BACKGROUNDKeratinocyte serum-free medium (K-SFM) is a defined medium used to support the growth of primary keratinocytes and embryonic stem cell. The aim of this research was to optimize enrichment of breast cancer stem cells (CSCs) using K-SFM.

METHODSA K-SFM was used to enrich CSCs from two breast cancer cell lines and a primary culture of breast cancer. RPMI-1640 supplemented with 10% fetal calf serum (FCS) was used as a control. CSCs were identified with flow cytometry using CD44(+)/CD24(-) as molecular markers. The expression of a variety of CSC markers (Oct-4, ABCG2, Nanog, N-cadherin, and E-cadherin) was analyzed with real-time PCR.

RESULTSMuch higher percentage of CSCs was achieved with K-SFM: 17.3% for MCF-7 cells, 17.4% for SKBR-3, and 20.0% for primary breast cancer culture. Less than 1% CSC was achieved using RPMI-1640 supplemented with 10% FCS. In comparison to the CSCs obtained with RPMI-1640, CSCs in the K-SFM expressed higher levels of Oct-4, ABCG2, Nanog and N-cadherin, and lower level of E-cadherin.

CONCLUSIONK-SFM is an optimal culture medium to maintain and to enrich breast CSCs.

ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; genetics ; Cadherins ; genetics ; Cell Culture Techniques ; methods ; Cell Line, Tumor ; Culture Media, Serum-Free ; Female ; Homeodomain Proteins ; genetics ; Humans ; Keratinocytes ; cytology ; Nanog Homeobox Protein ; Neoplasm Proteins ; genetics ; Neoplastic Stem Cells ; cytology ; metabolism ; Octamer Transcription Factor-3 ; genetics ; Real-Time Polymerase Chain Reaction

OBJECTIVETo investigate the value of technetium-99m methoxyisobutylisonitrile ((99)Tc(m)-MIBI) imaging in predicting the efficacy of neoadjuvant chemotherapy (NCT) and prognosis in patients with operable breast cancer.

METHODSSixty five patients with breast cancer underwent (99)Tc(m)-MIBI scintimammography before NCT, and static planar images were taken at 10 min and 180 min after scintimammography. The clearance rate was calculated in each patient, correlation between the clearance rate and efficacy of NCT, and the disease free survival rate were analyzed.

RESULTSThe mean clearance rate of 65 patients was (17.4 ± 6.8)%. The efficacy of NCT was 86.2% (CR 4 cases, PR 52 cases, SD 8 cases, and PD 1 case), and the mean clearance rate of patients with good response or poor response of chemotherapy were (15.5 ± 5.0)% and (29.2 ± 3.2)%, respectively. There was a significant difference between the two groups. The average disease free survival rate in the group with low clearance rate was (75.8%, P = 0.046), significantly higher than that in the group with high clearance rate (53.1%).

CONCLUSIONScintimammography of (99)Tc(m)-MIBI may be used to evaluate the efficacy and prognosis of NCT for patients with operable breast cancer.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; diagnostic imaging ; drug therapy ; Carcinoma, Ductal, Breast ; diagnostic imaging ; drug therapy ; Carcinoma, Lobular ; diagnostic imaging ; drug therapy ; Chemotherapy, Adjuvant ; Cyclophosphamide ; therapeutic use ; Disease-Free Survival ; Epirubicin ; therapeutic use ; Etoposide ; therapeutic use ; Female ; Fluorouracil ; therapeutic use ; Follow-Up Studies ; Humans ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Predictive Value of Tests ; Radionuclide Imaging ; Radiopharmaceuticals ; Remission Induction ; Taxoids ; therapeutic use ; Technetium Tc 99m Sestamibi

The teaching status of Molecular Pharmacognosy in 28 institutions in China was investigated by questionnaire and the survey data was analyzed by SPSS. Research contents included course beginning years, majors, class hours, characteristics of the course, teaching ways, the theory and practice contents, evaluation modes, selection of teaching material, teaching achievements, teachers and so on for undergraduates and graduates. Research results showed that with 20 years' development, Molecular Pharmacognosy had been offered for both undergraduate and graduate students in at least 20 colleges and universities and Molecular Pharmacognosy education in China showed good development momentum. At the same time, to promote the development of Molecular Pharmacognosy further, investment for it should be increased and practical teaching condition should be improved.
China ; Humans ; Molecular Biology ; education ; manpower ; methods ; trends ; Pharmacognosy ; education ; manpower ; methods ; trends ; Plants, Medicinal ; chemistry ; genetics ; Surveys and Questionnaires ; Teaching ; manpower ; methods ; trends

BACKGROUNDL-glutamate (L-GLU) is a major neurotransmitter in the nucleus ambiguus (NA), which can modulate respiration, arterial pressure, heart rate, etc. This study investigated the effects and mechanisms of L-GLU microinjected into NA on gastric motility in rats.

METHODSA latex balloon connected with a pressure transducer was inserted into the pylorus through the forestomach for continuous recording of the gastric motility. The total amplitude, total duration, and motility index of gastric contraction waves within 5 minutes before microinjection and after microinjection were measured.

RESULTSL-GLU (5 nmol, 10 nmol and 20 nmol in 50 nl normal saline (PS) respectively) microinjected into the right NA significantly inhibited gastric motility, while microinjection of physiological saline at the same position and the same volume did not change the gastric motility. The inhibitory effect was blocked by D-2-amino-5-phophonovalerate (D-AP5, 5 nmol, in 50 nl PS), the specific N-methyl-D-aspartic acid (NMDA) receptor antagonist, but was not influenced by 6-cyaon-7-nitroquinoxaline-2,3-(1H,4H)-dione (CNQX) (5 nmol, in 50 nl PS), the non-NMDA ionotropic receptor antagonist. Bilateral subdiaphragmatic vagotomy abolished the inhibitory effect by microinjection of L-GLU into NA.

CONCLUSIONSMicroinjection of L-GLU into NA inhibits the gastric motility through specific NMDA receptor activity, not non-NMDA receptor activity, and the efferent pathway is the vagal nerves.

2-Amino-5-phosphonovalerate ; pharmacology ; 6-Cyano-7-nitroquinoxaline-2,3-dione ; pharmacology ; Animals ; Gastrointestinal Motility ; drug effects ; Glutamic Acid ; administration & dosage ; pharmacology ; Male ; Medulla Oblongata ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Receptors, N-Methyl-D-Aspartate ; antagonists & inhibitors ; Vagotomy

OBJECTIVETo evaluate the relationship between the clearance rate of technetium-99m methoxyisobutylisonitrile ((99m)Tc-MIBI) in scintimammography and multidrug-resistant proteins expression in breast cancer tissues.

METHODSSeventy-six patients with breast cancer underwent (99m)Tc-MIBI scintimammography before treatment, and static planar images were taken at 10 min and 180 min after scintimammography. The clearance rate of (99m)Tc-MIBI was calculated in each patient. Immunohistochemical analysis was performed on pathological specimens of the 76 breast tumors to determine the expression of P-glycoprotein (P-gp), glutathione S-transferase Pi (GST-pi) and topoisomerase II (Topo II).

RESULTSThe clearance rate was significantly higher in 36 patients with positive P-gp expression when compared with that in 40 patients with negative P-gp expression. There was no significant relationship between GST-pi, Topo II and the clearance rate of (99m)Tc-MIBI.

CONCLUSIONThe clearance rate of (99m)Tc-MIBI in breast imaging may be used to evaluate the P-gp level in breast cancers.

ATP Binding Cassette Transporter, Sub-Family B ; analysis ; Adult ; Aged ; Breast Neoplasms ; chemistry ; diagnostic imaging ; Carcinoma, Ductal, Breast ; chemistry ; diagnostic imaging ; DNA Topoisomerases, Type II ; analysis ; Drug Resistance, Multiple ; Female ; Glutathione S-Transferase pi ; analysis ; Humans ; Immunohistochemistry ; Middle Aged ; Multidrug Resistance-Associated Proteins ; analysis ; Radionuclide Imaging ; Radiopharmaceuticals ; Technetium Tc 99m Sestamibi

Background The incidence of brain metastases in patients with breast cancer is approximately 10％-16％,and survival after diagnosis of brain metastases is usually short.This study was designed to evaluate the risk factors associated with brain metastases in advanced breast cancer patients,with a view to help predict patient groups with high risk of brain metastases.Methods In total,295 patients with advanced breast cancer were evaluated.All patients were pathologically confirmed and metastatic lesions were confirmed pathologically or by imaging.All patients were examined at least once every 6 months with head CT or MRI.Patients showing symptoms underwent immediate inspection,and brain metastatic lesions were confirmed by head CT and/or MRI.Results At a median follow-up of 12 months from the occurrence of metastases,brain metastases had occurred in 49 patients (16.6％).In our univariate analysis,variables significantly related to increased risk of brain metastases were hormone receptor-negative tumors,epidermal growth factor receptor 2 (HER2)-positive tumors,and multiple distant metastases.Patients with dominant tumor sites in soft tissue,or defined as Luminal A subtype,tended to have a lower risk of brain metastases than patients with visceral metastases,Luminal B subtype,triple-negative subtype or HER2-enriched subtype tumors.Conclusions Our results strongly suggest that factors such as Luminal B,triple-negative,and HER2-enriched subtypes are high risk factors for brain metastases.These data,therefore,provide pivotal clinical evidence towards a comprehensive understanding of the risk factors of brain metastases in advanced breast cancer patients.

BACKGROUNDRecent advances in real-time three-dimensional echocardiography (RT3DE) offer the potential to assess the left ventricular (LV) dyssynchrony simultaneously by analyzing the 17 segments time-volume curves. The purpose of this study was to test the feasibility and accuracy of RT3DE for quantitative evaluation of left ventricular systolic synchronicity.

METHODSTwenty-four patients with dilated cardiomyopathy (DCM) and twenty-five healthy volunteers were enrolled in this study. Full volume RT3DE was performed by using Philips IE33 with X3-1 probe. The global and 17-segmental time-volume curves were obtained by the on-line Qlab software (version 4.2). The time to minimal systolic volume in each segment (T(msv)) was taken to derive the following indexes of systolic asynchrony: T(msv) 16-SD, T(msv) 16-Dif, T(msv) 12-SD, T(msv) 12-Dif, T(msv) 6-SD and T(msv) 6-Dif, which meant the standard deviation or the maximal difference of T(msv) among the 16, 12 and 6 segments of the left ventricle respectively. The software also provided with each of the above parameters as a percentage of the cardiac cycle.

RESULTST(msv) 16-SD, T(msv) 12-SD and T(msv) 6-SD were all significantly larger in the DCM group than those of the control group [T(msv) 16-SD: (52.9 +/- 40.6) ms vs (8.8 +/- 6.2) ms; T(msv) 12-SD: (29.5 +/- 30.8) ms vs (6.9 +/- 4.0) ms; T(msv) 6-SD: (28.9 +/- 34.6) ms vs (7.0 +/- 4.7) ms, all P < or = 0.001]. T(msv) 16-Dif, T(msv) 12-Dif and T(msv) 6-Dif were also significantly larger in the DCM group. There were close negative relations between the LVEF determined by RT3DE and each of the indexes of systolic asynchrony, among which the indexes of T(msv)-16-SD% and T(msv)-16-Dif% correlated most closely (r = -0.703 and r = -0.701, respectively). The DCM patients had significantly larger EDV and ESV, with significantly reduced LVEF compared with the healthy subjects.

CONCLUSIONRT3DE provides a simple, useful and unique approach to assess the systolic synchronicity of all the left ventricular segments simultaneously.

Adult ; Aged ; Cardiomyopathy, Dilated ; diagnostic imaging ; physiopathology ; Echocardiography, Three-Dimensional ; Female ; Humans ; Male ; Middle Aged ; Stroke Volume ; Systole ; Ventricular Function, Left

Background Keratinocyte serum-free medium (K-SFM) is a defined medium used to support the growth of primary keratinocytes and embryonic stem cell. The aim of this research was to optimize enrichment of breast cancer stem cells (CSCs) using K-SFM.Methods A K-SFM was used to enrich CSCs from two breast cancer cell lines and a primary culture of breast cancer.RPMI-1640 supplemented with 10％ fetal calf serum (FCS) was used as a control. CSCs were identified with flow cytometry using CD44+/CD24-as molecular markers. The expression of a variety of CSC markers (Oct-4, ABCG2, Nanog,N-cadherin, and E-cadherin) was analyzed with real-time PCR.Results Much higher percentage of CSCs was achieved with K-SFM: 17.3％ for MCF-7 cells, 17.4％ for SKBR-3, and 20.0％ for primary breast cancer culture. Less than 1％ CSC was achieved using RPMI-1640 supplemented with 10％ FCS. In comparison to the CSCs obtained with RPMI-1640, CSCs in the K-SFM expressed higher levels of Oct-4,ABCG2, Nanog and N-cadherin, and lower level of E-cadherin.Conclusion K-SFM is an optimal culture medium to maintain and to enrich breast CSCs.

OBJECTIVETo estimate the effect of the lymph duct ligation on systemic inflammatory factors and endotoxins during intestinal ischemia-reperfusion (I/R).

METHODSMale SD rats underwent occlusion of superior mesenteric artery for 60 min followed by reperfusion for 120 min plus lymph duct ligation or not. Forty rats were randomly divided into 4 groups: group A (blank); group B (sham); group C (intestinal I/R); group D (intestinal I/R plus lymph duct ligation). Mesenteric lymph nodes were harvested for standard bacteriologic cultures. The endotoxin, D-lactate, diamine oxidase (DAO), and cytokines in serum were detected.

RESULTSThe rates of bacterial translocation to mesenteric lymph nodes were 40% in group C and 20% in group D. No positive lymph node cultures were encountered in any of group A and B. The serum cytokines (except for sICAM-1) , D-lactate, DAO and endotoxin levels were lower in group D than those in group C (P<0.05), but both were higher than those in group A and B (P<0.05).

CONCLUSIONDuring intestinal I/R injury, blockage the lymph flow from gut into bloodstream decreases the levels of cytokines, and significantly attenuates the increase in intestinal permeability.

Animals ; Disease Models, Animal ; Inflammation ; Intestinal Diseases ; metabolism ; microbiology ; pathology ; Intestines ; blood supply ; pathology ; Ligation ; Lymph Nodes ; pathology ; Lymphatic System ; surgery ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; microbiology ; pathology

BACKGROUNDEmerging evidence suggests that stem cells can be used to improve cardiac function in patients after acute myocardial infarction. In this randomized trial, we aimed to use Doppler tissue tracking and strain imaging to assess left ventricular segmental function after intracoronary transfer of autologous bone-marrow stem cells (BMCs) for 6 months' follow up.

METHODSTwenty patients with acute myocardial infarction and anterior descending coronary artery occlusion proven by angiography were [corrected] randomized into intracoronary injection of bone-marrow cell (treated, n = 9) or diluted serum (control, n = 11) groups. GE vivid 7 and Q-analyze software were used to perform echocardiogram in both groups 1 week, 3 months and 6 months after treatment. Three apical views of tissue Doppler imaging were acquired to measure peak systolic displacement (Ds) and peak systolic strain (epsilonpeak) from 12 segments of LV walls. Left ventricular ejection fraction (LVEF), end-diastolic volume (EDV) and end-systolic volume (ESV) were obtained by Simposon's biplane method.

RESULTS(1) 3 months later, Ds and epsilonpeak over the infract-related region clearly increased in the BMCs group [Ds: (4.49 +/- 2.71) mm vs (7.56 +/- 2.95) mm, P < 0.01; epsilonpeak: (-13.40 +/- 6.00)% vs (-17.06 +/- 6.05)%, P < 0.01], but not in the control group [Ds: (4.74 +/- 2.67) mm vs (5.01 +/- 3.23) mm, P > 0.05; epsilonpeak: (-13.84 +/- 6.05)% vs (-15.04 +/- 6.75)%, P > 0.05]. At the same time, Ds over the normal region also increased, but the Ds enhancement was markedly higher in the BMCs group than that in the control group [(3.21 +/- 3.17) mm vs (0.76 +/- 1.94) mm, P < 0.01]. Parameters remained steady from the 3rd to 6th month in either group (P > 0.05). (2) LVEF in treated and control groups were almost the same at baseline (1st week after PCI) [(53.37 +/- 8.92)% vs (53.51 +/- 5.84)%, P > 0.05]. But 6 months later, LVEF in the BMCs group were clearly higher than that in the control group [(59.33 +/- 12.91)% vs (50.30 +/- 8.30)%, P < 0.05]. (3) There were no evident difference in EDV or ESV between two groups at baseline [EDV: (113.74 +/- 23.24) ml vs (129.94 +/- 32.72) ml, P > 0.05; ESV: (57.12 +/- 18.66) ml vs (62.09 +/- 17.68) ml, P > 0.05]. Three months later, EDV and ESV in the control group were markedly greater than those in the BMCs group [EDV: (154.89 +/- 46.34) ml vs (104.85 +/- 33.21) ml, P < 0.05; ESV: (82.91 +/- 35.79) ml vs (49.54 +/- 23.32) ml, P < 0.05]. But EDV and ESV did not change much from 3rd to 6th month in either group (P > 0.05).

CONCLUSIONSEmergency transplantation of autologous BMCs in patients with acute myocardial infarction helps to improve global and regional contractility and attenuate post-infarction left ventricular remodeling. Tissue tracking and strain imaging provide quick, simple and noninvasive methods for quantifying left ventricular segmental function in humans.

Aged ; Double-Blind Method ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; physiopathology ; therapy ; Transplantation, Autologous ; Ventricular Function, Left ; Ventricular Remodeling