Polymorphism, Genetic ; Gene Frequency ; China ; Genetics, Population ; Microsatellite Repeats/genetics*

Objective To establish a method of isolation and purification of rat skeletal muscle satellite cells,and observe the characteristics of proliferation and myotube cell formation of skeletal muscle satellite cells cultured in vitro. MethodsPurified skeletal muscle satellite cells were obtained by improved two-step enzymatic digestion method and pre-plating technique.Immunohistochemical staining was employed to identify the skeletal muscle satellite cells cultured in vitro.The growth of skeletal muscle satellite cells cultured in vitro was examined by MTT assay.The differentiation of skeletal muscle satellite cells was observed by inverted microscopy. Results The skeletal muscle satellite cells with higher purity were obtained and confirmed by the high expression of desmin.When cultured in vitro,the latent phase of skeletal muscle satellite cells was the first to the second day,and the platform phase was the fifth to the sixth day.Myotube cells gradually formed when cell confluence was more than 60% to 70% or differential medium with lower fetal bovine serum was used.Conclusion The combination of improved two-step enzymatic digestion method and pre-plating technique serve as an easy and practical way to obtain skeletal muscle satellite cells with higher purity.Skeletal muscle satellite cells can form myotube cells with contraction characteristics without any special induction.

Aged ; Breast Neoplasms ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Mastectomy ; methods ; Metaplasia ; pathology ; Papilloma, Intraductal ; pathology ; surgery ; Sebaceous Glands ; pathology

BACKGROUND:The main treatment of lumbar isthmic spondylolisthesis is the surgery, in a broader attempt to decompression, reduction, fixation and fusion of the lesioned segments. The golden standard of the treatment is biological fusion, while internal fixation is a reliable assistance for fusion therapy. ＜br> OBJECTIVE:To discuss the clinical value and curative effect of intervertebral fusion cage combined with pedicle screw systems for the treatment of lumbar isthmic spondylolisthesis. ＜br> METHODS:From March 2010 to March 2013, 21 cases of isthmic spondylolisthesis were treated with intervertebral fusion cage combined with pedicle screw systems, including 18 cases of spondylolisthesis of degree II and 3 cases of spondylolisthesis of degree III. Al patients were fol owed up regularly, taking JOA lumbago score and visual analog scale score as the objective evaluation criteria of pain in postoperative fol ow-ups. The curative effect was assessed by Macrab standard, and the functional recovery was evaluated based on indicators such as Prolo, and the spinal fusion rate was assessed according to Lenke criteria. Changes of slippage rate, slippage angle, sacral inclination angle and intervertebral space post height in preoperative and postoperative periods were evaluated by iconography data. ＜br> RESULTS AND CONCLUSION:Al the 21 patients with isthmic spondylolisthesis were fol owed up for 12-16 months. JOA lumbago score and vasual analog scale score of al patients were improved after treatment, and the difference was statistical y significant compared with before treatment (P=0.000). According to Macrab evaluation criteria, there were 17 excellent cases and 4 good cases. Each indicator evaluated by preoperative Prolo activities and symptom grading showed significant differences in preoperative and postoperative periods (P=0.003). Postoperative lumbar spondylolisthesis was basical y reset, the slippage angle was significantly reduced, the sacral inclination angle was increased, and the height of the intervertebral space was recovered basical y. Intervertebral fusion cage combined with pedicle screw systems was one of the effective strategies to treat lumbar isthmic spondylolisthesis.

Objective To investigate the effects of early nasal intermittent positive pressure ventilation (NIPPV) compared with early continuous positive airway pressure (NCPAP) in low birth weight preterm infants with respiratory distress syndrome (RDS).Methods We performed a prospective,randomized controlled trial involving 364 low birth weight preterm infants with respiratory distress syndrome within 6 hours of birth.The infants were randomly assigned to NIPPV (n=188) or NCPAP (n=176) groups.Non invasive ventilation was initiated in the neonatal intensive care unit (NICU).The rate of mechanical intubation (within 3 days or 7 days),the use of pulmonary surfactant (PS),the rate of complications and mortality were compared between the two groups.Mann Whitney U,t and Chi-square tests were used for statistical analysis.Results The average time of invasive mechanical ventilation in NIPPV group were lower than that in NCPAP group[2.0 (1.0-4.0) d vs 7.0 (3.0-8.5) d,U=-3.457,P=0.001].The need for intubation and mechanical ventilation by day 3 and day 7 in the NIPPV group were less than those in the NCPAP group [day 3:4.8％ (9/188) vs 10.8％ (19/176),x2=4.621,P=0.032; day 7:9.0％ (17/188) vs 16.5％ (29/176),x2=4.551,P=0.033].In the NIPPV group,infants who got PS therapy was less than that in the NCPAP group [3.2％ (6/188) vs 8.5％ (15/176),x2=4.752,P=0.029].There was no significant difference in the fatality rate between the NIPPV and the NCPAP group [12.8％ (24/188) vs 10.8％ (19/176),P ＞ 0.05].There were no significant difference in the incidence of air leak,intracranial hemorrhage,periventricular leukomalacia,retinopathy of prematurity,necrotizing enterocolitis,patent ductus arteriosus,and bronchopulmonary dysplasia between the NIPPV group and the NCPAP group.Conclusion Among low birth weight prcterm infants with RDS,the early use of NIPPV reduces the need for PS,intubation and invasive ventilation compared with NCPAP.

0.05).(4) Significant differences between CC and CIN Ⅰ for p16,CDH1,RASSF1A and TIMP3 genes(P

We have constructed an immunotoxin(Ng76-TCS),which was composed of a monoclonalantibody directed against human melanoma and trichosanthin(TCS)——a single chain ribosomeinactivating protein.The cultured human melanoma cells(M21)were inhibited effectively byNg 76-TCS.The cytotoxicity of Ng76-TCS to M21 cells was 2,000-fold higher than that of free TCS and Ng76 mixture.A conjugate,which was prepared with normal mice immunoglobulinand TCS(NIgG-TCS),was 160-fold less cytotoxic to M21 cells.Meanwhile Ng76-TCS was125-fold less cytotoxic to nontarget cells Hela.These results showed that the immunotoxinNg76-TCS was a potent and specific anti-human melanoma agent.

Objective To evaluate the efficacy and safety of intravesical instillation of sodium hya- luronate in relieving the symptoms of refractory non-bacterial cystitis.Methods Totally,20 patients(1 man and 19 women;mean age,47 years;age range,22-68 years)with refractory non-bacterial cystitis were included.Their disease course ranged from 1 month to 30 years.Of them,16 patients had the disease for more than 1 year.Instillation of sodium hyaluronate(50 ml solution of 40 mg HA)was performed intravesically in them for a total of 12 weeks(once a week for the first 4 weeks and once every 4 weeks for the succeeding 8 weeks).Follow-up visit ended at 16 weeks,when the patients filled in the symptom assessment scores(VAS on pain,frequency and urgency)according to the subjective evaluation.Therapeutic results were classified as(1)total response:symptom disappearance or relief by at least 90% of the symptom scores;(2)partial response:symptom relief by 50%-89%;(3)mild response:symptom relief by 49% or less;(4)no re- sponse:no improvement in symptom scores.Results Nineteen patients completed the 16-week follow-up with only 1 withdrawing.The average general symptom score decreased from 22.32?5.53 to 9.47?5.88 (P＜0.001);frequency score from 8.21?1.75 to 3.89v2.31(P＜0.001);urgency score from 7.47?2.14 to 3.37?2.14(P＜0.001);pain score from 6.63?3.47 to 2.21?2.74(P＜0.001).After 16 weeks,overall positive response rate(total+partial response)was 53%(10/19).No serious adverse effect occurred except for urethral irritation(25%,5/20),worsening of distending pain of the bladder(5%,1/20) and vertigo(15%,3/20)caused by the procedure itself.Conclusions For patients suffering from refrac- tory non-bacterial cystitis,intravesical instillations of sodium hyaluronate can significantly relieve the symp- toms and is safe and of good patient compliance.

OBJECTIVETo construct the recombinant plasmid pcDNA3.0-RGC32 and evaluate the effect of the response gene to complement-32 (RGC32) on cell cytoskeleton in vitro.

METHODSThe full-length cDNA of RGC32 was obtained by RT-PCR and inserted into the eukaryotic expression vector pcDNA3.0 to generate the recombinant plasmid pcDNA3.0-RGC32. After transfection of the recombinant plasmid into SW480 cells, the expression of RGC32 in the cells was detected by Western blotting. The cytoskeleton of SW480 cells was visualized before and after the transfection, and the changes in the cell migration ability was assessed by wound-healing assay.

RESULTSThe recombinant plasmid pcDNA3.0-RGC32 was successfully constructed. The expression of RGC32 was significantly increased in SW480 cells after transfection with pcDNA3.0-RGC32. Before the transfection, the microfilaments of SW480 cells were few and short without obvious polarity, but after the transfection, the microfilaments were increased and elongated with also an obvious polarity, and the invasive structures of lamellae and lamellipodia occurred. The migration ability of the cells was enhanced after transfection with pcDNA3.0-RGC32.

CONCLUSIONOverexpression of RGC32 can cause the reorganization of cytoskeleton and promotes the cell migration, which can be an important mechanism of RGC32 in promoting cancer metastasis.

Cell Cycle Proteins ; biosynthesis ; genetics ; Cell Line, Tumor ; Cell Movement ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Cytoskeleton ; chemistry ; metabolism ; Genetic Vectors ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Neoplasm Metastasis ; genetics ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo investigate the effects of CD137-CD137L interaction on the nuclear factor of activated T cells c1 (NFATc1) in apolipoprotein E knockout (ApoE(-/-)) mice.

METHODSAtherosclerotic plaque model was produced by rapid perivascular carotid collar placement in ApoE(-/-) mice. In vivo, the expression of NFATc1 in mice plaque and lymphocytes was detected by immunohistochemical and flow cytometry, respectively. In vitro, the NFATc1 mRNA and protein expressions in cultured lymphocytes of ApoE(-/-) mice were measured by RT-PCR and flow cytometry, respectively.

RESULTSIn vivo, after stimulating CD137-CD137L signal pathway, the expression of NFATc1 was significantly upregulated in the atherosclerotic plaques and lymphocytes. In vitro, the mRNA and protein expressions of NFATc1 in cultured leukocytes of ApoE(-/-) mice were also significantly increased, the maximal effect appeared post 20 µg/ml anti-CD137 mAb-stimulation and reached maximum at 24 h at any concentrations. Anti-CD137L mAb significantly downregulated the mRNA and protein expressions of NFATc1 in lymphocytes of ApoE(-/-) mice, maximal effect appeared at 20 µg/ml anti-CD137L mAb and reached minimum at 24 h.

CONCLUSIONCD137-CD137L interactions can modulate the expression of NFATc1 in this ApoE(-/-) mice atherosclerotic plaque model.

4-1BB Ligand ; metabolism ; Animals ; Apolipoproteins E ; genetics ; Disease Models, Animal ; Mice ; Mice, Knockout ; NFATC Transcription Factors ; metabolism ; Plaque, Atherosclerotic ; metabolism ; RNA, Messenger ; genetics ; Tumor Necrosis Factor Receptor Superfamily, Member 9 ; metabolism