Objective To explore the efficacy of radiofrequency ablation (RFA) and ethanol ablation(EA) for treating partially cystic thyroid nodules.Methods One hundred and twenty-four patients with a single partially cystic thyroid nodules which caused pressure symptoms or cosmetic problems were treated with RFA (n =42),EA (n =39) or RFA + EA(n =43).The inactivation rate of the nodule,tumor volume reduction rate,symptom scores (0-10) and complications were evaluated before and after treatment.Results The mean inactivation rate of RFA,EA and RFA + EA groups was (87.50 ± 5.77) ％,(57.00 ± 6.12) ％,(90.03 ± 5.39) ％,respectively.The inactivation rate of the nodule was negatively related to the volume of the solid part of the nodule.As to nodules with the volume ranging from 30 to 45 ml,the inactivation rate of RFA group was lower than RFA + EA group.Nodule volume reduction ratio (percentage) at 12-month follow-up was 90.45％ for RFA group,69.88％ for EA group,and 93.28％ for RFA + EA group.Regarding mean volume reduction,there were no difference between RFA + EA and RFA group (P ＞0.05),while there were significant differences between RFA + EA and EA group (P ＜0.05),also RFA and EA group (P ＜0.05).The patients in EA subgroup with cystic part percentage less than 50％ had a unsatisfying outcome (the volume reduction ratio less than 50％),while there were no differences(P ＞0.05) on the volume reduction ratio of nodule based on cystic part percentage between RFA group and RFA + EA group.The rate of the complication was 7.14％,2.56％,2.33％,respectively.Conclusions Ultrasound-guided percutaneous aspiration combined with RF ablation yielded better results than EA ablation.But due to ethanol ablation being less expensive and simpler to perform than RFA,ethanol ablation should be the first line treatment technique for benign thyroid nodules with cystic part percentage more than 50％,while radiofrequency ablation as first-line treatment for benign thyroid nodules with cystic part percentage less than 50％.

Objective:To investigate the influence of different salt concentration on the renal fibrosis and macrophages infiltration in salt sensitive hypertension.Methods:Dahl salt sensitive rats were randomly divided into the normal salt (0.3 ％ nacl) group,4 ％ high salt,8 ％ high salt groups at six weeks continuously feeding for 8 weeks,each group contained 15 rats.Tail-cuffmethod was used to value rat blood pressure at 8 weeks,Masson trichromatic method was used to detect renal fibrosis of the three groups at 8 week.Immunohistochemistry and Western blot method were used to depict the renal macrophage infiltration at 8 week.Results:1) The blood pressure of 4 ％ salt and 8％ high salt group rats were significantly higher than those of the normal salt group at 8week,meanwhile the blood pressure of 8 ％ high salt was further increased than that of 4 ％ high salt group at 8 week.2) The relative kidney weight and renal fibrosis of 4 ％ salt and 8 ％ high salt group rats were obviously higher than that of normal salt group at 8week,meanwhile the relative kidney weight and renal fibrosis of 8 ％ high salt were further increased than those of 4 ％ high salt group at 8 week.3) The macrophage infiltration of 4 ％ salt and 8％ high salt group rats were higher than that of the normal salt group at 8week,meanwhile the macrophage infiltration of 8 ％ high salt was further increased than that of 4 ％ high salt group at 8 week.Conclusion:Different high salt concentrations had different effect on the renal fibrosis and macrophage infiltration in the salt sensitive hypertension,high salt concentration could exacerbate the renal fibrosis and macrophage infiltration.

Objective To evaluate the combination use of thyroid fine needle aspiration biopsy (FNAB)and V-Raf murine sarcoma viral oncogene homolog B1 (BRAF) V600E gene mutation testing in differentiating benign from malignant thyroid nodules.Methods A total of 64 patients with pathologically-proved thyroid nodules were included in this study.The clinical data,including preoperative ultrasound-guided thyroid FNAB and BRAF V600E gene mutation detection,were retrospectively analyzed.Taking postoperative histopathological results as diagnostic gold standard for the thyroid nodule,the diagnostic values of simple FNAB,simple BRAF V600E gene mutation testing,and combination use of FNAB and BRAF V600E gene mutation detection were separately assessed.Results In the 62 patients a total of 64 nodules were detected (2 patients having bilateral nodules) and treated with surgery.Of the 64 nodules,BRAF V600E mutation was detected in 44 nodules,and 43 nodules were proved to be thyroid papillary carcinoma by postoperative pathological examination.Among the 44 nodules showing BRAF V600E mutation,FNAB made malignant diagnosis in 28,benign diagnosis in 6,and uncertain diagnosis in 10.Of the 20 nodules showing no BRAF V600E mutation,FNAB made malignant diagnosis in 5,benign diagnosis in 3,and uncertain diagnosis in 12.The postoperative pathological examination confirmed that 14 lesions were thyroid papillary carcinoma,4lesions were nodular goiter,one lesion was subacute thyroiditis,and one lesion was thyroid adenoma.Among the 57 thyroid papillary carcinomas,BRAF V600E mutation was detected in 43,with the mutation rate being 75.4％.Compared with the gold standard based on pathological diagnosis,the sensitivity,specificity,positive predictive value,negative predictive value,correct diagnosis rate of FNAB for judging benign or malignant thyroid nodules were 78.9％,85.7％,97.8％,33.3％ and 79.7％ respectively,which of BRAF V600E gene mutation detection for judging benign or malignant thyroid nodules were 75.4％,85.7％,97.7％,30.0％ and 76.6％ respectively,and which of FNAB plus BRAF V600E gene mutation detection for judging benign or malignant thyroid nodules were 94.7％,71.4％,96.4％,62.5％ and 92.2％ respectively.By using McNemar paired data x2 test to compare FNAB with combination use of FNAB plus BRAF V600E gene mutation detection in diagnosing thyroid nodules,the results indicated that statistically significant deference in differentiating benign from malignant thyroid nodules existed between the two methods (P＜0.001).Conclusion For the qualitative diagnosis of thyroid nodules which nature cannot be determined by simple FNAB,FNAB combined with BRAF V600E gene mutation detection can improve the diagnostic accuracy for benign and malignant thyroid nodules.

BACKGROUND:It has been found that cel ular repressor of E1A-stimulated genes (CREG) is a lysosomal protein binding directly to the mannose-6-phosphate (M6P)/insulin-like growth factor II receptor (IGFIIR) and depends on the interaction with M6P receptors for efficient delivery to lysosomes OBJECTIVE:To study the interactions between the exogenous CREG protein and cathepsins and M6P/IGFIIR and to confirm the effect of CREG protein on expression and distribution of M6P/IGFIIR. METHODS:Double-stained immunofluorescence and coimmunoprecipitation were applied to observe the interactions between the exogenous CREG protein and cathepsin B, cathepsin L and M6P/IGFIIR. Using gain-of-function and loss-of-function approaches, the effect of CREG on expression and distribution of M6P/IGFIIR were studied by western blot assay and immunofluorescence staining. RESULTS AND CONCLUSION:Double-stained immunofluorescence and coimmunoprecipitation analyses confirmed the direct interactions between the exogenous CREG protein and cathepsin B, cathepsin L and M6P/IGFIIR. It was verified that CREG plays a critical role not in the expression but in the distribution of M6P/IGFIIR using gain-of-function and loss-of-function approaches. These findings provide evidence that exogenous CREG protein is located in lysosomes and has interactions with cathepsins and M6P/IGFIIR, also CREG plays a critical role in the distribution of M6P/IGFIIR.

Objective To investigate the role of pathogenic Leptospira interrogans lipopolysaccha-ride (L-LPS) and outer membrane proteins (L-OMP) in the apoptosis of mouse macrophages (J774A.1) and their association with Fas/FasL pathway .Methods Phenol-water extraction and Triton X-114 phase separation were used to extract L-LPS and L-OMP from L.interrogans serogroup icterohaemorrhagiae serovar Lai strain Lai, respectively.Polymyxin B ( PMB) and protease K ( PK) were used to treat L-LPS and L-OMP, respectively.J774A.1 cells were stimulated by L.interrogans strain Lai with or without ultraviolet inactivation.In parallel, the cells were stimulated by extracted L-LPS and L-OMP with or without PMB and PK treatments .The apoptosis and necrosis of J 774 A.1 cells before and after treatment were detected by flow cytometry.The siRNAs were used to silence the expression of Fas or FasL gene in J 774A.1 cells and their inhibitory effects were further validated by using real-time fluorescent quantitative RT-PCR.Flow cytometry was used to detect the effects of L-LPS or L-OMP on the apoptosis of J774A.1 cells with Fas or FasL gene-knockdown .Results L.interrogans strain Lai with or without ultraviolet inactivation could cause similar early apoptosis rates (47.1%and 55.6%) and late apoptosis/necrosis rates (7.6%and 7.9%).The ear-ly apoptosis rates of 1×105 J774A.1 cells were 40.4%and 34.0%after the treatment with 100 ng of L-LPS and 100 μg of L-OMP for 4 h.The late apoptosis/necrosis rates of the cells were 7.5%and 6.9%upon the treatments with L-LPS and L-OMP, respectively.However, the apoptosis or necrosis of the cells was not ob-served when using L-LPS and L-OMP pre-treated by PMB and PK, respectively.Silenced expression of Fas or FasL gene reduced the L-LPS-induced J774A.1 cells apoptosis (P<0.05), while decreased early apopto-sis rate of J774A.1 cells mediated by L-OMP was only observed in Fas gene-knockdown cells (P<0.05). Conclusion Both L-LPS and L-OMP can cause the Fas/FasL-associated apoptosis of macrophages , which is beneficial for L.interrogans to establish the productive infection in hosts .

0.05).The expression of TNF-? mRNA was significantly upregulated in the lower respiratory tract,C-7-T-5 spinal ganglia and corresponding spinal dorsal horn of the experimental asthmatic guinea pigs compared with the normal saline control group and the simple sensitized group(P

Objective To explore the expression of NF-?B in C-7-T-5 spinal ganglia and possible mechanism of treating asthma with dexamethsone. Methods The alterations of NF-?B activity and its mRNA expression were investigated by means of immunohistochemistry and in situ hybridization histochemistry combined with the micro-image analysis in C-7-T-5 spinal ganglia in the asthmatic and dexamethsone-treated guinea pigs. Results The NF-?B immunoreactivity was found mainly in cytoplasm and weakly in nucleus,its mRNA was expressed weakly in C-7-T-5 spinal ganglia in the control groups.The NF-?B immunoreactivity was more in nucleus and less in cytoplasm,the expression of its mRNA increased significantly in C-7-T-5 spinal ganglia in the asthmatic group compared with the control groups(P0.05).Conclusion The present results indicate that NF-?B in C-7-T-5 spinal ganglia might be involved in the pathogenesis of the bronchial asthma,and inhibiting the expression and activity of NF-?B in C-7-T-5 spinal ganglia might be one of the mechanisms of treating the bronchial asthma with dexamethsone.

Autologous red blood cells (RBC) labeled with fluorescence were immitted into microvessel of SD rat and observed under microscope. The movement of each individual labeled RBC was recorded by microscope video camera system. The recorded videotape is replayed to sample dark background fluorescent images through frame grabber. Sampled frame images were separated into odd and even field sequence images. Then these sequence images were analyzed to get the flow rate. The error between the actual flow velocity value and the flow velocity value of fluorescent globules in the chamber measured under the same system was below 7%. The upper limit was 9.6 mm/s. There are no obvious differences (P > 0.05). This system has been applied in the research of rat microcirculatory disturbance, and the temporal flow rate change in microvessel was obtained.
Animals ; Blood Flow Velocity ; physiology ; Image Processing, Computer-Assisted ; Microcirculation ; physiology ; Microscopy, Fluorescence ; Rats ; Rats, Sprague-Dawley ; Splanchnic Circulation ; physiology ; Videotape Recording

Objective To investigate whether chemokine CC motif 2 (CCL2) is involved in the high residual platelet response,and the mechanism of CCL2 being involved in the regulation of platelets.Methods Forty patients with ST elevation myocardial infarction (STEMI) were admitted.P2Y12 reaction unit (PRU) was detected by VerifyNow.Forty patients were divided into high platelet reactivity group (high reactivity group,n=24) and normal platelet reactivity group (normal reactivity group,n=16) according to the results of PRU detection.Plasma CCL2 concentration of the STEMI patients was examined by ELISA.The expressions of CCL2 and CCR2 in the platelets were detected by Western blotting.After CCL2 stimulation,the kinases of which phosphorylation was changed in the platelets were screened by ARY003B protein chips.The phosphorylation of p38MAPK and HSP27 in the platelets was tested by Western blotting after CCL2 stimulation in the presence or absence of CCR2 antagonist (RS 102895) or p38MAPK signal pathway inhibitor (SB 203580).Results The plasma CCL2 concentration of high reactivity group was markedly higher than that of normal reactivity group.Moreover,compared with normal reactivity group,the expressions of CCL2 and CCR2 in the platelets of high reactivity group significantly increased.After the platelets were stimulated by CCL2,the phosphorylation of p38α and HSP27 enhanced in the platelets by protein chips screening.When RS 102895 or SB 203580 was treated before CCL2 stimulation,the phosphorylation of p38MAPK and HSP27 decreased.Conclusions CCL2 participates in high residual platelet response in an autocrine/paracrine way.CCL2/CCR2 might affect the function ofplatelets through p38MAPKHSP27 signal pathway.

BACKGROUND:Embryonic stem cells (ESCs) serve as a major cell source for smooth muscle cells,but the heterogeneity of cells derived from ESCs result in difficulty to obtain high purity smooth muscle cells.OBJECTIVE:To construct a double expression vector of puromycin resistance (pac) gene and enhanced green fluorescence protein (EGFP) gene driven by smooth muscle specific SM22α promoter (pSM22α-PAC-IRES2-EGFP),in addition,to detect its availability and specificity in ESCs.DESIGN,TIME AND SETTING:The observational experiment of gene level was performed at the Cardiovascular Institute,General Hospital of Shenyang Military Region from April 2007 to September 2008.MATERIALS:ESCs line R1 with number SCRC-1011TM was purchased from American ATCC Company.The pSM22α-EGFP vector was constructed by our laboratory.And the pIRES2-EGFP,pSM2C and pSuper.basic vectors were purchased from Invitrogen Company.METHODS:SM22α promoter was cloned from pSM22α-EGFP by polymerase chain reaction.CMV promoter of pIRES2-EGFP vector was replaced by SM22 promoter to establish pSM22α-IRES2-EGFP.Pac gene,excised from pSM2C by HindⅢ/Clal digestion,was sub-cloned into pSuper.basic to establish pSuper-PAC.After BgⅢ/Accl enzyme digestion of pSuper-PAC,pac gene fragment was obtained,which was further sub-cloned into pSM22α-IRES2-EGFP to produce pSM22α-PAC-IRES2-EGFP.ESCs were transfected with pSM22α-PAC-IRES2-EGFP using lipofectamine.Positive clones were selected by G418 and induced to differentiate and further identified by amplification of pac gene by RT-PCR.Differentiated cells were immunostained by SM α-actin,and expression of SM α-actin and EGFP was observed simultaneously under fluorescence microscope.MAIN OUTCOME MEASURES:Sequencing result of pSM22α-PAC-IRES2-EGFP;Amplification of pac gene;EGFP expression;as well as SM α-actin immunostaining.RESULTS:Three segments of 261 bp,664 bp,and 5000 bp were obtained by HindⅢ/Clal digestion,which was coincident with expectation,and the sequencing results showed that pSM22α-PAC-IRES2-EGFP vector was successfully constructed.Amplification of pac gene identified 4 ESCs clones successfully transfected.After induction of differentiation,partial portion of differentiated cells expressed EGFP,accompanied by positively stained by SM α-actin antibody.CONCLUSION:pSM22α-PAC-IRES2-EGFP vector was successfully constructed.ESCs clones transfected with this vector expressed pac gene and EGFP gene,and the expression of EGFP is smooth muscle specific.