The effects of dexamethasone on the over -expression of TGF-β_1 in the cerebral spinal fluid of rats following SAH / 中国医师杂志

Objective To study the preventive and therapeutic effect of dexamethasone on communicating hydrocephalus following subarachnoid hemorrhage (SAH), and explore the mechanism of communicating hydrocephalus following SAH. Methods SAH rat model was constructed by injecting auto blood through foramen magnum into subarachnoid space. There were two groups ( one control group and one experimental group) in this study with 16 rats each group. Sufficient doses of auto blood were injected into subarachnoid space of the rats of the control group and sufficient dose of auto blood together with sufficient dose of dexamethasone into subarachnoid space of the rats of the experimental group. The changes of rats transforming growth factor bata-1 (TGF-β_1 )in the cerebrospinal fluid (CSF)and thickness of pia mater collagen fibrosis( CF) following SAH were observed after 1 day and 20 days. The levels of TGF-β_1, in the CSF were quantitatively detected by the way of ABC-ELISA. The brain tissues were dyed through Massions technique, and then the average thicknesses of meningina CF were measured by the software of micrograph collection and analysis computer system. Results In later stage after SAH, TGF-β_1, concentrations of the experimental group was lower than control group( P <0. 01). At the same time, the average thicknesses of meningina CF of experimental groups was thinner than control group( P <0. 01). Moreover, the relationship between the level of TGF-β_1 in the CSF and the thickness of the CF of the rat's cerebral pia mate had positive correlation. Conclusions In the later stage after SAH, dexamethasone can inhibit the over expression of TGF-β_1 in rat CSF following SAH and also can inhibit the accrementition of rat collagenous fibers. Therefore, this study provided experimental and theoretical evidence for preventing and treating communicating hydrocephalus following SAH.

Exploring a New Gene Containing ACP Like Domain in HumanBrain and Expression It in E. coli / 生物化学与生物物理进展

To look for new genes from human brain, get a fragment was obtained using adaptor primer and 3' anchor polymerase chain reaction (PCR) with the human adult whole brain cDNA as template. The fragment was cloned into T easy vector and automatically sequenced with 310 Genetic Analyzer. Later the whole length cDNA of this novel gene was got with the method of 3'rapid amplification of cDNA end (RACE). The whole length of cDNA of this novel gene is 2 024 bp. Chromsome location is at 14q11.2 including 16 extrons and 15 introns. After scanning the sequence against GenBank it is proved that the sequence is a new one. ORF analysis showed that there is a complete coding region in it,it can interprate a protein containing 357 amino acid residules. ProDom analysis result showed that there is an acyl carrier protein (ACP) like domain in it. The gene was banked into GenBank. Then, a pare of primers were designed and were used to amplify the coding region and cloned into pGEX-4T1 expressing vector to express it in E. coli . The Dot blotting and Northern blot showed that this novel gene is highly expressed in the normal adult human brain.

The Progress of the Methods for Screening DifferentiallyExpressed Genes and Proteins / 生物化学与生物物理进展

Cloning and identification of differentially expressed genes or proteins is helpful not only for finding the functions of genes and proteins, but also for discovery of the mechanism of some diseases. Some methods have been developed for screening differentially expressed genes, such as differential display RT-PCR (DDRT PCR), subtractive hybridization (SH), DNA chip technique, and serial analysis of gene expression (SAGE). In subtractive hybridization, there have advanced three improved methods which include representational difference analysis (RDA), suppression subtractive hybridization (SSH), and full-length-gene-obtainable subtractive hybridization. For obtaining differentially expressed proteins, scientists have only two choices so far. One is two-dimentional gel electrophoresis. The other is phage display antibody repertoire library technique. Since all of the methods above have their own advantages and disadvantages, they should be used according to different needs.screening, differentially expressed genes, differentially expressed proteins

Preventive effect of DNA vaccine based on xenogeneic homologous calciumactivated chloride channel on airway hyperresponsiveness in asthmatic mice / 中国病理生理杂志

AIM: To observe the preventive effect of DNA vaccine based on human calcium-activated chloride channel 1 (hCLCA1) on airway hyperresponsiveness in asthmatic mice.METHODS: The DNA vaccine was constructed by inserting the hCLCA1 gene into pSecTag-2A, and then BALB/c mice were vaccinated by im. once every two weeks. Serum antibody was checked with the antigen of mCLCA3 by ELISA analysis. Asthma was induced with ovalbumin in the vaccinated mice. The airway pressure time index (APTI), the contractile responsiveness of isolated tracheal rings and the number of eosinophil in bronchoalveolar lavage (BAL) were investigated. Mice injected with pSecTag-2A, saline or normal mice were regarded as control groups. RESULTS: The title of antiserum binding to mCLCA3 in vaccine group was 1∶800 to 1∶(1 000) after three times vaccination. Compared with normal group, APTI, contractile responsiveness and number of eosinophil in vaccine group, pSecTag-2A or saline group were increased markedly (P