Objective: To study expression of the gene of n-6 fatty acid desaturase fat-1 in human breast cancer cell, composition change of fatty acids of cell membrane, and effect of the gene on apoptosis of breast cancer cell. Methods: Construct recombinant adenovirus vector (Ad.GFP.fat-1) containing fat-1 gene, the recombinant adenovirus was produced in 293 package cell, then it infected the breast cancer cells MCF-7; Total RNA of the cells was isolated and hybridized with antisense RNA probe of fat-1 mRNA by Northern to analyze the expression of fat-1 in MCF-7;The effect of fat-1on the proliferation of MCF-7 cell was analyzed by MTT method,apoptosis of the cells were detected by apoptosis kit;Content of n-6 PUFAs/n-3 PUFAs was analyzed by Gas Chromatography. Results: The proposed recombinant virus was got through DNA recombinant technique; fat-1 gene effectively expressed in human breast cancer cell MCF-7; The fat-1 mRNA band appeared 2 days after infection of virus Ad.GFP.fat-1;Compared with the control cell (Ad.GFP), proliferation of MCF-7 cell was markedly inhibited by the gene fat-1( 23%, p

[To explore the effect of Humifuse Euphorbia Herb ( HEH) on alleviating insulin resistance in type 2 diabetic KK-Ay mice. Totally 40 KK-Ay mice fed with high-fat diet were divided into four groups: the metformin group, the model group, the HEH low-dose group and the HEH high-dose group, and orally administrated with metformin hydrochloride (250 mg x kg(-1)), distilled water, humifuse euphorbia herb 1 g x kg(-1) and 2 g x kg(-1). Besides, C57BL/6J mice with ordinary feed were taken as the normal control group and orally administrated with equal distilled water. The oral administration for the five groups lasted for eight weeks. Before and after the experiment, weight, fasting glucose and insulin tolerance were determined. The morphological changes in pancreas were observed through hematoxylin-eosin (HE) staining on pancreatic tissue sections. The serum insulin, TNF-α, IL-6, adiponectin (ADPN) and leptin (LEP) were detected by ELISA. The results showed that HEH could reduce weight and fasting glucose in KK-Ay mice, alleviate hyperinsulinemia, reduce blood glucose-time AUC, increase 30-min blood glucose decline rate, relieve insulin resistance, significantly ameliorate the pathomorphological changes in pancreas in each group, decrease serum TNF-α, IL-6 and leptin levels in KK-Ay mice and rise serum ADPN level. This study proved that humifuse euphorbia herb can ameliorate the insulin resistance in KK-Ay mice, and its mechanism may be related to the effect on inflammatory factors and adipocytokines.
Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Euphorbia ; chemistry ; Humans ; Insulin ; metabolism ; Insulin Resistance ; Interleukin-6 ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

OBJECTIVETo explore the role of Bmi-1 gene in the proliferation of squamous carcinoma cells and whether the silencing Bmi-1 can inhibit the growth of squamous cell carcinomas cells.

METHODSThe expressions of Bmi-1 in primary cultured Fibroblasts, karatinocyte cell line Hacat,squamous carcinoma cell line A431, and ECA109 were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. Recombinant plasmid inserted with Bmi-1 gene short hairpin RNA (shRNA) expression vector PGPU6/GFP/Neo-shBmi-1 was constructed and transfected into ECA109 cells with control set. After transfection for 48 and 72 hours,the mRNA and protein levels of Bmi-1 were examined with RT-PCR and Western blot analysis, respectively. The proliferation of the ECA109 cells was evaluated by MTT method and flow cytometry.

RESULTSBmi-1 was highly expressed in A431 and ECA109 cells than in Fibroblast cells and Hacat cells. The mRNA and protein expressions of Bmi-1 were significantly silenced in ECA109 cells after recombinant expression vector PGPU6/GFP/Neo-shBmi-1 transfection (P<0.05). Compared with the control groups,the proliferation of ECA109 transfected with PGPU6/GFP/Neo-shBmi-1 was significantly inhibited (P<0.05), and cells in G1 phase increased while in S phase decreased (P<0.05).

CONCLUSIONSBmi-1 is involved in the proliferation of squamous carcinoma cells. After the silencing of Bmi-1 expression,the proliferation ECA109 cells is suppressed due to the altered cell cycle.

Blotting, Western ; Carcinoma, Squamous Cell ; Cell Cycle ; Cell Line ; Cell Proliferation ; Fibroblasts ; Flow Cytometry ; Humans ; Plasmids ; Polycomb Repressive Complex 1 ; RNA, Messenger ; RNA, Small Interfering ; Transfection

OBJECTIVETo observe the regulative effect of opuntia powder on blood lipids in wistar rats and to explore its mechanism.

METHODForty normal rats were divided into four groups:control group (fed with basal feed), opuntia high, middle and low dosage groups (fed with basal feed and opuntia powder of high, middle and low dosage. The influence of opuntia powder on serum total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), arteriosclerosis index (AI), serum malondialdehyde (MDA), superoxide dismutase (SOD) were observed. (2) All of the hyperlipemia wistar rats for experiments were divided into four groups: model control group and other three groups (high, middle, low dosage groups respectively). Three weeks later, samples of blood were taken for survey of levels of TC, TG HDL-C, LDL-C, AI, MDA, SOD.

RESULTAfter opuntia powder treatment,the level of TC in nomal wistar rats was decreased. However, there was no significant difference comparing with control group (P > 0.05). The serum MAD level in the low, middle and high dosage groups were all obviously decreased, which were significantly lower than that in the control group. The SOD activities were all higher than that in the control group. The level of TC, LDL-C, AI (P < 0.01), TG (P < 0.05) were lower significantly in hyperlipemia wistar rats after treated by opuntia powder of high, middle and low dosage. The down-regulation of blood lipids was related with the dosage of opuntia powder.

CONCLUSIONThe opuntia powder may regulate the level of blood lipids in normal and hyperlipemia wistar rats. The effect is more obviously in hyperlipemia rats than that in normal rats.

Animals ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Hyperlipidemias ; blood ; Lipids ; blood ; Male ; Malondialdehyde ; blood ; Mice ; Opuntia ; chemistry ; Plants, Medicinal ; chemistry ; Powders ; Random Allocation ; Rats ; Rats, Wistar ; Superoxide Dismutase ; blood ; Triglycerides ; blood

OBJECTIVETo identify the localization of hair follicles stem cell (HFSC) in different stages of hair and explore the differentiating capacity of HFSC into epidermis in vitro.

METHODSHFSC were detected by K19 immunostaining in normal human skin. Then, the isolated HFSC through enzyme digestion were seeded on dermal equivalent (DE) and cultured between the air-liquid interfaces for 14 days. Skin-equivalents was harvested and used for evaluation.

RESULTSHFSC mainly located in outer root sheet in hair follicle and human anagen hair follicles containing two distinct reservoirs for K19-positive cells located in the bulge and bulb of the follicle. These two reservoirs fused in line of outer root sheets during the catagen-telogen transition phase and individualized again in the newly forming anagen hair follicle. Based on DE, growing HFSC built a multilayered and confined epidermis.

CONCLUSIONHFSC located in outer root sheets can promote hair cycle and differentiate into epidermis in vitro.

Cell Differentiation ; physiology ; Cells, Cultured ; Epidermis ; cytology ; Hair Follicle ; cytology ; Humans ; Stem Cells ; cytology

OBJECTIVETo evaluate the feasibility and efficacy of intraperitoneal chemotherapy for malignant ascites caused by different types of abdominal cancers guided by chemo-sensitivity methyl tetrojolium coloremetric (MTT) assay in vitro.

METHODSCancer cells in the malignant ascites were collected for MTT assay to determine the chemo-sensitivity. The drug producing the highest or the second highest inhibition rate was selected for intraperitoneal chemotherapy. The correlation between the results of MTT assay and the response of malignant ascites, the clinical features, Karnofsky performance score (KPS) and prognosis were analyzed.

RESULTSMTT assay indicated that Taxotere (TXT) and Hydroxycamptothecin (HCPT) were the most effective to cancer cells in malignant ascites, and HCPT was mostly frequently used for intraperitoneal chemotherapy (56.9%). Twenty-four patients showed response by intraperitoneal chemotherapy (complete response: 7; partial response: 17) with a slightly significant correlation between the results of MTT assay and response of malignant ascites (P = 0. 014). The KPS of the responders was improved significantly (P < 0.001), and the response of malignant ascites to intraperitoneal chemotherapy was demostrated as an independent prognostic factor by multi-variate analysis in this series.

CONCLUSIONIn vitro chemo-sensitivity MTT assay guided intraperitoneal chemotherapy for malignant ascites is simple, effective and safe, which can improve the KPS and prognosis of the responders.

Adenocarcinoma ; drug therapy ; pathology ; Adult ; Aged ; Antineoplastic Agents ; administration & dosage ; pharmacology ; therapeutic use ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; therapeutic use ; Ascites ; drug therapy ; pathology ; Camptothecin ; administration & dosage ; analogs & derivatives ; pharmacology ; therapeutic use ; Cell Survival ; drug effects ; Colorectal Neoplasms ; drug therapy ; pathology ; Female ; Humans ; Injections, Intraperitoneal ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Pancreatic Neoplasms ; drug therapy ; pathology ; Stomach Neoplasms ; drug therapy ; pathology ; Taxoids ; administration & dosage ; pharmacology ; therapeutic use ; Tumor Cells, Cultured

OBJECTIVETo develop a method to obtain and identify human coronary artery endothelial cells obtained during percutaneous coronary interventions (PCI).

METHODSCoronary guide wires were used to obtain endothelial cells from coronary arteries in 28 patients undergoing PCI. The cells were eluted from the wire tips and then purified by magnetic beads coated with anti-CD146 antibody. von Willebrand factor (vWF) was used as an immunocytochemical marker for endothelial cells. The cellular viability was evaluated by observing cell membrane integrity and energy-dependent uptake of DiI-labeled acetylated low-density lipoprotein.

RESULTSAn average of 96 coronary artery endothelial cells with good viability per patient were obtained by one guide wire. vWF identification showed their endothelial morphology and immunoreactivity.

CONCLUSIONThe viable coronary endothelial cells could be obtained during routine percutaneous coronary interventions combined with magnetic beads isolation technique. These cells may be used for further cellular functional analyses (such as immunocytochemistry and molecular biology) and expand our understanding on mechanisms of coronary artery diseases.

Biopsy ; methods ; Coronary Vessels ; cytology ; pathology ; Endothelium, Vascular ; cytology ; pathology ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo study the expression of integrin beta1 in squamous cell carcinoma (SCC) and explore the relationship between stem cell marker and SCC.

METHODSThe expressions of integrin beta1 in SCC tissues and SCC cell strain A431 were detected with immunohistochemical methods and cell staining method. The differentiation of SCC cells were induced with all-trans-retinoic acid (ATRA). The changes of integrin beta1 levels before and after induction were detected with RT-PCR.

RESULTSIn highly differentiated SCC tissues, integrin beta1 was constantly expressed in the basal-like cells in the edge of tumor; some cells inside arranged as island also showed positive integrin beta1 expression. In poorly differentiated SCC tissues, island-like integrin beta1-positive cells remarkably increased and distributed in a diffuse way. In SCC A431 cells, integrin beta1 was expressed unevenly in tumor cells. After treatment by ATRA, level of integrin beta1 mRNA in A431 cells significantly decreased compared with untreated control (P < 0.05), and the ratios between the intensity values of integrin beta1 to beta-actin were 0.071 +/- 0.025 and 0.029 +/- 0.018 at 24 h and 48 h, respectively, whereas in controls were 0.148 +/- 0.027 and 0.136 +/- 0.011 (P < 0.05).

CONCLUSIONSIntegrin beta1 is heterogeneously expressed in both SCC tissues and SCC A431 cells. The expression of Integrin beta1 decreases when the differentiation level of tumor cells increase, indicating that integrin beta1 is closely related with the initiation of SCC and potential cancer stem cells in SCC.

Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; Cell Differentiation ; Female ; Humans ; Integrin beta1 ; metabolism ; Male ; Middle Aged ; Skin Neoplasms ; metabolism

OBJECTIVETo explore the regulative efficacy of Pu'er tea () extract on metabolic syndrome.

METHODSNinety patients with metabolic syndrome were randomly divided into two groups, the intervention group administered with Pu'er tea extract, and the placebo group with placebo capsules. After 3 months' treatment, body mass index, waist hip ratio, blood lipids, blood sugar, immune and inflammatory index, and oxidation index of the patients with metabolic syndrome were tested and analyzed.

RESULTSIn the intervention group, the body mass index, waist-hip ratio, fasting and 2 h postprandial blood glucose, serum total cholesterol, triglycerides, low density lipoprotein and apolipoprotein B-100 all decreased in the patients with metabolic syndrome, and also the high-density lipoprotein level increased and apolipoprotein A-1 showed the tendency to increase. Serum C-reactive protein, tumor necrosis factor-α, and interleukin-6 were decreased in the intervention group. Interleukin-10 level was increased, MDA was decreased and superoxide dismutase was increased. Compared with before treatment and the placebo group, there were significant differences (P<0.05, P<0.01).

CONCLUSIONSPu'er tea demonstrated excellent potential in improving central obesity, adjusting blood lipid, lowering blood sugar, regulating immunity and resisting oxidation. It can adjust the metabolic syndrome of different clinical phenotypes to different degrees, and is ideally fit for early prevention of metabolic syndrome.

Adult ; Aged ; Blood Glucose ; metabolism ; Body Mass Index ; Double-Blind Method ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Inflammation ; blood ; complications ; Lipids ; blood ; Male ; Malondialdehyde ; blood ; Metabolic Syndrome ; blood ; complications ; drug therapy ; Middle Aged ; Oxidation-Reduction ; Placebos ; Plant Extracts ; therapeutic use ; Superoxide Dismutase ; blood ; Waist-Hip Ratio

BACKGROUNDWhite blood cell (WBC) counts and differentials performed using an automated cell counter typically require manual microscopic review. However, this last step is time consuming and requires experienced personnel. We evaluated the clinical efficiency of using flow cytometry (FCM) employing a six-antibody/five-color reagent for verifying automated WBC differentials.

METHODSA total of 56 apparently healthy samples were assessed using a five-color flow cytometer to verify the normal reference ranges of WBC differentials. WBC differentials of 622 samples were also determined using both a cell counter and FCM. These results were then confirmed using manual microscopic methods.

RESULTSThe probabilities for all of the parameters of WBC differentials exceeded the corresponding normal reference ranges by no more than 7.5%. The resulting WBC differentials were well correlated between FCM and the cell counter (r > 0.88, P < 0.001), except in the case of basophils. Neutrophils, lymphocytes, and eosinophils were well correlated between FCM and standard microscopic cytology assessment (r > 0.80, P < 0.001). The sensitivities of FCM for identification of immature granulocytes and blast cells (72.03% and 22.22%, respectively) were higher than those of the cell counter method (44.92% and 11.11%, respectively). The specificities of FCM were all above 85%, substantially better than those of the cell counter method.

CONCLUSIONThese five-color FCM assays could be applied to accurately verify abnormal results of automated assessment of WBC differentials.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Infant ; Leukocyte Count ; methods ; Leukocytes ; cytology ; Male ; Middle Aged ; Young Adult