Objective To investigate the effect of up-regulation of tetraspanin-5 (Tspan-5) expression on the metastatic ability of colorectal cancer (CRC) cells. Methods The coding sequence of Tspan-5 mRNA was amplified from total RNA of LoVo cells by RT-PCR and cloned into pEGFP-C1 to construct Tspan-5/pEGFP-C1,a eukaryotic expression vector. The expression of Tspan-5 in LST-R1 cells was up-regulated by gene tranfection. Western blotting was used to detect the expression level of Tspan-5/EGFP fusion protein,and laser scanning confocal microscopy (LSCM) was used to observe the distribution of Tspan-5/EGFP in LST-R1 cells. The changes in adhesion,migration and invasion ability of LST-R1 cells were determined by adhesion,migration and invasion experiment in vitro,respectively. Liver metastasis model of nude mice was used to test the changes in liver metastasis ability of LST-R1 cells in vivo. Results The Tspan-5/EGFP fusion protein could be detected by Western blotting in Tspan-5/pEGFP-C1 transfectants. LSCM showed that Tspan-5/EGFP protein located in cell membrane. Adhesion and migration assays showed that up-regulation of Tspan-5 expression level significantly promoted the adhesion and migration ability of LST-R1 cells on a series of extracellular matrix (ECM) components,including collagen IV,FN,LN and VN (P

Objective To investigate the radioprotective function and its mechanism of Sulforaphane (SF) in mice acute radiation-induced lung injury.Methods Totally 40 female C57BL/6J mice were equally divided into 5 groups randomly.Group A,treated by SF 3 mg/kg plus radiation;group B,treated by SF 5 mg/kg plus radiation;group C,treated by SF 10 mg/kg plus radiation;radiation group with a single dose of 12 Gy in 6 MV X-ray by a linear accelerator,and control group with sham radiation.The mice in drug group were administered intraperitoneally with different concentration of SF every other day from 7 d before irradiation to 7 d after irradiation,while the same volume of DMSO plus physiological saline solvent was given in the control and radiation groups.After being sacrificed at 14 d of SF administration,the pathomorphological changes of mice were observed in trauma lung tissue,the positioning and expression of NLRP3 was observed by immunohistochemical staining,the levels of IL-6,TNF-α and TGF-β1 in bronchoalveolar lavage fluid (BALF) were measured by ELISA,the expressions of NLRP3 and IL-1β mRNA in lung tissue were assayed by qRT-PCR,the expressions of NF-κB p65,NLRP3 and IL-1β proteins in lung tissue were assayed by Western blot,the activity of NF-κB was detected by EMSA.Results In comparison with radiation group,there was an obvious amelioration in pathological injury of lung tissue in the treatment groups:the expression of NLRP3 in lung tissue decreased;the concentration of NLRP3 in the drug intervention group (SF 10 mg / kg) markedly decreased (F =42.750,P ＜ 0.05).the IL-6,TNF-a and TGF-β1 levels in BALF decreased (tIL-6 =-62.65-21.00;tTNF-α =-32.18-16.57;tTGF-β1 =-58.22-46.11,P ＜ 0.05);the expressions of NLRP3 and IL-1β mRNA markedly decreased (tNLRP3 =-6.56-5.68;tIL-1β =-29.75--21.20,P ＜ 0.05),and the expressions of NF-κB p65,NLRP3 and IL-1β proteins decreased (tNF-κB p65 =-34.00--1.71,tNLRP3 =-25.01--16.91,tIL-1β =-73.70--55.14,P ＜ 0.05);the relative expressions of NF-κB p65 and NLRP3 were reduced in a dose-dependent manner (r =0.945,0.926);and the activity of NF-κB were obviously reduced (tNF-κB =-38.68,-614.82,-2 831.40,P ＜ 0.05).Conclusions Sulforaphane effectively alleviates the RILI in lung of mice by downregulating the expressions of inflammatory factor NLRP3.

Altitude ; Animals ; CA1 Region, Hippocampal ; metabolism ; physiology ; Male ; Neural Cell Adhesion Molecules ; metabolism ; Rats ; Rats, Sprague-Dawley ; Time Factors

Objective To determine the median effective dose ( ED50 ) of 0?5% ropivacaine for ul?trasound?guided adductor canal saphenous nerve block. Methods Twenty?three patients of both sexes, aged 18-45 yr, weighing 50-85 kg, of American Society of Anesthesiologists physical statusⅠ or Ⅱ, scheduled for elective arthroscopic meniscectomy, received an ultrasound?guided adductor canal saphenous nerve block. After successful location, 0?5% ropivacaine was injected. The sensory block of the area in?nervated by the saphenous nerve was assessed by pin?prick test. The dose of ropivacaine was determined by up?and?down technique. The initial dose of ropivacaine was set at 18 ml. Each time the dose increased∕de?creased in the next patient according to whether or not the sensory block of the area innervated by the saphe?nous nerve was complete, and the ratio between the two successive doses was 1 ∶ 1?2. The ED50 and 95%confidence interval of 0?5% ropivacaine for adductor canal saphenous nerve block were calculated by probit analysis. Results The ED50 (95% confidence interval) of 0?5% ropivacaine for ultrasound?guided adduc?tor canal saphenous nerve block was 10?7 ( 8?8-12?2) ml. Conclusion The ED50 of 0?5% ropivacaine for ultrasound?guided adductor canal saphenous nerve block is 10?7 ml.

Background There are several methods for postoperative analgesia for knee surgery.The commonly utilized method is multimodal analgesia based on continuous femoral nerve block.The aim of this study was to investigate the application of continuous adductor canal block for analgesia after total knee replacement and compare this method with continuous femoral nerve block.Methods Sixty patients scheduled for total knee replacement from June 2013 to March 2014 were randomly divided into a femoral group and an adductor group.Catheters were placed under the guidance of nerve stimulation in the femoral group and under the guidance of ultrasound in the adductor group.Operations were performed under combined spinal and epidural anesthesia.After the operations,0.2％ ropivacaine was given at a speed of 5 ml/h through catheters in all patients.Visual analogue scale (VAS) pain scores at rest and while moving were noted at 4,24,and 48 hours after the operation,and quadriceps strength was also assessed at these time-points.Secondary parameters such as doses of complementary analgesics and side effects were also recorded.Results There were no significant differences between the groups in VAS pain scores at rest or while moving,at 4,24,or 48 hours after the operation (P ＞0.05).At these time-points,mean quadriceps strengths in the adductor group were 3.0 (2.75-3.0),3.0 (3.0-4.0),and 4.0 (3.0-4.0),respectively,all of which were significantly stronger than the corresponding means in the femoral group,which were 2.0 (2.0-3.0),2.0 (2.0-3.0),and 3.0 (2.0-4.0),respectively (P ＜0.05).There were no significant differences between the groups in doses of complementary analgesics or side effects (P ＞0.05).X-ray images of some patients showed that local anesthetic administered into the adductor canal could diffuse upward and reach the femoral triangle.Conclusions Continuous adductor canal block with 0.2％ ropivacaine could be used effectively for analgesia after total knee replacement.Compared with continuous femoral nerve block,this analgesic method has similar analgesic effects and is associated with less weakness of quadriceps muscle.

Aged ; Carcinoma, Renal Cell ; pathology ; Carcinosarcoma ; pathology ; Chondroma ; pathology ; Chondrosarcoma ; complications ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Kidney Neoplasms ; complications ; metabolism ; pathology ; secondary ; surgery ; Lung Neoplasms ; secondary ; Nephrectomy ; Pleural Effusion, Malignant ; etiology ; S100 Proteins ; metabolism ; Soft Tissue Neoplasms ; pathology ; Vimentin ; metabolism

Objective To investigate the correlation between plasma homocysteine (Hcy) and high sensitivity C-reactive protein (hs-CRP) ,Hcy and von Willebrand factor(vWF) in patients with acute cerebral infarction ,and to analyze their clinical significance . Methods 150 patients with acute cerebral infarction were enrolled in the study ,and were divided into 3 groups according to their plasma Hcy concentrations .Meanwhile 50 healthy people were recruited as control group .Plasma hs-CRP ,vWF and Hcy concentra-tions were determined and compared between groups .Then data correlation analysis was performed .Results In patients with acute cerebral infarction plasma hs-CRP ,vWF and Hcy concentrations were significantly higher than those in healthy control group (P<0 .05) .Plasma Hcy concentrations in patients with acute cerebral infarction were positively correlated with plasma hs-CRP and vWF concentrations .Conclusion Hs-CRP ,vWF and Hcy are biomarkers of endothelial injury ,the combined detection of the three indica-tors might contribute to the early diagnosis and treatment of acute cerebral infarction .

Objective To investigate the level change and the significance of serum homocysteine(Hcy),endothelial nitric oxide synthase(NOS)and soluble thrombomodulin(STM)in the patients with acute cerebral infarction(ACI).Methods The values of serum Hcy,NOS and STM were detected in the 150 patients with ACI and the 50 healthy individuals and the detection results were performed the significance testing and the correlation analysis.Results Compared with the normal control group,the levels of ser-um Hcy,NOS and STM in the ACI patients were significantly increased with statistical differences(P <0.05).The increased level of NOS and HCY in the ACI patients was positively correlated with the increased Hcy level.Conclusion Hcy,NOS and STM are the markers of endothelial injury occurrence and their detection has certain significance for judging the disease condition and guiding treatment.

Objective To investigate the clinical and molecular pathological features of limb-girdle muscular dystrophy 2A (LGMD2A) of Chinese patients. Methods Thirty cases of LGMD with excluding LGMD2B were included in this study. The muscle specimens were performed by a standard series methods of histochemistry, enzymohistochemistry, immunohistochemistry and Western blot. The clinical and molecular pathological features of LGMD2A were retrospective analyzed. Results Five cases with no or only trace expression of calpain-3 protein were diagnosed as calpainopathy (LGMD2A) by Western blot analysis. The age of onset of these 5 patients ranged from 10 to 45 years and the duration of the disease were about 2-10 years. Proximal muscles weakness and atrophy of lower limbs were predominantly involved. In all patients,symptoms progressed slowly. The ambulation could be retained for many years but running and jumping were impaired early. The serum creatine kinase level was elevated moderately to markedly. Electromyography showed myopathic patterns in all cases. Two siblings had similar symptoms indicating autosomai recessive inherited pattern. Pathologically, there was marked variation in fibre size and most small fibres were round. Some necrotic and regenerating fibers were seen. Fibres with centrally placed nuclei can be found frequently. No infiltrations of inflammatory cells were seen. Lobulated fibers were observed in 2 patients by NADH-TR stain. The expression of dystrophin, caveolin-3, α-, β-, γ- and δ-sarcoglycan protein were normally staining of 5 LGMD2A patients' specimens by immunohistochemistry. Two patients had reduced staining of dysferlin by immunohistochemistry study. Conclusions Clinical and pathological characteristics of our 5 LGMD2A patients are consistent with typical muscular dystrophy features reported in other countries. Identification of calpian-3 deletion by Western blot is essential for the diagnosis of calpainopathy.

Objective:To verify the fracture risk assessment tool ( FRAX) to estimate the probability of osteoporotic fracture in patients with rheumatoid arthritis ( RA ) with or without bone mineral density (BMD), and identify associated risk factors of osteoporosis .Methods: In the study, 200 patients with rheumatoid arthritis aged more than 40 years in Peking University First Hospital from Dec .2009 to Dec. 2012 were recruited.Clinical information was obtained from a questionnaire of their case history and medical records.FRAX tool was administered.Their lumber spine and left femoral BMD were determined by dual energy X ray absorptiometry.The gender, age, disease duration, menopause status, body mass index ( BMI) and accumulative dose of glucocorticoid were obtained in retrospect .Correlation analysis was conducted between the BMD and clinical information .Results:The study population ( female, 77.5%) had a mean age of 59.4 years, in which 10 (13%) patients showed a normal BMD, 67 (87%) were osteopenia or osteoporosis , while 32 patients (16%) had fragile fracture.Compared with the patients with normal BMD, the subjects with low BMD had significantly older age , longer period for corticoids usage , higher day dose and accumulated dose of corticoids .The 10-year fracture risk of sustai-ning major osteoporotic fractures and hip fracture was higher .No significant difference was observed be-tween the 10-year fracture risks calculated with BMD and without BMD .The values of the different area under the receiver operating characteristic ( ROC) curve ( AUC) for major and hip fractures calculated in three ways:without BMD, with the femoral neck BMD, and with T-score.The best result was for FRAX tool for hip fracture with the T-score ( AUC 0 .899 ) .A stepwise multivariate linear regression model was constructed to explore the relationship between the different clinical factors studied and a low BMD . Three statistically significant variables for lumber BMD were pain on visual assessment scale ( VAS ) (P=0.02), fracture history (P=0.003) and a higher steroid accumulated dose (P=0.008).Three statistically significant variables for left hip BMD were age (P<0.001), fracture history (P=0.05) and lower BMI ( P=0.03) .Conclusion:Low BMD is a common complication in RA patients .Risk factors for major fracture and hip fracture are increased .There is a positive correlation between FRAX calculated with and without BMD or T score .FRAX with the femoral neck T score or BMD presents a discriminatory capacity better than FRAX without BMD , according to the AUC ROC .