Objective To observe the effects of mesenchymal stem cells (BMSCs) on the expression of Bcl2 and Bax in atrophied muscles.Methods Immature rats (80～ 100 g) were anesthetized to collect marrow from their femurs and tibias.BMSCs were isolated from the marrow,cultured and purified using the whole bone marrow adherence method.Their right hindlimbs were immobilized fiom the thigh to the paw with the knee in extension and the ankle in plantar flexion.After the modeling,24 of the male rats were divided into a sham-operation group,a BMSC group and a phosphate buffer solution (PBS) group,each of 8.The BMSC and PBS groups were injected with either approximately 106 BMSCs or an equal volume of PBS into the belly of the soleus muscle after they had been immobilized for 48 hours,while the control group did not undergo any treatment except for the injection of PBS.All of the rats were sacrificed for analysis after 14 days.Results The BMSCs were mainly spindle cells,showing radial colony arrangement.They grew vigorously and could passage in a continuous and stable manner over 10 passages.At the 4th passage the BMSCs were positive for CD44 (95.84％) and CD90 (96.00％),but negative for CD34 and CD45.Western blotting assay demonstrated that the expression of Bax protein as measured in grey-scale value (0.41±0.08)in the BMSC group was significantly lower than in the PBS group (0.63±0.10),but significantly higher than in the control group (0.14±0.11) on average.The expression of Bcl-2 (0.47±0.14) was also significantly higher in the BMSC group than in the PBS group (0.22-± 0.13),but significantly lower than in the control group (0.81 ± 0.06).Conclusion Bone marrow mesenchymal stem cells can upregulate the expression of anti-apoptotic Bcl-2 protein and downregulate that of the apoptotic Bax protein when injected early into the belly of a muscle in an immobilized limb.

Objective To observe the effects of treadmill training at different intensities on the expression of p-AKT in rats' gastrocnemius muscles after focal cerebral ischemia,and to investigate whether intensive training is beneficial for the recovery of motor function.Methods Left middle cerebral artery occlusion (MCAO) was induced in 120 male Wistar rats using the intraluminal thread method,and they were divided into an MCAO group (no training),a normal training group (treadmill training once a day for 30 min) and an intensive training group (treadmill training twice a day for 60 min),each of 30 rats.There was also a sham control group with 30 members not given MCAO or training.The four groups were further divided into 3 day,7 day and 14 day subgroups.Five rats randomly selected from each subgroup were sacrificed for hematoxylin-eosin staining after 4％ paraformaldehyde treatment.Neurological function was evaluated using Zausinger scores,and the expression of p-Akt was detected by western blotting.Results No significant differences in Zausinger scores were observed between the intensive training group and the normal training group after 1,3 or 7 days of training.However the average Zausinger score in the intensive training group was significantly higher than in the normal training group after 14 d of treadmill training.After 7 d and 14 d of treadmill training the average cross-sectional area of the gastrocnemius muscles in the sham group was significantly higher than in the other three groups.The average area of the intensive training group was significantly larger than that of the normal training group.The expression of p-Akt in the gastrocnemius was significantly increased in the intensive training group compared with the normal training group in the 7 day and 14 day subgroups.Conclusion Treadmill training can improve the expression of p-Akt in atrophied gastrocnemius muscles caused by MCAO.Intensive training is more effective for the recovery of muscle function.

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Phosphoglycerate mutase (PGAM) is a key enzyme in glycolytic pathways. With PCR technique based on an EST identified in our lab, a novel gene named SjPGAM (GenBank Accession No. EU374631) was cloned. Sequence analysis revealed that the ORF of SjPGAM gene contained 753 nucleotides, encoding 250 amino acids, and the molecular weight was about 28.26 kD. Real-time PCR analysis showed that the mRNA level of SjPGAM was much higher in the 14 days and 19 days schistosomula than other stages, suggesting that the gene was a schistosomula stage differential expression gene. The SjPGAM cDNA fragment was subcloned into an expression vector pET-28a (+) and transformed into Escherichia coli BL21 cells. In the presence of IPTG, the 31 kD fusion protein was expressed in included bodies. Western blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation. The study provides important basis for investigating the mechanism of the PGAM in the glycolytic pathways of Schistosoma japonnicum.
Animals ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Male ; Phosphoglycerate Mutase ; genetics ; immunology ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Schistosoma japonicum ; enzymology ; genetics ; Schistosomiasis japonica ; immunology ; parasitology

Objective To evaluate the efficacy and safety of tigecycline-based treatment approach on severe infection of patients with hematological diseases. Methods The clinical data of 64 patients who were treated with tigecycline-based treatment approach for severe infection were retrospectively reviewed. The curative effect was evaluated, meanwhile the drug side effects were observed. Results A total of 51 strains of bacteria were isolated from 64 patients, including 12 extended-spectrum β-lactamase(ESBL)and 15 multi-drug resistant strains and the total effective rate was 59.4%(38/64). Five patients diagnosed as carbapenem resistant infection and were treated with the addition dose of tigecycline and 3 patients relieved. Main adverse events were nausea, vomiting, diarrhea and hepatic dysfunction, but all events were slight. Conclusions Tigecycline-based treatment approach has a good clinical efficacy in treating severe infection of patients with hematological diseases, and the side effect is few.Tigecycline-based treatment approach could be used as a new choice for patients non-responding favorably to conventional anti-infective treatment or multiple resistant bacteria.

OBJECTIVETo explore the relationship between plasma microRNA126 (miR-126) level and coronary collateral circulation (CCC) formation and to determine whether the miR-126 in plasma could serve as a blood-based biomarker for CCC in patients with severely narrowed coronary arteries (CAD).

METHODSIn this prospective study, a total of 120 consecutive CAD patients with ≥ 95% stenosis in one epicardial coronary artery were enrolled. Thirty healthy people served as normal control. They were divided into two groups according to Rentrop grades: patients with grade 2 and 3 collateral development (good CCC group, n = 64) and patients with grade 0 and 1 collateral development (poor CCC group, n = 56). Plasma miR-126 was measured by RT-PCR and serum VEGF was evaluated by ELISA method.

RESULTSFasting plasma glucose (FPG) was significantly lower in patients with good CCC than in patients with poor CCC ((5.99 ± 1.48) mmol/L vs. (6.40 ± 2.50) mmol/L). Plasma miR-126 levels and VEGF levels were significantly lower in CAD patients than in healthy people (0.04 ± 0.01 vs. 0.07 ± 0.02, P = 0.023 and (2 110 ± 455) ng/L vs. (2 574 ± 450) ng/L, P = 0.011, respectively). miR-126 and VEGF levels were significantly higher in good CCC group than in poor CCC group (miR-126: 0.06 ± 0.02 vs. 0.03 ± 0.01, P = 0.021;VEGF:(2 549 ± 614) ng/L vs. (1 759 ± 452) ng/L, P = 0.008) . In CAD patients with good CCC, the miR-126 level was positively correlated to the VEGF expression (r = 0.712, P = 0.005) while there was no correlation between miR-126 level VEGF in CAD patients with poor CCC (r = 0.342, P = 0.483) . Multivariate analysis revealed that plasma miR-126 (OR = 2.145, 95% CI 1.691-2.988, P = 0.001) and VEGF (OR = 1.279, 95% CI 1.068-2.295, P = 0.013) were independent predictors of collateral formation in patients with severely narrowed coronary arteries. In CAD patients, the area under the miR-126 ROC curve is 0.951 (P = 0.002).

CONCLUSIONPlasma miR-126 level is positively correlated to the CCC formation and is an independent predictor of CCC development in patients with severely narrowed coronary arteries, suggesting that plasma miR-126 might be a useful new, stable blood biomarker for predicting CCC formation in patients with severely narrowed coronary arteries.

Biomarkers ; Collateral Circulation ; Coronary Angiography ; Coronary Artery Disease ; blood ; Coronary Circulation ; Coronary Disease ; Heart ; Humans ; MicroRNAs ; blood ; Multivariate Analysis ; Plasma ; Prospective Studies ; ROC Curve