Based on link control concepts, the life cycle of science and technology projects was divided into five stages,, such as project application, selection, implementation, output and assessment.All the science misconducts that might occur among these five stages were listed.Finally, different precautions were made to prevent different misconducts during different stage..

Objective To characterize and quantify the CD4 +CD25 + regulatory T (Treg) cell population in peripheral blood of patients with ankylosing spondylitis (AS) and to determine the influence of treatment with tumor necrosis factor (TNF)-a inhibitors on them.Methods Peripheral blood mononuclear cells (PBMC) were isolated from 25 patients with active AS,in which 10 patients were treated with 12 weeks of etanercept,and 21 healthy subjects.CD4+CD25high T cells were analyzed using flow cytometry,and mRNA expression of FOXP3 was determined by real-time polymerase chain reaction (PCR).Proliferation of T cells to PHA was measured by WST-1 assay using depleted CD25+ cells by immunomagnetic sorting.Results There was no significant difference in the percentage of CD4+CD25high cells in peripheral blood between patients with active AS and controls (P＞0.05).However,PBMC from patients with active AS expressed reduced levels of FOXP3 mRNA (P＜0.01) which were inversely correlated with C-reactive protein (CRP)(P＜0.01).CD4+CD25+ cells in peripheral blood of both active AS patients and controls exhibited suppressive capacity on the proliferation of effector T cells in vitro (both P＜0.01).Treatment with etanereept increased significantly CD4+CD25high cells and FOXP3 mRNA expression (both P＜0.01),with negative correlations between these increases and decrease in CRP levels (P＜0.05 and P＜0.01,respectively).Conclusion In AS patients,peripheral FOXP3-expressing CD4 +CD25 + Treg cells are abnormal,and are up-regulated by etanercept treatment.This suggests a possible pathogenesis of AS and a potential mechanism for clinical efficacy of TNF-α inhibitors.

Objective To investigate the effects of cold exposure with different time courses on cardiovascular function and skeletal muscle contraction in Sprague-Dawley (SD) rats.Methods The male SD rats were randomly divided into control,12-h cold,24-h cold,and 48-h cold groups.The rats in all cold group were maintained in 4 ℃ environment for indicated time and allowed free access to standard rat chow and water ad libitum.After the animal model was established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Lanendorff heart persusion was used to detect some parameters of cardiac function,such as coronary flow (CF),mean left ventricular systolic pressure (mLVSP),mean left ventricular diastolic pressure (mLVDP) and maximum ascending and declining rate of left ventricular pressure (± dp/dt max) ; and then the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes of above cardiac function indexes were recorded again.Organ bath method was employed to detect contraction response to potassium chloride (KCl) or phenylephrine (PE) and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings.The twitch tension,titanic tension,and fatigue test of soleus and EDL were examined by electrical stimulation to reflect contractive function of skeletal muscle.Results The coronary flow (CF) were significantly reduced in 24-h and 48-h cold groups compared to control group [(9.0 ± 1.7) ml/g and (9.8 ± 1.3) ml/g vs (11.6 ± 1.2) ml/g,P ＜ 0.05] ; 48-h cold exposure also showed significantly increased mLVSP (P ＜0.01) and mLVP (P ＜ 0.05) of rats.After ischemia-reperfusion,the heart re-beat time was shorter in all cold exposure groups (all P ＜ 0.01),mLVP and mLVDP were higher in 24-h and 48-h cold group than control group (all P ＜ 0.01).The contraction of aortic rings responses to 60 mmol/L KCl [(0.63 ±0.13) g and (0.79 ±0.11)g vs (1.28±0.17)g,P ＜0.01] and 1 μmol/L PE [(0.62 ±0.18)g and (0.68 ±0.13)g vs (1.25 ± 0.18)g,P ＜ 0.01] were decreased while the endothelium-dependent relaxation responses to low concentration ACh (3 × 10-8 ～ 3 × 10-7) were increased in 24-h and 48-h cold groups relative to control group (all P ＜ 0.05).The 48-h cold also reduced titanic tension of SOL and EDL in comparison with control [(4.14 ± 0.62) N/cm2 vs (5.50 ± 0.75) N/cm2,P ＜ 0.05 and (3.00 ± 0.57) N/ cm2 vs (4.70 ± 0.85) N/cm2,P ＜ 0.01].Conclusions 24 ～ 48 h cold exposure could reduce CF but enhance the tolerance of heart from rats to ischemia reperfusion injury and also decrease the contraction of aortic rings response to KCl or PE but increase the sensitive responses to ACh of aortic rings from rats.The 48-h cold could decrease the contraction of skeletal muscle elicited by electrical stimulation.

Objective To investigate the effects of fasting with different time courses on the contraction of skeletal muscle and cardiovascular function in Sprague-Dawley (SD) rats.Methods The healthy and male SD rats were randomly divided into (1) control,(2) 12 h fasting,(3) 24 h fasting,and (4) 48 h fasting groups.After animal models of fasting with different time courses were established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Langendorff heart perfusion was used to detect some parameters of cardiac function,including left ventricular systolic pressure (LVSP),left ventricular end-diastolic pressure (LVEDP),and maximum ascending and declining rate of left ventricular pressure (± dP/dt max).Then,the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes in above cardiac function parameters were recorded again.The contraction response to potassium chloride or phenylephrine and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings were measured with organ bath.The changes of twitch tension and titanic tension of soleus and EDL were examined to reflect contractive function of skeletal muscle.Results The + dP/dt max was significantly reduced in 24 h and 48 h fasting groups compared to the control (P ＜ 0.01).After ischemia-reperfusion,the heart re-beat time was significantly longer [(24.3 ± 7.0)min vs (14.0 ± 2.9)min,P ＜0.05],both mLVDP and mLVP were significantly higher in 48 h and 24 h fasting groups than control group (P ＜ 0.05),and those changes did not normalized until re-perfusion 60 min.The 48 h fasting increased significantly endothelium-dependent relaxation of aortic rings response to low concentration acetylcholine (P ＜ 0.05),but did not affect the maximum relaxation of aortic rings response to high concentration acetylcholine and contraction of aortic rings responses to potassium chloride and phenylephrine.The 48 h fasting also enhanced significantly titanic tension of soleus compared to the control [(7.01 ± 1.22) N/cm2 vs (5.5 ± 0.75) N/cm2 for48 h-Fasting vs Control,P ＜0.05].Conclusions The 24 ～48 h fasting may induce inhibition of cardiac function,while fasting 48 h can increase the sensitivity of aortic rings responses to ACh and contraction function of soleus in healthy and male rats.

BACKGROUND:It has been recently indicated that nervous factors are able to adjust and dominate bone fracture healing. Type Ⅰ collagen is a major factor to promote the differentiation of osteoblasts and enhance the adhesion of osteoblasts; while, it is also a matrix protein for composing bone framework. Type Ⅱ collagen is derived from chondrocytes. OBJECTIVE: To study changing law of type Ⅰ and Ⅱ collagen expression during denervated bone fracture healing. DESIGN, TIME AND SETTING: Randomized controlled anima study was performed at the Animal Laboratory and Cell Biology Laboratory, the Second Military Medical University of Chinese PLA between May and December 2005. MATERIALS: Forty 3-month-year healthy male SD rats were randomly divided into fracture group (tibial fracture alone) and combination group (spinal cord injury combined with tibial fracture), with 20 rats at each group. METHODS: A φ 0.8 mm Kirschner wire was inserted into anterior border of left tibial plateau to establish tibial fracture models in the fracture group. A 0.3-cm spinal cord transection was cut at T10 segment to establish tibial fracture models with entire spinal cord injury. MAIN OUTCOME MEASRUES: Type Ⅰ and Ⅱ collagen protein expressions of callus were detected using Western blot technique in week 1, 2, 4, and 5 post-injury. RESULTS: One week after injury, type Ⅰ and Ⅱ collagen was represented in callus in the two groups, while the expressions in the combination group were significantly higher than fracture group (P<0.05); two weeks after injury, type Ⅱ collagen expression reached at the peak in the combination group, and the expression was significantly higher than the fracture group (P<0.05); four weeks after injury, type Ⅰ collagen expression reached at the peak in the fracture group, and the expression was significantly higher than the combination group (P<0.05), while type Ⅱ collagen still highly expressed in the combination group; five weeks after injury, type Ⅰ and Ⅱ collagen expressions were decreased in the two groups. CONCLUSION: Secretory law of type Ⅰ and Ⅱ collagen during denervated bone fracture healing is similar to normal bone fracture healing; however, the differences at time points, in particular expression at peak, are remarkable between them.

Objective According to the characteristics of skin defects in ankle and foot select the posterior lateral leg pedide skin flaps supplyed by different blood to repair,providing advice and reference.Methods Between January,2001 and December,2015,163 cases of soft tissue defects at the foot and ankle were treated in our department.①The sural neurovascular flap was used in 93 cases in ankle and foot defect.②The sural nerve nutritional vessel flap pediele with the perforating branch of the peroneal artery in 36 cases in ankle and foot defect.③The sural nerve nutritional vessel flap pediele with the perforating branch of the peroneal artery including sensory reconstruction in 16 cases in heel rejion defect.④Peroneal artery perforator flap in 10 cases in anterior ankle,lateral malleolus and posterior malleolus defect.⑤The use of the sural fasciocutaneous flap alonged with peroneal artery and perforators in 8 cases in dorsal foot defect.The donor site with skin graft.Results Of the 93 sural neurovascular flaps,8 had partial loss,which were cured after dressing.All the sural nerve nutritional vessel flaps pediele with the perforating branch of the peroneal artery survived.Sensory grading standard by UK Medical Research Council was used to evaluate the recovery of sensory function on the last follow-up.The sensory function recovery of heel region flaps with reconstruction of the sensory was between S0 and S1.All peroneal artery perforator flaps survived.One of the sural fasciocutaneous flap alonged with peroneal artery and perforators occurred distal epidermis,which were cured after dressing.All patients were followed up 6-50 months (mean 20 months).All patients had recoveryed walking function,and infection wound had no recurrence after surgery.Conclusion According to the location,size,severity and the injury of peripheral vascular,select the most simple,safe,minimal damage flap for the soft tissue defects at the foot and ankle.The right choice and the exact design can improve flap survival rate,and recieve good clinical results.

Objective To study the evaluation between the X-ray findings and ultrasound(US) manifestations of upper digestiveobstruction in infancy.Methods X-ray and US manifestations of 20 cases of infant with upper digestive obstruction proved by operationwere analysed respectively.Results In 20 cases,12 patients had pyloric stenosis,3 duodenal stresia,3 midgut malformation,2 hiatalhernia.Pyloric stenosis was diagnosed by X-ray in 11 cases,by US in 9,misdiagnosed by US in 1 case.Duodenal stresia was observed by X-ray and US in 2 cases separately,not observed by X-ray and US in 1 case separately.Midgut malformation was demonstrated by X-ray and US in 1 case separately,not demonstrated in 2 cases separately.Hiatal hernia was found by X-ray in 2 cases,misdiagnosed by US in 2 cases.Conclusion The two methods can't substitute with each other but complement.X-ray is better than US in the diagnosis of pyloric stenosis and hiatal hernia.US is better than X-ray in the diagnosis of midgut malformation.

Objective To examine the effect and mechanism of different targets of glucose control on liver damage in rats with sepsis .Methods The rat sepsis model was established by cecal ligation and puncture (CLP) .Forty Sprague‐Dawley rats were randomly divided into five groups (eight rats to each group):sham operation (sham group) ,sepsis (CLP group) ,glycemic control A group (glucose target 4 .6‐6 .1 mmol/L ) ,glycemic control B group (glucose target 6 .2‐8 .3 mmol/L ) and glycemic control C group (glucose target 8 .4‐10 .0 mmol/L) .The animals were sacrificed 12 hours after CLP .Venous blood was sampled for testing alanine transaminase (ALT ) , aspartate transaminase (AST ) and free fatty acid (FFA ) . Peroxisome proliferator activated receptor‐α (PPAR‐α) and liver carnitine palmitoyltransferase 1 (CPT‐1 ) protein were determined by immunohistochemistry .The pathological changes of liver tissue was observed under an optical microscope .Results The levels of ALT ,AST and FFA in venous blood and the pathological tissue injury score in sepsis groups were higher than those in sham group and all glycemic control groups (P<0 .05) .However ,the level of these markers significantly decreased in group A than those in group B or group C (P<0 .05) ,and lower in group B than those in group C (P< 0 .05) .PPARα and liver CPT‐1 expression levels were lower in sepsis group than those in sham group and all glycemic control groups except group C (P>0 .05) .The levels of PPARαand liver CPT‐1 were significantly higher in group A than in group B or group C (P<0 .05) ,and lower in group C than in group B(P<0 .05) .Conclusions The lowest target of glucose control(4 .6‐6 .1 mmol/L)shows better protective effects on liver damage in rats with sepsis ,the mechanism of which may be related to upregulation of PPARα and liver CPT‐1 expression .

Objective To investigate the relationship between the serum apolipoprotein B (ApoB)/ ApoA-Ⅰ ratio and intracranial artery stenosis in young patients with ischemic stroke.Methods Patients with ischemic stroke aged from 18 to 45 were enrolled in the study.Brain CT angiography was used to evaluate the degree of intracranial artery stenosis,and the concentrations of serum total cholesterol,triglyceride,highdensity lipoprotein cholesterol,low-density lipoprotein cholesterol,ApoA-Ⅰ,and ApoB were detected.The ratio of ApoB/ApoA-Ⅰ was calculated.The Demographic and clinical characteristics of the intracranial artery stenosis group and the non-intracranial artery stenosis group were compared.Multivariate logistic regression analysis was used to identify the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke.Results A total of 161 young patients with ischemic stroke were enrolled,including 89 in the intracranial artery stenosis group and 72 in the non-intracranial artery stenosis group.The constituent ratios of diabetes mellitus (20.2％ vs.6.9％;x2 =4.641,P =0.032),smoking (47.5％ vs.15.2％;x2 =15.121,P=0.001),hyperlipidermia (56.1％ vs.48.6％;x2 =4.197,P=0.040),as well as the radios in serum high-density lipoprotein cholesterol (1.29 ± 0.30 mmol/L vs.1.65 ± 0.34 mmol/L;t =7.131,P=0.002),ApoA-Ⅰ (1.49 ± 0.65 g/L vs.1.63 ± 0.23 g/L;t =2.751,P =0.001),ApoB (1.49 ± 0.65 g/L vs.1.63±0.23 g/L;t=2.751,P=0.001),and ApoB/ApoA-Ⅰ ratio (1.49±0.65 vs.1.63± 0.23;t =2.751,P=0.001) had significant differences between the two groups.Multivariate logistic regression analysis showed that diabetes (odds ratio [OR] 3.052,95％ confidence interval [CI] 1.186-7.856;P =0.021),smoking (OR 2.997,95％ Cl 1.456-6.172;P =0.003),hyperlipidemia (OR 4.745,95％ CI 2.108-10.668;P =0.001),ApoB (OR 4.861,95％ CI 3.029-7.802;P=0.001),and ApoB/ ApoA-Ⅰ ratio (OR 5.684,95％ CI 2.215-14.584;P=0.002) were the independent risk factors for intracranial artery stenosis in young patients with ischemic stroke,while HDL-C (OR 0.561,95％ CI 0.354-0.888;P=0.014) and ApoA-Ⅰ (OR 0.065,95％ CI 0.010-0.409;P=0.004) were the independent protective factors.After adjustment for hypertension,diabetes,smoking,hyperlipidemia,HDL-C,ApoA-Ⅰ,and ApoB,ApoB/ApoA-Ⅰ ratio was still an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke (each increase of 1 standard deviation,OR 4.255,95％ CI 2.348-7.711;P=0.001).Conclusion ApoB/ApoA-Ⅰ ratio is an independent risk factor for intracranial artery stenosis in young patients with ischemic stroke.

Objective To investigate the effect of Yiqi Huoxue Huatan Decoction(YHHD) on gene expression of vascular endothelial growth factor(VEGF), intercellular adhesion molecule 1(ICAM-1) in the colonic mucosa of ulcerative colitis rats. Methods Sixty rats were divided into six groups, namely normal control group, model control group, low-, middle- and high-dose Y HHD group(gastric gavage of YHHD in the dosage of 13, 26, 52 g·kg-1·d-1 respectively) , and Mesalazine(0.5 g·kg-1·d-1 ) group. T he rat model of ulcerative colitis was established by enema with 2, 4, 6-trinitrobenzene sulfonic acid(TNBS)mixture(2.5%TNBS and 50% ethanol). The changes in rat body weight were monitored. After treatment, pathological features of the colonic mucosa were observed under light microscope, and VEGF and ICAM-1 gene expression levels in the colonic mucosa were measured by reverse t ranscripti on polymerase chai n reaction(RT-PCR). Results Body weight was lighter, and relative gene expression levels of VEGF and ICAM-1 in the model group were significantly higher than those in the normal control group, and pathological features of the colonic mucosa was obvious, the differences being significant between the two groups(P<0.01). Compared with the model group, body weight was increased , pathological features of the colonic mucosa were relieved, and relative gene expression levels of VEGF and ICAM-1 were significantly decreased in YHHD groups(P<0.01), and high-dose YHHD showed the strongest effect. Conclusion YHHD can down-regulate the gene expression of VEGF and ICAM-1, which may be one of its therapeutic mechanisms for ulcerative colitis.