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

BACKGROUND: Focal adhesion kinase (FAK) phosphorylation influences the proliferation, migration and apoptosis of cells. However, there are no reports about the role of FAK phosphorylation in migration and proliferation of vascular smooth muscle cells during the process of vascular restenosis.OBJECTIVE : To analyze the correlation between FAK phosphorylation and vascular remodeling after balloon endothelial denudation in rats.DESIGN: Random controlled experiments in animals.SETTING: Cardiology Institute of Xianning College; Shenzhen Center for Chronic Disease Prevention and Control;Department of Cardiology at the Affiliated Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology.MATERrALS: The experiment was carried out in the Laboratory of Cardiology at the Affiliated Tongji Hospital from March to May in 2005. Forty male Wistar rats were used in the study, and they were managed according to the ethical standard of animals.METHODS: ①Forty rats were randomized into five groups (n =8): non-balloon endothelial denudation control group, and 4-day, 8-day, 16-day and 24-day model groups after balloon endothelial denudation. ②The rat models of restenosis following balloon endothelial denudation were built. At days 4, 8, 16 and 24 postoperatively, the denudated vessels were harvested for the detection of the morphological changes, which were taken as the judgment of successful models;Western blot method was used to detect the expression and activity of FAK. Abdominal aorta of left common carotid in rats was served as controls.MAIN OUTCOME MEASURES: The expressions of FAK and phosphorylation FAK in rat artery.RESULTS: Totally 40 rats were all involved in the result analysis.①A small quantity of FAK without phosphorylation could be detected in the controlled rats. Compared with the control group, the expression and activity of FAK in the model group rapidly increased at 4 days after balloon injury (P ＜ 0.05), and they changed in a similar manner with the increase of injury duration. The expression of FAK reached a peak at 8 days and began to descend at 16 days. The expression of phosphorylation FAK reached a peak at 16 days and then rapidly descended. ②Focal hyperplasia of intima was found at 8 days after balloon endothelial denudation, and diffuse hyperplasia of intima was found at 16-24 days after denudation.CONCLUSION: FAK can take part in vascular remodeling after balloon endothelial denudation in rats. The level of intimal hyperplasia is closely associated with the level of FAK and FAK phosphorylation.

Objective To study the effect of new barrel theory in improving evaluation of hospitalized patients. Methods Eight hundred hospitalized patients from September 2012 to August 2013 were randomized equally into the control group and the observation group.The last one or two items affecting patient satisfaction from the control group were used as objectives to be improved.Causal effect analysis was done pertinent to the items and the worksheet was bettered and improved and then enforced.The two groups were compared after intervention with new barrel theory in terms of satisfaction of patients during admission and discharge. Result The satisfaction of patients in the observation group during admission and discharge was significantly better than that in the control group(P<0.05).Conclusion The new barrel theory used to detect the flaws in nursing service and improve the workflow can improve assessment from the patients so that the management quality can be enhanced.

Objective To explore the features, diagnosis and treatment of pulmonary infection after kidney transplantation. Methods The clinical data of 31 pulmonary infection cases among 150 patients underwent kidney transplantation were analyzed retrospectively. Results The 31 patients with pulmonary infection after kidney transplantation included 9 cases of simple bacterial infection, 3 cases of fungus infection, 5 cases of CMV infection, 1 case of TB, 10 cases of mixed infection, and 3 cases of infection with unclear pathogen. 27 cases of the patients(27/31,87.1%) were cured, while 4 cases died of pulmonary infection. Conclusion Pulmonary infection is a common and severe complication after kidney transplantation. Early etiological diagnosis, the prompt treatment of antibacterium,antivirus and antifungus, adjustment of immunosuppression regime, and strengthening the support therapy would improve the curative rate.

To investigate the potential role of hepatitis B virus(HBV) preS1 protein in mediating HBV adhesion to liver cell, we prepared recombinant proteins of HBV preS1 in yeast. PCR was performed to amplify the gene of HBV preS1 from the plasmid pCP10/HBV ayw subtype containing the whole fragment of HBV and the PCR product was cloned into pGEM T vector. The gene of HBV preS1 was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, the pGBKT7 plamids containing preSl were transformed into yeast cell AH109. The yeast protein was isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. The results showed that the presence of HBV presl proteins in yeast cells was confirmed by Western slot analysis. the molecular weight of the expressed product was about 30000 Da. The findings indicated that HBV preS1 was successfully expressed in yeast system.

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.