Objective To explore the role of protein kinase(PKC)in the nosogenesis of hypoxic-ischemic encephalopathy(HIE).Methods Twenty-four seven-day-old SD rats were divided randomly into normal group (N),model control group(C)and HIE group(HIE).The biochemistry techniques of SDS-PAGE and western bolt were used to examine the level of PKC membrane translocation of cortex and hippocampi.Results The level of PKC membrane translocation of cortex and hippocampi in N group was 0.28±0.02,0.29±0.02 respectively,and 0.29±0.02,0.30±0.02 in C group,and 0.38±0.04,0.40±0.06 in group HIE.The level of PKC membrane translocation of cortex and hippocampi was higher in HIE group as compared with that in N,C group(P＜0.01),but there was no different between N group and C group(P＞0.05).Conclusion PKC are activated in HIE,which indicates that PKC may involve in the pathophysiologic mechanism of HIE.

BACKGROUND: Gene transfer to study the effect of a exogenous gene on cells is a commonly used technique for molecular biology. Compared with viral vector,pcDNA4.0-TGF-?1 eukaryotic expression plasmid transfection has no inherent toxicity or cytotoxicity,with high safety. OBJECTIVE: To construct and identify eukaryotic expression plasmid carrying recombined TGF-?1 by cloning transforming growth factor (TGF)-?1 gene. DESIGN,TIME AND SETTING: The in vitro cell experiment was performed at the Opening Laboratory of Key Clinical Medical Sciences,Henan Provincial High-learning School from March 2007 to February 2008. MATERIALS: Clean healthy Sprague Dawley rats aged 2 months,of both gender,were used for isolating RNA from cells. METHODS: The TGF-?1 mRNA was extracted from the rat bone marrow cells,and amplified by reverse transcription-polymerase chain reaction (RT-PCR),and cloned into the pGEM-T-vector and identified by PCR. Then TGF-?1 gene was subcloned into pcDNA4.0 plasmid vector. The inserting DNA sequences were analyzed. MAIN OUTCOME MEASURES: Expression of TGF-?1 cDNA,pGEM-T-TGF-?1 and recon pcDNA4.0-TGF-?1. RESULTS: There was an obvious electrophoresis strip of TGF-?1 cDNA by RT-PCR. The obtained pGEM-T-TGF-?1 was confirmed correct with PCR amplification and sequence identification. Recombinant eukaryotic expression plasmid pcDNA4.0-TGF-?1 was obtained and identified by digestion. The inserted sequences were conformity to the designed sequences. CONCLUSION: Rat TGF-?1 gene has been successfully cloned,and recombinant TGF-?1 eukaryotic expression vector has been constructed successfully.

Objective To explore the early diagnosis and treatment of acute superior mesenteric arterial embolism (ASMAE).Methods The clinical data of 25 patients with acute superior mesenteric arterial embolism were retrospectively analyed.Results All 25(100%) patients had severe abdominal pain with abdominal signs,21(84%) patients had history of valvular heart disease or atrial fibrillation,and 17(68%) patients had hematochezia.All the cases were misdiagnosed preoperatively,including 11 patients were misdiagnosed as abdominal pain of unknown causes,3 patients as acute pancreatitis,3 patients as acute gastroenteritis,2 patients as necrotizing enteritis.1 patient as acute appenditis,1 patient as acute myocardial infarction,and 1 patient as acute cholecystitis;but 3 patients were preoperatively suspected to have impairment of mesenteric blood flow.All of the 25 patients underwent necrotic bowel resection,6 patients died,and 7 patients abandoned treatment because of serious complications such as short bowel syndrome etc.Mortality rate was 51%(13/25).Conclusions Early diagnosis of patients with acute superior mesenteric arterial embolism is difficult;the main cause of misdiagnosis is lack of recognition of its clinical signs.Fully grasping the characteristics of patients with early stage disease and effective early intervention are the fundamental measures for reducing mortality.

Objective To investigate the influence of perioperative continuing aspirin therapy on tooth extraction surgery in elderly patients with coronary heart disease.Methods A total of 926 patients with coronary heart disease who required tooth extraction were enrolled in this study.Among them,241 patients discontinued aspirin therapy before tooth extraction,685 patients continued aspirin therapy before tooth extraction.The entering rate of operation was evaluated.100 patients continuing aspirin and 100 patients discontinuing aspirin were randomly selected.Systemic pressure,diastolic pressure and heart rate before and after tooth extraction were detected.100 matched healthy elderly patients were selected as normal control group.Multivariate logistic regression analysis was used to estimate the risk for tooth extraction.Results The entering rate of operation was 67.9％ (131 cases) in 193 patients with angina pectoris discontinuing aspirin therapy and 82.1 ％ (312 cases) in 380 patients with angina pectoris continuing aspirin therapy (x2 =14.77,P＜0.01).The entering rate of operation was 41.7％ (20 cases) in 48 patients undergoing coronary revascularization and discontinuing aspirin therapy,and 80.7％ (312 cases) in 305 patients undergoing coronary revascularization and continuing aspirin therapy (x2=33.95,P＜0.01).The changes in systolic blood pressure before versus after dental extraction had a significant difference between the discontinuing aspirin group and control group [(15.9±5.5) mmHg vs.(12.2±4.7)mmHg,P＜0.05,1 mmHg=0.133 kPa],while had no significant difference between the continuing aspirin group and control group [(13.6±4.5) mmHg vs.(12.2±4.7) mmHg,P＞0.05].There were no significant differences in changes in diastolic blood pressure and heart rate in the intraoperative period between the patients and controls (P＞0.05).The bleeding rate in patients continuing aspirin therapy was not significantly different as compared with control group (P＞ 0.05).Multivariate logistic regression analysis revealed that perioperative discontinuing aspirin therapy remained the second significant risk factor for tooth extraction.Conclusions Preoperative administration of aspirin should be continued in dental extraction surgery in elderly patients with coronary heart disease.

Objective To investigate the impacts of γ-ray radiation on gene expressions of peripheral blood lymphocytes in SD rats,and to screen differential expression genes and biological pathways closely related to radiation injury.Methods The differential gene expression of peripheral blood lymphocytes in SD rats was selected by microarray at 6 h after 2 Gy 60Co γ-ray exposure in vitro and in vivo.Bioinformatics analysis of the differentially expressed genes was performed by using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases.Real-time quantitative PCR was applied to verify the screen results of the microarray.Results Fifty-five genes with three times over-expressed level were screened out from the radiation groups both in vitro and in vivo and they were involved in 6 signaling pathways.There were two differentially expressed genes of microarray assay were verified by PCR assay.Conclusions Differential expressed genes in the peripheral blood lymphocytes of SD rats could be induced by γ-ray radiation and they cooperatively contributed to a variety of biological processes.

BACKGROUND: The application of adipose derived stromal cells to tissue engineering has been more and more popular around the world. Compatibility of scaffold material is the key point for its further research.OBJECTIVE: To determine the growth rules of rabbit adipose-derived stromal cells with polylactic-co-glycolic acid (PLGA).DESIGN, TIME AND SETTING: An in vitro study was performed at the Institute of Ophthalmology, Otolaryngology Hospital,Fudan University and Shanghai Tissue Engineering Center from September 2007 to March 2009.MATERIALS: Six female New Zealand rabbits aged six months were used for extraction of adipose-derived stromal cells. PLGA was provided by Sigma, USA.METHODS: Adipose tissue was harvested from the nape fat pad of the rabbits following anesthesia. Primary cultured cells were established using type I collagenase and cell cultures were maintained with DMEM containing 10% volume fraction of fetal bovine serum. Cells were passaged when 80% was confluent. The fourth passages of cells were utilized for the study. PLGA consisted of polylactic acid and polyglycolic acid as the ratio of 7:3, and the relative molecular mass was 104900.MAIN OUTCOME MEASURES: Initially, the adherent rate of cells to scaffold was detected. After one week co-culture, the scaffold bearing adipose-derived stromal cells labeled with Dio agent were investigated by fluorescence inverse microscope,scanning electron microscopy and laser scanning microscope.RESULTS: The best adherent rate of adipose-derived stromal cells with PLGA reached 99%. After one-week-incubation in vitro,cells of fiber-shaped or ovule-shaped proliferated well and exhibited stratified growth on the surface of PLGA scaffold. In addition,it also secreted visible extracellular matrices, which could be examined by scanning electron microscopic examination.Meanwhile, the adipose-derived stromal cells grew well and distributed equably inside the PLGA in terms of the investigation with laser scanning microscope.CONCLUSION: The compatibility of adipose-derived stromal cells to PLGA in vitro was well.

Objective To discuss the surgical management of von Hippel Lindau(VHL) syn drome.Methods The clinical data of four VHL patients who were clinically diagnosed from March1999 to October 2006 were reviewed.The first patient was a 56 year old man with the chief complaintof hypodynamia and low serum glucose as 2.37 mmol/L.CT scan showed three masses in the the leftkidney.He had a history of cerebral haemangioblastoma ten years before.The second case was a 57 year old woman with the chief complaint of lumbago in the left side.CT scan showed masses in theleft kidney,adrenal gland and panerease.The third case was a 39 year old woman with the chief com plaint of an accident mass in the left adrenal gland.She had the history of cerebellar hemangioblastoma9 years before,spinal hemangioblastoma resection 7 years before.The fourth case was a 41-year oldwoman,she was found brain tumors and cured by gamma radiation abalation.Bilateral renal masseswere found by B ultrasound one month later,CT scan showed four masses in the both kidneys,leftadrenal gland and pancreas.Results All 4 cases underwent surgical approach.The first case under went radical nephrectomy which pathological report was PEComa of kidney.The blood glucose wasnormal one week later.The second case underwent resection of the left adrenal gland,kidney,pancre atic body and tail and spleen.Pathological report was clear cell carcinoma,islet cell tumor and adrenal cyst.Three months later she was found spinal hemangioblastoma and refused treatment.The thirdcase underwent adrenalectomy in the left side and pathologieal report was adrenal pheoehromocytoma.There was not tumor reeurrenee during 2 years' follow up.The nephrectomy and adrenalectomy wasperformed for the last ease whose pathological report was clear celt carcinoma and pheochromoeytoma.Three weeks later,tumor enueleating of the right kidney was undertaken; the result was clear cellcancer.During the follow up for one year there was no relapse of tumor.Conclusions For VHL ac companied with multiple organ tumors,surgery resection is the proper approaeh when tumors of centralnervous system is large.Different approaches could be taken to deal with multiple tumors of VHL such aswatchful waiting,nephron sparing surgery.

Abstract? AlM: To explore the effect of Danshen injection iontophoresis combined with lumbrukinase on early patients with retinal vein occlusion ( RVO ) , and observe the prognosis to provide objective basis for clinical?METHODS: Eighty-two patients ( 117 eyes ) with early RVO were randomly selected in our hospital from January, 2011 to April, 2014, 41 cases (59 eyes) of control group treated with Lumbrokinase. Forty-one cases ( 58 eyes) of the observation group, treated with Danshen injection iontophoresis combined with lumbrokinase. The main indexes were the treatment effect, visual acuity and retinal circulation time.?RESULTS: The total effective rate of the observation group was 85. 4% ( 35/41 ), more than that of control group 68. 3% ( 28/41 ), the difference was statistically significant (P<0. 05). The invalid rate of the observation group was 14. 6% (6/41), lower than that of the control group 31. 7% ( 13/41 ), the difference was statistically significant (P<0. 05). The vision of the observation group 4. 8 ~5. 0 was 53. 7% (22/41), more than that of control group 41. 5% ( 17/41 ), the difference was statistically significant ( P < 0. 05 ). The restore vision of the observation group 4. 3 ~ 4. 7 was 31. 7% (13/41), lower than that of the control group 41. 5% ( 17/41 ), the difference was statistically significant ( P<0. 05 ). After treatment, retinal circulation time of two groups were lower than before treatment, the difference was statistically significant (P<0. 05). After treatment of retinal vein circulation time of the observation group was 8. 15 ± 1. 30s, 9. 70 ± 1. 28s lower than that of the control group, the difference was statistically significant (P<0. 05).?CONCLUSlON: There is better effective on Danshen injection iontophoresis combined with Lumbrokinase for early retinal vein occlusion, and can improve the patient's visual acuity, promote the rehabilitation of patients.

Objective:To detect mutations of exons 5 and exons 8 of PTEN gene in oral squamous cell carcinoma(OSCC),and to explore the relationship between gene mutations and development of OSCC.Methods: Mutations of exons 5 and exons 8 of PTEN gene were detected by polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP).Results: The whole sequences of exons 5 and exons 8 of PTEN gene of all cases of OSCC were expanded.There was no mutation in exons 5 and exons 8 of PTEN gene of all cases of OSCC.Conclusion: There is no mutation in exon 5 and exon 8 of PTEN gene in OSCC.This might show that there is no correlation between mutations in exon 5 and exon 8 of PTEN gene and the development of OSCC.