Objective　Influence of the function of coagulation and anticoagulation in the elderly patients with thrombotic diseases by activated protein C resistance(APCR) was studied. 　Methods　 Prothrombin fragments 1 and 2 (F1+2),fibrinopeptide A (FPA)，soluble fibrin monomer complex(SFMC) were examined by enzyme linked immunoassay(ELISA); fibrinogen(FIB),antithrombin III antigen(AT-Ⅲ:Ag) and its activity(AT-Ⅲ:A), protein C antigen(PC:Ag) and its activity(PC:A) and APCR were determined using CA 530 blood coagulation analyzer. 　Results　 Compared the elderly patient group with thrombotic diseases with the healthy elderly group, the parameters of coagulation system were obviously increased 〔F1+2(5.4±0.6)nmol/L vs (2.6±1.3)nmol/L,FPA (6.9±1.9)μg/L vs (4.8±2.3)μg/L,SFMC (72.4±8.7)g/L, vs (56.3±11.7)g/L,FIB (5.18±0.68)g/L　vs (3.82±0.74)g/L〕 and that of the anticoagulation system was significantly decreased〔AT-Ⅲ:Ag (212±19)mg/L vs (255±26)mg/L，PC:Ag (63.5±9.2)mg/L vs (82.7±20.1)mg/L〕.The above parameters changed more obviously in the elderly patients with positive APCR than those elderly patients with normal APCR.　Conclusions　APCR accelerated the activation of coagulation system and led to the weakening of the anticoagulation system function in the elderly patients with thrombotic diseases. The positive APCR may be one of the risk factors in the elderly patients who suffered from thrombosis.

Objective To investigate the value for the level mensuration of von Willebrand factor antigen (vWF:Ag) in stroke risk assessment in the patients with non-valvular atrial fibrillation (AF).Methods 180 non-valvular AF patients were selected from the Tianjin medical university general hospital from the 2009 to 2011 for retrospective cohort study,112 males and 68 females in the group,age 61-87 years.Using the IL ACL-9000 blood coagulation instrument assay the level of vWF:Ag.Using ROC curve to analyze the diagnosis performance of vWF:Ag,using Cox regression analysis model to evaluate the of vWF:Ag effect on prognosis,using x2 test to analyze the relevance between vWF:Ag and clinical pathological factors.Compared the patients group with CHADS2 score with the patients group with CHA2DS2VASc score date using t test.Results vWF:Ag levels were control group (112 ± 34)％,paroxysmal AF group (119 ±31)％,the persistent AF group (179 ± 47)％,permanent AF group (217 ± 56)％,atrial fibrillation associated with stroke group (235 ± 104)％ respectively.There was no difference between the paroxysmal AF group and control group (q =1.75,P ＞ 0.05) ; vWF:Ag level was higher in persistent atrial fibrillation group than in paroxysmal AF group (q =10.10,P ＜ 0.01); permanent atrial fibrillation group was higher than that of the persistent AF group (q =5.21,P ＜ 0.01).The optimum cut-off point with vWF:Ag for stroke diagnosis was 188.5％,the area under ROC curve =O.843 (95％ confidence interval:0.785-0.901).In Cox regression multianalysis,the vWF:Ag (HR =0.405; 95％ CI =0.268-0.716; P =0.026),the congestive heart failure(HR =2.901 ; 95％ CI =1.837-3.951 ; P =0.001),stroke/transienl ischemic attack (HR =4.665 ; 95 ％ CI =2.837-7.291 ; P =0.000),age (HR =0.474 ; 95 ％ CI =0.211-0.765; P =0.039),the Cox analysis showed that vWF:Ag was the independent prognosis factor for stroke in AF patients.Inx2 analysis,there was the relationship between the level of the vWF:Ag and the congestive heart failure/LVdysfunction (x2 =8.227,P ＜ 0.01),hypertension (x2 =3.305,P ＜ 0.05),age (x2 =7.581,P ＜ 0.01),diabetes mellitus (x2 =6.730,P ＜ 0.01),stroke/ transient ischemic attack/thromboembolism (x2 =4.825,P ＜ 0.05),vascular disease (x2 =4.126,P ＜ 0.05).Compared the subjects with CHADS2 (score =1) with the CHA2DS2VASc(score =1),the level of the vWF:Ag was higher in patients with CHADS2 score =1 (t =4.283,P ＜ 0.01).Conclusion There was relationship between the level of vWF:Ag and main pathologic factors in patients with AF,and changed with the condition,high vWF:Ag level was an independent predictor of stroke risk,and had superior reference value for in assessment of stroke in patients with atrial fibrillation.

Objective To characterize the alteration in peripheral blood endothelial progenitor cells (EPCs) of patients with systemic lupus erythematosus (SLE).Methods Venous blood samples were obtained from 82 female patients with SLE aged (35 ± 10) years and 50 healthy female controls aged (35 ± 13) years.ACL 9000 automated coagulation analyzer was used to determine the level of Von Willebrand factor antigen (vWF Ag).Flow cytometry was performed to detect peripheral blood EPCs and circulating endothelial cells (CECs).Analysis of variance was performed to assess the differences in these parameters between patients with active and stable SLE and the controls,and Pearson correlation analysis was conducted to evaluate the relationship between these parameters.Results The number of CD34+ cells,CD133+ cells and CD34+CD133+cells per 200 000 peripheral blood mononuclear cells was 35.4 ± 16.7,86.5 ± 32.1 and 361.3 ± 176.4 in patients with active SLE,significantly higher than that in the patients with stable SLE (17.1 ± 10.9,28.7 ± 21.5,107.2 ±44.3,respectively,all P ＜ 0.01)) and the controls (13.8 ± 9.6,11.2 ± 5.5,92.3 ± 50.5,respectively,all P ＜0.01).The patients with active SLE exhibited an elevated level of vWF Ag (438.9％ ± 205.3％ vs.130.2％ ±51.5％,P ＜ 0.01),an increased number of EPCs (361.3 ± 176.4 vs.107.2 ± 44.3,P ＜ 0.01) but a similar number of CECs (127±51 vs.118 ± 39,P ＞ 0.05) per 200 000 peripheral blood mononuclear cells compared with the healthy controls.No significant differences were observed in these parameters between the patients with stable SLE and the controls (all P ＞ 0.05).The number of EPCs was positively correlated with the level of vWF Ag (r =0.67,P ＜ 0.01),but uncorrelated with the number of CECs (P ＞ 0.05) in patients with active SLE.Conclusions The quantity of EPCs in peripheral blood is closely correlated with the level of the vascular injury marker vWF Ag,hinting that the number of EPCs can serve as a useful marker of disease severity.

Objective To study the mechanism of effects of estrogen receptor (ER) on T and B lymphocytes in patients with SLE and synergistic effect of T and B lymphocytes in the pathogenesis of SLE.Methods Real-time fluorescence quantitative PCR was performed to detect the mRNA expressions of ER-α,interleukin 10 (IL-10) and B lymphocyte stimulator (BLyS) in peripheral blood mononuclear cells (PBMCs)from 40 SLE patients and 40 normal human controls. The clinical and laboratory correlation with the levels of these parameters was analyzed. Results A significant increase was observed in the relative expression levels of ER-α, IL-10 and BLyS mRNA in SLE patients compared with the normal human controls (P ＜ 0.05 or 0.01 ), in active SLE patients compared with inactive SLE patients (P ＜ 0.05 or 0.01 ). Additionally, the mRNA expression levels of the 3 parameters were significantly correlated with the presence of renal damage, proteinuria, arthritis, etc. No statistical difference was observed in the mRNA expression levels of these parameters between female and male patients or between female and male normal controls. Conclusions IL-10 and BLyS appear to be correlated with the disease activity and severity of SLE, and ER-α may play an important role in the action mechanism of T and B lymphocytes in SLE.

Objective To study the effects of parecoxib sodium combined with continuous femoral nerve block on the balance of Th1/Th2 in elderly patients after total knee arthroplasty. Methods Fifty elderly patients,33 males and 1 7 females,aged 65 to 80 years,with ASA Ⅰ or Ⅱtreated with total knee arthroplasty were randomly divided into two groups (n =25 each):parecoxib sodium combined with continuous femoral nerve block group (group A)and parecoxib sodium com-bined with intravenous analgesia group (group B).Quick induce laryngeal mask anesthesia and intra-venous parecoxib sodium 40 mg at 30 min before skin incision were adopted in both groups.After the first 8 h,parecoxib sodium of 40 mg was intravenously injected again.The patients in group A re-ceived femoral nerve puncture and continuous electronic analgesia pump.The patients in group B re-ceived postoperative intravenous electronic analgesia pump.Visual analogue score(VAS)during rest and movement at 6,12,24,36,48 h after operation,side effects in two groups were recorded,venous blood samples were taken before anesthesia (T0 ),at the end of operation (T1 ),and 24 h(T2 ),48 h (T3 ),72 h (T4 )after operation for determination of plasma IFN-γ,IL-10 and cortisol (Cor). Results The values of VAS were significantly lower in group A compared with group B in rest at 6, 12 h after operation,the values of VAS were significantly lower in group A compared with group B in moving at every time points after surgery (P <0.05 ).The plasma Cor concentrations were signifi-cantly increased at T1-T3 compared with baseline value at T0 in two groups(P <0.05 ).The plasma Cor concentrations in group A were significantly decreased compared with group B at T1-T3 (P <0.05).The plasma IFN-γconcentrations were significantly lower at T2 and T3 compared with baseline value at T0 in group A,the plasma IFN-γ concentrations were significantly lower at T2-T4 compared with baseline value at T0 in group B (P <0.05),the plasma IFN-γ concentrations were significantly increased in group A compared with group B at T2-T4 (P <0.05 ).The plasma IL-10 concentrations were significantly increased compared with baseline value at T1-T4 in two groups (P < 0.05 ),the plasma IL-10 concentrations were significantly decreased in group A compared with group B at T1-T4 (P <0.05).Compared with group B,the incidence of postoperative nausea,vomiting and pruritus in group A was lowered significantly (P <0.05).Conclusion Parecoxib sodium combined with continu-ous femoral nerve block on postoperative pain in elderly patients after total knee arthroplasty can a-chieve good effect of postoperative analgesia and fewer complications, lower cortisol secretion, slowing down the decrease of IFN-γ secretion and the increase of IL-10 secretion.This method could effectively protect the immune function of patients undergoing arthroplasty.

Objective:To observe the effects of ischemic postconditioning (I-postC)on the lung injury after limb ischemia reperfusion (LIR)in the rats,and to investigate the protective effect and the possible mechanisms. Methods:24 Wistar rats were randomly divided into control group, ischemia-reperfusion group (IR group)and I-postC group (n=8 ). Referring to routine method in our department, the model rats which underwent 4 h ischemia and 4 h reperfusion of hind limbs were made.In control group,the rubber band around the limb was loose and the blood flow was not blocked. In I-postC group, before reperfusion, ischemia 5 min and reperfusion 5 min were performed in the rats,repeated for 3 times and then perfusion 4 h was taken,The blood and lung tissue from every rat were taken accurately. The percentages of CD1 8 positive cells in peripheral blood,the levels of soluble intercellular adhesion molecule-1 (sICAM-1)and P-selectin in plasma,the myeloperoxidase (MPO)activities in lung tissue,the levels of intercellular adhesion molecule-1 (ICAM-1)and P-selectin in lung tissue of the rats in various groups were detected. The partial pressure of oxygen (PaO2 ) and partial pressure of carbon dioxide (PaCO2 )were measured.The morphological changes of lung tissue under light and electron microscopes were observed.Results:Compared with control group,the percentage of CD18 positive cells and the levels of sICAM-1 and P-selectin of the rats in IR groups were increased (P<0.01);PaO2 and PaCO2 were decreased significantly;the MPO activity in lung tissue was also significantly increased (P<0.01).The HE staining results showed lung interstitial vascular dilation, congestion, PMN infiltration, the increased gap blood vessel, alveolar septal thickening,alveolar exudation, bronchial epithelial cell shedding and necrosis of the rats in IR group. Compared with IR group,the values of biochemical indicators mentioned above were decreased obviously (P<0.01);PaO2 and PaCO2 were increased significantly (P<0.01);the activities of inflammatory factors in plasma and lung tissue were decreased (P < 0.01 ); the pathological changes of lung damage were improved significantly. Conclusion:I-postC can reduce the lung injury after LIR in the rats,and the mechanism may be related to inhibiting the inflammatory reaction.

Objective To observe the effects of ischemic postconditioning (I-postC) on lung injury after limb ischemia reperfusion (LIR) in rats, and to investigate the protective effect and the mechanisms. Methods Twenty-four Wistar rats were divided into three groups:control group (group Control), ischemia-reperfusion group (group IR) and ischemic postcondi?tioning group (group I-postC). Referring to routine method in our department, the model rats underwent 4-hour ischemia and 4-hour reperfusion of hind limbs were made. In group Control, the rubber band around the limb was loose,which did not block the blood flow. Rats in group I-postC were given repeated 3 times of 5 min ischemia-5 min reperfusion, and then did perfusion 4 h before reperfusion. The blood and lung samples were collected for detecting arterial gas of partial pressure of oxygen [p(O2)] and partial pressure of carbon dioxide [p(CO2)]. The plasma and lung tissue levels of malondialdehyde (MDA), superoxide dismutase (SOD) and xanthine oxidase (XOD) were detected. The morphological changes of lung tissue were ob?served under light microscope and electron microscope. Results It was found that after suffering from ischemia-reperfu?sion, levels of p(O2) and p(CO2) decreased significantly. The activity of SOD in plasma and lung tissues decreased, but XOD and MDA increased significantly (P<0.05). With microscope, lung interstitial vascular dilation, infiltration of neutrophils, the width of the alveolar space, alveolar septal thickening and alveolar exudate were found. Compared with IR group, it was found that p(O2) and p(CO2) increased significantly in group I-postC. The activity of SOD in plasma and lung tissues in?creased, but XOD and MDA decreased significantly(P<0.05). The mild damage of pathological changes were found. Conclu?sion Ischemic postconditioning can reduce the lung injury after limb ischemia reperfusion in rats, which may be related to the inhibition of lipid peroxidation.

Objective:To observe the inhibitory effect of ischemic postconditioning (I-postC) on the apoptosis of renal cells after limb ischemia reperfusion(LIR) in the rats, and to investigate the possible mechanisms. Methods:Thirty SD rats were randomly divided control group, ischemia-reperfusion group(IR group) and I-postC group(n=10).4 h ischemia and 4 h reperfusion with the rubber band in two hind limbs of the rats were performed to establish the models.In control group, the rubber band around the limb was loose and the blood flow was not blocked.As for I-postC group, before perfusion, 5 min ischemia and 5 min reperfusion were performed in the rats and repeated 3 times named I-post C.The levels of blood creatinine (Cr), blood urea nitrogen(BUN) and C-reactive protein (CRP) in plasma of the rats in various groups were measured by automatic biochemistry analyzer.The expressions of Bcl-2 protein and Bax protein in renal were detected by immunohistochemical method,and its quantitative results were observed with automatic image analysis system and the ratio of Bcl-2/Bax was calculated.The apoptotic cells in kidney tissue were determined by terminal-deoxynucleotidy1 transferase-mediated d-UTP nick end labeling (TUNEL) under laser scanning confocal microscope(LSCM).The ultrastructures of kidney tissue were observed under electron microscope.Results:Compared with control group, the levels of Cr,BUN and CRP in plasma of the rats in IR group and I-postC group were increased(P<0.05 or P<0.01);compared with IR group, the levels of Cr, BUN and CRP in plasma of the rats in I-postC group were decreased(P<0.01).Compared with control group,the expression levels Bax and Bcl-2 in kidney tissue of the rats in IR group and I-postC group were significantly increased (P<0.05 or P<0.01),and the ratios of Bcl-2/Bax were reduced(P<0.05);compared with IR group,the expression level of Bax in kidney tissue of the rats in I-postC group was decreased (P<0.05),the expression level of Bcl-2 was increased(P<0.01),and the ratio of Bcl-2/Bax was increased(P<0.05).Under laser confocal microscope,the number of apoptotic cells in kidney tissue of the rats in IR group was increased compared with control group;the number of apoptotic cells in I-postC group was decreased compared with IR group.Under transmission electron microscope,the changes in IR group were found as follows: renal proximal convoluted tubule epithelial cell nucleus vacuoles,increased lysosome and dense particle deposition, some mitochondria crest fracture or fuzzy;irregular and fusion, glomerular podocyte protuberance,mitochondrial cristae fracture and reducetion with vacuoles, rough endoplasmic reticulum expansion;the damage levels of renal tubular epithelial cells and glomerulus in I-postC group were improved compared with IR group.Conclusion: Limb ischemia reperfusion can induce the apoptosis of renal cells, I-postC can inhibit the apoptosis of renal cells,and it would be helpful to improve the kidney function.

Objective To detect the expressions of osteopontin (OPN) and inerleukin-18 in patients with bullous pemphigoid (BP),and to analyze their relationship with clinical and laboratory indices.Methods Enzyme linked immunosorbent assay (ELISA) was performed to quantify the serum levels of osteopontin (OPN) and inerleukin (IL)-18 in 30 patients with BP and 30 health controls.Results The serum level of OPN was statistically higher in the patients than in the healthy controls ((8.29 ± 2.76) vs.(3.88 ± 1.41 ) ng/ml,P ＜ 0.01 ),and was positively correlated with the severity of BP (r =0.658,P ＜ 0.01 ) and with some laboratory indices in the patients.Increased serum IL-18 level was observed in patients complicated by cardiovascular diseases compared with those without cardiovascular diseases ((37.49 ± 6.43) vs.(31.10 ± 5.40) pg/ml,P ＜ 0.01).Moreover,the BP patients with diabetes,tumor,hepatic and renal impairment displayed an enhanced level of serum OPN and IL-18 than those without (all P ＜ 0.05).Conclusions OPN may be positively correlated with the severity of BP,while IL-18 may be involved in the development of complications of BP.

This study was designed to explone whether catalpol exerts a protective effects on kidney of diabetic rats and its possible mechanism.Insulin-like growth factor- I ( IGF- I ) and protein kinase B (PKB/Akt)mRNA and protein expression in kidney of diabetic rats were up-regulated as results of assessments by RT-PCR,immunohistochemistry and Western blot.Catalpol could partially reduce IGF- Ⅰ and Akt expressions in kidney of diabetic rats,and thus alleviate the damage of kidney.