Objective To investigate the influence of Smac to the chemosensitivity of cyclophosphamide (CTX) and doxorubicin (DOX) in MCF-7 cells.Methods MCF-7 cells were exposed to CTX,DOX and the combination of both.3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2 H-tetrazolium bromide (MTT) assay was used to estimate the cell viability.Apoptosis was measured by acridine orange staining and Ho.33342/PI dou-ble staining.The mRNA and protein expressions of Smac were determined by RT-PCR and Western blotting.The study also analyzed the changes of pro-apoptotic proteins active caspase-3 and active caspase-9.Results CTX,DOX and the combination of both drugs reduced the cell survival rates in a concentration-dependent manner.The cell viability after being treated with 4.0 μg/ml CTX or 0.2 μg/ml DOX or 2.0 μg/ml CTX and 0.1 μg/ml DOX for 48 hours was (52.90 ± 8.78) ％,(53.35 ± 6.29) ％ and (34.19 ± 5.43) ％,respectively.The drug combination developed a stronger inhibitory effect compared to the single drugs (t =9.051,P=0.014;t =9.074,P =0.014).The Smac mRNA and protein levels in 2.0 μg/ml CTX and 0.1 μg/ml DOX group were 7.47 ± 0.82 and 4.13 ± 0.36,which were higher than those in 4.0 μg/ml CTX group (3.27 ± 0.40 and 2.28 ± 0.27;t =-50.120,P =0.000;t =-42.588,P =0.000) and 0.2 μg/ml DOXgroup (3.34±0.62and2.45±0.40;t=-46.233,P=0.000;t=-39.541,P=0.000).Furthermore,pro-apoptotic proteins active caspase-3 and active caspase-9 increased activity was confirmed by Western blotting.Conclusion Smac plays a vital role in enhancing the sensitivity of chemotherapeutic drugs CTX and DOX in MCF-7 cell line.

BACKGROUND: Presently, the biomaterial used in ligament tissue engineering such as collagen protein, polylactic acid, polyglycolic acid, small intestinal submucosa, glycan and nanomaterial are characterized by rapid degradation, resulting in inflammatory reaction after applying in host.OBJECTIVE: To investigate mechanical strength and in vitro degradation of silk scaffold and explore the reaction to macrophages.DESIGN: Controlled experiment.SETTING: Experiments were performed at the Department of Orthopaedics, Union Hospital, Huazhong University of Science and Technology from September 2004 to January 2005.MATERIALS: White raw Bombyx mori silkworm fibers of size 20/22 (according to the manufacturer) were obtained from the market. Bundles of 30 parallel fibers were prepared for a bundle of scaffold, which was put into fervens 5g/L Na2CO3for degumming. Ratio of Na2CO3 solution (Ml) to raw silk (g) was 1000.METHODS: In vitro degradation: 8cm long silk scaffold was weighed after drying. Subsequently, the silk scaffold was separately dipped into phosphate buffer saline (PBS) and 1.0g/L collagenase prepared with PBS. Twelve weeks later, silk scaffold was weighed to calculate weight loss rate. Simultaneously, tensile test was performed to detect the ultimate tensile strength (UTS) of samples. Culture of monocyte strain RAW264.7:2×108L-1 macrophage suspension (1mL) were separately added in a silk scaffold group, a control group and a lipopolysaccharide (LPS) group. At days 1 and 7, cell supernatant was collected from each group. Tumor necrosis factor-α(TNF-α) levels were measured by enzyme linked immunosorbent assay (ELISA).MAIN OUTCOME MEASURES: ① Changes in weight loss rate and UTS of the silk matrices after incubated with collagenase and the PBS. ②TNF-αlevels in the supernatant of each groups at days 1 and 7.RESULTS: Mass of silk matrices reduced by over 50% after incubated with collagenase for 8 weeks, but no change was found in PBS. UTS decreased by over 50% 8 weeks after incubated with collagenase, but no change was detected in PBS. At days 1 and 7, TNF-α levels in the supernatant was less in the silk scaffold group; TNF-α levels in the supernatant was significantly higher in the LPS group than in the silk scaffold group (P<0.01), but no significant difference in TNF-α levels was measured between the silk scaffold group and the control group (P>0.05).CONCLUSION: After 12-weeks degradation, silk scaffold still has good mechanical properties. Macrophages possess immunological inertia at days 1 and 7 after inoculated with macrophages.

Objectives To investigate the factors influencing long-term quality of life (QOL)of patients with first-episode ischemic stroke(IS)and partial side limb dysfunction.Methods 126 cases of the first ischemic stroke patients with partial side limb dysfunction admitted to hospital from November 2012 to July 2013 in Shanghai First People's Hospital were selected.General information of patients was collected and evaluated using NIH Stroke Scale(NIHSS)and Modified Rankin Scale(MRS).Following-up was conducted using NIHSS,MRS,stroke specific quality of life (SS-QOL) and Hamilton Depression Scale (HAMD) during 2-3 years after stroke Factors related to long-term QOL were screened by single factor analysis,then factors influencing QOL by multiple stepwise regression analysis.Results The average QOL score of patients with first IS partial side limb dysfunction was(181.5 ± 46.4)points scale.In each single scoring,the scale was(10.4±4.3)points in energy,(10.8±4.2)points in family roles,(21.8±6.2)points in language,(22.5 ±7.6) points in activity,(18.5±6.3)points in mood,(10.1±4.4)in personality,(19.2±5.9)points in self-care ability,(16.1 ± 7.0) points in social roles,(11.65 ± 3.31) points in thinking,(19.1 ± 6.6) points in upper limb function,(14.0 ± 2.0) points in visual acuity,(8.7 ± 4.1) points in work.The multiple regression analysis showed that the influencing factors of life quality included long-term NIHSS evaluation(β=-0.376,P＜0.01),HAMD evaluation(β=-0.375,P＜0.01),hypertension history(β=-0.178,P＜0.05).HAMD evaluation affected 9 fields of QOL,and NIHSS as long-term evaluation affected 6 fields of QOL.Conclusions Predictive factors for declined QOL of first-episode ischemic stroke patients with partial side limb dysfunction include higher NIHSS score,severer depression after long-term stroke and combined hypertension in the early stage.These indicate that recovering of body structure and function injury,control of hypertension,effective prevention and treatment of post-stroke depression maybe important measures for improving life quality of ischemic stroke patients with partial side limb dysfunction.

Objective The present study investigated the expression of heat shock cognate protein 70 (Hsc70) in the synovial tissues and blood samples of patients with rheumatoid arthritis (RA) to determine the pathological role of this protein in the pathogenesis of the disease.Methods The expression of Hsc70 in synovial membranes was quantitatively analyzed by immunohistochemistry,real-time quantitative PCR and western blotting.The samples from osteoarthritis (OA) and ankylosing spondylitis (AS) were used as controls.The levels of Hsc70 in blood of patients with RA were determined using enzyme linked immunosorbent assay (ELISA) with the samples of the healthy subjects as controls.Statistical analysis was conducted with one-way ANOVA,LSD test and Spearmen's correlation.Results Immunohistochemistry showed that Hsc70 had significantly increased expression in synovial tissues of RA than in the samples of OA and AS.Real-time PCR and western blotting confirmed the above findings.ELISA detected significantly elevated level of Hsc70 in blood of patients with RA as compared with samples from the controls (P＜0.01).Conclusion The study suggests that the up-regulation of Hsc70 may be involved in the pathogenic process of RA.

Objective To investigate the effects of intervertebral bridging ossifications in patients of osteoporotic vertebral compression fracture (OVCF) on bone fracture healing.Methods A total of 170 patients of thoracolumbar vertebral endplate fracture who were admitted into our hospital were selected.Divided these patients into the observation group,namely 60 patients with nonunion of vertebral endplate after 3 months of conservative treatment,and the control group, including 110 patients with well healed vertebra after 3 months of conservative treatment.Compared the distribution of intervertebral bridging ossifications of the two groups 3 weeks after injury.Results The incidence of bridging ossification at levels of T9 to T10,T10to T11,T11to T12 in the observation group were significantly higher than that in the control group.And it showed a significantly higher incidence of bridging ossification at the second proximal intervertebral segment in the observation group than that of the control group.There was a significantly greater sagittal wedge angle in the observation group compared with the control group.Conclusion Conservative treatment may increase the risk of nonunion of osteoporotic vertebral compression fractures when there is a bridging ossification at the second proximal intervertebral level or the sagittal wedge angle was greater than 14.2°in a fresh osteoporotic vertebral compression fracture.It should be a careful choose whether to take conservative treatment or surgical intervention.

Objective To investigate the expression of the interleukin-1 receptor(IL-1R)Ⅰ,IL-1RⅡand IL-1R accessory protein(IL-1RAcP)in osteoarthritis and analyse their biological significance.Methods Immunohistochemistry and reverse transcription-polymerase chain raction(RT-PCR)were adopted to detect the expression of IL-1RⅠ,IL-1RⅡand IL-1RAcP on the synovium of 107 OA patients.Results Immunohis- tochemistry showed strong positive expression of IL-1RⅠand IL-1RAcP,and positive expression of IL-1RⅡ. The expression was distributed in lining cells,monocyts and vascular endothelial cells of the sublining area, but all of them were negative or weak positive in normal synoviums.RT-PCR showed the expression of IL-1RⅠ,IL-1RⅡand IL-1RAcP in OA synoviums was significantly enhanced than normal synoviums (P＜0.05),and the expression of IL-1RⅠwas significantly enhanced than IL-1RⅡ(P＜0.05),but no sig- nificant difference with IL-1RAcP(P＞0.05).In stageⅡandⅢOA synoviums,the expression of IL-1RⅠand IL-1 RAcP had no significant difference with normal synoviums(P＞0.05).The expression of IL-1RⅡin stageⅢOA synoviums was significantly enhanced than normal(P＜0.05).Conclusion IL-1RⅠ,IL-1RⅡand IL-1RAcP play significant roles in the pathogenesis of OA,especially IL-1RⅠand IL-1RAcP.But their increase is only observed in the early stage of OA.These suggest that they may have no association with the development of OA and have no direct association with the severity of OA.OA can be cured by interrupting the signal transduction path in which IL-1 has played biological roles.

Objective To establishing an immortal cell line of familial papillary thyroid carcinoma (FPTC), and explore a new approach for studying familial non-medullary thyroid carcinoma (FNMTC). Methods The specimen from a patient with FPTC was selected, separated, and the primary cells were cultured using DMEM/F12 medium (with TSH, T3, EGF and hydrocortisone). To inducing cell immortalization, the exogenous genes SV40T/TERT were transfected into cells by two ways. RT-PCR was used to detect the expressions of thyroid peroxidase (TPO), thyroid globulin (TG), thyroid stimulating hormone receptor (TSHR) and sodium/iodide co-transporter (NIS). Immunofluorescence method was used to detect the expressions of TPO and GPC3. In order to detect the genomic mutations, the peripheral blood DNA of the patient was extracted. The cell genome was detected. Results The FPTC cells adhered to the plate and showed an irregular polygon shape. The cells can stably grow for six months, FPTC-S (with SV40T transfected) passaged to p26, FPTC cells passaged to p23 and FPTC-ST (with SV40T/TERT transfected) passaged to p19. Both FPTC-S and FPTC-ST can stably express TPO, TG and TSHR in mRNA level. MLH1 R217C mutation existed in the peripheral blood of the patient, and BRAF V600E mutation existed in the primary cultured cells. Either the primary or the immortal cells showed MLH1 R217C mutation. Conclusion This study preliminarily established an immortal cell line of familial papillary thyroid carcinoma with MLH 1 R217C and BRAF V600E mutations. This cell line provides a research model for studying these mutations in FPTC.

The incidence of solitary fibroma of seminal vesicle is low, and the source of seminal vesicle is rare. A 38-year-old patient was admitted to hospital because of intermittent gross hematuria for more than one month. Seminal vesicle space occupying lesions can be seen in pelvic MRI. Laparoscopic resection of right seminal vesicle tumor was performed, and the pathological diagnosis tended to solitary fibroma. During the 5-month follow-up, the symptoms of hematuria disappeared and no recurrence.

Objective:To explore the differences of leukocyte subtypes and thyroid function of the patients with different thyrotoxicosis diseases,and to clarify the practical significance of leukocyte subtypes and thyroid function tests in the differential diagnosis of Graves disease thyrotoxicosis and destructive thyrotoxicosis.Methods:A total of 33 patients with Graves disease thyrotoxicosis and 30 patients with destructive thyrotoxicosis confirmed by clinical and laboratory examination were selected; the levels of neutrophils (Ne),lymphocyte (Ly),basophils (Ba), eosinophil (Eo)and mononuclear cells (Mo),serum free thyroxine (FT4),three free iodine thyroid former glycine (FT3),and thyroid stimulating hormone (TSH)of the patients in two groups were analyzed and the receiver operator characteristic curve (ROC)was used to evaluate the values of the indicators with statastical significance in the differential diagnosis of Graves disease thyrotoxicosis and destructive thyrotoxicosis.Results:The levels of serum Eo,FT4 and FT3,and the modified data Eo/Mo,Eo × FT3 / Mo of the patients in Graves disease thyrotoxicosis group were significantly higher than those in destructive thyrotoxicosis group (P <0.05);the levels of TSH and Mo of the patients were lower than those in destructive thyrotoxicosis group (P < 0.05).The ROC curve analysis results showed that the sensitivities and specificities of Eo,Eo/Mo,Eo×FT3/Mo in the differential diagnosis of two diseases were good, and the best diagnostic boundaries were 1.54, 0.34, and 3.94. Conclusion:Eo,Mo,TSH,FT3,FT4,Eo/Mo,and EoxFT3/Mo could be regarded as the basis in the differential diagnosis of Graves disease thyrotoxicosis and destructive thyrotoxicosis ,and the practical significances of Eo,Eo/Mo,and Eo×FT3 /Mo in the differential diagnosis of Graves disease thyrotoxicosis and destructive thyotoxicosis are bigger.

Intraoperative ultrasound assisted localization is routinely used in the surgical treatment of completely endogenous renal cell tumor, however the localization and guidance ability of conventional ultrasound will decline to a certain extent for isoechoic renal tumor. A 62 years old female patient with right renal tumor was reported. The tumor diameter was about 2.3 cm× 1.7 cm, equivalent to the isoechoic of kidney. Laparoscopic partial nephrectomy was performed under the real-time guidance of contrast-enhanced ultrasound. The tumor was found to be lack of blood supply during the operation, and the tumor contour was clearly developed by contrast agent.The operation was successfully completed, and the pathological diagnosis was polycystic renal tumor with low malignant potential.The incisional margin was negative.The patient recovered well after operation without complications.No recurrence or metastasis was found after 6 months of follow-up.The renal function was good.