The template-independent teminal transferase activity of Taq DNA polymerase results in an overhanging dA at the 3′end of its PCR products. The pGEMX vector constructed in this experiment forms a single overhanging dT at its 3′end as the result of cleavage with Xcm I restriction enzyme. This vector is very efficient for direct cloning of PCR product obtained by using Taq DNA polymerase.Recombinant colonies can be selected by Blue/white screening. Moreover,insertion fragment can be easily released from the vector simply with either BamH I or Hind III digestion.

Objective The primary purposes of this article were to examine the distributions and correlation between conditions of sleep and the quality of life,so as to explore the risk factors of abnormal sleep among elderly in the rural areas of Anhui province.Methods Elderly who aged 60 years or more (n= 1680) were randomly selected from rural areas in Anhui province.Conditions of sleep and quality of life of the subjects were assessed independently,using the pittsburgh sleep quality index (PSQ1) scale and SF-36 scale.A generalized logistic model analysis was performed,using sleep-related factors as explanatory variables.Results The average PSQI score was 7.43 + 4.13.Persons whose sleep conditions were graded as well,common or poor were accounted for 29.4%,24.3%,46.3%,respectively.Sex and sleep quality were different in terms of quality of life among all the factors (P＜0.05).Men had better quality than women,and poor sleepers had poorer quality of life as compared to the one with better conditions.Factors as being married/living alone/economically independent,having better social function,often eating meat or wheat were significant predictors for having good sleep conditions.However,factors as better education received,living alone,with poor vitality/general health,having chronic diseases (back pain,coronary heart or stomach disease),less meat intake,rice as major foods etc.were predictors for poorer sleeping condition.Conclusion Our data showed that it was unsatisfactory on the quality of sleep among the rural elderly in Anhui province.Conditions on general,health and nutrition should be improved in order to prevent the poor sleep condition.

OBJECTIVETo observe the neurological protective effects of progesterone (PROG) on focal cerebral ischemia/reperfusion injury in rats and to explore its possible mechanism.

METHODSOne handred and twenty male SD rats were divided into three groups randomly: sham-operated group, middle cerebral artery occlusion ( MCAO ) group and PROG + MCAO group( n = 40). The right temporary MCAO model was established by the line-embolism method. The PROG + MCAO group rats were according to 8 mg/kg intraperitoneal injection PROG, after that 30 min, the rats were suffered ischemia/reperfusion. After rats were suffered ischemia for 2 h and reperfusion 0, 24, 48, 72 h stress, the nervous functional defect degree were evaluated by longe scoring, and the expression of two-pore domain K channel 3 (TASK3) mRNA in brain tissue were detected by the real-time PCR.

RESULTSPROG (8 mg/kg) could significantly reduced the nervous functional defect degree in rats after ischemia/reperfusion 24, 48, 72 h (P < 0.05). The results of real-time PCR showed that the TASK3 mRNA expression in the brain tissue at all time points significantly decreased in MCAO group compared with sham-operated group (P < 0.05). However, compared with MCAO group, the expression of TASK3 mRNA in brain tissue at all time points dramatically increased in PROG + MCAO group (P < 0.05).

CONCLUSIONPROG can improve the nervous functional defect degree after focal cerebral ischemia/reperfusion injury in rats, and the mechanism might be associated with up-regulating the expression of TASK3 mRNA in brain tissue.

Animals ; Brain ; drug effects ; pathology ; Brain Ischemia ; drug therapy ; Infarction, Middle Cerebral Artery ; Male ; Neuroprotective Agents ; pharmacology ; Potassium Channels, Tandem Pore Domain ; metabolism ; Progesterone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Reperfusion Injury ; drug therapy

Objective To retrospectively analyze the treatments of nonvalvular atrial fibrillation (nvAF) in elderly patients aged 80 years and over,and to investigate the influencing factors for occurrence of stroke and transient ischemic attack(TIA)and relationships between antithrombotic therapy and stroke or TIA.Methods 101 elderly patients with nvAF were enrolled and grouped according to the occurrence of stroke/TIA and antithrombotic-correlated bleeding.The influencing factors were retrospectively analyzed and antithrombotic schemes were compared.Results Incidence rate of stroke/TIA was 28.7％ (29/101).Among all patients,70 cases were treated with antiplatelet therapy,19 cases were treated with anticoagulation therapy,while 12 cases received no antithrombotic (antiplatelet or anticoagulation) therapy before stroke.Both the nvAF time course and the antithrombotic strategy were significantly different between post-AF stroke/TIA group and non-postAF stroke/TIA group(both P＜0.05).The difference was reflected in ratios of antiplatelet therapy/anticoagulation therapy.The proportion of anticoagulation therapy was higher in non stroke/TIA group(x2 =5.778,P =0.016).Different antiplatelet therapy scheme significantly affected occurrence of stroke/TIA(P＜0.05).There was no significant effect of antithrombotic schemes on hemorrhagic events(x2=0.708,P =0.702).Multiple logistic regression analysis showed that hypertension,coronary heart disease,cancer,diabetes and previous stroke history,as well as nvAF duration were the independent risk factors for post-AF stroke/TIA(OR=1.351,95 ％CI:1.129-1.617).Conclusions Currently,the proportion using anticoagulation therapy is low,and single antiplatelet therapy is the main regimen in the elderly patients with nvAF.For elderly patients with nvAF,anticoagulation therapy has a protective effect against the occurrence of post-nvAF stroke/TIA,meanwhile there is no significantly increased risk of bleeding,which makes anticoagulation therapy advisable in the elderly.The nvAF time course is one of the risk factors,which is worth experts' attention in risk evaluation of thrombus in elderly patients.

Objective To investigate the expression and distribution of cellular FLICE-inhibitory protein (c-FLIP) in peripheral blood and lesions of psoriatic patients. Methods Peripheral blood and skin samples were obtained from 30 patients with psoriasis vulgaris and 20 normal controls. Flow cytometry was used to detect intracellular c-FLIP protein in peripheral T and B lymphocytes, immunohistochemistry to examine the expression of c-FLIP in lesional tissue. Results Based on the positivity rate of c-FLIP, there was a significant increase in T lymphocytes in active psoriasis compared with regressive psoriasis and normal controls (6.32%±1.17% vs 2.64%±0.74% and 2.28%±0.54%, P＜0.01 and 0.05, respectively), while no significant difference was found in B lymphocytes among these three groups (0.78%±0.16%, 0.71%±0.32%, 0.69%±0.18%, respectively, P＞0.05). The expression intensity of c-FLIP in keratinocytes was also higher in active psoriasis than in regressive psoriasis and normal controls (89.73±5.24 vs 117.40±7.50,121.58±7.93, P＜0.01 and 0.05 respectively), and there was no difference between regressive psoriasis and normal controls (P＞0.05). Conclusions c-FLIP is highly expressed in lesions and peripheral T lymphocytes of patients with active psoriasis, suggesting the possible involvement of c-FLIP in the proliferation of T lymphocytes in psoriasis.

The article probes into the approach to the interns' capability of clinical thinking and analyzing and solving problems by disscussing causes of gastrointestinal hemorrhage. Correct clinical thinking always comes from solid basis and clinical theory and knowledge, careful history taking, meticulous physical examination, rigorous logical thinking and constant summing up experiences and lessons. As a teacher, we should help interns develop clinical thinking ability during their whole peroid of medical education.Besides, theory should be combined with practice to prevent and correct vicious clinical thinking

Adult ; Female ; Humans ; Lead ; toxicity ; MMPI ; Male ; Middle Aged ; Occupational Exposure ; Personality ; drug effects

Nitrobenzene is one of the toxic compounds. Much work had focused on biodegradation of it sofar. Two main pathways for nitrobenzene biodegradation, oxidative and partial reductive pathways, were reviewed in this article. The mechanism of these pathways including involved enzymes and genes was introduce in details. Comparative analysis of the pathways would provide basis for the development and application of biodegradation technology for nitrobenzene and other organic pollutants.

The role of hydrogen sulfide (H2S) in portal hypertension (PH)-induced esophagus-gastric junction vascular lesions in rabbits was observed.The rabbit PH models were established.The animals were randomly divided into the following groups:normal,PH,PH+sodium hydrosulfide (PH+S),PH+propargylglycine (PH+PPG).The plasma H2S levels,apoptosis of esophageal-gastric junction vascular smooth muscle cells,and the expression of nuclear transcription factor-κB (NF-κB),p-AKT,Iκ Ba and Bcl-2 were detected.The cystathionine γ lyase (cystathionine-gamma-splitting enzyme,CSE) in the junction vascular tissue was measured.The results showed that the plasma H2S levels and the CSE expression levels had statistically significant difference among different groups (P＜0.05).As compared with PH group,plasma H2S levels were declined obviously (11.9±4.2 vs.20.6±4.5,P＜0.05),and CSE expression levels in the junction vascular tissue were notably reduced (1.7±0.6 vs.2.8±0.8,P＜0.05),apoptosis rate of vascular smooth muscle cells per unit area was significantly decreased (0.10±0.15 vs.0.24±0.07,P＜0.05),and the expression levels of p-AKT and NF-κB were significantly decreased (2.31±0.33 vs.3.04±0.38,P＜0.05;0.33±0.17 vs.0.51±0.23,P＜0.05),however,Iκ Ba and Bcl-2 expression increased obviously (5.57±0.17 vs.3.67±0.13,P＜0.05;0.79±0.29 vs.0.44±0.36,P＜0.05) in PH+PPG group.As compared with PH group,H2S levels were notably increased (32.7±7.3 vs.20.6±4.5,P＜0.05),the CSE levels in the junction vascular tissue were significantly increased (6.3±0.7 vs.2.8±0.8,P＜0.05),apoptosis rate of vascular smooth muscle cells per unit area was significantly increased (0.35±0.14 vs.0.24±0.07,P＜0.05),and the expression levels of p-AKT and NF-κB were significantly increased (4.29±0.49 vs.3.04±0.38,P＜0.05;0.77±0.27 vs.0.51±0.23,P＜0.05),yet Iκ Ba and Bcl-2 expression decreased significantly (3.23±0.24 vs.3.67±0.13,P＜0.05;0.31±0.23 vs.0.48±0.34,P＜0.05) in PH+S group.It is concluded that esophagus-gastric junction vascular lesions happen under PH,and apoptosis of smooth muscle cells is declined.H2S can activate NF-κB by the p-AKT pathway,leading to the down-regulation of Bcl-2,eventually stimulating apoptosis of vascular smooth muscle cells,easing PH.H2S/CSE system may play an important role in remission of PH via the AKT-NF-κB pathway.

0.05).CART vaccine at 10?g significantly depressed the analgesic effect of morphine analgesia (P