Objective To investigate the combined effect of exemestane and low-dose methotrexate on exemestane-resistant MCF-7 human breast cancer cells( MCF-7/EXE). Methods Antiproliferative effects of exemestane and low-dose of methotrexate, alone and in combination on growth of MCF-7/EXE cells were assessed by using the MTT assay. Synergistic interaction between the two drugs was evaluated in vitro by using the combination index ( CI) method. The cell cycle distribution was analyzed by flow cytometry in a half inhibitory concentration of exemestane and low-dose of methotrexate . The changes of apoptosis on MCF-7/EXE cells exposed to two drugs alone or in com-bination were observed by fluorescence microscope. The expression of Bcl-2,AKT,P-AKT and cyclooxygenase-2 was investigated by Western blot. Results MTT assays indicated that the combination treatment apparently decreased the viability of MCF-7/EXE cells compared to single drug treatment (CI<0. 9). In addition, the combination of exemestane and low-dose methotrexate exhibited a synergistic inhibition of cell proliferation, arrested the cell cycle in the S phase significantly and produced a stronger inhibitory effects on P-AKT, Bcl-2 and cyclooxygenase-2 ex-pression than control or individual drug treatment. Conclusion The combination of the two inhibitors significantly increases the response as compared to single agent treatment, suggesting that combination treatment which can re-verse the resistance of exemestane could be a more effective approach to breast cancer.

Background The pathogenic mechanism of thyroid associated ophthalmopathy (TAO) is still unclear,which is considered to be an autoimmune disease.It is confirmed that interleukin-17A (IL-17A) plays an important role in the occurrence and development of many autoimmune diseases.It is unclear that whether IL-17A participates in the pathogenesis of TAO.Objective This study was to explore whether IL-17A secreted by coculture system of peripheral blood mononuclear cells (PBMCs) and orbital fibroblasts (OFs) participates in the pathogenesis of TAO and its possible mechanism.Methods Periphery blood and orbital connective tissue were obtained from 12 patients with TAO and 8 patients who received prosthesis implantation for eyeball atrophy in Xiangya Hospital during April to December 2014.PBMCs were isolated by density gradient centrifugation,and OFs were cultured by explant culture method.The purity of T leukomonocyte in PBMCs was tested by flow cytometry,and OFs were identified by Giemsa staining and immunochemistry.OFs and PMBCs were incubated into 96-well plate in a 1:20 proportion to establish co-culture system.Different concentrations of phytagglutinin (PHA) (0,1.0,2.5,5.0,10.0 μg/ml) was added for 72 hours,and IL-6,IL-17A levels in the co-culture system supernatant and IL-17A receptor (IL-17RA) of the total cell membranes in the co-culture system were assayed by ELISA.The differences of IL-6,IL-17A,IL-17RA levels in co-culture system were compared between the TAO group and control group.Results The mean purity of T leukomonocyte in PBMCs was (81.10±0.21)％ in the TAO group and (80.05 ±0.38)％ in the control group respectively,with no significant difference between them(t =0.923,P＞0.05).Cultured OFs showed the positive response for Vimentin expression and Giemsa staining.After stimulated by 1.0 μg/ml PHA,the proliferation of both PBMCs and OFs were increased in the co-culture system.Apoptosis exist in PBMCs and the number of OFs decreased when PHA was higher than 1.0 μg/ml.The growth of PBMCs and OFs was faster in the TAO group than that in the control group in the same concentration of PHA.The contents of IL-6,IL-17A and IL-17RA in co-culture system were significantly different among various concentrations of PHA subgroups (IL-6:Fgroup =12.561,P=0.000;F ion =23.356,P =0.001.IL-17A:Fgroup =12.037,P =0.000;Fconcentration =19.206,P=0.000.IL-17RA:Fgroup =16.216,P=0.000;Fconcentraction =4.627,P=0.018).The production of IL-6,IL-17A and IL-17RA reached peak in both TAO group and the control group after 1.0 μg/ml PHA stimulated.However,the concentrations of IL-6,IL-17A and IL-17RA reduced with the increase of PHA concentration.The concentrations of IL-6,IL-17A and IL-17RA in co-culture system were significantly higher in the TAO group than those in the control group under the stimulation of the same concentration of PHA (all at P＜0.05).Conclusions The co-culture system of PBMCs and OFs stimulated with PHA can be the imitation of TAO pathogenesis in vitro,and PHA can amplify its immune reaction to imitate TAO pathogenic processes intuitively.The IL-6,IL-17A and IL-17RA secreted by PBMCs and induced by PHA are increased in TAO patients,implying that IL-17A participates in the pathogenesis of TAO through magnifying cellular immune response and inflammatory reaction.

BACKGROUND:Because of its advantages, Endobutton has been widely used in clinic. Currently, its shortcomings are increasingly recognized. Tightrope that overcomes the shortcomings of Endobutton has been gradualy accepted by a doctor skiled in sports injuries.
OBJECTIVE: To analyze and compare the differences in the effects of Tightrope and Endobutton in the reconstruction of the cruciate ligament.
METHODS:Totaly 60 cases of anterior cruciate ligament rupture were selected and subjected to anterior cruciate ligament reconstruction under arthroscopy, of which 30 cases were randomly assigned to reconstruction by Endobutton device and 30 cases underwent reconstruction by Tightrope device. Al operations were performed by the same surgeon. Al patients were subjected to regular functional exercise and were folowed up regularly after operation. Effects of Tightrope and Endobutton in the cruciate ligament reconstruction were evaluated by comparing various indexes in the two groups.
RESULTS AND CONCLUSION:Compared to the Endobutton fixation system, the Tightrope fixation system could shorten the operation time, reduce the length of tendon incision, and decrease the loss of bone mass in the femoral bone tunnel. There were no significant differences in the maximum knee flexion, knee joint score and Tegner movement level score between the two groups at 3 and 6 months after operation. These findings indicate that the Tightrope fixation system is superior to the Endobutton fixation system, because it is more simple and convenient to operate and has less bone loss. However, their clinical efficacy has no difference after 6 months.

Objective To compare the MR features of small hepatocellular carcinoma (sHCC) and small intrahepatic mass-forming cholangiocarcinoma (sIMCC) (≤ 3 cm) and improve the accuracy in diagnosis.Methods Imaging studies of 318 patients with 351 sHCCs and 42 patients with 47 sIMCCs were retrospectively reviewed.Signal intensity on T2WI and on each phase of dynamic enhanced imaging,enhancement patterns,and accompanying findings of the two groups were compared by qualitative and quantitative studies.The qualitative imaging features of tumors were compared by the x2 test and the Fisher exact test,and the statistically significant variables obtained from univariate analysis were used to conduct multivariate stepwise logistic regression analysis.Enhancement pattern between sHCCs with elevated AFP and normal AFP were assessed by thex2 test.Results Multivariate analysis indicated that signal features of enhancement pattern and the accompanying findings of tumor capsule and hepatic capsule retraction were key predictors in differentiating sHCC from sIMCC:Wash-out pattern (276/351) with enhanced tumor capsule (275/351) highly indicated sHCC,while progressive enhancement pattern (27/47) with hepatic capsule retraction (8/47) indicated the diagnosis of sIMCC (P ＜ 0.01).Compared with sHCC with normal AFP,atypical patterns as progressive (n =4) and stable enhancement (n =32) were more commonIy seen in lesions with elevated AFP,dynamic enhancement pattern was different between the two subgroups (x2 =8.241,P ＜ 0.05).Conclusions Enhancement patterns and the accompanying tumor capsule and hepatic capsule retraction were the most important predictors in differentiating sHCC from sIMCC.In addition,enhancement pattern was relatively more typical in sHCCs with normal AFP which provided valuable information in diagnosis for clinicians.

BACKGROUND: Although there are many studies on acute patellar dislocation in adolescents, the optimal treatment strategy is under discussion.OBJECTIVE: To summarize the clinical efficacy of reconstruction of the medial patellofemoral ligament (MPFL) in the treatment of adolescent acute patellar dislocation using double-pulley technique.METHODS: Thirty cases of acute patellar dislocation were enrolled and received reconstruction of the MPFL using double-pulley technique. We proceeded to prepare the bone bed for anchor placement at the medial point of the superior edge of the patella and at the upper 1/3 patella along the long axis of the patella using arthroscopic grinding. Two double-loaded 5.0 anchors were respectively placed at the medial point of patella and at the upper 1/3 patella parallel to the patellar articular surface, and then tied using the double-pulley technique. Long longitudinal incisions were performed at the medial condyle and at the adductor tubercle to expose these two structures, the deep fascias tunnel between two incisions were penetrated to pull the tendon weave end through the tunnel until the lateral side along a femoral tunnel; finally the tendon was fixed using screws in appropriate tightness. All patients were followed for more than 10 months, and the patellar-related parameters and knee function parameters were compared before and after operation. RESULTS AND CONCLUSION: There were 12 cases of intra-articular cartilage injury and 4 cases of meniscus injury. The patella title angle, Q-angle and outward shift distance at the last follow-up were less than those before operation. Lysholm and International Knee Documentation Committee scores at the last follow-up were higher than those before operation. The mean operation time was (90±10.5) minutes. There was one patient with knee movement angle from 0° to 30° and one patient with a patellar fracture caused by slipping and treated with internal fixation. None of patients appeared with patellar dislocation, positive apprehension test infection or poor wound healing. These results indicate that the simple and mini-invasive double-pulley technique is a good treatment strategy for acute patellar dislocation, because it is consistent with patellar anatomical stability, holds patellar trajectory visibility, quick recovery and good curative effect. Additionally, osteotomy is recommended when the simple efficacy is not satisfactory.

The process of exercise regulating bone metabolism is complicated, which involves a number of signaling pathways. A large number of studies in vitro have indicated that mechanical stress regulates bone metabolism by Wnt, bone morphogenetic protein (BMP), and osteoprotegerin (OPG)/receptor activator of NF-κB ligand (RANKL)/receptor activator of NF-κB (RANK) signaling pathways. Both the in-tensity and frequency of mechanical stress have varing impact on bone tissue and cells. Plenty of studies in vivo also have shown that exer-cise regulates bone metabolism by key factors in bone metabolism signaling pathways. This paper reviewed the effects of exercise on bone metabolism pathways and their mechanisms.

Objective To study the clinicopathological features of primary and metastatic hepatic neuroendocrine carcinoma.Methods The records of 35 patients with primary hepatic neuroendocrine carcinoma and 35 patients with metastatic hepatic neuroendocrine carcinoma were retrospectively reviewed.These patients served as the primary group(priNET,n=35)and the metastasis group (metNET,n=35),respectively.Results There were significant differences between the two groups of patients in gender,site,size and number of tumor(P＜0.05).Although there was no significant difference between the two groups in the distribution of the tumors in the two lobes of liver (P＞0.05),priNET had more tumors localized to one lobe of liver while metNET had more tumors involving both lobes of liver(P＜0.05).Conclusions Gender,size,site and number of tumor may play an important role in the differentiation of primary or metastatic hepatic neuroendocrine tumor.

[ ABSTRACT] AIM:To investigate the effect of silencing cell division cycle 25a ( CDC25a) gene on the prolifera-tion of human hepatoma HepG2 cells.METHODS:CDC25a gene in human hepatoma HepG2 cells was silenced by RNA interference.Real-time PCR was applied to detect the expression of CDC25a, cyclin E and CDK2 at mRNA levels in the HepG2 cells.Western blotting was applied to detect the expression of CDC25a at protein level.In addition, MTT assay, Giemsa staining and flow cytometry were used to measure the proliferation of human hepatoma HepG2 cells.RESULTS:The expression of CDC25a at mRNA and protein levels in RNA silence group was lower than those in negative control group and normal control group (P<0.05).The mRNA expression of cyclin E and CDK2 in silence group was lower than that in negative control group and normal control group (P<0.05).The cell proliferation in silence group was lower than that in negative control group and normal control group ( P<0.05) .The results of flow cytometry revealed that the cells in silence group were blocked in G1 phase.CONCLUSION:Infection of LV-CDC25a-RNAi recombinant to the HepG2 cells effec-tively inhibits the CDC25a gene expression and the proliferation of human hepatoma cells, and arrests the cells in G1 phase, suggesting that CDC25a gene may be a key target for the treatment of liver cancer.

OBJECTIVE:To study the effects of Yupingfeng powder on immune function of rats with lung-qi deficiency syn-drome. METHODS:60 SD rats were randomly divided into normal group,model group,positive group [Renshen beiqi tablet 0.6 g (crude drug)/kg] and Yupingfeng powder high-dose,medium-dose and low-dose groups [24,12,6 g (crude drug)/kg],with 10 rats in each group. Except for normal group,those groups were given SO2+comprehensive cold stimulation to induce lung-qi defi-ciency syndrome model. After modeling,treatment groups were given relevant drug solutions intragastrically,and normal group and model group were given constant volume of normal saline intragastrically,once a day,for consecutive 13 d. After administra-tion,serum levels of IL-3 and IL-6,the proportion of CD3+,CD4+,CD8+in T lymphocyte were determined,and pathological chang-es of lung and bronchus were observed. RESULTS:Compared with normal group,serum levels of IL-3 and IL-6 and the propor-tion of CD8+ in T lymphocyte increased significantly in model group,while the proportions of CD3+ and CD4+ in T lymphocyte and the ratio of CD4+/CD8+ decreased(P<0.01);obvious lung and bronchus injury and inflammatory reaction were observed. Compared with model group,serum level of IL-3 decreased significantly in positive group and Yupingfeng powder high-dose group;serum level of IL-6 and the proportion of CD8 + in T lymphocyte decreased significantly in positive group and Yupingfeng powder high-dose and medium-dose groups,while the proportion of CD3+ in T lymphocyte increased significantly;the proportion of CD4+and the ratio of CD4+/CD8+increased significantly in treatment groups(P<0.05 or P<0.01);lung and bronchial injury,and inflam-mation reaction were relieved. CONCLUSIONS:Yupingfeng powder can enhance immune function of rats with lung-qi deficiency syndrome.