Objective:To investigate the expression of protein kinase C (PKC) isoforms ?,? in diabetic rats and evaluate the relation between altered PKC ?,? expression and the pathogenesis of diabetic retinopathy (DR).Methods:Wistar rats were randomly divided into control group and diabetic group induced by streptozotocin;osmotic shock method was used to isolate the retinal vessels of rats;and PKC isoforms ?,? espression were determined after 2 weeks,2 months,4 months and 6 months follw-up.DR was assessed by quantitative morphometry of capillary bed.Results:Diabetes could induce the translocation of PKC isoform ? from cytosol to membrane pool after 2 weeks,2 months,4 months and 6 months,but ? isoform translocated from membrane to cytosol pool in retinal vessels of diabetic rats after 2 weeks,2 months,4 months and 6 months.Diabetic rats showed pericyte decrease and capillary basement thickening in deep capillary bed of retina after 6 months follow-up.Conclusion:The expression of PKC isoforms ?,? varies in different stages of DR,and the effect of PKC isoforms on the pathogenesis of DR is different.

Objective:To construct the phage display library of randomized P2/NC protease cleavage sequences in HIV-1 Gag protein,so as to provide evidences for studying the effect of drug resistance-associated mutations on susceptible cleavage sequences of HIV-1 PR.Methods: CAP2 fragments and NC fragments of HIV-1 gag were generated by PCR.StuⅠ restriction site was introduced to the 5′ terminal of CAP2 fragments and the protease cleavage sequences of 3′ terminal of CAP2 fragments were randomized.The overlapping CAP2 sequence was introduced to the 5′ terminal of NC fragments and Sal Ⅰ restriction site was introduced to the 3′ terminal.CAP2 and NC fragments were linked by overlapping PCR method and the fused CAP2 and NC fragments were cloned into pCANTAB5S-LD3 to construct the phage display library of the P2/NC cleavage site in HIV-1 Gag protein.Results: As much as 2.1?10~6 clones were obtained in the phage library and the titer was 3.0?10~(12)TU/ml.There were 52.1% clones containing inserted CAP2/NC fragments.Sequence analysis of 12 samples showed that nucleotide acids and amino acids were randomly distributed at randomized PR cleavage sites.Nine of 10 positive monoclonal phages specifically bound to human IgG.Conclusion: CAP2/NC protease cleavage sequences of HIV-1 Gag protein have been successfully randomized and its phage display library has been constructed,which may help to screen the phages containing susceptible cleavage sequences of PR with drug resistance-associated mutations and establish the phage model for in vitro screening of PR inhibitor.

[Objective]To explore the efficacy of combination of TCM and WM on venous ulcer. [Method] Administer oral taking and external application of Chinese medicine decoction on the base of operation to 73 cases. [Result] All cases had effect, 72 limbs were cured, 5 recurred. [Conclusion] Combination of TCM and WM can improve cure effect, reduce recurrence rate, much better than pure operation or TCM therapy.

Systemic review of all the relevant randomized controlled trials (RCTs) was performed. RCTs were identifiedfrom Cochrane Library, MEDLINE, EMBASE. Limited evidence showed that problem-based learning in continuingmedical education increased participants' knowledge, performance and patients' health. Moderate evidence also showedthat doctors are more satisfied with problem-based learning. Problem-based learning may work in continuing medicaleducation, however,the evidence is too weak to be recommended. Rigorously large sample, well-designed randomizedcontrolled trials are required.[

Based on analyzing problems that traditional medical eduction mode faces,the author puts forward the construction of education mode of evidence-based medicine in medical students. And at the same time ,the article also makes research and discussion in relation to the following aspects: how to introduce evidence-base medicine into medical education; how to effectively implement teaching of evidence-based medicine.

180% elevation of PKC activity in the retinal vasculature.????????except ??? distributed in the membrane and cytosolic fraction in retinal vessels in normal or diabetic rats,the expression of ?????-PKC significantly increase in the membrane pool OD Value 282%?32%?340%?26%?187%?14% respectively ,but ?????-PKC expression significantly decrease in cytosolic pool in retinal vessels of diabetic rats compared with nondiabetic rats,OD value 71%?11%?63%?8%?84%?6%,respectively,however ? isoenzym expression in cytosolic pool significantly increase 208%?6%,compared with normal rats.Conclusion:Diabetes can induce the translocation of ?????-PKC from cytosol to membrane pool,but ? isoform translocated from membrane to cytosol pool in retinal vessels of diabetic rats.

ObjectiveTo evaluate the impact of hyperglycemia in patients admitted to medical intensive care units(MICU)on their prognosis.MethodsMedical records of 1016 patients,including 126 cases of known diabetes,359 cases with newly diagnoses hyperglycemia,and 531 normoglycemic cases,admitted to MICU in Wuhan University People's Hospital during September 2006 to September 2007 were reviewed to compare their case-fatality,length of hospital stay,length of MICU stay,mechanical ventilation,blood transfusion and antibiotics use.ResultsIn.hospital case-fatality was 22.8%(82/359)in patients with newly-diagnosed hyperglycemia.significantly higher than that in patients with known diabetes of 12.7% (16/126)and in normoglyeemic cases of 5.1%(27/531)(P＜0.01),and that Was higher in those with diabetes than those with normal blood glucose level(P=0.002).Proportions of patients who needed mechanical ventilation,blood transfusion,antibiotics use was higher in newly diagnosed hyperglycemia group than those in the other two groups.ConclusionsHyperglycemia represents an important mal'ker of poor clinical outcomes and mortality in patients admitted tO MICU.Patients with newly diagnosed hyperglycemia had a significantly higher case-fatality and poorer outcomes than those with known diabetes or normoglyeemia.

Objective To study the clinical characteristics of silent paraganglioma ( PGL) .Methods A total of 21 patients with silent PGL confirmed by surgical pathology and hospitalized in the First Affiliated Hospital of chongqing Medical University from Mar .2008 to Dec.2013 were enrolled in this study .The clinical manifesta-tions, hormone level, imaging findings, pathologic characteristic, treatment and follow-up data were reviewed and analyzed retrospectively .Results Among the 21 cases of silent PGL , 8 were male and 13 were female, with 47.05 years as the mean age(ranging from 26 to 67 years old).PGL tumors in silent group were mainly located in the right side, and left to right ratio was 2∶5.Silent PGL with tumor diameter <3 cm, 3-5 cm, >5 cm were 5, 7,and 9 cases respectively.Its constituent ratio was 23.8%, 33.3% and 42.9% respectively, significantly higher compared with that of symptomatic PGL (9.8%,43.9%,and 46.3%respectively).Silent PGL can be i-dentified from other adrenal incidentaloma based on enhanced CT value in each period .Biochemical examination or functional imaging further could facilitate the diagnosis of silent PGL .All the 21 cases were conducted with surgical treatment , and hypertensive crisis occurred to 4 cases during operation .Conclusions Silent PGL is not easy to be detected due to its non-specific clinical manifestations .The diagnosis needs the combined analysis on anatomical imaging characteristics , biochemical examination , functional imaging and pathological findings .Sur-gery is the most effective treatment .Postoperative pathological tips and long-term follow-up are both required for PGL, no matter benign or malignant .

Objective To investigate the clinical outlook of hepatic stem cells and its relations with liver cancer. Methods The literatures of recent years on the studies of hepatic stem cells were reviewed. Results Liver cancer may consist of cells of various differentiation grades and it may result from the perodifferentiated hepatic stem cells or abnormal differentiated cells. Conclusion The hypothesis of hepatic stem cells has been identified extensively. Further study maybe helpful for revealing the origin, carcinogenesis of hepatic cancer, and may also be useful for the understanding of the mechanism of metastasis.

Aim To investigate the effects of human telomerase reverse transcriptase(hTERT)antisense phosphorothioate oligodeoxynucleotide(ASODN)on telomerase activity and apoptosis induced by crisplatin(DDP)in HeLa cell lines.Methods HeLa cells were collected after transfection of 0.05,0.1,0.2 ?mol?L-1of ASODN and the total DNA was extracted.Telomeric repeat amplification protocol(TRAP)-AgNO3 stain was used to detect the activity of telomerase.Apoptosis of HeLa cells was detected by AO staining morphological observation and flow cytometry(FCM)analysis technology.Results The telomerase activity of HeLa cells was expressed in high level,which was significantly reduced dose-dependently and time-dependently after treatment with ASODN of hTERT.Two days after transfection of ASODN of 0.05 ?mol?L-1,0.1 ?mol?L-1,0.2 ?mol?L-1,the telomerase activity decreased by 21.77%,52.42% and 71.10% respectively.After treatment with ASODN for 24,48 h and 72 h,the telomerase activity decreased by 12.48%,38.27% and 71.10% respectively.In morphological observation using acridine orange(AO)staining method,HeLa cells displayed typical apoptotic figure after treated with DDP plus ASODN of hTERT.The percentage of apoptosis of HeLa cells treated with DDP of 1.5 mg?L-1,3.0 mg?L-1,for 48 hours after 24 hours exposure to ASODN of 0.2 ?mol?L-1(50.35%,29.67%)was significantly higher than that treated with DDP alone(19.67%,11.38%,P