In this study,the VP2 gene of feline panleukopenia virus(FPV) isolated from tiger was successfully cloned and expressed in Pichia pastoris yeast.The VP2 gene was amplified by PCR with a pair of specific primer.Then PCR products was purified and cloned into pGEM-T for sequencing.The interesting gene fragment was recovered after the double enzyme digestion of EcoRI/NotI,then subcloned into pPICZ?A for secretory expression.The recombinant pPICZ?AVP2 was linearized with SacI and then transformed into competence yeast GS115 for expression under the induction of 1% methanol.The positive recombinants were screened by PCR method.The expression product was identified by SDS-PAGE and western-blotting.The results showed that there was a molecular weight of 32 kD,which could be specifically recognized by polyclonic antibody against FPV.It revealed that the recombinant VP2 protein had correct three-dimensional structure,which would be used as a antigen protein for the diagnosis and prevention of FPV infection in tigers.

Varicella is an acute,highly contagious respiratory and caused by Varicella-zoster virus(VZV),After the primary infection,VZV induce Varicella,then the virus remains dormant in sensory nerve roots in life.Upon the reactivation,shingles can occur.Most of patients can recovery very soon,but kinds of potentially sever complication can induce death.Nowadays the VZV vaccination isn’t included in the expanded immunization program in our country. Varicella outbreaks take place at times in the middle of children of our country.The paper offers concise description about pathogen characteristics,diagnosis methods,clinical character,epidemiology knowledge and strategy of prevention.

Anti tumor angiogenesis offers new opportunity for tumor treatment;Chinese medicine is an effective angiogenesis inhibitor with advantages of cheapness and innocuity.With its anti tumor and angiogenesis research being further continuously,Chinese herbal medicine is hopeful to be ideal and general preventive and treatment remedy for tumor.

Bone morphogenetic protein is a key regulation factor in osteoplast cell differentiation and bone formation.It takes a role of differentiation inducting and orientating by modulating the transcription factor and through some unknown signal transduction pathway,and its expression and function can be adjusted by signal transducting system and specific acceptor inhibiter.This article reviews the research progresses on the aspects.

The effects of high frequency jet ventilation(HFJV) on brain resuscitation had been investigated in 28 cats in which simple brain functional failure was produced by occlusion of both carotid and both vertebral arteries. The results suggested that HFJV could improve endurance of cats when subjected to brain ischemia, raise the survival rate of cats with reversible brain functional failure, and prolong the survival time of cats with permernent brain functional failure. The mechanisms of how HFJ would exert beneficial effect on brain resuscitation was discussed.

Induced pluripotent stem cells (iPSCs) are a type of pluripotent stem cells that can be generated from adult somatic cells.They have similar characteristics and function to embryonic stem cells (ESCs).Over the past decade,iPSCs were widely concerned in regenerative medicine and stem cell field.Especially the patient specific iPSCs have several advantages over ESCs,such as convenient source,do not exist immune rejection and ethical issues,even keep certain individual genotype.At present,tremendous progress have been made about the application of iPSCs in a variety of retinal diseases.Here,this article reviews pluripotent stem cell sources of RPE,photoreceptors and retinal ganglion cells and current transplantation strategies,the safety problems and prospects.

AIM: To study the endothelial progenitor cell markers and biological characteristics of human umbilical cord blood CD133+ cells isolated by immunomagnetic bead (IMB) method. METHODS: The experiment was carried out in the Laboratory of Burn Surgery, Xijing Hospital of the Fourth Military Medical University of Chinese PLA from November 2002 to August 2003.①CD133+ cells were enriched from human umbilical cord blood of placenta through spontaneous delivery or cesarean delivery (offered by Department of Gynecology and Obstetrics, Xijing Hospital of the Fourth Military Medical University of Chinese PLA), with the informed consents of parturients.②Human umbilical cord blood was anticoagulated by ACD-B and then diluted, 7 mL of cord blood combined with 3 mL lymphocyte isolation liquid were centrifuged for 30 minutes at 2 100 r/min. Low-density mononuclear cells coloring buffy were selected to wash off platelet and inoculated for 30 minutes by the IMB and FITC-CD133 antigen at room temperature to wash excessive antigens. The FITC-CD 133-coated IMB was added into mononuclear cells which were cultured for 30 minutes at room temperature and placed in magnetic field for 1 minute to remove the cell suspension. IMB-cell isolation liquid was supplemented to release CD133+ endothelial progenitor cells, which were cultured with EGM-2MV media in dishes coated with collagen I. And 24 hours later those unadherent cells were detached, culture liquid was removed half after 3 hours, and total culture duration was 3-4 weeks.③The morphologic change of CD133+ cells isolated with IMB were observed; The percentage of CD133+ cells in cord blood monocytes and sorting efficiency of IMB were analyzed by fluorescence activated cell sorting (FACS). The cells cultured in vitro were identified by fluorescence immunoassay and enzymo-histochemistry while transmission electron microscope (TEM) was used to observe the W-P body of mature vascular endothelial cells. RESULTS: ①Morphologic observation of CD133+ cells culture in vitro: The adherent cells expanded pseudopods on days 4 and 5, and grew rapidly in clonal manner 7 days later. The cells showed a typical spindle-shaped morphology during 2-3 weeks and cluster areas of cobblestone-like cells after 2-week culture and confluence.②Isolation rate of CD133+ cells: The percentage of CD133+ cells in cord blood monocytes was 0.91%. And after IMB sorting, the percentage of CD133+ cells was 85.52%.③On the fifth day, few cells were positive for CD133 while most cells were positive for CD 34 and vWF; On the tenth day, all cells were negative for CD133 and most cells were still positive for CD 34 and vWF.④The cells after 10-day culture had typical W-P body in TEM. CONCLUSION: Immunomagnetic sorting can efficiently select enriched CD133+ endothelial progenitor cells from human umbilical cord blood. Our data of CD133, CD34, vWF antibodies and TEM support the hypothesis that endothelial progenitor cells may contribute to mature vascular endothelial cells.

Objective To explore the effect s of nimodipine therapy and prognosis in patients with traumatic subarachnoid hemorrhage.Methods 138 patients were randomly divided into treatment group and control group.The treatment group(70 cases)received early nimodipine therapy and the control group(68 cases)was supported with common method.At the same time,the complications with cerebral infarction and hydrocephalus during treatment were observed and the prognosis were judged.All data were statistily analyse.Results The incidence rate of cerebral infarction and hydrocephalus in nimodipine treatment group was lower obviously than control group.And prognosis in treatment gruoup were better than those of control group.Conclusions It is useful that the cases of cerebral injury with traumatic subarachoid hemorrhage receive the treatment of nimodipine.The early nimodipine therapy can be beneficial to depress the occurrence of cerebral infarction and hydrocephalus and improve the prognosis.

Objective: To establish the quality standard for Ganmaoling Tablets (Radix Ilex asprella, Flos Chrysanthem Indici, Ramulus et Radix Evodia leptae, acetaminophe, etc.). Methods:The acetaminophen, chlorphenamini maleas, caffeine, Radix Ilex asprella, Flos Chrysanthem Indici, Ramulus et Radix Evodia leptae were identified by TLC. The acetaminophen and chlorogenic acid were determined by HPLC. Results:The average recovery of acetaminophe was 101.36%, and RSD was 0.93%, respectively. The average recovery of chlorogenic acid was 100.8%, and RSD was 0.9%, respectively. Conclusion:The method developed is accurate and the reproducibility is good. The method can be used for quality control of the Ganmaoling Tablets.

Objective To explore the value of left parasternal anterior mediastinaotomy (Chamberlain procedure) in the diagnosis of mediastinal lymph node enlargement with unknown causes and anterior mediastinal space-taking lesions. Methods By using the Chamberlain procedure, biopsy was performed in 32 cases of enlarged mediastinal lymph nodes with unknown causes or mediastinal space-taking lesions, which were found by CT scans. Results All of the 32 cases were pathologically diagnosed, with a diagnostic accuracy of 100%. Three patients with pericardial effusion received concurrent pericardial fenestration and then their symptoms relieved. Four patients underwent concurrent lung biopsy. The operating time was (48?15) min, the blood lose was ( 40.6?23.5) ml, and the postoperative hospital stay (3.6?1.4) days. No deaths or postoperative complications occurred. Conclusions Chamberlain procedure is a safe and valuable method in the diagnosis of mediastinal space-taking lesions with unknown causes or enlargement of the fifth and sixth groups of mediastinal lymph nodes, which routine mediastinoscope cannot reach. Some other simple therapies, such as lung biopsy or pericardial fenestration, can also be employed at the same time.