BACKGROUND:Cel cryopreservation is required for clinical use of stem cel s, and the current process of cryopreservation however may be harmful to cel viability, pluripotency and differentiation capacity.
OBJECTIVE:To explore the effect of fructose and dithiothreitol on pluripotency and osteogenesis of cryopreserved bone marrow mesenchymal stem cel s.
METHODS:Bone marrow mesenchymal stem cel s were isolated from the bone marrow of Sprague-Dawley rats and pretreated with fructose (200μmol/L), dithiothreitol (500μmol/L) or combined components before cryopreservation. Then the cel s were cryopreseved for 6 months and the morphology of cel s was observed by inverted microscopy. The cel viability was evaluated by MTT, and real-time PCR was used to detect the mRNA expression of Nanog, OCT4 and Sox2. Alkaline phophatase activity assay and alizarin red staining were utilized to detect the osteogenic capacity of bone marrow mesenchymal stem cel s.
RESULTS AND CONCLUSION:Images captured by inverted microscopy showed no significant difference in cel morphology between groups. The MTT results indicated that fructose and combined pretreatment could promote the cel viability of bone marrow mesenchymal stem cel s after cryopreservation, while the real-time PCR results demonstrated that dithiothreitol significantly facilitated the expression of Naogo and Sox2 in bone marrow mesenchymal stem cel s. Moreover, ALP activity assay and alizarin red staining confirmed the positive effects of fructose, dithiothreitol and combined pretreatment on osteogenic capacity of bone marrow mesenchymal stem cel s after cryopreservation, and the best effects were found after pretreatment with dithiothreitol and combined components. Overal , these findings indicate that fructose pretreatment is beneficial for cel viability of cryopreseved bone marrow mesenchymal stem cel s, and dithiothreitol contributes to maintaining the pluripotency and osteogenesis capacity of cryopreseved bone marrow mesenchymal stem cel s.

Objective To assess the feasibility and curative effects of laser surgery in treatment of early-stage glottic carcinoma under self-retaining laryngoscope with nasal endoscopic assistance.Methods Thirteen cases of glottic carcinoma(TisN0M0:2 cases;T1aN0M0:7 cases;T1bN0M0:3 cases;T2N0M0:1 case) extirpated with Nd:YAG laser or Diomed laser from June 1999 to March 2003 were retrospectively analyzed.Because of the poor glottic exposure with self-retaining laryngoscope under general anesthesia,various angular nasal endoscopes were used.The tumor was distinctly displayed on monitor screen with nasal endoscopic assistance and were completely removed through vapourization or removed with Nd:YAG laser or Diomed laser.Results The follow-up period was three to five years.Two patients were found to have local recurrence.One patient received treatment with Nd:YAG laser surgery under electronic laryngoscope with local and superfacial anesthesia,and the patient was followed up for three years without showing recurrence.The other patient underwent vertical hemilaryngectomy and was followed up for three and half years without showing recurrence.All 13 cases were followed up for three years,and all of them were alive.Three-year survival rate was 100%(13/13).When they were followed up for five years,one patient lost track(counted as death),therefore the five-year survival rate was 92.3%(12/13).Conclusions With the assistance of various angular nasal endoscope,the glottic tumors can be exposed distinctly under self-retaining laryngoscope and removed completely.Nd:YAG laser or Diomed laser is an effective and safe procedure with minimal invasion for the treatment of early-stage glottic carcinoma.The advantages of this technique include lower complication rate and high rate of preservation of laryngeal functions,avoidance of tracheotomy and the sequela of laryngeal fissure.

Objective To study the effects of osteopontin (OPN) on the nuclear mechanics of bone marrow-derived mesenchymal stem cells (BMSCs) as well as its involved mechanisms. Methods The BMSC migration was evaluated using the Transwell assay. An atomic force microscope (AFM) was used to determine the elastic modulus of the BMSC nucleus and analyze the changes in the nuclear mechanics of the BMSCs after treatment with OPN. The activation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase1/2 (ERK1/2) was measured by Western blot. The role of the FAK-ERK1/2 signaling pathway in mediating the OPN-affected BMSC nuclear mechanics was investigated by employing a specific inhibitor. RT-PCR and Western blot were used to detect the expression of Lamin A/C at mRNA and protein levels in the BMSCs, respectively. Results The elastic modulus of the BMSC nucleus exhibited a significant decrease after OPN treatment compared with that of the control group. OPN could upregulate the phosphorylation level of FAK and ERK1/2, but the inhibitor of FAK or ERK1/2 restored the OPN-decreased elastic modulus of the BMSC nucleus and inhibited the BMSC migration significantly. After treatment with OPN, the expression of Lamin A/C in the BMSCs reduced significantly, and such a reduced expression could be suppressed by the inhibitor of FAK or ERK1/2. Conclusions OPN could probably downregulate the expression of Lamin A/C of the BMSCs via the FAK-ERK1/2 signaling pathway, decrease the stiffness of the BMSC nucleus, and promote the migration of the BMSCs. The research outcomes provide the experimental evidence for further understanding the mechanism of the OPN-regulated BMSC migration and its potential clinical application.

PURPOSE: Several studies have compared the effects of coronary stenting and coronary-artery bypass grafting (CABG) on left main coronary artery (LMCA) disease. However, there are limited data on the long-term outcomes of these two interventions in diabetic patients. MATERIALS AND METHODS: We evaluated 56 patients with LMCA stenosis who underwent drug-eluting stent (DES) implantation and 116 patients who underwent CABG in a single hospital in China between January 2004 and December 2006. We compared long-term major adverse cardiac events (death; a "serious outcome" composite of death, myocardial infarction, or stroke; and target-vessel revascularization). RESULTS: In-hospital (30-day) mortality was 0% for the DES group and 3.4% for the CABG group (p=0.31). There was no difference between the two groups in terms of risk of death [hazard ratio for stenting group, 0.49; 95% confidence interval (CI), 0.13-1.63; p=0.55] or risk of serious outcome (hazard ratio for DES group, 1.11; 95% CI, 0.39-1.45; p=0.47). The target-vessel revascularization rate was higher in the DES group than in the CABG group (hazard ratio, 3.67; 95% CI, 1.24-11.06; p=0.018). CONCLUSION: In this cohort of diabetic patients with LMCA stenosis, there was no difference in composite endpoints between patients receiving DESs and those undergoing CABG. However, stenting was associated with higher rates of target-vessel revascularization than CABG. DES implantation in diabetic patients with LMCA disease was found to be at least as safe as CABG.
Angioplasty, Balloon, Coronary/*methods ; Coronary Stenosis/*therapy ; Diabetes Mellitus ; *Drug-Eluting Stents ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome