Pluronic modified polyamidoamine (PAMAM) conjugate (PF127-PAMAM) was prepared and the inhibiting effect of MDR against MCF-7/ADR was investigated with doxorubicin (DOX) as model drug. 1H NMR and FTIR spectra showed that the conjugate was synthesized successfully. Element analysis accurately measured that 27.63% amino of per PAMAM was modified by pluronic (PAMAM : PF127, 1 : 35.37 mole ratio). PF127-PAMAM showed an increased size and a reduced zeta potential compared to PAMAM. PF127-PAMAM had lower hemolytic toxicity and cytotoxicity due to the reduced zeta potential and the protection of PF127. Each PF127-PAMAM molecular could load 19.58 DOX molecules, and the complex exhibited sustained and pH-sensitive release behavior. PF127-PAMAM/DOX exhibited weaker cytotoxicity than free DOX in MCF-7 cells; while the complex showed much stronger reverse effect of drug resistance in MCF-7/ADR cells, and resistance reversion index (RRI) was as high as 33.15.
Dendrimers ; pharmacology ; Doxorubicin ; pharmacology ; Humans ; MCF-7 Cells ; drug effects ; Poloxamer ; pharmacology

OBJECTIVETo study the prevalence and drug resistance of extended-spectrum-β-lactamases (ESBLs)-producing bacteria in blood culture isolated from children with hematological malignancy after chemotherapy.

METHODSBlood samples taken from 3264 children with hematological malignancy and severe infection following chemotherapy between 2002 and 2008 were cultured using the Bact/ALTER 3D blood culture system. VITEK 60 automicroscan was used to identify viral species and to conduct drug resistance tests. The results were indentified according to National Committee for Clinical Laboratory Standard guidelines.

RESULTSFifty-eight strains of Escherichia coli and fifty-one strains of Klebsiella pneumoniae were isolated. Thirty-eight strains of Escherichia coli and nineteen strains of Klebsiella pneumoniae were ESBLs-producing and these ESBLs-producing strains were less susceptible than those that were non-ESBLs-producing to most antibiotics. Both ESBL- and non-ESBL-producing strains were susceptible to imipenem, piperacillin/tazobactam and amikacin.

CONCLUSIONSThe prevalence of ESBLs-producing bacteria is high in childrn with hematological malignancy and infection following chemotherapy. ESBLs-producing bacteria are resistant to common antibiotics, suggesting that antibiotic treatment based on the result of antimicrobial susceptibility test is necessary in these children.

Adolescent ; Bacteremia ; microbiology ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; enzymology ; isolation & purification ; Female ; Hematologic Neoplasms ; drug therapy ; microbiology ; Humans ; Klebsiella pneumoniae ; drug effects ; enzymology ; isolation & purification ; Male ; Microbial Sensitivity Tests ; beta-Lactamases ; biosynthesis

OBJECTIVETo investigate the distribution of ER isoforms in endometriosis and eutopic endometrium.

METHODSTissue samples of patients with ovarian endometriosis, treated in People's Liberation Army General Hospital from January 2004 to December 2006, were retrieved. A total of 60 cases of ovarian endometriotic cysts with their corresponding eutopic endometrium (30 cases of proliferation phase and 30 of secretary phase eutopic endometrium) and 30 cases of normal endometrium (15 proliferative and 15 secretary phase endometrial samples respectively) were included. Expressions of ERalpha and ERbeta were analyzed using immunohistochemistry and the expression ratio was statistically analyzed by using SPSS 12.0 software.

RESULTSExpressions of both ERalpha and ERbeta in epithelial cells were positively correlated with that of the stromal cells. The expression of ERalpha in eutopic endometrium (73.3% in epithelium and 76.7% in stroma) was significantly higher than that in ovarian endometriotic cysts (43.3% in epithelium and 46.7% in stroma), or normal control (56.7% in epithelium and 50.0% in stroma, respectively, each P < 0.05. However, the expression of ERbeta (90.0% in epithelium and 76.7% in stroma) was higher in ovarian endometriotic cysts than that in the eutopic endometrium (68.0% in epithelium and 63.3% in stroma respectively, P < 0.05), and ERbeta expression in eutopic endometrium was higher than that in the normal control endometrium (36.7% in epithelium and 26.7% in stroma, respectively, P < 0.05). The expressions of both ERalpha and ERbeta changed periodically in eutopic and normal endometrium, whereas ERalpha and ERbeta level were less variable in the ectopic endometrium. The expression of ERbeta was statistically higher than that of ERalpha (P < 0.05) in ectopic endometrium, whereas no significant difference was seen between the two isoforms in the eutopic or normal endometrium.

CONCLUSIONSBoth ERalpha and ERbeta have higher expression levels in eutopic endometrium of patients with ovarian endometriotic cysts. ERbeta is predominantly expressed in endometriotic cysts, where the expression of ERalpha is limited. The different distribution of ERalpha and ERbeta may play an important role in the development of ovarian endometriosis.

Adult ; Choristoma ; pathology ; Endometriosis ; metabolism ; pathology ; Endometrium ; metabolism ; pathology ; Epithelium ; Estrogen Receptor alpha ; analysis ; Estrogen Receptor beta ; analysis ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Protein Isoforms ; analysis ; Receptors, Estrogen ; analysis ; Stromal Cells ; metabolism

Objective To explore the effect of high-frequency, repeated transcranial magnetic stimulation (rTMS) together with mirror neuron training on the cognition of persons with vascular cognitive impairment but without dementia (VCIND). Methods Thirty-three persons with VCIND were randomly divided into an rTMS+MNS group (n =17) and an rTMS group (n =16) using a random number table. Both groups were given conventional rehabilitation training and rTMS over the left dorsolateral prefrontal cortex at 10 Hz, 2000 pulses per day at their individual motor thresholds on weekdays for 4 weeks using a CCY- I stimulator. The rTMS+MNS group was additionally given mirror neuron training. The Montreal cognitive assessment (MoCA), the mini-mental state examination (MMSE) and the modified Barthel Index (MBI) were administered before and after the treatment. The P300 latency and amplitude of both groups were also measured. Results Before the treatment, no significant differences were found in any of the measurements. After the treatment, the average MoCA, MMSE and MBI scores had increased significantly in both groups, with those of the rTMS+MNS group increasing significantly more than those of the rTMS group. After the treatment, the average P300 latency and amplitude of both groups were also significantly better than before the treatment. Compared with the rTMS group, the average P300 latency of the rTMS+MNS group was significantly shorter, while the average amplitude was significantly greater. Conclusion Mirror neuron training combined with rTMS is more effective than rTMS alone in improving the cognition and ADL performance of VCIND patients. It is worth applying in clinical practice.

Objective To investigate the protective effect of prostaglandin E1 on renal injury in rats with obstructive jaundice.Methods The Wistar rats were randomly divided into three groups (n =15):sham operation group,model group and experimental group.The rat model of obstructive jaundice was established by double ligation of common bile duct.The experimental group rats were injected intraperitoneally with prostaglandin E1 0.1 μg · kg-1 on the seventh postoperative day and treated for 14 d.The other two groups were given the same dose of normal saline.The expression of nuclear factor-κB (NF-κB),B lymphocyto tumor-2 (Bcl-2) and intercellular adhesion molecule-1 (ICAM-1) protein were detected by immunohistochemistry.The tubular epithelial apoptosis index was detected by TdT-mediated dUTP nick-end labeling method.Results The expression of NF-κB protein in sham operation group,model group and experimental group were 1.16 ±0.24,9.28 ± 0.48 and 6.52 ± 0.72;the expression of Bcl-2 protein in the three groups were 0.86 ± 0.54,11.28 ± 0.27 and 8.36 ± 0.48;the expression of ICAM-1 protein in the three groups were 1.04 ± 0.52,18.80 ± 3.26 and 11.18 ± 4.62;the renal tubular epithelial cell apoptosis index in the three groups were O,(5.58 ± 0.46)％ and (1.28 ± 0.24) ％.Comparison between sham operation group and model group,the expression of NF-κB,Bcl-2,ICAM-1 protein and cell apoptosis index was significant (all P ＜ 0.05);comparison between model group and experimental group,the diffrence of the factors was significant (all P ＜ 0.05).Conclusion Prostaglandin E1 had a protective effect on renal injury in rats with obstructive jaundice.The mechanism of the protective effect is related to inhibit the activation of NF-κB and decrease the expression of inflammatory factors.

OBJECTIVETo explore the anatomic features of mesoesophagus in combined thoracoscopic and laparoscopic esophagectomy with three-fields lymphadenectomy.

METHODSClinical data of 67 patients undergoing thoracoscopic and laparoscopic esophagectomy with three-fields lymphadenectomy from July 2011 to September 2012 were analyzed retrospectively. All the patients underwent three-fields lymphadenectomy. Proper surgical planes were selected according to anatomy of mesoesophagus. Thoracoscopic surgical space was bounded on azygotic vein and divided into upper and low esophageal triangle. Pancreas was the key anatomical mark for laparoscopic gastric dissection, and peripancreatic space was the natural laparoscopic surgical plane. Prevertebral fascia was bottom surface of neck dissection and carotid sheath was the boundary of two sides.

RESULTSThe median operative time was 251.6 min (range, 220 to 320 min). The median operative blood loss was 105.6 ml (range, 40 to 320 ml). The median number of lymph nodes dissected was 29.1 (range, 13 to 46, totally 1949). There was no perioperative death. Sixty-six patients were followed up with a mean follow-up time of 8.2 months (range, 2 to 14 months). Postoperative complications included reflux esophagitis in 10 and anastomotic stenosis in 3 cases.

CONCLUSIONIt is safe and more radical for minimally invasive esophagectomy that overall concept of minimally invasive anatomy of mesoesophagus is applied to identify the anatomic plane and landmark during operation.

Aged ; Esophageal Neoplasms ; pathology ; surgery ; Esophagectomy ; Esophagus ; anatomy & histology ; pathology ; Female ; Humans ; Lymph Node Excision ; Male ; Middle Aged ; Retrospective Studies

Background and Objective:Intensity-modulated radiotherapy (IMRT)has recently gained popularity in the treatment of nasopharyngeal carcinoma(NPC)and improved the local-regional control rate.This study was to explore whether IMRT could improved the survival rate while reduce the radiation-related injury for primary NPC patients compared with conventional radiotherapy (CRT). Methods: From Nov.2003 to Dec. 2005,190 patients with NPC treated with IMRT in a single hospital were retrospectively analyzed.Another 190 patients treated with conventional radiotherapy at the same period were matched by prognostic factors respectively. The survival status and treatment-induced adverse effects were investigated. Treatment results, the occurrence and severity of adverse effects of two groups were compared. Results: In the treatment of NPC,IMRT was superior to CRT in term of 4-year local regional control rate,relapse-free survival rate without reducing the overall survival rate. But there were no significant differences in the 4-year progress-free survival rate and distant metastasis-free survival rate between the two groups. Significant reductions of the occurrence rates and severity of acute skin reaction, neck fibrosis, trismus and xerostomia were noted in IMRT arm. But there were no differences in mucositis,hematological toxicity,hearing loss and radiation induced cranial neuropathy between IMRT arm and CRT arm. Conclusions:IMRT could improve the local regional control rate and relapse-free survival rate while reduce some radiation-related complications in patients with NPC.But the improvement of overall survival rate did not reach significant level.

BACKGROUND: Bone morphogenetic protein 2 (BMP-2) has a strong ability to induce and promote the osteogenic differentiation of mesenchymal stem cells. OBJECTIVE: To evaluate the BMP-2 effect on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) on an injectable nano-hydroxyapatite/chitosan (nHA/CS) composite scaffold. METHODS: (1) Experiment 1: Passage 3 BMSCs were divided into two groups and cultured with the nHA/CS scaffold or cultured alone. Cell counting kit-8 was used to detect cell proliferation at 1, 3, 5, 7, 14 days of culture. (2) Experiment 2: Passage 3 BMSCs were seeded onto the nHA/CS scaffold and cultured in culture medium containing BMP-2 or not. Alkaline phosphatase activity in cells was detected at 3, 6, 9, 12, 15 days of culture. Cell counting kit-8 was used to detect cell proliferation at 1, 3, 5, 7, 14 days of culture. Alizarin red staining was used to observe the osteogenic differentiation of cells at 1 and 2 weeks of culture. RESULTS AND CONCLUSION: (1) Experiment 1: With the prolongation of culture time, the absorbance values in the two groups were gradually increased, but there was no significant difference between the two groups. At 7 days of culture, the BMSCs adhered tightly to the scaffold surface. (2) Experiment 2: With the prolongation of culture time, the alkaline phosphatase activities in the two groups were gradually increased, and moreover, the alkaline phosphatase activity in the experimental group was higher than that in the control group at different culture time (P < 0.05). The absorbance values in the two groups were also gradually increased, and the value in the experimental group was higher than that in the control group at different culture time (P < 0.05). At 1 and 2 weeks of culture, the number of calcified nodules was higher in the experimental group than the control group. To conclude, BMP-2 has a promotion role in the proliferation and differentiation of BMSCs cultured on the injectable nHA/CS scaffold.

The tumor suppressor gene p53 and p16, both of which play an important role in inhibition of tumorigenesis, are homozygously deleted in human myeloid leukemia cell line K562. To explore the inhibition of K562 cell proliferation by wild type p16 and p53 genes, both p16 and p53 genes were co-transfected into K562 cells mediated by liposome. The expression of the two genes was measured by immunocytochemical method, the cell cycle was analysed by flow cytometry, and the number of recovered viable cells was assessed after transfection. After co-transfection, the p53 and p16 positive cells were 23% and 28%, respectively. The results showed that co-transfection of p16 and p53 genes significantly inhibits cell proliferation comparing with transfection either by p16 gene or by p53 gene (P < 0.05). Expression of p16 and p53 proteins increased the cell number in G(1) phase but decreased the cell number in S phase. It is concluded that co-transfection of p16 and p53 genes has a stronger growth-inhibitory effect on K562 cell growth than that of transfection only by p16 gene or by p53 gene, may be a pathway for gene therapy in leukemia.
Cell Division ; Genes, p16 ; physiology ; Genes, p53 ; physiology ; Humans ; K562 Cells ; Plasmids ; Transfection

OBJECTIVETo investigate the protective effects of estrogen on the mitochondria in human umbilical vascular endothelial cells (HUVECs).

METHODSHUVECs were exposed to H2O2 at 250 micromol/L for 4 h with or without pretreatment with 17-estradiol (E2) and ICI182780. Complex IV activity of the cells was measured with chromometry, and 2, 7-dichlorofluorescein diacetate (DCFH-DA) was used to determine intracellular reactive oxygen species (ROS). Intracellular adenosine triphosphate (ATP) level was quantified with a luciferin- and luciferase-based assay.

RESULTSCompared to the blank control group, H2O2 caused a decrease in complex IV activity, intracellular ATP level, and the cell viability, but elevated intracellular ROS. E2 pretreatment of cells significantly attenuated these effects of H2O2 exposure. ICI182780 administered prior to E2 pretreatment antagonized the protective effects of E2 against H2O2 exposure.

CONCLUSIONE2 offers mitochondrial protective effects on HUVECs, which is mediated by the estrogen receptors.

Cells, Cultured ; Cytoprotection ; drug effects ; Electron Transport Complex IV ; metabolism ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Estrogens ; pharmacology ; Female ; Humans ; Hydrogen Peroxide ; pharmacology ; Mitochondria ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Pregnancy ; Reactive Oxygen Species ; metabolism ; Umbilical Veins ; cytology