Aim To study the effect of pectin-adriamy-cin conjugate ( PAC) on cardiac toxicity .Methods 50 female SD rats were randomly divided into 5 groups with 10 animals in each group .Adriamycin ( ADM ) group received 3 mg? kg -1 , ip, every other day for 6 times.PAC group received ADM equivalent 1.5,3 and 6 mg? kg -1 , ip, every other day for 6 times.Control group received normal saline parallel to ADM .Rats were sacrificed and the echocardiogram , cardiac en-zymes , the oxidative stress levels in myocardial cells and histopathological changes after 48 h administration were detected.S180 ascites tumor bearing mice models were established to investigate the antitumor activity of PAC.Results The survival rate of ADM group was 50% and that of PAC each group was 100%.PAC could significantly increase body weight ,heart index and immune index and increase HR ,EF,FS,reduce LVIDd, LVIDs.PAC could also significantly increase the AST , LDH, CK, CK-MB level in serum .GSH-Px and SOD activities of PAC group were significantly increased and MDA contents were reduced , and histopathological changes decreased .PAC could effectively inhibit the growth of tumor cells and extend the survival period of mice.Conclusion PAC induces a significant reduc-tion in cardiotoxicity by increasing survival rate , im-mune and cardiac function , improving cardiac en-zymes ,oxidative stress and myocardial cell injury , and also PAC has obvious antitumor effect .

Purpose To investigate the expression level of MED27 in lung cancer tissue samples and lung cancer cell lines and to further study the biological function of MED27 in lung cancer cells.Methods Immunohistochemistry and Western blot were used to detect MED27 expression in 70 lung cancer tissues and 5 different lung cancer cell lines,and the correlation between MED27 expression and gender,age as well as PTNM was also analyzed.The silence sequence of MED27 was designed by the siRNA technique.Western blot was used to detect the silence efficiency of MED27.The proliferation,migration and invasion ability of cells were assessed by CCK-8 assay,Scratch assay and Transwell assay after the MED27 was knocked down.Western blot was used to detect the expression of protein involved in the cell proliferation,migration and invasion.Results The results of immunohistochemistry and Western blot showed that MED27 expression was higher in lung cancer tissues and cells (P ＜ 0.05).The expression of MED27 was positively correlated with lymph node metastasis (x2 =9.438,P =0.002,P ＜ 0.05).However,it was not related with gender,age,tumor size and distant metastasis (P ＞ 0.05).The knockdown of MED27 by MED27 specific siRNA could inhibit the proliferation,migration and invasion of H460 cells (P ＜ 0.05).The expression of MMP-2 and MMP-9 involved in the cell migration that were significantly inhibited in H460 cells transfected by MED27 siRNA,and the expression of E-cadherin,related with cell invasion was also decreased,while E-cadherin negative regulatory protein Snail was increased.Conclusion MED27 is highly expressed in lung cancer tissues and cells and high expression of MED27 predicts poor prognosis in lung cancer patients.The knockdown of MED27 inhibits the proliferation,migration and invasion ability of lung cancer cells.All of the above results suggest that MED27 is expected to be a candidate target of lung cancer gene therapy.

OBJECTIVETo study the effects of interferon-gamma and interferon-gamma combined with interferon-alpha on HCV RNA replication and the possible mediators of interferon-gamma anti-HCV in vitro.

METHODSAn HCV replicon cell culture system was established and the cells were treated with interferon-gamma or interferon-gamma combined with interferon-alpha. HCV RNA levels in the cells were evaluated by semi-quantitative RT-PCR and real-time PCR, and the levels of NS5A protein were examined by Western blot.

RESULTSInterferon-gamma could inhibit HCV RNA replication and NS5A protein expression effectively; The anti-HCV effects of interferon-gamma were both in time-dependent and does-dependent manners; Pretreating the cells with interferon-gamma could significantly enhance the antiviral effects of interferon-alpha; The expressions of IRF-1, 2'5'-OAS1(p46), 2'5'-OAS2(p69), ISGF3gamma and STAT1 were significantly increased after interferon-gamma treatment.

CONCLUSIONInterferon-gamma has a direct inhibitory effect on HCV replicon RNA replication and NS5A expression, and both are in a dose and time dependent manner; Interferon-gamma has a synergistic anti-HCV effect with interferon-alpha. IRF-1, 2'5'-OAS1(p46), 2'5'-OAS2(p69) and ISGF3gamma may mediate the anti-HCV effects of interferon-gamma.

Antiviral Agents ; pharmacology ; Female ; Hepacivirus ; drug effects ; Humans ; Interferon-alpha ; pharmacology ; Interferon-gamma ; pharmacology ; Liver Neoplasms ; pathology ; Male ; RNA, Viral ; biosynthesis ; drug effects ; Replicon ; drug effects ; Tumor Cells, Cultured ; Virus Replication ; drug effects

To investigate the effects of 2-(4-methoxycarbonyl-2-tetradecyloxyphenyl)carbamoylbenzoic acid (CX09040) on protecting pancreatic β cells, the β cell dysfunction model mice were induced by injection of alloxan into the caudal vein of ICR mice, and were treated with compound CX09040. Liraglutide was used as the positive control drug. The amount and the size of islets observed in pathological sections were calculated to evaluate the β cell mass; the glucose stimulated insulin secretion (GSIS) test was applied to estimate the β cell secretary function; the oral glucose tolerance test (OGTT) was taken to observe the glucose metabolism in mice; the expressions of protein in pancreas were detected by Western blotting. The effects on the target protein tyrosine phosphatase 1B (PTP1B) were assessed by the PTP1B activities of both recombinant protein and the intracellular enzyme, and by the PTP1B expression in the pancreas of mice, separately. As the results, with the treatment of CX09040 in alloxan-induced β cell dysfunction mice, the islet amount (P<0.05) and size (P<0.05) increased significantly, the changes of serum insulin in GSIS (P<0.01) and the values of acute insulin response (AIR, P<0.01) were enhanced, compared to those in model group; the impaired glucose tolerance was also ameliorated by CX09040 with the decrease of the values of area under curve (AUC, P<0.01). The activation of the signaling pathways related to β cell proliferation was enhanced by increasing the levels of p-Akt/Akt (P<0.01), p-FoxO1/FoxOl (P<0.001) and PDX-1 (P<0.01). The effects of CX09040 on PTP1B were observed by inhibiting the recombinant hPTP1B activity with IC50 value of 2.78x 10(-7) mol.L-1, reducing the intracellular PTP1B activity of 72.8% (P<0.001), suppressing the PTP1B expression (P<0.001) and up-regulating p-IRβ/IRβ (P<0.01) in pancreas of the β cell dysfunction mice, separately. In conclusion, compound CX09040 showed significant protection effects against the dysfunction of β cell of mice by enlarging the pancreatic β cell mass and increasing the glucose-induced insulin secretion; its major mechanism may be the inhibition on target PTP1B and the succedent up-regulation of β cell proliferation.
Alloxan ; Animals ; Benzoates ; pharmacology ; Biological Assay ; Disease Models, Animal ; Glucose ; metabolism ; Glucose Tolerance Test ; Insulin ; secretion ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; Liraglutide ; pharmacology ; Mice ; Mice, Inbred ICR ; Molecular Weight ; Pancreas ; drug effects ; enzymology ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; antagonists & inhibitors ; Signal Transduction

Arm ; Fibroma ; Histiocytoma, Malignant Fibrous ; Humans ; Soft Tissue Neoplasms

OBJECTIVETo identify the survival prognostic factors and clinical outcome of the patients with spinal metastatic tumors and to discuss the surgical treatment strategy of spinal metastatic tumors.

METHODSThe patients with spinal metastatic tumors who received surgeries during January 2003 to June 2012 were enrolled. The survival was analyzed by Kaplan-Meier survival curve. The prognostic factors, divided into patient-related factors, tumor-related factors and therapy-related factors, were analyzed univariately and multivariately by Cox comparative hazard model.

RESULTSThere were 453 patients were enrolled in research including 263 male and 190 female patients with an average age of (56 ± 13) years (10-86 years). The median postoperative survival was 9 months. Local recurrences and peri-operative complications were found in 78 (17.2%) and 72 (15.9%) patients, respectively. Univariate analysis showed the significant prognostic factors for postoperative survival included poor preoperative general condition (χ(2) = 4.16), severe preoperative neurologic deficit(χ(2) = 10.23), not receiving bisphosphonate therapy(χ(2) = 10.47), short disease-free interval before spinal metastasis (χ(2) = 23.31), spinal metastasis as the first manifestation (χ(2) = 10.94), rapid-growth primary tumor(χ(2) = 15.45), visceral metastasis (χ(2) = 4.10), not receiving postoperative radiotherapy(χ(2) = 18.10) and not receiving post-operative sensitive systemic therapy(χ(2) = 11.20) (P < 0.05). Multivariate analysis showed the independent prognostic factors include severe preoperative neurologic deficit (P = 0.012, 95%CI: 1.11-2.30), short disease-free interval before spinal metastasis (P = 0.023, 95%CI:1.05-1.83), rapid-growth primary tumor (P = 0.000, 95%CI:1.74-3.06), visceral metastasis (P = 0.008, 95%CI: 1.08-1.68), not receiving postoperative radiotherapy (P = 0.000, 95%CI:1.38-2.35) and not receiving post-operative sensitive systemic therapy (P = 0.045, 95%CI:1.01-1.58).

CONCLUSIONThe prognostic factors for survival are useful for determining the indication of operation and improving survival and clinical outcome for patients with spinal metastatic tumors.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Spinal Neoplasms ; diagnosis ; secondary ; surgery ; Treatment Outcome ; Young Adult

This study was aimed to establish a model for detecting the donor chimerism rate following the multi-donor hematopoietic stem cell transplantations, and simplify its calculation method. Patients with hematologic disease receiving allogeneic hematopoietic stem cell transplantation including single-donor and multi-donor were selected in this study and the donor cell chimerism rates were detected, using STR-PCR combined with capillary electrophoresis. The results indicated that the peaks of the sister alleles coming from the same individual were confirmed to have the approximate areas and can be replaced each other in the situation of mixed chimerism. In the calculation model, the value between reference chimerism and approximate chimerism have no significant difference using the hypothetical peak areas, and the result was confirmed to be accepted basing on typical measurement error between sister alleles (5% - 20%). It is concluded that the areas of share peaks can be replaced by non-share peaks and this conclusion can be used to calculate the double-donor CHM (DD-CHM)(%). Compared to the D alleles, R alleles show more strategic importance because it can lead to more accurate result and allowed simplifying the arithmetic calculations for DD-CHM(%).
Alleles ; Electrophoresis, Capillary ; Hematopoietic Stem Cell Transplantation ; Humans ; Polymerase Chain Reaction ; Postoperative Period ; Tissue Donors ; Transplantation Chimera ; genetics ; Transplantation, Homologous

OBJECTIVETo observe the effect of vascular endothelial growth factor (VEGF) on bone marrow-derived mesenchymal stem cell (MSC) proliferation and explore the signaling mechanism involved.

METHODSMSC culture was performed following the classical whole bone marrow adhering method. The characteristics of MSC were identified by induction of multi-lineage differentiation and flow cytometry for surface marker analysis (CD34, CD45, CD29, and CD90). Following the addition of 50 nmol/L wortmannin, 50 µmol/L PD98059, 30 µmol/L SB203580, 10 µmol/L H89, 20 µmol/L Y27632, 1 µmol/L rapamycin, 10 µmol/L straurosporine, 6 nmol/L Go6976, or 50 µmol/L Pseudo Z inhibitors in the cell culture, the MSC were treated with 20 ng/ml VEGF and the changes of the cell proliferation rate was measured with MTT assay.

RESULTSCultured MSC were capable of multi-linage differentiation and did not express VEGF-R, CD29 or CD90. Treatment with 20 ng/ml VEGF obviously promoted MSC proliferation, and this effect was inhibited partially by p38 mitogen-activated protein kinase (MAPK) inhibitor rapamycin, PD98059, SB203580, Go6976, and straurosporine.

CONCLUSIONSVEGF promotes MSC proliferation in close relation to the AKT-PKC pathway, in which PKC signal pathway may play the central role.

Animals ; Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Protein Kinase C ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Vascular Endothelial Growth Factor A ; pharmacology

This study was conducted to produce pectin-doxorubicin conjugate (PDC) nanosuspensions by high-pressure homogenization, and investigating the physico-chemical properties, the cumulative release rate in vitro and in vivo, and the anti-tumor activity. The major production parameters such as pressure, cycle numbers and types of stabilizers on the mean particle size and polydispersity index (PI) of PDC nanosuspensions were investigated. The cumulative release rate in phosphate buffer saline (PBS) at pH 5.1 or 7.0 were studied. The concentration of doxorubicin (DOX) in plasma of rabbit were recorded after intraperitoneal injection of PDC nanosuspensions (DOX was equivalent to 10mg·kg^-1) or DOX (10mg·kg^-1). We established an animal model of the nude mice with SKOV₃ cell, and injected the PDC nanosuspensions (DOX was equivalent to 10, 5, 2.5 mg·kg^-1) in the first day, and observed the growth state of nude mice. The particle size of PDC nanosuspensions was 118.8±6.93 nm, PI was 0.14±0.03, as well as the zeta potential was -27.2±0.36mV. It shows that no drug release was found in PBS at pH 7.4. About 40% cumulative release was determined in PBS at 5.1 after 30 h. The concentration of DOX in plasma of PDC group was 60 ng·mL^-1, and was lower than that of DOX group. Compared with control group, high-dose-group decreased the weight of nude mice's ascites tumor and burrknot. PDC nanosuspensions can inhibit the growth of SKOV₃ cell line in nude mice. In summary, PDC nanosuspensions are target-specific drugs with high efficiency and low toxicity in the ascites cancer model.

Objective To investigate the effects of microRNA( miR)-29a-3p on the proliferation and invasion of gastric cancer cells and analyze its related molecular mechanism. Methods The expression level of miR-29a-3p in gastric cancer cells was detected, and the role of miR-29a-3p in the proliferation, migration, and invasion of gastric cancer cells was evaluated. Western blotting and luciferase analysis showed that miR-29a-3p was directly bound to Serpinhl 3 ' -untranslated region(3' UTR). In addition, the effects of the miR-29a-3p/Serpinhl axis on the proliferation, migration, and invasion of gastric cancer cells were detected by MTT assay, colony formation assay, and Transwell assay in vitro. Results After transfection, the expression of miR-29a-3p in the miR-29a-3p mimic group was significantly higher than that in the miR-29a-3p negative control and blank group. After transfection, the proliferation of BGC823 cells decreased significantly. Luciferase analysis showed that miR-29a-3p inhibited the expression of Serpinhl by targeting the 3 ' UTR of Serpinhl. In addition, overexpression of miR-29a-3p significantly inhibited the proliferation, invasion, and migration of gastric cancer cells by targeting Serpinhl. Conclusion MiR-29a-3p can target Serpinhl and regulate the proliferation and invasion of gastric cancer cells.