Notch Signaling Promotes Proliferation and Migration of SW982 Synovial Sarcoma Cells / 中国医科大学学报

Objective To investigate the effect of the Notch signaling pathway on the proliferation and invasion of human SW982 synovial sarcoma cells. Methods SW982 cells and normal human synovial cells were routinely cultured, and the expression of proteins related to the Notch pathway was compared. The Notch signaling pathway was manipulated by NICD1 overexpression, CFB1 shRNA lentivirus, and the γ-secretase inhibitor, DAPT. CCK-8 and wound healing assays were carried out to investigate the role of the Notch signaling pathway in SW982 cells. Results The Notch signaling pathway clearly showed higher activity in human SW982 synovial sarcoma cells than in normal human synovial cells (P ＜ 0.05). The proliferation and invasion of SW982 cells were significantly upregulated by overexpressing NICD1; however, were suppressed by downregulating the Notch signaling pathway using CFB1 shRNA or DAPT (P ＜ 0.05). Conclusion Our findings demonstrate that the proliferation and invasion of human SW982 synovial sarcoma cells are dependent on Notch signaling pathway activity.

Notch signaling pathway regulates osteosarcoma stem cell characteristics by inhibiting Eph pathway / 中华骨科杂志

Objective To investigate the role of Notch signaling pathway to maintain the stem cell-like characteristics of osteosarcoma and its underlying mechanism.Methods Lentiviral NICD1 or Numb-shRNA was transduced into MG63 osteosarcoma cells to activate Notch activity in vitro.The impact of Notch on osteosarcoma stem cells were assessed by the tumor sphere formation assay and flow cytometry analysis of cell surface markers STRO-1/CD117.The expression of stem cell related genes (Sox2,Oct4) were evaluated by Western blot and qPCR.The nude mice were randomly divided into 3 groups:the NICD1 overexpression (NICD-OE) group,the DAPT group and the control (CON) group.The tumor growth was monitored for 8 weeks and the tumor volume and weight were recorded weekly.To investigate whether Notch regulates Eph pathway,Eph pathway related protein EphB,pEphB was measured by Western blot.The impact of ephrinB 1 on NICD overexpression cell were assessed by tumor sphere formation assay.The expression of Sox2 and Oct4 was evaluated by Western blot.Results NICD1 overexpression or Numb-shRNA increased the activity of Notch pathway.The Notch-activated osteosarcoma showed enhanced in vitro tumor spheroid formation capacity,increased Stro-1/CD117double positive ratio,and upregulated expression of Sox2 and Oct4 in vitro.In animal experiments,it was found that activation of Notch pathway promoted tumor formation in vivo and Notch inhibition decreased it.The primary osteosarcoma cells were obtained from mice xenograft treated with DAPT and its tumor sphere formation capacity was significantly reduced.Finally,The Notch pathway inhibits the phosphorylation of EphB,as well as the downstream signal pathway of EphB,but there is no significant change in total EphB.The activation of Eph pathway inhibited Notch induced up-regulation of tumor sphere formation and Sox2 and Oct4 expression.Conclusion Notch signaling pathway maintains the stem cell-like characteristics of osteosarcoma probably by inhibiting the Eph pathway.

Anlotinib hydrochloride capsules for advanced soft tissue sarcoma: single-center data analysis of a stageⅡmulticenter clinical trial / 中国肿瘤临床

Objective: To investigate the efficacy and safety of anlotinib hydrochloride capsules for the treatment of advanced soft tissue sarcoma based on the data from Department of Bone and Soft Tissue Tumor, Peking University Cancer Hospital&Institute. Methods: Patients were randomized allocated at 2:1 ratio for the anlotinib treatment and placebo group. The treatment group received 12 mg/day of anlotinib for 14 consecutive days in a 21-day cycle. The primary end-point was progression-free survival (PFS), and the secondary end-points were disease control rate (DCR), overall survival (OS), and adverse event rate. Results: A total of 46 patients were enrolled in this study; 7 of them were excluded from per protocol set (PPS). Among the remaining 39 patients, 28 were included in the anlotinib group and 11 in the placebo group. In the anlotinib group, 4 patients had partial remission and 13 had stable disease (SD), whereas in the placebo group, 3 patients had SD. The difference in DCR between the 2 groups was statistically significant (60.7% vs . 27.3%, P=0.082). The DCR of the advanced soft tissue sarcoma in the anlotinib group was 78.6% (11/14). The median PFS in the anlotinib group was 12.4 (95% confidence interval [CI]: 7.6 to 17.2) months, which was significantly longer than 4 months in the placebo group (95% CI: 1.7 to 6.3 months, P=0.043); however, the difference in OS between the 2 groups was not significant (19.4 vs . 17.6 months, P=0.961). Regarding the safety, 2 patients had severe adverse events (7.14%) possibly related with treatment in the anlotinib group; one of them had pneumothorax. The other adverse events were grade 1 to 2. Conclusions: Soft tissue sarcoma is highly responsive to anlotinib, with prolonged PFS. Anlotinib is well tolerated and can be used as a treatment option for advanced soft tissue sarcoma.

Clinical features and prognosis of 25 cases of soft tissue sarcoma with soft tissue me-tastasis / 中国肿瘤临床

Objective:To explore the outcome of soft tissue sarcoma (STS) on patients with soft tissue metastasis. Methods:We ana-lyzed 25 STS patients with soft tissue metastasis primarily localized on extremity and trunk. The study was conducted from June 2010 to June 2016 by retrospective analysis of the clinical and pathological characteristics of the patients. The assessed endpoints were overall survival. Results:Six patients (24%) had synchronous soft tissue metastasis, and 19 patients (76%) had metachronous metasta-sis. The average time for primary tumor recession of metastatic lesions was 45.3 months. Metastases were most common in parts of the trunk in 18 patients (72%), followed by the head and neck in 5 patients (20%). Eleven patients (44%) with lung metastasis had poor prognosis. Conclusion:STS occurred more rarely in soft tissue metastasis than in pulmonary metastasis. Neoadjuvant chemotherapy and surgical treatment were the major therapies employed. Targeted therapy as a new treatment rendered good results.

Low-dose methotrexate combined with vinorelbine for inoperable desmoid tumor: efficacy and the prognostic factors / 南方医科大学学报

<p><b>OBJECTIVE</b>To characterize the clinical features of desmoid tumor, assess the efficacy of conservative chemotherapy for inoperable desmoid tumor and analyze the prognostic factors.</p><p><b>METHODS</b>From August 2009 to December 2013, 52 patients with inoperable desmoid tumor were treated in our department and received chemotherapy with vinorelbine combined with low-dose methotrexate. The clinical data of the patients were analyzed retrospectively.</p><p><b>RESULTS</b>The patients studied included 22 male and 30 female patients with the age of disease onset ranging from 2 to 46 years (mean 18.7 years). The lesions occurred most frequently in the lower limbs (36.5%, 19/52) and the tumor size ranged from 2.7 to 37 cm (mean 9.5 cm). The patients were followed up for a median of 29 months (7 to 64 months). The chemotherapy lasted for 4 to 30 months (median 12 months). After completion of the chemotherapy, 1 patient had a complete response (CR), 18 showed partial responses (PR), 27 cases had stable disease (SD), and 6 had progressive disease (PD), with an overall response rate (ORR) of 88.5%. The progression-free survival (PFS) time of the patients ranged from 4 to 63 months (median 26.5 months) with a 2-year PFS rate of 76.7% and 5-year PFS rate of 41.9%. A longer chemotherapy duration (over 12 months) was associated with a more favorable prognosis. No significant differences in PFS were found between the patients stratified by gender, age of disease onset, age when receiving chemotherapy, tumor site, or tumor size.</p><p><b>CONCLUSION</b>For recurrent, inoperable and progressive desmoid tumor, long enough cycles of vinorelbine combined with low-dose methotrexate can be an effective and safe option for tumor control.</p>

Analysis of prognostic factors and the effect of adjuvant radiother-apy and chemotherapy for liposarcoma of the extremities / 中国肿瘤临床

Objective:To analyze the clinical prognostic factors of liposarcoma on the extremities and trunk, as well as to retrospectively analyze the effect of adjuvant radiotherapy and chemotherapy on liposarcoma of the extremities. Methods:Patients with liposarcoma of the extremities treated in our hospital from July 1, 2007 to December 31, 2012 were followed up. The relationship of clinical prognostic factors with gender, age, location, depth, and size of the tumors, as well as the histological grade and admission status, were statistically analyzed. The effects of adjuvant radiotherapy or chemotherapy on overall survival (OS) and disease free survival (DFS) were evaluated. Results:A total of 82 patients with extremity liposarcoma received surgery-based comprehensive treatment in our hospital. Of the total patients, 73 received a 24-month to 88-month satisfied follow-up;the median follow-up time was 47 months. The OS rate was 83.6%(61/73), and the DFS rate was 68.5%(50/73). Multivariate Cox regression analysis showed that the tumor location, histological grade, and admission status were the independent correlative factors influencing DFS, and the age and pathologic grading were the independent correlative factors influencing the OS. Kaplan-Meier survival analysis showed that radiation therapy can significantly improve the DFS and OS of the G2 and G3-grade liposarcoma (DFS:59.1 months vs. 28.4 months, P<0.01;OS:70.8 months vs. 55.1 months, P<0.05). Significant difference was not found in the effect of chemotherapy on OS and DFS. Conclusion:The prognosis of liposarcoma was significantly associated with the pathologic grades and subtypes. Auxiliary radiotherapy could improve the survival and prognosis of G2 and G3 liposarcoma of the extremities, but the role of chemotherapy in treating liposarcoma remained unclear.

Xenograft and calcium sulphate in treating benign bone tumor / 中国组织工程研究

BACKGROUND:Autologous bone graft is the best method to repair bone defects after tumor curettage, but its shortcomings are as folows: increased surgical trauma, sequelae at bone graft site such as infection and pain, and a limited amount of autologous bone. OBJECTIVE:To analyze the effectiveness of xenograft and calcium sulphate artificial bone in treating bone defects after benign bone tumor removed. METHODS:Totaly 26 cases of benign bone tumor were selected, including 8 cases of giant celltumor, 5 of enchondroma, 4 of fibrous histiocytoma, 3 of bone fibrous dysplasia, 2 of non-ossifying fibroma, 2 cases of bone cysts, 1 of aneurysmal bone cyst and 1 of aneurysmal bone cyst and 1 case of chondroblastoma. Of the 26 cases, 12 cases underwent calcium sulphate pelets alone to fil bone defects after benign bone tumor removed, 6 cases were subjected to xenograft alone, and 8 cases were treated with calcium sulphate pelets combined with xenograft. The X-rays were taken at 1 week, 3 months, and 1 year after the operation in al patients to assess the bone healing process. RESULTS AND CONCLUSION:Al the patients were folowed up for 36-72 months. The absorption of calcium sulphate appeared to be absorbed earlier, the earlier absorption appearance could be observed as earlier as 1 month after the implantation, and most calcium sulphate was absolved and replaced by new bone at 3 months after the operation. The xenograft bone was degraded at 3 months post implantation and new bone formed. Osseo integration of the graft was observed at the periphery of the implant at 6 months post implantation. One year post implantation, trabecular bone was observed at the site with uniform bone density. In the combined group, thecalcium sulphate pelets were absorbed earlier and new bone formed earlier than the calcium sulphate alone group, and the xenograft absorbed later than the calcium sulphate pelets. Generaly, bony union was detectable 1 year after operation. These findings indicate that xenograft and calcium sulphate in treating benign bone tumor have acquired good results, which can be used as a substitute of autologous bone.

Changes of renal pathomorphology,transforming growth factor-β1 and connective tissue growth factor in type 1 diabetes mellitus / 中国综合临床

Objective To investigate changes ane clinical significance of renal pathomorphology ane transforming growth factor-β1(TGF-β1 ),connective tissue growth factor(CTGF)of rats with eifferent stages of eiabetes mellitus(DM). Methods Thirty-six SD rats were raneomly eivieee into control ane DM group. Rats in DM group were given streptozotocin(STZ)at a eose of 65 mg/ kg boey weight ane rats in control group were given same amount citric acie/ soeium citrate buffer solution. Rats were killee at 4th,12th ane 24th weeks after DM moeels establishment. The renal hyeroxyproline concentration(HPC)was measuree. Masson′s trichrome was performee to observe perivascular collagen area(PVCA). Glomerular collagen eeposition score(GCDS),tubular interstitial eisease score(TIDS)in kieney interstitial tissue were measuree. Immunohistochemical staining were usee to measure the expression of type Ⅰ ane Ⅲ collagen,TGF-β1 ,CTGF. Renal path morphology change were observee with light microscope ane transmission electron microscope. Results (1)HPC levels of rats in DM group at 4th,12th,24th weeks were(216. 50 ± 10. 05)μg/ g,(267. 67 ± 11. 29)μg/ g,(340. 67 ± 39. 22)μg/ g,higher than that of control group((123. 00 ± 6. 00)μg/ g,(158. 50 ± 8. 89)μg/ g,(201. 00 ± 7. 69)μg/ g). The PVCA score in DM group were higher than that of control group( 4 weeks:(0. 45 ± 0. 07)vs (0. 96 ± 0. 11);12 weeks:(0. 60 ± 0. 06)vs(1. 65 ± 0. 18);24 weeks:(0. 72 ± 0. 07)vs(2. 63 ± 0. 0. 40);P< 0. 05). Meanwhile the scores of GCDS ane TILS in DM group)at 4th,12th,24th weeks increasee comparee with control group(4 weeks:((0. 34 ± 0. 03)vs(0. 14 ± 0. 01))× 103 / μm2 );12 weeks:((0. 49 ± 0. 03)vs (0. 26 ± 0. 04))× 103 / μm2;24 weeks:((0. 62 ± 0. 06)vs(0. 33 ± 0. 03))× 103 / μm2 )). The expression of CollagenⅠane Collagen Ⅲ in DM group were higher than that of control at 4,12,24 weeks after DM moeel establishment(Collagen Ⅰ:4 weeks:((4. 56 ± 0. 75)vs(1. 34 ± 0. 34))× 103 / μm2;12 weeks:((6. 07 ± 0. 74)vs(2. 17 ± 0. 12))× 103 / μm2;24 weeks:((7. 22 ± 0. 19)vs(3. 00 ± 0. 33))× 103 / μm2;CollagenⅢ:4 weeks:((3. 44 ± 1. 41)vs(1. 58 ± 0. 24))× 103 / μm2;12 weeks:((5. 60 ± 1. 34)vs(2. 95 ± 0. 41)) × 103 / μm2;24 weeks:((6. 22 ± 0. 18)vs(3. 27 ± 0. 27))× 103 / μm2 )),ane the eifferences between the groups all significant(P < 0. 05). Diffuse thickening of glomerular capillary basement membrane ane interstitial collagen hyperplasia in eiabetic rats were observee by electron microscope.(2)TGF-β1 expression in DM group were(2. 16 ± 0. 50)× 103 / μm2 ,(5. 31 ± 0. 25)× 103 / μm2 ,(6. 44 ± 0. 74)× 103 / μm2 respectively at 4th, 12th,24th weeks after DM moeel establishment,higher than that of control group((1. 18 ± 0. 15)× 103 / μm2 , (2. 99 ± 0. 25)× 103 / μm2 ,(3. 28 ± 0. 66)× 103 / μm2 )ane the eifferences were significant(P < 0. 05). The same trene was also seen in terms of CTGF expression((6. 02 ± 1. 39)vs(2. 32 ± 0. 87))× 103 / μm2 ,((6. 51 ± 0. 51)vs(3. 96 ± 0. 92))× 103 / μm2 ,((7. 89 ± 1. 46)vs(4. 58 ± 1. 02))× 103 / μm2;P < 0. 05) Conclusion Early eiabetic rats show renal fibrosis ane it may be relatee with overexpression of CTGF ane TGF-β1.

Application of sentinel lymph node biopsy assisted by 99Tcm-IT-Rituximab in cutaneous malignant melanoma of limbs / 中国肿瘤临床

Objective:To evaluate the accuracy and feasibility of sentinel lymph node biopsy (SLNB) marked by 99Tcm-IT-Ritux-imab and to discuss the clinical value of the method in diagnosis and treatment of cutaneous melanoma. Methods:A total of 67 patients with cutaneous malignant melanoma received 99Tcm-IT-Rituximab-tagged SLNB from March 2008 to March 2012. Lymphoscintigra-phy was conducted 30 min to 1 h after intra-dermal injection of 99Tcm-IT-Rituximab. Subsequently, the surgery of SLNB was carried out using gamma probe. The detection and positive rates of SLNB were counted. The relationship between the status and the clinical features of the sentinel lymph node (SLN) was analyzed, such as the T stage, ulceration, age, gender, and location. The influence of SLN status on overall survival (OS) and disease-free survival (DFS) was evaluated. Results:SLNs were detected in all the 67 patients by SPECT and gamma detector, with detection rate of 100%. Fifteen patients had SLN metastasis, and the positive rate was 22.4%. Chi-square indicates that SLN metastasis is associated with age, T stage, and ulceration (P<0.05). A total of 63 patients were followed up for 24-69 months, and the median follow up time was 43 months. Kaplan-Meier survival analysis shows that both OS and DFS in the SLN-negative group are better than those in the SLN-positive group (OS:93.9%vs. 57.1%, P<0.01;DFS:79.6%versus 28.6%, P<0.01). Cox-regression multiple factors analysis suggests that both SLN status and T stage are independent factors that affect the DFS of malignant melanoma. Conclusion:SLNB assisted by 99Tcm-IT-Rituximab can well reflect the state of lymph node metastasis and is es-sential for accurate staging, prognosis judging, and treatment guiding. Its operation procedure is simple with high accuracy, and the im-aging status is stable. Therefore, it is convenient and feasible as a means of SLNB.

Echocardiographic valuation of left ventricular diastolic function and the dynamic observation of ultrastructure and argyrophilic fiber in dabetic rats / 中国综合临床

Objective To explore the relationship between left ventricular diastolic function and the dynamic changes of myocardial ultrastructure and argyrophilic fiber in diabetic rats on the different periods of lesions(week 4,12 and 24).Methods The diabetes mellitus(DM)in healthy male Sprague-Dawley rats was induced by a single injection of streptozotocin(STZ,Sigma)into intraperitoneal at a dose of 65 mg/kg body weight.The left ventricular diastolic function was measured by Color Doppler Flow Imaging-Pulsed Wave(CDFIPW)and Doppler Tissue Imaging(DTI)echocardiography.Heart tissue at the apex was obtained rapidly for transmission electron microscope study.Argyrophilic staining was used in the study of argyrophilic fiber volume fraction(APFVF)in heart interstitial tissue.Results Diastolic dysfunction of left ventricle was detected in STZinduced diabetic rats by CDFI-PW(E/A ＜ 1)at week 4,and progressed gradually.Pseudonormal filling (E/A ＞ 1)was found in diabetic rats at week 24,which could be identified by DTI(Ea/Aa ＜ 1).Diastolic function of normal rats was not impaired(E/A ＞ 1 and Ea/Aa ＞ 1).Transmission electron microscopy revealed a spectrum of subcellular remodeling in myocardium which was characterized by myofibril content decrease,disorganization,mitochondrial degeneration,sarcoplasmic reticulum,structural disorder.Compared with the control group,APFVF in myocardium was increased significantly in diabetic rats(P ＜ 0.05).Conclusion The diastolic dysfunction in STZ-induced diabetic rats correlates with damage of ultrastructure and increase of myocardial argyrophilic fiber.

Influence of glucose-lowering rate on left ventricular function in type 2 diabetes mellitus patients with coronary heart disease / 中华内分泌代谢杂志

Objective To explore the influence of glucose-lowering rate on left ventricular function in patients with type 2 diabetes mellitus (T2DM). Methods One hundred and thirty-two cases of type 2 diabetes mellitus and 135 cases of type 2 diabetes mellitus with coronary heart disease (T2DM+CHD)received intensive glucose lowering therapy. Then, after measuring left ventricular ejection fraction (LVEF) and E/A ratio, the variation was analyzed. Results LVEF was significantly higher than that before intensive therapy in T2DMsubgroup with glucose-lowering rate less than 6 m mol · L-1 · d-1( P＜0.05 ). So was T2DM+CHD subgroup with glucose-lowering rate less than 4 mmol· L-1 · d-1 (P＜0.05). LVEF was significantly lower than that before intensive therapy in T2DM+CHD subgroup with glucose-lowering rate greater than 4 mmol · L-1 · d-1( P＜0. 05 ),while by the end of following up for 3 months, LVEF stepped up and no significant difference was observed between subgroups ( P ＞ 0. 05 ). The E/A ratio stepped up in both subgroups after intensive therapy ( P ＜ 0. 05 ).Conclusions For T2DM patients with coronary heart disease, excessively fast glucose-lowering rate may impair left ventricular function. Long-term good control of blood glucose restores the impaired left ventricular function causes by excessively fast glucose-lowering rate. After intensive therapy, left ventricular diastolic function finally improves in both subgroups regardless of the glucose-lowering rate and coronary heart disease.

Post-operative radiotherapy improves survival in patients with soft tissue malignant fibrous histiocytoma / 中华放射肿瘤学杂志

stage, post-operative radiotherapy are prognostic factors in patients with soft tissue MFH. Post-operative radiotherapy may be the best modality in improving the prognosis of MFH.

The echoeardiographic study on diabetic cardiomyopathy of rats / 中华超声影像学杂志

Objective To valuate the left ventricular function and myocardial fibrosis using Dopple tissue imaging(DTI) and integrated backscatter(IBS) in diabetic cardiomyopathy(DCM) rats. Methods Diabetes mellitus(DM) in healthy male SD rats was induced by single injection of streptozotocin (STZ,Sigma) into intraperitoneal at a dose of 65mg/kg body weight. Left ventricular fractional shortening (LVFS),ratio of peak early to late diastolic mitral inflow velocity(E/A) by color Doppler flow imaging-pulsed wave (CDFI-PW), ratio of peak early to late diastolic mitral annulus velocity (Ea/Aa) by DTI,myocardial calibrated IBS (IB%), cyclic variation of IBS(CVIB) by IBS, were determined in rats of 4,12 and 24 weeks after DM was induced. Heart weight index(HWI), Masson's trichrome staining, expression of connective tissue growth factor(CTGF) and activity of sarcoplasmic reticulum Ca2+ -ATPase(SERCA2a) in heart were measured, while the age-matched health rats served as control group. Results ① Diastolic dysfunction of left ventricular was found in diabetic rats detected by CDFI-PW(E/A<1) at the 4th week,and developed gradually. Pseudonormal filling(E/A>1) was found in major diabetic rats at the 24th week,which could be identified by DTI(Ea/Aa<1). The IB% of left ventricular posterior wall was significantly higher in diabetic rats than those in the control group( P<0.05), but the CVIB of the 24th week diabetic rats was lower(P<0.05). The LVFS had no great alteration in six groups(P>0.05).②Compared with the control group, the activity of SERCA2a was reduced, while HWI, collagen volume fraction(CVF),perivascular collagen area(PVCA) and the expression of connective tissue growth factor(CTGF) protein in heart were increased in diabetic rats (P<0.05), and correlation between IB% and HWI, CVF, PVCA,CTGF was significant (P<0.05). Conclusions DTI and IBS can assess early left ventrieular diastolic function and fibrosis of diabetic cardiomypathy in STZ-induced diabetic rats. DTI echoeardiography can identify "pseudonormal" mitral inflow patterns. DCM may be relatived with overexpression of CTGF and decreasion of SERCA2a activity.