Breast cancer is a heterogeneous disease.Its invasion and metastasis,recurrence and part of the chemotherapy drug tolerance are currentlv difficulties of treatment.Breast cancer stem cells have the ability of selfrenewal,proliferation and differentiation potential,which may correlate closely with breast cancer recurrence,treatment failure.This assay aims to briefly analyse markers and significance of breast cancer stem cells,and the relationship between the treatment of breast cancer and breast cancer stem cells.

To investigate the effects of Tianqi Pingchan (TQPC) Granule, a compound traditional Chinese herbal medicine with antitremor activity, on levodopa-induced dyskinesia and the expression of G protein-coupled receptor kinase 6 (GRK6) in rats with Parkinson disease (PD).

ObjectiveTo investigate the effect of MK-801 on levodopa-induced dyskinesia (LID)in Parkinson' s disease (PD). MethodsRat models ( n = 25) of Parkinsonism related motor complications were established and were randomly divided into levodopa-induced dyskinesia (LID) group (n = 10), MK801 treatment group (n = 10) and PD group (n =5). Another 5 rats were served as control group. The behaviors of LID rats treated with MK-801 were observed. Immunohistochemistry and Western blot analysis were used to determine the expression of β-arrestin1 in the striate of rats.Results After MK-801 treatment, abnormalinvoluntarymovementscores and peakturning weredecreasedin LIDrats.Immunohistochemistry showed that β-arrestin1-positive cells of the lesioned side ((2. 95 ± 0. 44) × 104) in LID rats were decreased compared to the contralateral side ( ( 3.78 ± 0. 37 ) × 104, t = 5. 415, P ＜ 0. 05 ).Western blot showed that the levels of β-arrestinl in PD group ( presented as lesioned side/contralateral side) were ( 81.02％ ± 2. 23％ ). The levels of β-arrestin1 (64. 88％ ± 3. 10％ ) were deceased in LID rats compared to PD rats ( t = 9.47, P ＜ 0. 01 ). However, the levels of β-arrestin1 ( 89. 26％ ± 1.90％ )were increased in MK-801-treated rats (t = 14. 82, P ＜0. 01). ConclusionsMK-801 reduces LID in PD rats. The beneficial effect of MK-801 may be mediated through the increased expression of β-arrestinl which in turninhibits the overactivation of glutamate receptors.

Objective To investigate the relationship between G protein-coupled receptor kinase 6 (GRK6) and N-methyl-D-aspartate (NMDA) receptor in the mechanism study underlying motor complications in Parkinson's disease (PD).Methods The rat models (n= 25) of Parkinsonism related motor complications were established and were randomly divided into levodopa-induced dyskinesia (LID) group (n= 10,intraperitoneal injection of levodopa for 23 d),MK-801 treatment group (n= 10,intraperitoneal injection of MK-801 at day23 after intraperitoneal injection with levodopa for 22 days) and PD group (n= 5,intraperitoneal injection of vitamin C).Another 5 rats were served as controls (sham-operation group).The behavior changes of rats in MK-801 treatment group were observed,and the expression of GRK6 in the striate of rats was detected by immunohistochemistry and Western blot.Results After the chronic treatment with levodopa methyl ester,PD rats displayed abnormal involuntary movements,which was similar to levodopainduced dyskinesia in PD patients.Immunohistochemistry showed that GRK6-positive cells of lesion side were decreased in LID rats as compared with PD rats [(3.23±0.41 ) × 103 vs.(4.81 ± 1.31 ) ×103,P＜0.01].Rats in MK-801 treatment group displayed the decreased AIM scores and increased peak rotation,and the increased GRK6-positive cells of lesion side as compared with LID rats (P＜0.05).Western blot showed that the levels of GRK6 was 83.7% ±2.1% in PD group (presented as lesion side/unlesion side),76.8% ± 2.2% in LID group and 91.1% ± 2.7% in MK-801 treatment group (intergroups comparison:all P＜0.05).These results were in accordance with the results of immunohistochemistry.Conclusions Antagonist of NMDA receptor can be used to reduce the motor complications in rats.It may be due to increased GRK6 which inhibits the overactivation of glutamic acid receptors.

Objective To investigate the effects of levodopa/benserazide-loaded poly-lactide-coglycolide (PLGA) microspheres on motor deficits and levodopa-induced dyskinesia in a rat model of Parkinson' s disease (PD) and to explore the mechanisms underlying this effects.Methods The content of levodopa/benserazide released from the microspheres was determined by high-performance liquid chromatography.The rat model of PD was induced by 6-OHDA injections.Then the valid PD rats were treated with levodopa ( 12 mg/kg,s.c.)/benserazide ( 15 mg/kg,s.c.) or microspheres.Forepaw adjusting steps were measured on 1,4,7,10 and 14 days after treatment.After 2 weeks of treatment,the abnormal involuntary movements (AIM) were measured.Phosphorylated dopamine,cAMP-regulated phosphoprotein of 32 kDa (DARPP-32) at threonine 34 levels were determined by immunohistochemistry and Western blot respectively.In addition,the levels of △FosB were measured by Western blot.Results In vivo release test showed that 76.2％ of levodopa and 83.6％ benserazide were released from the microspheres on day 7.Forepaw adjusting steps showed that the scores of forepaw adjusting steps in microspheres-treated PD rats were ( 5.8 ± 1.6 ) and ( 5.2 ± 1.5 ) respectively on 10 and 14 days of treatment,which were increased compared to levodopa-treaded PD rats (2.4 ± 1.1 and 1.2 ± 0.5 ; t =4.12,5.43,all P ＜0.01 ).The AIM scores of microspheres-treated rats ( 16.0 ±2.1 ) were decreased significantly compared to levodopa-treated rats ( 26.0 ± 3.2 ) on day 14 ( t =6.59,P ＜ 0.01 ).Immunohistochemistry indicated that the phosphorylated levels of DARPP-32 in microspheres-treated rats ( (3.7 ± 1.3 ) × 104 )were decreased significantly compared to levodopa-treated rats ( (7.9 ± 2.2) × 104 ; t =2.95,P ＜ 0.05 ).In addition,Western blot showed that the levels of phosphorylated DARPP-32 and △FosB were 119.4％ ± 11.3％ and 149.3％ ± 12.3％ respectively,which were decreased significantly compared to levodopatreated rats ( 184.8％ ± 13.7％ and 300.4％ ± 14.2％ ; t =4.12,2.91,all P ＜ 0.05 ).Conclusions Microspheres can be used to improve the motor deficits and reduce the expression of dyskinesia in PD rats.This may be due to the continuous release of levodopa/beserazide from the microspheres,which leads to continuous stimulation of PD rats and reduces the levels of phosphorylated DARPP-32 and △FosB in the striata of these rats.

Objective To investigate the potential benefits of hepatic resection for patients with hepatic metastasis of gastroenteropancreatic neuroendocrine tumors.Methods A search in Cochrane Library,PubMed,Medline,Embase,CBM,Wanfang Databases for identifying randomized trails.Criteria for inclusion were established based on validity criteria by Cochrane Methods Group on screening and diagnostic tests,and correlated indexes of 10 articles including 1 567 patients were extracted for Meta-analysis.Results Compared with non-surgical treatment group,the 3-year survival rate (OR:0.07,95 ％ CI:0.02,0.22 ; Pheterogeneity =0.93,P ＜ 0.00001),the 5-year survival rate (OR:0.19,95 ％ CI:0.15,0.24 ; Pheterogeneity ＝ 0.11,＜ 0.00001) were significantly increased in the surgical group.Conclusions Surgical resection is the preferred treatment of gastroenteropancreatic neuroendocrine tumors in patients with liver metastasis.Surgery is safe and effective,furthermore it takes the advantages of significantly extend the surgical live time.

Objective To explore the clinical feature,suitable treatment and prognosis of soft tissue leiomyosarcoma of the trunk and extremities.Methods Clinical data of 18 cases of pathologically confirmed soft tissue leiomyosarcoma of the trunk and extremities from January 1999 to December 2012 were analyzed retrospectively.Primary tumors in 7 cases were marginally excised before admitted to our hospital,2 had open biopsy before admission to our hospital,and 8 cases had local relapse at admission; Only one patient took our institute as the first visit.Seventeen cases were performed extended excision of tumors and 1 case underwent marginal resection.All the patients were followed up and the follow-up period was from 16 to 158 months.Results During follow-up period,9 patients developed lung metastasis,and local recurrence occurred in 5 patients.Ten patients died and 8 survived.Of the ten dead cases,seven died of tumor progression and 3 died from non-tumor factors.In the 8 patients survived,2 survived with tumor.The 5-year overall survival rate was 59.2％.Conclusion Soft tissue leiomyosarcoma of the trunk and extremities are rare malignant tumors,mostly occurred in elder patients and presenting soft tissue mass.Local recurrence and distant metastasis are common and associated with a poor prognosis.Surgical excision combined with adjuvant radiation is the common treatment strategy.

Objective To observe the protection of human lens epithelial cells(HLEC) by different polar alkaloids extracted from Herba Dendrobii(HD).Methods We extacted the Herba Dendrobii powder with ethanol,and then treated the extract with falling-film concentration,acidification,salting out,chloroform extraction,and washing with water.Different polar alkaloids were extracted from HD after the above treatment.The protective effect of HD alkaloids was observed on HLEC,which were cultured with DMEM medium containing 10% fetal calf serum.Ten groups were set up for the experiment: normal control group,model group,high-and low-dose water-soluble alkaloids groups,high-and low-dose fat-soluble alkaloids groups,high-and low-dose low-polar alkaloids group,and high-and low-dose weak-polar alkaloids groups.The high-dose dosage of the alkaloids was 25.0 ?g/L and low-dose dosage was 12.5 ?g/L.Methyl thiazolyl tetrazolium(MTT) assay was used to evaluate the proliferation of HLEC under the different conditions of interventions.Results The single-factor experiments showed that the highest extracting rate of HD alkaloids was obtained under the conditions of extracting the powder with 80% ethanol for 3 times and for 3 hours.The results of protective experiment showed that the proliferation of HLEC in the model group was inhibited by hydrogen peroxide(H2O2),and the inhibitive rate was lower in low-dose fat-soluble alkaloids group than that in the model group(P

Objective To investigate the mechanism of tumor necrosis factor-α (TNF)-α inhibiting osteo blastdifferentiation of mesenchymal stem cells (BMMSCs) in the pathogenesis of osteoporosis in the mouse model of systemic lupus erythematosus (MRL/lpr). Methods The femurs of MRL / lpr and C3He/HeJ mice were isolated, the bone structure were examined by hematoxylin-eosin (HE) staining. The proteins of TNF-α, NF-κB P50, bone morphogenetic protein -2 (BMP-2) and PSmad1/5/8 were measured by immunohistochemical stain. Bone marrow mesenchymal stem cells (BMMSCs) were isolated. After BMMSCs grew on the cover slips, the proteins on top of it were evaluated by immunohistochemistry stain. Moreover, the alkaline phosphatase (ALP) staining was employed for the measurement of the early osteogenic differentiation. BMMSCs together with hydroxyapatite were embedded subcutaneously in the nude mice and eight weeks later, the ectopic bone formation was evaluated. The recombinant human tumor necrosis factor receptor type Ⅱantibody fusion protein (etanercept) or normal saline was subcutaneous injected to the mice with lupus. After four weeks, the expression of these proteins was observed and the ectopic bone formation was investigated. Image-Pro plus 6.0 software was employed for imagine analysis, and Studentˊs t-test was used to test the differences between 2 independent groups. Results MRL/lpr mice showed decreased volume of cortex and the percentage of cortex to the volume of bone of MRL/lpr mice was significantly lower compared to control groups and with C3He/HeJ mice (13.96±0.25 vs 23.61±0.71, n=3, P<0.01). The protein levels of both TNF-αand NF-κB P50 on the femur of MRL/lprl mice were higher than those of the control group (0.643±0.051 vs 0.405±0.022, 0.917±0.023 vs 0.650±0.032, n=3, P<0.01). The expressions of BMP-2 on the femur of MRL/lpr mice were lower than those of the C3He/HeJ mice (0.52 ±0.03 vs 0.72 ±0.03, n=3, P<0.01). There was no difference in the expression of PSmad1/5/8 on the femur between the two groups by immunohistochemistry detection (1.264 ±0.021 vs 1.301± 0.044, n=3, P>0.05). The expressions of TNF-α and NF-κB P50 in BMMSCs of MRL/lprl mice were higher than those of the C3He/HeJ (0.184±0.021 vs 0.136±0.013, 0.132±0.021 vs 0.097± 0.014, n=3, P<0.01), while BMP-2 and PSmad were lower than those of the control group (0.128±0.013 vs 0.216±0.221, 0.115±0.023 vs 0.196±0.034, n=3, P<0.01). After 7 days of BMP-2 stimulation, the activities of ALP of BMMSCs from MRL/lprl mice were reduced detected by ALP staining and the osteoblast differentiation of these cells were decreased than BMMSCs from the control mice by HE and Masson staining. The percentage of the cortex to the volume of bone of the etanercept injection MRL/lpr mice was higher than that of the control group (21.8±1.0 vs 14.3 ±0.6, n=3, P<0.01). Moreover, the proteins of TNF-α and NF-κB P50 on the femurs of such injected mice were lower than those of the control group (0.540±0.024 vs 0.682±0.031, 0.857±0.023 vs 1.098±0.044, n=3, P<0.05), while the expressions of BMP-2 were higher than the control group (0.99±0.04 vs 0.85±0.04, n=3, P<0.05). There was no difference in the PSmad1/5/8 expression on the bone of the two group of lupus mice (0.88 ±0.08 vs 0.84 ±0.04, n=3, P>0.05). The ectopic bone formation of BMMSCs of the etanercept injected MRL/lpr mice was higher than that of the normal saline injected mice, however, it was lower than that of the C3He/HeJ mice. Conclusion TNF-α inhibits osteoblast differentiation of mesenchymal stem cells by depressing Smad signaling which may contribute to the osteoporosis of the lupus mice.

Purpose Recently the incidence of bone and joint degeneration is increased,with the increasing of cardiovascular disease complication.This study aims to utilize realtime three-dimensional echocardiography (RT-3DE) in evaluating left ventricular remodeling index (LVRI),to guide treatment and improve the survival rate.Materials and Methods Fifty-seven cases of healthy volunteers (normal control group) and sixty-seven patients with bone and joint degeneration (research group) were respectively divided into two subgroups according to the age.Both groups underwent conventional echocardiography and RT-3DE.The LVRI,left ventricular end diastolic volume index (LVEDVI),left ventricular mass index (LVMI),left ventricular end systolic volume index (LVESVI) and left ventricular ejection fraction (LVEF) measured by different methods were analyzed,and the differences and correlations between groups and within each group were compared.Results The age of normal control group and research group was not significantly different (P＞0.05).The LVMI and LVRI in research group were significantly higher than those in the normal control group with same age (P＜0.05).The LVEDVI,LVMI and LVRI measured by two-dimensional echocardiography and M-mode echocardiography were higher than those measured by RT-3DE,and the difference was statistically significant (P＜0.05).Bland-Altman agreement analysis showed good agreement of measured LVRI using 2D echocardiography and RT-3DE.Conclusion Patients with bone and joint degeneration have left ventricular remodeling.RT-3DE can accurately assess LVRI with the advantages of convenience and lack of radiation,which can be widely used in clinic.