Objective:To introduce a "Four-step method" for extraction and separation of Candida albicans total proteins.Methods: Proteins of C.albicans were extracted step-by-step with 4 kinds of solutions with different solubilities.After quantification,the protein samples were separated by isoelectric focusing electrophoresis and then by SDS-PAGE.Results: Proteins with different solubilities were successfully extracted step-by-step from C.albicans and were separated by two-dimensional gel electrophoresis. Conclusion: The "Four-step method" for extraction of C.albicans proteins is an effective approach to study C.albicans membrane proteome and lays a foundation for further investigation of the mechanisms of antifungal agents and drug resistance in C.albicans.

Objective:To investigate the mechanisms of genistein (GEN) affecting the chemosen- sitivity of human breast cancer cell line MDA-MB-453 to paclitaxel (PTX) in vitro. Method:HER2/neu- overexpressing breast cancer cells MDA-MB-453 were treated by GEN, PTX alone or combined in vitro. Cell cycle was measured by flow cytometry. The expression of HER2/neu protein was observed by immunocytochemistry and. Akt, p-Akt, cyclin B1 and CDK1 protein by Western blot. Results:Cell cycle of MDA-MB-453 cells was blocked at G1/S after treatment of GEN, while at G2/M after treatment with PTX alone. Both GEN and PTX did not change the expression of HER2/neu, total Akt and CDK1 in MDA-MB-453 cells, but GEN significantly decreased p-Akt and cyclin B1 level, and PTX obviously increased cyclinB1 level. GEN antagonized the effects of PTX on level of cyclin B1 protein and blockage of G2/M in MDA-MB-453 cells after treatment with GEN and PTX in combination. Conclusion:The antagonism effects of GEN on the increase of cyclin B1 and blockage of G2/M induced by PTX may be one of the mechanisms of GEN affecting the chemosensitivity of MDA-MB-453 cells to PYX.

Objective To summarize the imaging characteristics of the hepatic epithelioid hemangioendothelioma (EHE).Methods The clinical data of 6 patients with hepatic EHE who were admitted to the Yantaishan Hospital (3 patients), Zhangzhou Hospital of Traditional Chinese Medicine (2 patients) and Zhangqiu Hospital of Traditional Chinese Medicine (1 patient) between March 2007 and June 2014 were retrospectively analyzed.All the patients underwent plain scan and dynamic enhanced scan of computed tomography (CT), and the number,shape, size, location, density or signal, level and method of enhancement of the lesions were observed and analyzed.Six patients were followed up by outpatient imaging examination up to June 2014, and the changes of lesions were observed.Results Among the 6 patients, 1 solitary lesion and 5 multiple lesions were detected, and the total lesions were 125 including 1 patient with 75 lesions.The lesions were round or round-like and originated commonly from the right lobe of liver and hepatic subcapsular with a maximum diameter of 0.5-3.5 cm.Plain scan of CT showed that the lesions in 6 patients had low density with the clear boundary.MRI showed that low T1 WI signal and high or slightly high T2WI signal of the lesions were detected in 4 patients.Two patients had liver capsular retraction sign.The ring-like enhancement of 1 lesion and homogeneous enhancement of 5 lesions were found by dynamic enhanced scan of CT in 6 patients and enhanced scan of MRI in 4 patients.Enhanced signal in all the lesions was detected in the delayed phase.Veins into or through lesions were found in 3 lesions, with normal or narrowing vascular cavity.One patient had visible lollipop sign.Of the 6 patients, 5 were confirmed as with metastatic carcinoma of liver, and 1 was suggested as with cholangiocarcinoma.Six patients were diagnosed with hepatic EHE by pathological examination using hepatic biopsy technique.Among the 2 patients with hepatic EHE who didn't receive antineoplastic treatment after the diagnosis, 1 patient received CT examination at year 2 after first visit, which showed capsular retraction sign, and then was diagnosed as with secondary hepatic cirrhosis by MRI at 4 years after first visit.Another patient was diagnosed as with hepatic cirrhosis by CT examination at year 6.5 after first visit.One patient was diagnosed with tumor recurrence of hepatic left lobe by CT reexamination at postoperative year 4, and underwent ultrasound-guided radio frequency ablation (RFA) treatment based on no enlargement of tumor during 1-year follow-up, and then returned a normal condition after half year follow-up.Other 3 patients undergoing operation were followed up at postoperative year 1 , 4, 5 with no recurrence and metastasis.Conclusions Intrahepatic single or multiple nodules and delayed reinforcement by dynamic enhanced scan of CT and MRI are the typical imaging performances of hepatic EHE.There are certain characteristics in the liver the lollipop sign, capsular retraction sign and veins into or through the lesions.Mutual fusion and fibrosis of lesions leading ultimately to secondary liver cirrhosis may be characteristics of EHE growth.

Objective　To investigate the expression of TGF-β and TGF-β receptor in human breast cancer cell Bcap-37 inhibited by soybean isoflavones. Methods　mRNA and protein of TGF-β1、TGF-βRⅠ in Bcap-37 cells were examined with reverse transcription ploymerase chain reaction(RT-PCR) and immunohistochemistry after cells were treated with daidein or genistein for 1-4 d.The expression of TGF-β1 and TGF-β2 was determined with TGF-β resistance test. Results　The TGF-β1, TGF-β2 and TGF-β recepor increased in Bcap-37 cells at a concentration of 3×10-5 mol/L of genistein. No changes was found when treated with daidzein. Conclusion　Genistein may inhibit the proliferation of Bcap-37 cells and accompany with increasing expression of TGF-β1, TGF-β2 and TGF-β receptor.

BACKGROUND: In the treatment of senile distal tibia fractures, locking compression plate (LCP) combined with minimally invasive percutaneous plate osteosynthesis (MIPO) exerts a satisfactory repair effect, and contributes to the function recovery of lower limbs.OBJECTIVE: To retrospectively analyze the efficacy of LCP combined with MIPO versus intramedullary interlocking nailing for senile distal tibia fractures.METHODS: Fifty-six elderly patients with distal tibia fracture were allotted to minimally invasive and intramedullary nailing groups (n=28 per group), and received the treatment of LCP combined with MIPO and intramedullary interlocking nailing fixation, respectively. The operation time, intraoperative blood loss, postoperative AOFAS ankle-hind foot scale scores, postoperative ambulation time, healing time, postoperative complications and the excellent and good rate in Johner-Wruhs' criteria were compared between two groups.RESULTS AND CONCLUSION: (1) The operation time, AOFAS ankle-hind foot scale scores, ambulation time, and healing time in the minimally invasive group were significantly superior to those in the intramedullary nailing group (P < 0.05). (2) The minimally invasive group showed a significantly higher excellent and good rate (96%) than the intramedullary nailing group (79%) (P < 0.05). (3) Compared with the intramedullary nailing group, the incidence of complications was significantly reduced in the minimally invasive group (P < 0.05). (4) Our findings suggest that the combination of LCP and MIPO not only preserves the most of blood supply, and soft tissues surrounding the fracture end as suggested by the BO principle, but also is conducive for fracture healing, and holds good efficacy.

Objective To investigate the effect of heme oxygenase-1 ( HO-1 ) on the acute radiation-induced skin injury by gene transfer.Methods Thirty-three male SD rats were randomly divided into three groups as PBS-injected group,Ad-EGFP-injeeted group and Ad-HO-1-injected group ( n =11 ).In each group,three rats were used for determining the expression of target gene and the other rats were irradiated on the buttock skin with 40 Gy electron beam generated by a linear accelerator.Immediately after irradiation,rats were administered with a subcutaneous injection of PBS,Ad-EGFP or Ad-HO-1,respectively.Subsequently,the skin reactions were measured twice a week using the semi-quantitative skin injury scale.Results The strong positive expression of HO-1 was observed in subcutaneous dermal tissue after injection of Ad-HO-1.Compared to the PBS-injected group or the Ad-EGFP-injected group,a significant mitigation of skin injury was observed in Ad-HO-1-injected mice 14 d after irradiation (q =0.000-0.030,P ＜ 0.05 ).Conclusions HO-1 could significantly mitigate radiation-induced acute skin injury and Ad-HO-1 could be used to treat radiation-induced skin injury.

Objective:We aimed to analyze the clinicopathological characteristics of differentiated thyroid carcinoma (DTC) in women. Methods:The clinical data of 1,034 female patients with thyroid nodules between January 2003 and December 2012 were retrospectively analyzed. These patients were from Yunnan Province, China. A database was established in Excel. Univariate and multivariate conditional logistic regression analyses were conducted by using SPSS 17.0. Results:Female patients with DTC were younger than those with thyroid nodule disease or benign thyroid disease (BTD). The results of univariate conditional logistic regression analysis showed that the preoperative mean level of serum thyrotropin was higher in patients with DTC than in patients with BTD (P=0.034). The positive ratios of thyroid peroxidase antibody, thyroglobulin antibody (TGAb), and thyrotrophin receptor antibody (TRAb) were higher in patients with DTC than in patients with BTD (P<0.001). The positive ratio of the coexistence of DTC with Hashimoto's thyroiditis (HT;13.3%) or with lymphocytic thyroiditis (LT;4.2%) was higher in patients with DTC than in patients with BTD and HT/LT (P<0.001). The ratio of the patients whose age of menarche was≤13 years, with≤2 of births, or were in pre-menopausal condition in the DTC group was higher than that in the BTD group. The results of multivariate conditional logistic regression analysis showed that age<45 years, nodal size<1 cm, and thyroglobulin increase were protective factors of DTC with odds ratios (ORs) of 0.06, 0.377, and 0.431, respectively. An abnormal increase in TGAb and TRAb was an independent risk factor of female patients with DTC with ORs of 4.949 and 23.001, respectively. Conclusion:Female patients aged 35 years to 44 years and with thyroid nodules were included in a high-risk group of DTC. Serous thyroid-stimulating hormone 1evel and coexistence with HT were positively correlated with the risk of DTC in females. Early menarche, late menopause, and low number of births were associated with the incidence of DTC in females. Age<45 years, nodal diameter<1 cm, and increase in thyroglobulin were protective factors of DTC in female. An abnormal increase in TGAb and TRAb was an independent risk factor of female DTC.

Objective To compare clinicopathological characteristics and prognosis between triple-negative and HER-2-overexpressed breast cancer patients.Methods From Jan.1997 to Jan.2007,data of 725 cases of primary breast cancer patients undergoing surgical treatment were analyzed.The patients were classified into triple-negative and HER-2-overexpressed phenotypes based on immunohistochemistry results.The clinicopathological characteristics and survival of the 2 groups were compared.Results Of the 725 cases,triple-negative and HER-2-overexpressed phenotypes accounted for 12.29%and 24.96%respectively.Compared with HER-2-overexpressed group,the triple-negative group had significantly higher proportion of familial history of malignancy (18.4% vs 5.5%,P=0.001)and higher proportion of patients'histological grade reaching grade 3(54.0% vs 42.0%,P=0.01).There were more lymph node metastasis in triple-negative group than that in HER-2-overexpression group(N1+N2+N3:74.7%vs 64.6%,P=0.045).The recurrence and metastasis rate within 2 years and brain metastasis rate in triple-negative group were higher than those in HER-2-overexpressed group (25.3%,8.0%vs 8.8%,2.2%,respectively,P<0.05).The 5-year disease-free survival rate in triple-negative group was significantly lower than that in HER-2-overexpressed group(55.6%vs 69.8%,P=0.041).There was no statistical difference between the 2 groups in terms of age,menopausal status,tumor size,pathological stage,surgcal procedure,pathological type,adjuvant radio-chemotherapy,the proportion of metastasis to liver,lung and bone as well as overall survival rate(P>0.05).Conclusion Compared to HER-2-overexpressed group,patients with triple-negative breast cancer show higher rate of familial history of malignancy,and they are associated with higher histological grade and poor prognosis.