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Qi Y.,Zhejiang University | Wang Y.,The First Affiliated Hospital | Yan W.,Zhejiang University | Li H.,Zhejiang University | And 2 more authors.
Cell Transplantation | Year: 2012

Repair of segmental bone defects remains a major challenge for orthopedic surgeons. This study aimed to investigate whether recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded calcium sulfate (CS) combined with mesenchymal stem cell (MSC) sheets could accelerate bone regeneration in ulnar segmental defects of rabbits. In vitro, the osteogenic differentiation of MSCs cultured on rhBMP-2-loaded CS was investigated. Forty complete 1.2-cm bone defects were treated with CS (group A), rhBMP-2-loaded CS (group B), MSC sheet-wrapped CS (group C), and MSC sheet-wrapped rhBMP-2-loaded CS (group D). At 4 and 8 weeks after implantation, the samples were treated by X-ray, microcomputed tomography, and histological observation. The rhBMP-2 could be released from the rhBMP-2-loaded CS scaffolds and maintain its bioactivity. The alkaline phosphatase (ALP) of MSCs cultured on rhBMP-2-loaded CS was significantly higher than that of CS at both 7 and 14 days (p < 0.05). The defects treated with MSC sheet-wrapped rhBMP-2-loaded CS showed significantly higher scores by X-ray analysis and more bone formation determined by both histology and microcomputed tomography than the other three groups at both 4 and 8 weeks after implantation (p < 0.05). No significant difference in X-ray score and bone formation was found between groups B and C, both significantly higher than group A (p < 0.05). The results suggested that MSC sheetwrapped rhBMP-2-loaded CS may be an effective approach to promote the repair of segmental bone defects and has great potential for repairing large segmental bone defects in clinic. © 2012 Cognizant Comm. Corp.

Li H.B.,The First Affiliated Hospital
Zhong yao cai = Zhongyaocai = Journal of Chinese medicinal materials | Year: 2011

To study the effects of berberine on hypertensive renal injury model in rats fed by enriched high fat-salt-fructose diet . hypertensive renal injury model was esteblished by feeding enriched high fat-salt-fructose diet for 8 weeks. On the basis of animal blood pressure, hypertensive rats were randomly divided into model group (10 rats, distilled water), captopril group (10 rats,25 mg/kg), berberine high dose group (10 rats, 300 mg/kg) and low dose group (10 rats, 100 mg/kg). These rats were fed by enriched high fat-salt-fructose diet and treated by intragastric administration with drugs for 4 weeks. And normal control group (10 rats) was set Blood pressure was determined at 0, 4, 8, 10, 12 weekend,and after 4 weeks of drugs treatment, getting urine to determine urine protein, taking blood serum to determine blood urea nitrogen, serum creatinine,GHb,MDA and activity of SOD. The content of H2O2 and GSH-Px and activity of CAT in kidney tissues were determined also. Compared with normal control group, blood pressure, urine protein, blood urea nitrogen, serum creatinine and MDA and GHb in serum of model rats obviously increased (P < 0.01), the activity of SOD decreased (P < 0.01), higher content of H2O2 and lower content of GSH-Px and activity of CAT (P < 0.01) in the kidney tissues. Treated with berberine for 4 weeks, elevated blood pressure and heightened levels of urine protein, blood urea nitrogen and serum creatinine in model rats were depressed significantly (P < 0.01), and elevated the activity of SOD, lowed the levels of MDA and GHb in blood serum (P < 0.01). At the same time, berberine increased the activities of GSH-Px and CAT (P < 0.01) and slightly lowed the content of H2O2 in the kidney tissues. Berberine has protecting effects on the hypertensive renal impairment model rats fed by enriched high fat-salt-fructose diet, which are concerned with elevated antioxidant capability in body and kidney tissues.

Sui R.,The First Affiliated Hospital | Zhang L.,Liaoning Medical College
Neurological Research | Year: 2011

Objective: To explore the risk factors for stroke-associated pneumonia (SAP). Methods: A retrospective research study was carried out to investigate the clinical data of 1435 patients admitted to the neurological intensive care unit at our university hospital between 1 January 2000 and 31 December 2009. Results: A multi-factorial analysis produced the following results: (1) SAP is 1.113 times more likely to occur for each 1-year increase in age; (2) diabetic patients are 1.612 times more likely to develop SAP than nondiabetic patients; (3) the incidence of SAP decreases by a factor of 0.890 with a one-point increase in the Glasgow coma scale score; (4) nasal feeding patients are 4.981 times more likely to develop SAP than nonnasal feeding patients; (5) patients who use H2-receptor blocking agents are 2.837 times more likely to develop SAP than those who do not; (6) patients who preventively use antibiotics are 2.675 times more likely to develop SAP than those who do not; (7) patients whose hospitalization periods are.> 20 days are 0.500 times more likely to develop SAP than those who do not; (8) patients who suffer from tracheal intubation are 2.980 times more likely to develop SAP than those who do not; and (9) patients who suffer from tracheal incision are 2.190 times more likely to develop SAP than those who do not. Conclusions: SAP was more closely related with diabetes, age, consciousness, days of hospitalization, tracheal intubation, tracheal incision, nasal feeding treatment, and the application of H2-receptor blocking agents and antimicrobials. © W. S. Maney & Son Ltd 2011.

Wang Q.,University of Sichuan | Wang Q.,Lovelace Respiratory Research Institute | Shi S.,University of Sichuan | Shi S.,Lovelace Respiratory Research Institute | And 6 more authors.
Oncotarget | Year: 2014

The elucidation of chemoresistance mechanisms is important to improve cancer patient survival. In this report, we investigated the role and mechanism through which receptor-interacting protein 1 (RIP1), a mediator in cell survival and death signaling, participates in cancer's response to chemotherapy. In lung cancer cells, knockdown of RIP1 substantially increased cisplatin-induced apoptotic cytotoxicity, which was associated with robust JNK activation. The expression of the JNK inactivating phosphatase, MKP1, was substantially reduced in RIP1 knockdown cells. Although MKP1 protein stability was not altered by RIP1 suppression, the synthesis rate of MKP1 was dramatically reduced in RIP1-suppressed cells. Furthermore, we found that the expression of miR-940 was substantially increased in RIP1 knockdown cells. Knockdown of miR-940 restored MKP1 expression and attenuated cisplatin-induced JNK activation and cytotoxicity. Importantly, ectopic expression of MKP1 effectively attenuated cisplatin-induced JNK activation and cytotoxicity. In addition, activation of the JNK upstream signaling kinase, MKK4, was also potentiated in RIP1 knockdown cells. Altogether, our results suggest that RIP1 contributes to cisplatin resistance by suppressing JNK activation that involves releasing miR-940-mediated inhibition of MKP1 and suppressing activation of MKK4. Intervention targeting the RIP1/miR-940/ MKP1/JNK pathway may be used to sensitize platinum-based chemotherapy.

Luo J.,The First Affiliated Hospital
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology | Year: 2012

To construct a lentiviral vector expressing small-hairpin RNA(shRNA) targeting SPARC gene and investigate its silenced effect on SPARC in human myelodysplastic syndromes(MDS) cell line SKM-1. The targeting sequence of SPARC gene which can be effectively silenced in RNA interference was confirmed in our previous study. The designed and synthesized single-stranded primers were annealed to double-stranded oligo sequences and subcloned into linear pGCSIL-GFP lentiviral plasmid digested by enzyme Age I and EcoR I to produce GC-shSPARC lentiviraL vector. After being identified by PCR and sequencing, plasmids GC-shSPARC with pHelper 1.0 and pHelper 2.0 were cotransfected into 293T cells to package lentiviral particles. The recombinant lentiviral vector was transfected into human SKM-1 cells, transfection efficiency was evaluated with expression of green fluorescent protein(GFP) determined by fluorescent microscope. Expression of SPARC in SKM-1 cells was detected using RT-PCR and Western blotting. A recombinant lentiviral vector, GC-shSPARC, expressing shRNAs targeting SPARC gene was constructed and confirmed by DNA sequencing. The recombinant lentivirus was harvested from 293T cells with a viral titer of 1×10(9); TU/mL. GFP was observed in the 70% of SKM-1 cells after transfection. Expression of SPARC mRNA and protein was significantly reduced in the GC-shSPARC transfected group than that in the control group (P<0.05). The lentivirus RNAi vector targeting SPARC has been successfully constructed, and can effectively inhibit the expression of SPARC in SKM-1 cell line, which shed light on the foundation for researching the inhibition of SPARC siRNA target against human MDS cells proliferation, induction apoptosis and gene therapy.

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