Objective To analyze the prevalence and diversity of genotype resistant mutations in 123 HIV/AIDS patients experiencing failure of high antiretroviral therapy (HAART) in Yunnan province.Methods A cross-sectional survey was conducted among 151 HIV/AIDS patients experiencing failure of antiretroviral therapy form January 2011 to January 2012 in Yunnan province.The HIV-1 pol gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR),the sequences were then submitted to the Stanford University HIV drug resistance database to analyze the prevalence of resistant mutations.The resistant mutations were statistically analyzed by gender,ethnic groups,transmission route and subtype,respectively.The chi-square or fisher's exact test was used for statistical test.Results Of the 151 cases,123 plasma samples were amplified successfully for protease PR (codon 1-99) and reverse transcriptase RT (codon 1-272) fragments.The most common mutation was M184 (72.4％),followed by the mutation at position K103 (47.2％),T215 (26.0％),D67 (15.4％),G190 (34.1％),Y181 (29.3％),K101 (17.1％).The frequency of mutations at position V75,A62 and M230 was higher in male population than that in female population (x2 =7.001,6.975,5.446,P ＜ 0.05).The frequency of variants at position Tl215,K70 and T69 was higher in the Han population than in the other ethnic population(x2 =5.290,4.060,3.860,P＜ 0.05).It was interesting that the variant M41L was rare in the other ethnic groups.The significant difference existed at various transmission routes.Frequencies of variants at position T215 and T69 were significantly higher among people infected HIV-I through sexual contact than the intravenous drug users (x2 =10.431,7.952,P ＜ 0.05).Frequencies of variants at position G190 were significantly higher among the intravenous drug users than the population infected HIV-1 through sexual contact(x2 =6.669,P ＜ 0.05),but the variant M230L never occurred in intravenous drug users.The RT mutations V75,T69,M230 were more frequently occurred in patients infected with CRF01_AE than in patients with subtype B (P＜ 0.05).The mutation L74 was never seen in patients infected with CRFOI_AE (P ＜ 0.05).Conclusions The HIV/AIDS patients with failure of high antiretroviral therapy (HAART) were attributed to HIV-1 genotype resistance mutations.The mutation sites among the HAART failure patients from the regions of Dehong,Gejiu,Wenshan and Yuxi were significant difference accordance among the gender,ethnicith,transmission route and subtype,respectively.

Objective This pre-DIC rat models were constructed by injecting LPS in order to reveal the relationship and significance among tissue factor pathway inhibitor (TFPI),platelet, fibrinogen, prothrombin time and activated partial thromboplastin time. Methods Enzyme-labeled immunosorbent assay was used to detect blood plasma TFPI in pre-DIC state rats(n=16),DIC state rats(n=8)and health rats(n=8).Platelet, fibrinogen, prothrombin time and activated partial thromboplastin time in the same rat were detected. Results TFPI increased first(pre-DIC state)and then decreased(DIC state)in the blood plasma of rata, but the levels of TFPI were always higher than that in the normal state. Conclusions TFPI is more sensitive and accurate to monitor the pro-DIC state than routine examination, which is very useful in the early diagnosis of pre-DIC state.

0.05). The median duration of survival was 11.5 months for TEP gr oup versus 8.5 months for EP group (P0.05,no statistical difference). The main toxicity wa s myelosuppression for the two groups.The incidence rate of Ⅲ～Ⅳ degree neutro p enia and thrombocytopenia was higher in the TEP group than that in the EP group( P

BACKGROUND: Nuclear factor erythroid-2-related factor-2 (NF-E2-related factor-2) is an important transcription factor to regulate anti-oxidative stress reaction. Some researches indicate that NF-E2-related factor-2 can be phosphorylated by numerous members of protein kinase C family. In order to investigate generant mechanism of microwave radiation on oxidative stress injury, whether microwave radiation can influence on anti-oxidative regulating system through NF-E2-related factor-2 or not should be further studied.OBJ ECTIVE: To analyze the effect of microwave radiation on phosphorylation of NF-E2-related factor-2 and activity of protein kinase C in vascular endothelial cells.DESIGN: Observational-contrast study.SETTING : Department of Labor Hygiene, the Third Military Medical University of Chinese PLA.MATERIALS: Vascular endothelial cell strain; H332PO4; Protein-A Sepharose (Sigma Company); mono-antibody of NF-E2-related factor-2 (H-300, Santa Cruz); α-mono-antibody of protein kinase C (Santa Cruz); glass microfiber filters(Whatman Company); gel scanning system (Gel Doc 2000, Bio-Rad); liquid scintillation spectrometer (LKB-117, Sweden).METHODS: The experiment was carried out in Laboratory of Electromagnetic radiation and Biological Effect, Department of Labor Hygiene, the Third Military Medical University of Chinese PLA from March to July 2003. ① Analysis of phosphorylation of NF-E2-related factor-2: Vascular endothelial cells were cultured with DMEM medium till the period of productive growth and incubated with 32Pi for 2 hours. And then, cultured bottle was maintained in water bath at 37℃ and performed with microwave radiation in dark chamber, whose reflectivity was about zero. It was regarded as radiation group, and the average power density of radiation was 30 mW/cm2; in addition, the duration of radiation was 30 minutes.Cells did not deal with microwave radiation were regarded as control group. Phosphorylation level of NF-E2-related factor-2 was measured at 2, 4, 8 and 24 hours after radiation with immune coprecipitation-autoradiography technique and dealt with semi-quantitative analysis with gel scanning system. Cells in the control group were analyzed directly. ②Active analysis and expressional measurement of protein kinase C: Cells in the radiation group and the control group were dealt with the same cultured method, condition, radiation styles, dosage and environment as mentioned above. At 2, 4, 8 and 24 hours after radiation, cells were split to extract plasma and membrane protein. Furthermore, activity of protein kinase C was measured with r-32P-ATP labeled liquid scintillation spectrometer; gray value of protein strap was dealt with semi-quantitative analysis with gel scanning system; staining degree of plasma was observed after immunocytochemical staining of protein kinase C. In addition, cells in the control group were measured and observed directly.MAIN OUTCOME MEASURES: ① Phosphorylation level of NF-E2-related factor-2 in radiation group and control group; ②Results of active analysis and expressional measurement of protein kinase C in radiation group and control group.RESULTS: ① Phosphorylation level of NF-E2-related factor-2 in radiation group and control group: Gray value of NF-E2-related factor-2 was higher in radiation group than that in control group at 2, 4 and 8 hours after radiation.Phosphorylation level of NF-E2-related factor-2 reached the peak at four hours after radiation. In addition, results of semi-quantitative scanning analysis showed that, at 2, 4 and 8 hours after radiation, phosphorylation level of NF-E2-related factor-2 was increased 33%, 261% and 141% in radiation group as compared with that in control group,respectively (t = 2.974, 4.209, 4.047, P ＜ 0.05), and then, fallen down to normal value 24 hours later. ② Results of active analysis and expressional measurement of protein kinase C in radiation group and control group: At 2, 4 and 8 hours after microwave radiation, expression of protein kinase C in radiation group was higher than that in control group,especially at the 4 hour. In addition, at 24 hours after radiation, expression of protein kinase C recovered the normal value. Results of immunocytochemical staining showed that staining of plasma was deeper in radiation group than that in control group at 4 hours after radiation. Moreover, results of r-32P-ATP labeled liquid scintillation spectrometer also suggested that, at 2, 4 and 8 hours after radiation, activity of protein kinase C was increased 36%, 93% and 47% in radiation group as compared with that in control group, respectively (t =2.801, 3.654, 3.035, P ＜ 0.05). And then, activity of protein kinase C was decreased after 24 hours, otherwise, activity of protein kinase C reached the peak at 4 hours after radiation.CONCLUSION: Microwave radiation can strengthen the phosphorylation level of NF-E2-related factor-2 in vascular endothelial cells during a special period; meanwhile, it can also cause the increase of expression of protein kinase C.Time effect of activity of protein kinase C is coincidence with phosphorylation level of NF-E2-related factor-2.

Objective To study the application of the array comparative genomic hybridization (Array-CGH) for the detection of chromosomal disorders in newborns.Method The Array-CGH technique was used to analyze the whole genome of the patients who were suspected of chromosomal disease in neonatal ward of our hospital from January to December in 2014,and further verification in genomic unbalanced ectopia was carried out by FISH (fluorescence in situ hybridization,FISH).Result Among 514 patients,104 were found carrying chromosomal abnormalities with a detection rate of 20.2％.The most common chromosomal disease is the Down syndrome syndrome (24 cases),followed by the chubby Willy and Angel syndrome(17 cases),while the Wolf-Hirschhorn syndrome in 5 cases,Williams syndrome in 5 cases and the Criduchat syndrone in 5 cases.The results of FISH were consistent with Array-CGH.Conclusion The technique of Array-CGH can be used to scan the whole genome of children with unknown disease.As a high-throughput and rapid research method,this technique has important clinical significance in the screening of chromosomal diseases.

Objective To evaluate the effect of sufentanil on autophagy during myocardial ischemia-reperfusion (I/R) in rats.Methods Eighteen adult male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n =6 each) using a random number table:sham operation group (group SH),I/R group,and sufentanil group (group S).Myocardial I/R injury was produced by occlusion of the anterior descending branch of the left coronary artery for 30 min followed by 120 min reperfusion in the rats anesthetized with chloral hydrate in I/R and S groups.Sufentanil 3 μg/kg was injected via the caudal vein at 30 min prior to ischemia in group S.The equal volume of normal saline was given in SH and I/R groups.At 120 min of reperfusion,arterial blood samples were collected for determination of serum creatine kinase-MB (CK-MB) and lactic dehydrogenase (LDH) concentrations,and myocardial specimens were obtained for examination of the ultrastructure of myocardial cells (using transmission electron microscopy) and for determination of autophagy-related proteins microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ),Beclin-1 and Bcl-2 expression (by Western blot).Results Compared with group SH,the serum CK-MB and LDH concentrations were significantly increased,the expression of LC3 Ⅱ and Beclin-1 was significantly up-regulated,and Bcl-2 expression was significantly down-regulated in group I/R,and the serum CK-MB concentration was significantly increased,the expression of Beclin-1 and Bcl-2 was significantly down-regulated in group S (P＜0.05).Compared with group I/R,the serum CK-MB concentrations were significantly decreased,the expression of LC3 Ⅱ and Beclin-1 was significantly down-regulated,Bcl-2 expression was significantly up-regulated (P＜ 0.05),no significant change was found in serum LDH concentrations (P＞0.05),and the pathological changes were reduced in group S.Conclusion The mechanism by which sufentanil reduces myocardial I/R injury may be related to decreased autophagy in rats.

Objective To evaluate the effect of remifentanil preconditioning on myocardial ischemia-reperfusion (I/R) injury in aged rats.Methods Twenty-four healthy male Sprague-Dawley rats,aged 15-18 months,weighing 465-580 g,were randomly divided into 3 groups (n =8 each) using a random number table:sham operation group (group S),group I/R and remifentanil preconditioning group (group RP).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.Myocardial I/R was induced by 30 min occlusion of anterior descending branch of left coronary artery followed by 120 min reperfusion in group I/R.In group RP,remifentanil 10 μg · kg-1 · min-1 was infused intravenously for 20 min followed by 10 min washout before myocardial I/R.In group S,the anterior descending branch was only exposed but not ligated.At 30 min before ligation and 120 min of reperfusion,the activities of serum lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) were determined.The hearts were removed at 120 min of reperfusion for determination of myocardial infarct size,the percentage of myocardial infarct size was calculated,and myocardial specimens were obtained for observing myocardial ultrastructure.Results Compared with group S,the activities of serum LDH and CK-MB were significantly increased at 120 min of reperfusion in I/R and RP groups.Compared with group I/R,the activities of serum LDH and CKMB were significantly decreased,the percentage of myocardial infarct size was decreased,and the pathological changes were attenuated in group RP.Conclusion Remifentanil preconditioning can attenuate myocardial I/R injury in aged rats.

Objective To investigate the effects of tropicamide and esculin-digitalisglycosides combination eye drops on low myopia of children (age ranging from 6 to 14). Methods Eighty children with 160 eyes of low myopia in the outpatient department of ophthalmology of our hospital were chose from July 2010 to April 2013, then the patients were equally divided into four groups:the control, esculin-digitalisglycosides eye drops,tropicamide,and tropicamide and esculin-digitalisglycosides combination groups. The naked vision and myopia correction of each group were compared before and after treatment for two weeks, and the treatment effects were compared among four groups. Results (1) The results showed there were no significant changes before and after treatment in the control, esculin-digitalisglycosides eye drops and tropicamide groups ( >0.05); (2) On the contrast, the tropicamide and esculin-digitalisglycosides combination have significantly improved naked vision and myopia correction after 2 weeks ( <0.05);(3) Prior to the treatment,there was no significant difference among four groups (<0.05);(4) After treatment,group under tropicamide and esculin- digitalisglycosides combined was statistical different from the other groups treated with other three eye drops ( <0.05);However, there was no statistical difference among groups in these three eyedrops. Conclusion Esculin-digitalisglycosides combined with tropicamide was the most effective treatment for low myopia of children.

BACKGROUND: Nuclear factor erythroid-2-related factor-2 (NF-E2-related factor-2) is an important transcription factor to regulate anti-oxidative stress reaction. Some researches indicate that NF-E2-related factor-2 can be phosphorylated by numerous members of protein kinase C family. In order to investigate generant mechanism of microwave radiation on oxidative stress injury, whether microwave radiation can influence on anti-oxidative regulating system through NF-E2-related factor-2 or not should be further studied. OBJECTIVE: To analyze the effect of microwave radiation on phosphorylation of NF-E2-related factor-2 and activity of protein kinase C in vascular endothelial cells.DESIGN: Observational-contrast study. SETTING: Department of Labor Hygiene, the Third Military Medical University of Chinese PLA.MATERIALS: Vascular endothelial cell strain; H332PO4; Protein-A Sepharose (Sigma Company); mono-antibody of NF-E2-related factor-2 (H-300, Santa Cruz); ?-mono-antibody of protein kinase C (Santa Cruz); glass microfiber filters (Whatman Company); gel scanning system (Gel Doc 2000, Bio-Rad); liquid scintillation spectrometer (LKB-117, Sweden).METHODS: The experiment was carried out in Laboratory of Electromagnetic radiation and Biological Effect, Department of Labor Hygiene, the Third Military Medical University of Chinese PLA from March to July 2003. ① Analysis of phosphorylation of NF-E2-related factor-2: Vascular endothelial cells were cultured with DMEM medium till the period of productive growth and incubated with 32Pi for 2 hours. And then, cultured bottle was maintained in water bath at 37℃ and performed with microwave radiation in dark chamber, whose reflectivity was about zero. It was regarded as radiation group, and the average power density of radiation was 30 mW/cm2; in addition, the duration of radiation was 30 minutes. Cells did not deal with microwave radiation were regarded as control group. Phosphorylation level of NF-E2-related factor-2 was measured at 2 , 4, 8 and 24 hours after radiation with immune coprecipitation-autoradiography technique and dealt with semi-quantitative analysis with gel scanning system. Cells in the control group were analyzed directly. ② Active analysis and expressional measurement of protein kinase C: Cells in the radiation group and the control group were dealt with the same cultured method, condition, radiation styles, dosage and environment as mentioned above. At 2, 4, 8 and 24 hours after radiation, cells were split to extract plasma and membrane protein. Furthermore, activity of protein kinase C was measured with r-32P-ATP labeled liquid scintillation spectrometer; gray value of protein strap was dealt with semi-quantitative analysis with gel scanning system; staining degree of plasma was observed after immunocytochemical staining of protein kinase C. In addition, cells in the control group were measured and observed directly.MAIN OUTCOME MEASURES: ① Phosphorylation level of NF-E2-related factor-2 in radiation group and control group; ② Results of active analysis and expressional measurement of protein kinase C in radiation group and control group.RESULTS: ① Phosphorylation level of NF-E2-related factor-2 in radiation group and control group: Gray value of NF-E2-related factor-2 was higher in radiation group than that in control group at 2, 4 and 8 hours after radiation. Phosphorylation level of NF-E2-related factor-2 reached the peak at four hours after radiation. In addition, results of semi-quantitative scanning analysis showed that, at 2, 4 and 8 hours after radiation, phosphorylation level of NF-E2-related factor-2 was increased 33%, 261% and 141% in radiation group as compared with that in control group, respectively (t = 2.974, 4.209, 4.047, P

Objective To investigate the effect of remote ischemic preconditioning on myocardial infarction and the involved protective mechanisms. Methods Twenty-four male SD rats were randomly divided into four groups: ischemia-reperfusion (I/R) group, rote ischemic preconditioning (RIPC) group, remote ischemic preconditioning + ischemia-reperfusion (RIPC+I/R) group, and RIPC+AG490+I/R group. The blood samples and myocardial specimens were collected and prepared for tests. The related enzymes were detected and the size of myocardial infarction was measured. The cardiac cells were determined by electron microscopy and light microscopy. Results The size of myocardial infarct and myocardial enzymes were significantly reduced in RIPC+I/R group compared to those in I/R group (P < 0.05). The size of myocardial infarction and myocardial enzymes were significantly increased in AG490 group compared to those in RIPC+I/R group (P < 0.05), but were significantly reduced in AG490 group compared to those in I/R group (P < 0.05). Conclusions Remote ischemic preconditioning may be effective in cardioprotection. The JAK/STAT pathway is involved in the cardioprotection of remote ischemic preconditioning.