To investigate effect of the transplantation of mesenchymal stem cells (MSCs) in combination with nerve growth factor (NGF) on the repair of spinal cord injury (SCI) in adult rats, spinal cord of adult rats (n= 32) was injured by using the modified Allen's method. One week after the injury, the injured cords were injected with Dubecco-modified Eagles medium (DMEM, Group I), MSCs (Group II), NGF (Group III), and MSCs plus NGF (Group IV). One month and two months after the injury, rats were sacrificed and their injured cord tissues were sectioned for the identification of the transplanted cells. The axonal regeneration and the differentiation of MSCs were examined by immunocytochemical staining. At the same time, rats were subjected to behavioral tests by using the open-field BBB scoring system. Immunocytochemical staining showed that axonal regeneration and the transplanted cells partially expressed neuron-specific nuclear protein (NeuN) and glial fibrillary acidic protein (GFAP). At the same time, significant improvement in BBB locomotor rating scale (P<0. 05) were observed in the treatment group. More importantly, further functional improvement were noted in the combined treatment group. MSCs could differentiate into neurons and astrocytes. MSCs and NGF can promote axonal regeneration and improve functional recovery. There might exist a synergistic effect between MSCs and NGF.

To investigate effect of the transplantation of mesenchymal stem cells (MSCs) in combination with nerve growth factor (NGF) on the repair of spinal cord injury (SCI) in adult rats, spinal cord of adult rats (n= 32) was injured by using the modified Allen' s method. One week after the injury, the injured cords were injected with Dubecco-modified Eagles medium (DMEM , Group Ⅰ), MSCs (Group Ⅱ), NGF (Group Ⅲ), and MSCs plus NGF (Group Ⅳ). One month and two months after the injury, rats were sacrificed and their injured cord tissues were sectioned for the identification of the transplanted cells. The axonal regeneration and the differentiation of MSCs were examined by immunocytochemical staining. At the same time, rats were subjected to behavioral tests by using the open-field BBB scoring system. Immunocytochemical staining showed that axonal regeneration and the transplanted cells partially expressed neuron-specific nuclear protein (NeuN) and glial fibrillary acidic protein (GFAP). At the same time, significant improvement in BBB locomotor rating scale (P＜0.05) were observed in the treatment group. More importantly, further functional improvement were noted in the combined treatment group. MSCs could differentiate into neurons and astrocytes. MSCs and NGF can promote axonal regeneration and improve functional recovery. There might exist a synergistic effect between MSCs and NGF.

Immortalized human precartilaginous stem cells (IPSCs) were established to provide stable cell resource for the study of the molecular mechanism of gene targeting on the differentiation of PSCs. Plasmid pCMVSV40T/PUR containing simian virus 40 large T antigen gene (SV40Tag) was transfected into human PSCs by using lipofectin transfection. Colonies were isolated by puromycin selection and expanded by multiple passages. Immunohistochemistry, RT-PCR and Southern blotting were used to identify the transfected cells and to detect the expression and integration of SV40Tag in expanded cell lines. The positive colonies were isolated and subcultured, designated immortalized precartilaginous stem cells (IPSCs), which were confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive cells by immunohistochemistry and RT-PCR. SV40Tag cDNA was found in cultured IPSCs of passage 8 by Southern blotting, and the expressions of SV40Tag mRNA and protein were confirmed by RT-PCR. These findings suggested that IPSCs strain with SV40Tag was constructed successfully.
Cartilage/*cytology ; Cell Proliferation ; Cell Transformation, Viral ; Cells, Cultured ; Fetus ; Simian virus 40/*genetics ; Stem Cells/*cytology ; Transfection

Objective To investigate the effect of sodium arsenite by Wnt signaling pathway on proliferation and apoptosis of oral squamous cell carcinoma.Methods Cell proliferation was detected after 1.25,2.5,5,10,20μmol/L sodium arsenite treatment human oral squamous cell carcinoma cell line Tca8113 for 24,48,72 hours by CCK8 experiment.0 and 14μmol/L sodium arsenite was used to treatment Tca8113 cells with 48h,cell apoptosis were detected by flow cytometry,Cleaved Caspase3,β-catenin,Cyclin D1 protein expression were detected by Western blot.Tca8113 cells were divided into control group,sodium arsenite group,activating agent+sodium arsenite group,all treated for 48hour,cell proliferation,apoptosis and Cleaved Caspase3,β-catenin,Cyclin D1 protein expression were detected by CCK8 assay,flow cytometry and Western blot.Results Tca8113 cell proliferation was inhibited significantly with the increase of treatment time and sodium arsenite concentration,and has a time and concentration dependent manner(P<0.05 or P<0.01).10μmol/L sodium arsenite as a follow-up study according to the IC50.Cell inhibition rate,apoptosis rate and Cleaved Caspase3 protein expression in 10μmol/L group were significantly higher than that of 0 mol/L group,the expression of β-catenin,Cyclin D1 protein was significantly lower than that of 0 mol/L group(P<0.01).Apoptosis rate,cell inhibition rate and Cleaved Caspase 3 protein expression in sodium arsenite group and activating agent+sodium arsenite group were significantly higher than control group,the expression of β-catenin and Cyclin D1 protein were significantly lower than control group(P<0.01).Apoptosis rate,cell inhibition rate and Cleaved Caspase 3 protein expression in activating agent + sodium arsenite group were significantly lower than that of sodium arsenite group,the expression of β-catenin and Cyclin D1 protein were significantly higher than that of sodium arsenite group(P<0.01).Conclusion Sodium arsenite can inhibit the proliferation of oral squamous cell carcinoma and promote apoptosis,and the mechanism was related to regulation of Wnt signaling pathway.

Objective To study the biological effects of pulsed electromagnetic fields (PEMFs) on the pro-liferation of immunomagnetically separated human precartilaginous stem cells (PSCs) in vitro. Methods The cells from an aborted fetus's metaphysis were digested using collagenase. The PSCs were isolated by magnetic cell sorting (MACS), then subcultured and amplified. Flow cytometry, immunohistochemistry, immunofluorescence and RT-PCR analysis were performed to identify the purified PSCs. The PSCs were stimulated by PEMFs at 50 Hz frequency and 1 mT intensity. Cell proliferation was measured at different time points using methyl thiazolyl tetrazolium ( MTT), and the cell growth curve was plotted. Flow cytometry was applied to measure the cell cycle and apoptosis. Results The PSCs were successfully cultured. There was fibroblast growth factor receptor-3 (FGFR-3) on their sur-face. Cell proliferation was promoted after 4 and 6 days of PEMF stimulation. The percentage of cells at the S phase was higher than in a control group. Early, late and total rates of apoptosis in the experimental group decreased signifi-cantly. Conclusion PEMFs can enhance the proliferation and inhibit the apoptosis of human PSCs, and it is possi-ble to cultivate the high density human PSCs in vitro.

Objective To explore the possibility of immortalized human precartilaginous stem cells (IPSCs) differentiating into nucleus pulposus-liked cells induced by transforming growth factor-β1 (TGF-β1)and examine its biological characters.Methods The IPSCs were seeded on the thermosensitive chitosan/glycerophosphate (C/GP) scaffolds and induced into nucleus pulposus-like cells in culture medium with the adding of TGF-β1 under hypoxia condiction.The growth and differrentiation of IPSCs on C/GP scaffolds were observed.Seven days later,Alcian blue staining was used to detect the formation of glycosaminoglycans (GAG) of extracellular matrix by the differentiating cell.RT-PCR was carried out to identify the expression of characteristic genes of nucleus pulposus-liked cells,including collagen Ⅱ and Aggrecan.Western blot were used to examine the expression of β-catenin.Results IPSCs grew well on the thermosensitive C/GP scaffolds.After 7 days,Alcian blue staining exhibited more formation of GAG in experimental group as compared with control group.RT-PCR manifested that the gene expression of collagen Ⅱ and Aggrecan were upregulated.Likewise,Western blot manifested that the expression of β-catenin was upregulated.Respectively,all of the content in the induction group obviously increased compared with that of the control group.Conclusion IPSCs can be differentiated into nucleus pulposus-like cells under the induction of TGF-β1,the differentiating cells have a favourable secretory function,which can secrete extracellular matrix effectively.Differentiation of IPSCs to nucleus pulposus-liked cells may be through upregulating the expression of β-catenin in cells.

Objective To investigate the variation of multivoxel 1 H magnetic resonance spectroscopy(1 H-MRS)before and after continuous positive airway pressure(CPAP)treatment in obstructive sleep apnea hypopnea syndrome(OSAHS).Methods Brain multivoxel 1 H-MRS examinations were performed in 25 cases of moderate or severe OSAHS patients before and after CPAP treat-ment,and 25 cases of healthy.The ratios of brain metabolites of the frontal lobe were recorded and analyzed respectively.To observe whether the lactate(Lac)peak appeared or not.Results In the frontal lobe,the NAA/Cr,NAA/Cho of the patients before treatment (2.021 2±0.231 2 and 1.608 8±0.257 1,respectively)was decreased compared with the healthy (2.726 8±0.607 1 and 2.445 6± 0.437 5).The NAA/Cr,NAA/Cho of the patients after treatment (2.314 0±0.312 8 and 2.01 6 4±0.424 0,respectively)was in-creased compared with the patients before treatment (2.021 2±0.231 2 and 1.608 8±0.257 1).The NAA/Cr,NAA/Cho of the pa-tients after treatment (2.314 0±0.312 8 and 2.01 6 4±0.424 0,respectively)was decreased compared with the healthy (2.726 8± 0.607 1 and 2.445 6±0.437 5),and the difierences were statistically significant (P <0.01).The Cho/Cr of the patients before treatment (1.293 2±0.261 5)was increased compared with the healthy (1.129 2±0.157 7),the difference was statistically significant(P <0.05).Lac peak was not detected in all.Conclusion Multivoxel 1 H-MRS can demonstrate sensitively the changes of brain metabolism in pa-tients with OSAHS before and after CPAP treatment,and may provide imaging evidence for clinical therapeutic effect and prognostic evaluation.

OBJECTIVE: To study whether negative-pressure septal drainage could be an alternative to packs after septoplasty. METHOD: This was a randomized controlled trial. The study involved 60 patients who underwent septoplasty. Patients were randomly divided into two groups, one with anterior nasal packs and the other with negative-pressure septal drainage. Patients were asked to record pain levels using a visual analogue scale (VAS). Postoperative symptoms and complications were compared during 24 h and 48 h postoperative period including pain, drying sensation of mouth, sleep difficulty, conjunctival congestion, haemorrhage. VAS scores and incidence were evaluated during 1 week and 6 weeks postoperative period including pain, bleeding, haematoma, septal perforation, synechiae and septal perforation. RESULT: Patients of negative-pressure septal drainage suffered from less pain than patients of nasal packs during the first 24 h and 48 h postoperative period. The results for pain, drying sensation of mouth, sleep difficulty, conjunctival congestion, haemorrhage were different between groups (P < 0.05), especially the amount of bleeding during 48 h postoperatively in patients undergoing negative pressure drainage [(0.52 ± 0.63)ml] was significantly less than the group who received anterior nasal packs [(21.03 ± 5.88) ml] (P < 0.01). On the other hand, haematoma, synechiae and perforation were not statistically different between groups during 1 week and 6 weeks follow-up period (P > 0.05). CONCLUSION Using negative-pressure drainage instead of nasal packs after septoplasty seems a more reasonable option. The negative-pressure drainage technique may be the preferred option to provide higher patient satisfaction and has the same level of postoperative complica.tion to nasal packs as for septoplasty surgery.
Drainage ; Humans ; Nasal Septum ; surgery ; Nasal Surgical Procedures ; Negative-Pressure Wound Therapy ; methods ; Nose ; Pain Measurement ; Patient Satisfaction ; Postoperative Period ; Tampons, Surgical

Objective To analyze the etiological factor and genetic feature of a familial hemophagocytic lymphohistiocytosis patient with PRF1 mutation (FHL2) with human herpesvirus 7 (HHV7)infection and its family constellation. Methods Clinical characteristics, laboratory examinations of a FHL2 case with HHV7 infection were reported. HHV1-HHV8 virus DNA was screened by PCR; NK cell function was analyzed by flow cytometry; PRF1 gene mutations were analyzed by PCR and direct sequencing, structure of mutant PRF1 proteins were analyzed using ExPasy and I-TASSER server and genetics pedigree were analyzed. Results The patient's HHV7 viral was detected positive with DNA copy number of 350/106 peripheral nucleated cells. Flow cytometry analysis showed decrease both in proportion of perforin positive NK cells and perforin protein expression. Genetic testing showed PRF1 biallelic heterozygote mutations (c. 503G ＞ A/p. S168N and c. 1177T ＞ C/p. C393R) and pedigree analysis showed they were inherited. The patient was then treated with antivirus therapy, dexamethasone and VP16 therapy, but only achieved partial response. The patient was then followed by human leukocyte antigen 10/10 allele identical nonconsanguinity allogeneic hematopoietic stem cell transplantations (allo-HSCT) and soon the successful implantation of donor hematopoietic cells and persistent recovery was achieved. The patient was now surviving without recurrence for 9 months after allo-HSCT. Conclusions FHL is prone to be misdiagnosed as lymphoma. Genetic analysis of related gene mutation and herpes simplex virus detection will help in early and accurate diagnosis. Allo-HSCT is a fundamental treatment of FHL.

Objective To investigate the potential role of disc repositioning and condyle restoration in the treatment of type Ⅲ traumatogenic temporomandibular joint (TMJ) ankylosis. Methods Eight patients including four females and four males at age range of 7-22 years (mean 13.6 years) were enrolled in this study. The patients suffered from traumatogenic TMJ ankylosis for 1-12 years. The preoperative interincisal opening distances ranged from 2 mm to 10 mm. During surgery, the traumatogenic callus of the lateral condyle process was removed, the condyle process was formed, and then the dislocated disc was sutured to the articular capsule or soft tissues around. Results All patients were followed up for 6-38 months and the last follow-up examination showed that the average interincisal opening distance was 30 mm. No recurrence or TMJ symptoms were found during the period of follow-up. Conclusions Disc repositioning and condyle restoration has the advantages of simple procedures, minor trauma and little recurrence and proves to be a feasible and effective method for the treatment of type Ⅲ traumatogenic TMJ ankylosis.