Secondary brain injury is closely associated with brain edema, inflammation response and other injury factors after intracerebral hemorrhage, such as the complement system activation, excitatory amino acid toxicity, the release of vasoactive substances and free radical damage. The main mode of neuronal death during secondary brain injury after intracerebral hemorrhage are necrosis and apoptosis.

Spermatogenesis is a complex process. Current knowledge about human spermatogenesis is mainly based on the mouse model while little is known about the initial stage of this fundamental process in humans. The establishment of the model of spermatogenesis in vitro may contribute to an overall understanding of male germ cell development, an insight into the mechanisms of infertility, and clinical management of male infertility. This review summarizes current knowledge about the generation of germ cell-like cells from induced pluripotent stem cells (iPSCs) in vitro and discusses the potential application of iPSCs in the treatment of male infertility.
Animals ; Cell Differentiation ; Germ Cells ; Humans ; Induced Pluripotent Stem Cells ; Infertility, Male ; therapy ; Male ; Mice ; Spermatogenesis

Objective To observe the clinical efficacy and safety of propafenone on paroxysmal supraventricular tachycardia (PSVT).Methods 37 patients with PSVT were injected with 70 mg propafenone intravenously in 5 minutes.The unrecovered patients in 20 minutes were injected with 70 mg propafenone intravenously again,who were given 70 mg propafenone intravenously once again if not controlled in late 30 minuted.Blood pressure,heart beat,12 lead electrocardiogram were recorded before and after use of drug.Results The significant effective rate was 77.5% and effective rate was 21.6% with overall effective rate of 97.3% and effective time of 1～55 (7.1?2.8) minutes.The average accumulated dose of propafenone was 105 mg.Some side effects were observed in part of patients,including slight reduction of blood pressure,P R interval.QRS wave and Q T interval prolonged,temporary type sino atrial block,vertigo and nausea,etc whice could vanish gradually if not treated.Conclusion Propafenone by intravenous injection can quickly contro PSVT with saftey and effectiveness.

Objective To investigate the regulatory effects of bombesin on the growth of human immortalized gastric epithelial cell line(GES-1), and its mechanisms. Methods ① The expression of gastrin releasing peptide receptor(GRP-R) mRNA in GES-1 was detected. ② The expression of GRP-R protein was tested by cross-linking experiment and the location of the receptors in the cells were investigated by cytochemistry. ③GES-1 cell line was incubated with varying concentrations of bombesin with or without its antagonist and growth of the cell line was determined. ④The effect of protein kinase C (PKC) inhibitor on cell growth induced by bombesin was studied. ⑤After treated with bombesin, the intracellular IP 3 and translocation of PKC activity were measured in GES-1. ⑥Semiquantification of GRP-R mRNA in this cell line treated with bombesin was performed. Results ①Expression of mRNA of GRP-R was demonstrated in GES-1 cells. ②The GRP-R protein was about 75?103 as revealed by cross-linking study, and the receptors were identified on the cell membranes by cytochemistry. ③Bombesin stimulated the growth of GES-1 significantly, which could be inhibited by specific antagonist of bombesin. ④Bombesin-induced growth of GES-1 was also inhibited by PKC inhibitor. ⑤Bombesin induced an increase of IP 3 generation in GES-1 as well as remarkable translocation of PKC activity from cytoplasm to the cell membranes. ⑥An increase in GRP-R mRNA was induced by treatment of cell line with bombesin. Conclusions Bombesin stimulates the growth of this GES-1 via its receptor GRP-R and through IP 3, PKC signal pathway. The increase in expression of GRP-R mRNA in GES-1 induced by bombesin indicates that bombesin might upregulate the GRP-R in the GES-1 cells.

Protein protein binding is the basis of virus and host cell interactions. With the application of technology of studying of protein interactions, more knowledge of replication and pathogenesis of hepatitis C virus (HCV) could be acquired. Non structure protein NS5A is one of the important regulatory factors in virus replication , transcription and signal transduction, but there are controversy in effect on HCV pathogenesis and resistance to interferon ?(IFN?). In order to describe the relationship between NS5A and host proteins, we use yeast two hybrid system 3 to screen the gene encoding proteins that could interact with NS5A from human hepatocyte library. Thirty five clones were obtained including apo A1, apo A2, apo B100, haplotype mitochondrion complete genome, phosphatidic acid phosphatase type 2B, albumin similar to tumor endothelial marker 5 precursor, matrix metalloproteinase 14, and three of the Homo sapiens hypothetical proteins. The study paved a way for further studies on the pathogenesis of HCV NS5A

To clone the genes of hepatocyte protein interacting with hepatitis C virus NS3 protein, "bait" plasmids of hepatitis C virus NS3 were constructed. After verifying that hepatitis C virus NS3 protein could be steadily expressed in AH109 yeast strain, yeast two hybrid assay was berformed by mating AH109 with Y187 that pre transformed with liver cDNA library plasmids pACT2, and the diploidy yeast cells were plated on quadruple dropout (QDO) medium and assayed for X ? gal activity. Nineteen yeast colonies that could grow on QDO and had ? gal activity were obtained, then the library plasmids were extracted and sequenced. The gene sequences from the 19 positive colonies were aligned with the genes deposited in GenBank. It was found hepatitis C virus NS3 protein could interact with some proteins which have different functions.

The nonstructural protein 5A (NS5A) of the hepatitis C virus (HCV) has been shown to interact with a variety of cellular proteins and implicated in the regulation of cell growth, interferon resistance, and other cellular signaling pathways. Using the yeast-two hybrid method, we have isolated a clone that encodes a novel NS5A--associated binding protein: NS5ABP37. Reverse transcription polymerase chain reaction (RT-PCR) method was employed to amplify the full fragment,and the plasmid pGADT7-NS5ABP37 with the Saccharomyces cerevisiae vector pGADT7 was constructed. To prove the interaction, yeast cell Y187 transformed with pGADT7-NS5ABP37 was mated with yeast cell AH109 containing pGBKT7-NS5A to verify the interaction between the novel protein coded by the new gene NS5ABP37 and NS5A.

Objective HCV NS3 protein plays an important role in disease caused by HCV. We investigate the gene expression of HCV NS3 in yeast for future study of the function of the protein. Methods PCR was performed to amplify the gene of HCV NS3 from the plasmid pBRTM/HCV containing the whole fragment of HCV and the gene was cloned into pGEM T vector. Thereafter, HCV NS3 gene was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, and recombinant pGBKT7∶NS3 was transformed into yeast AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. Results HCV NS3 gene was successfully cloned into pGBKT7. The results of SDS PAGE and Western blotting assay showed that the molecular weight of the expressed product was about 22000 Da and HCV NS3 protein was existed within yeast cells.Conclusions HCV NS3 was successfully expressed in yeast expression system.

When Po. Wang Xingkuan treating long cough, he is good at treating from the liver, clearly identifying mechanism, getting good effect in treating long cough of the type of Attack of the Lung by Liver Fire with revised Sit-blood Decoction.

Objective To observe the specific humoral and cellular immune response in BALB/c mice injected with pS and p18S. Methods pS and p18S were constructed separately by inserting HBsAg gene fragment and the fusion gene fragment of HBsAg and mouse interleukin 18(IL 18) into the reading frame of pcDNA3.1+. Mice were injected with either plasmid intramuscularly in a total dose of 300 ?g per mouse. Every serum sample was detected for anti HBs using enzyme linked immunosorbent assay(ELISA). Furthermore, HBsAg specific cytotoxic T lymphocytes activity was measured. Results The expression of HBsAg was demonstrated by ELISA in p815 cells transfected with pS and p18S. pS can stimulate a positive antibody response. The average level was 135 mIU/ml, with the highest level of 530 mIU/ml. p18S could elicit relatively lower antibody response which was 20 mIU/ml. HBsAg specific CTL activities were 37.1% and 34% separately in pS and p18S immunized mice. It was only 13.2% when detected in pcDNA3.1+ immunization. Conclusion pS is effective to stimulate a humoral and cellular response in H 2d mice. IL 18 gene can not enhance the immune response when fused with HBsAg gene. Conversely, it seems to inhibit an immune response.