated in this study. The motility, membrane and acrosome integrity and capacitated percentage of goat spermatozoa were assessed after different heparin treatments, and rates of fertilization and embryo cleavage were compared after in vitro insemination of oocytes with spermatozoa capacitated by different heparin treatments. The major results are summarized as follows: 1) When spermatozoa were capacitated with heparin at 5, 10, 25, 50 and 100 microg/mL for 45 min, 50 and 100 microg/mL heparin treatments produced the highest capacitated percentages of 55% and 56%, respectively, but the percentage of spermatozoa with intact acrosomes in the 100 microg/mL heparin treatment decreased significantly (P < 0.05) in comparison with that in the control group, indicating that the optimal heparin concentration for goat sperm capacitation would be 50 microg/mL. 2) Capacitated percentage of spermatozoa increased with extension of treatment time when goat sperm were treated with 50 microg/mL heparin for 0, 10, 20, 30, 45, 60 or 120 min. Although heparin treatments for 45 to 120 min did not differ significantly (P > 0.05) in capacitated sperm percentages, sperm motility and membrane integrity decreased significantly when treated with heparin for 120 min. This suggested that the optimal exposure time of heparin at 50 microg/mL for goat sperm capacitation would be 45 to 60 min. 3) Significantly higher capacitated percentages of spermatozoa were obtained when goat sperm were treated at 42 and 38.5 degrees C than at 15 and 37 degrees C, but sperm motility and acrosome integrity were significantly lower when spermatozoa were treated at 42 degrees C than they were treated at other temperatures. Temperature of 38.5 degrees C would, therefore, be the optimal temperature for goat sperm capacitation. 4) The capacitated percentage of spermatozoa was significantly higher when goat sperm were co-cultured with oviductal epithelial cells than when treated with heparin alone or co-cultured with cumulus cells, but sperm motility and membrane and acrosome integrity did not differ significantly among the three treatments. Rates of fertilization (91.3%) and cleavage (72.2%) were significantly higher in the oviductal epithelial cell co-culture group than those in the heparin alone group. This indicated that co-culture with oviductal epithelial cells significantly enhanced goat sperm capacitation by heparin treatment.