Objective To explore the immunosuppression effects,outcomes and clinical significance of mesenchymal stem cells (MSCs) transplantation in burn treatment by comparing the levels of WBC,C-reaction protein (CRP),interferon-γ(IFN-γ),interferon-o (TNF-α),interleukin-6 (IL-6),interleukin-10 (IL-10) between control burn models and models conducting MSCs transplantation.Methods After stripping Wharton's Jelly from the neonatal umbilical cord,MSCs was cultured and expanded.Burn models were constructed in male SD rats weighted at (200± 5) g,and randomly grouped for control and MSCs transplantation.The rats in transplantation group were injected subcutaneously with hUCMSCs (1.5× 106) at 24 h after burning.The content of WBC,CRP,IFN-γ,TNF-α,IL-6,IL-10 in blood samples at 0,1,2,3,5 and 7 d after burning were detected by enzyme-linked immunosorbent assay (ELISA).The ELISA results,the wound healing rate at 7,14,21 and 28 d,as well as wound healing time were compared and analyzed statistically by ANOVA between the two groups.Results WBC in control group increased significantly at 1 and 2 d,and CRP in control group increased substantially at 2 and 3 d.IFN-% IL-6 and IL-10 levels in serum showed increase on 5 d,and TNF-α arrived at its peak value at 7 d.By contrast,WBC,CRP,TNF-α,IL-6 and IL-10 of the MSCs transplantation group showed slightly increase after burning and the differences were convinced by statistical analysis,while IFN-γ showed little difference among the two groups.The MSCs transplantation group also showed significantly higher wound healing rate at 14,21,28 d and shorter wound healing time than that of the control.Conclusions MSCs transplantation can suppress the over-inflammatory response by significantly mediating the inflammatory cytokines as WBC,CRP,TNF-α,IL-6,IL-10 in burn.IFN-γseems not affected by MSCs transplantation.MSCs would be suitable to promote wound healing and repair in burn,which is achieved not only by differentiation and paracrine,but also by subtle immunoregulation.

Objective To observe the differentiation of bone marrow multipotent adult progenitor cells(MAPCs)into skin tissue cells of rats in vivo. Methods Magnetic activated cell sorting (MACS)was used to remove MAPCs from bone marrow of male rats through negative screening.Tail vein injection with combined with MAPCs Was done in C57BL/6 mice with skin wound and nude mice with immunodeficieney.Immunohistochemical staining was used to examine the expression of MHCI antigen in the healed skin of donor SD rats. Results Positive MHCI cells were found in the epidermal fundus and Some hair follicle-like structures of the healed skin of C57BL/6 mice.Hair follicle-like structure appeared in the healed skin of nude mice group,in which positive MHCI cells were found in the basal epidermal and some hair follicle-like structures. Conclusions During skin damage,MAPCs can migrate to the injured skin area and skin adnexa hair foilicle area,transform into epidermal cells and hence participate in the healing of the wound skin.

Stem cells have self-renewal and differentiation potential.Cyclin-dependent kinase (CDK) plays an important role in promoting pluripotency and self-renewal.The crucial activity of S-phase,DNA replication,presents a unique opportunity during the cell cycle for the genetic and epigenetic regulation that may be involved in stabilizing the pluripotent state.It is also clear that Myc acts to coordinate both the cell cycle and the pluripotency transcription network in stem cells.Here we review the regulating mechanisms of stem cell cycles and pluripotency by CDK and Myc to help researchers obtain a better understanding of mutual regulation of the cell cycle and the pluripotent state by CDK and Myc which may be exploited in regenerative medicine.

0.05). Conclusion The technique of PEP-MN-PCR from single-cell for simultaneous detection of HLA-A, B, DR types were efficient and accurate. It was possible to apply the techniques of PEP-MN-PCR to detect HLA-A, B, DR typing for preimplantational diagnosis, preliminary selection of HLA matching embryos and the patient sibling who needs cord blood stem cell transplantation.

Objective To investigate the protective effect of agmatine (AGM) against peritoneal inflammatory response and neutrophil (PMN) infiltration induced by zymosan (ZYM) in mice. Methods Thirty-six adult male C57BL/6 mice were randomly divided into sham group, model group, and AGM treatment group. Peritonitis model was reproduced by intra-peritoneal injection of 1 mg/mL ZYM (0.5 mL), while equivalent phosphate buffer saline (PBS) was given to sham group. 200 mg/kg AGM was injected into peritoneal cavity after ZYM challenge in AGM treatment group. Six mice in each group were sacrificed at 2 hours and 6 hours, respectively, after reproduction of the model. Blood sample and peritoneal lavage fluid (PLF) were collected. The levels of keratinocyte-derived chemokine (KC), macrophage inflammatory protein 2 (MIP-2), tumor necrosis factor-α (TNF-α), interleukins-6 (IL-6) in serum and PLF were determined by enzyme linked immunosorbent assay (ELISA). The number of leukocytes and PMN in PLF were determined by hemocytometer and flow cytometry, respectively. Results Compared with sham group, all serum and PLF levels of KC, MIP-2, TNF-α and IL-6 were greatly elevated at 2 hours after ZYM injection in model group, while AGM treatment could dramatically reduce the levels of the above-mentioned cytokines in serum and PLF as compared with those of the model group [serum KC (ng/L): 990.7±137.9 vs. 2 053.2±262.7, MIP-2 (ng/L): 642.2±124.4 vs. 1 369.7±146.5, TNF-α (ng/L): 608.6±38.1 vs. 1 044.7±101.0, IL-6 (ng/L): 1 058.2±129.1 vs. 1 443.3±190.1; PLF KC (ng/L): 7 462.3±839.6 vs. 12 723.5±1 515.7, MIP-2 (ng/L): 1 570.8±193.4 vs. 3 471.4±384.7, TNF-α (ng/L): 1 115.8±156.7 vs. 1 499.2±231.2, IL-6 (ng/L): 2 646.5±223.2 vs. 3 126.7±291.4; all P < 0.05]. The expressions of KC, MIP-2 and TNF-α at 6 hours were significantly lower than those at 2 hours in model group and AGM treatment group, but IL-6 levels were further increased. The levels of KC and MIP-2 in serum and PLF at 6 hours were decreased to the levels of sham group. At 6 hours after the reproduction of the model, the number of total inflammatory cells and PMN of PLF in the model group was significantly higher than those of the sham group. In contrast, AGM notably lowered the number of inflammatory cells and PMN in peritoneal fluid after ZYM attack [total inflammatory cells (×109/L): 14.7±1.1 vs. 2.0±0.4, 10.1±1.2 vs. 14.7±1.1; PMN (×109/L): 11.37±1.22 vs. 0.18±0.05, 7.69±0.57 vs. 11.37±1.22, all P < 0.05]. Conclusion AGM can effectively alleviate acute peritoneal inflammatory injury induced by ZYM, mainly through reducing the secretion of inflammatory mediators and chemokines, and inhibiting the infiltration of leukocytes and neutrophils.

Objective: To evaluate the effects of 650 nm-10.6 μm combined laser in patients with knee Osteoarthritis (OA) and to determine whether the combined laser provides greater pain relief and improved function compared with red light. Methods: Forty-eight patients with knee OA were randomly allocated to two groups (24 per group), receiving 20 rain irradiation with 650 nm -10.6 μm combined laser or red light emitting diode respectively on point Dubi (ST 35) 3 times a week for the first course (2 weeks) and twice a week for the second one (4 weeks). The main outcome measures were WOMAC (Western Ontario and McMaster Universities Osteoarthritis Index) scores. In addition, patients' global assessment, adverse effects and validation of patient blinding were analyzed. Results: All the patients completed the first course, but 12 were lost during the second one. Due to the high dropout rate by the second course, only the data acquired from the first course could be analyzed. No differences of general data of patients and WOMAC scores were found in between-group comparison before treatment (P>0.05). The WOMAC scores of patients in both combined laser group and red light group reduced significantly compared to baseline by the end of the first course (P<0.01). There were no significant differences on the reduction rate of WOMAC scores between two groups (P>0.05). Neither the patients' global assessment nor the dropout rate showed statistical differences between two groups (P>0.05). There was no difference between two groups in patients correctly guessing the treatment assignment (P>0.05). There was no significant difference in the reduction rate of WOMAC scores and the patients' global assessment between patients who guessed their assignment (P>0.05). Conclusion: Both combined laser and red light irradiation are beneficial to patients with knee OA. But as the statistical indifferences between two groups, the authors can't conclude from this study whether the combined laser is more effective.

Proteasome is a multi-subunit protease complex in eukaryocytes, and plays an important role in ubiquitin-proteosome pathway. Recombinant proteasome can be used to screen proteasome inhibitors. In this study, recombinant plasmid of pET28a-PSMB1 was constructed by inserting human proteasome catalytic subunit (PSMB1) cDNA (726 bp) into the prokaryotic expression vector pET28a(+), and transforming the plasmid into E. coli BL21(DE3) cells for expression. After overnight induction (1 mmol/L IPTG, 20 degrees C), an expected protein band with molecular weight of 27 kDa was observed on SDS-PAGE gel. The recombinant protein was then purified through affinity chromatography, and the purity is more than 95%. The amino acid sequence of the recombinant protein was validated by NanoLC-MS/MS. The data from in vitro BIAcore analysis showed that the recombinant PSMB1 could bind to celastrol. The binding affinity between PSMB1 and 10 micromol/L celastrol was more than 27RU.
Binding Sites ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Proteasome Endopeptidase Complex ; biosynthesis ; genetics ; isolation & purification ; Proteasome Inhibitors ; isolation & purification ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; Triterpenes ; metabolism ; Ubiquitin

OBJECTIVETo seek possible effect targets of bufalin in HeLa cells by studying the impact of bufalin on cell protein expression profile after treatment on human cervical carcinoma cell lines HeLa.

METHODBufalin's ICs0was measured by MTr assay. The apoptosis of cells was observed by FCM (flow cytometry) and Hoechst 33342 staining assay. Differentiated expression protein spots were founded and identified using proteomic techniques, which could induce HeLa cell apoptosis.

RESULTBufalin showed remarkable cytotoxic effect on HeLa cells. IC50 (154 +/- 21.5) nmol X L(-1) indicated the possibility of inducing cell apoptosis. The protein expression profile showed 11 differentiated expression protein spots. Among the 11 proteins, nudix-type motif 5, vimentin, hnRNP C1/hnRNP C2 variant, HNRPK, HNRPK isoform a variant (two spots are the same protein), heat shock protein 27, macrophage-capping protein, SELENBP1 protein were down-regulated, while ribosomal protein, large, P0 and S-adenosylmethionine synthetase 2 were up-regulated by bufalin treatment. They may be effect targets of bufalin in HeLa cells. Western blotting showed consistent results in heat shock protein 27, vimentin and HNRPK between expression after treatment with bufalin and two-dimensional electrophoresis.

CONCLUSIONBufa-Lin can induce apoptosis in human cervical carcinoma cells HeLa and the effect of bufalin may be related to the joint intervention with multiple protein targets.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Female ; HeLa Cells ; Humans ; Neoplasm Proteins ; metabolism ; Proteomics ; methods ; Uterine Cervical Neoplasms ; drug therapy ; metabolism ; pathology

A model of fluid dynamics related to the myocardial bridginged and mural coronary artery was designed and manufactured according to the physical principle and characteristic of the mural coronary artery. The model can imitate systematically well the effect of myocardial bridging on hemodynamic change of the mural coronary artery under different controlled experimental parameter. The methodology is proved to be feasible and has good prosperity of experimental study.
Coronary Vessels ; physiology ; Hemodynamics ; Humans ; Models, Cardiovascular ; Myocardial Bridging

Hepatocellular carcinoma (HCC) is one of leading causes of death in the world. Although various treatments have been developed, the therapeutic side effects are far from desirable. Chinese medicines (CMs, including plants, animal parts and minerals) have drawn a great deal of attention in recent years for their potential in the treatment of HCC. Most studies have shown that CMs may be able to retard HCC progression with multiple actions, either alone or in combination with other conventional therapies to improve quality of life in HCC patients. Additionally, CMs are used for preventing HCC occurrence. The aim of this study is to review the potential prophylactic and curative effects of CMs on human HCC and the possible mechanisms that underlie these pharmacological actions. Publications were collected and reviewed from PubMed and China National Knowledge Infrastructure from 2000 to 2014. Keywords for literature searches include "Chinese medicine", "Chinese herb", "traditional Chinese Medicine", "hepatocellular carcinoma" and "liver cancer". CMs in forms of pure compounds, isolated fractions, and composite formulas are included. Combination therapies are also considered. Both in vitro and in vivo efficacies of CMs are being discussed and the translational potential to bedside is to be discussed with clinical cases, which show the actions of CMs on HCC may include tumor growth inhibition, antimetastatic activities, anti-inflammation, anti-liver cancer stem cells, reversal on multi-drug resistance and induction/reduction of oxidative stress. Multiple types of molecules are found to contribute in the above actions. The review paper indicated that CMs might have potential to both prevent HCC occurrence and retard HCC progression with several molecular targets involved.
Carcinoma, Hepatocellular ; drug therapy ; prevention & control ; Drug Resistance, Multiple ; Humans ; Liver Neoplasms ; drug therapy ; prevention & control ; Medicine, Chinese Traditional ; NF-E2-Related Factor 2 ; physiology ; Neoplastic Stem Cells ; drug effects ; Reactive Oxygen Species ; metabolism