ObjectiveTo determine the effect of acute and chronic spinal cord injury (SCI) resulted from thoracic cord transection on the urinary bladder spinal neural pathway.Methods76 adult Sprague-Dawley rats were randomly divided into 4 groups, non-SCI, SCIa, SCIb and SCIc respectively. The non-SCI rats underwent no surgical procedure except Pseudorabies virus (PRV) tracer injection into the bladder tissue, while the rats of other groups were spinalized and given PRV injection at different time after SCI. Transcardiac perfusion fixation was done in appropriate survival periods after PRV injection. Then sections of dorsal root ganglion (DRG) and spinal cord were processed for visualization of virus by the Streptavidin-Peroxidase (SP) immunohistochemical procedure. All sections were measured with the Olympus Cue-2 image analysis system.ResultsThe bladder weights in SCIb and SCIc groups markedly increased (P<0.001). The time-ordered flow charts of PRV tracing were similar in the non-SCI rats and in the SCI rats. The cross-sectional area of the labeled DRG cell profiles increased significantly after SCI (P<0.001). The number of labeled cells in dorsal horn in L6 and S1 segments 3 days after PRV injection markedly increased in chronic SCI rats, and so did the number of labeled motor neurons 4 days post-injection. ConclusionThe acute and chronic SCI have little effect on the process of virus transneuronal transport below the level of lesion. Subsequent to chronic SCI, marked reorganization of the micturition reflex pathways occurs.

Background Marrow mesenchymal stem cells (MSCs) has been successful induced to differentiate into corneal cells,retinal ganglion cells (RGCs) and retinal neuron-like cells in recent years,but there are few studies about MSCs induced into photoreceptor cells and their microenvironment.Objective The aim of this study was to explore the induce and differentiation of bone marrow mesenchymal stem cells (BMSCs) into photoreceptor-like cells in vitro and microenvironment.Methods The second generation of human BMSCs strain and RPE cells strain were cultured and passaged,respectively,and the fourth generation of BMSCs and the third generation of RPE cells were used in the experiment.BMSCs were cocultured using the mesenchymal stem cells medium (MSCM) contained 20 μg/L basic fibroblast growth factor (bFGF),20 μg/L epithelial growth factor (EGF)and 20 μg/L brain derived neurotrophic factor (BDNF) with RPE cells to induce the differentiation of BMSCs in the induced group,and BMSCs were cultured in MSCM only in the control group.The morphology of induced and differentiated cells were observed under the inverted microscope.Inmmunocytochemistry was used in induced for 3-,5-,7-day cells to detect the expression rate of rhodopsin protein for the identification of phenotype of the differentiated cells.RT-PCR was used in induced for 5-,7-day cells to detect the expressions of rhodopsin mRNA and recoverin mRNA.Results Cultured BMSCs grew well with the spindle shape,and passaged RPE cells showed the uniform size and polygon shape with the abundant pigment in the cells.Some induced cells appeared to be the neuron-like cells with round shape and long prominence and the secondary reticular branches.The expression rates of rhodopsinin the cells were (5.83±0.29)％,(20.36±0.32)％ and (29.80±2.30)％ in the third,fifth and seventh day after induce,which were significantly higher than (0.71 ±0.35) ％,(2.56±0.24) ％ and (2.32±0.42) ％ of control cells (t3 d =41.510,t5d =107.290,t7 d =30.036,P＜0.01).The grey scales of rhodopsin mRNA and recoverin mRNA were significantly elevated in the induced and differentiated cells compared with control cells in the fifth and seventh day (rhodopsin mRNA:t5 d =103.506,t7 d =122.584,P＜0.01 ; recoverin mRNA:t5 d =106.674,t7 d =189.992,P＜0.01).Conclusions BMSCs can be successfully induced to differentiate into photoreceptor cells after cocultured by conditioned medium with RPE cells.

Objective To report the results of the surgical treatment of cervical spondylotic myelopathy(CSM) secondary to athetoid cerebral palsy and clarify the characteristics and the principle of treatment of the disease. Methods The age of the patient, segments of vertabrae involved and operated upon, results and complications of 32 cases with CSM secondary to athetoid cerebral palsy were statistically analyzed and compared with 93 patients who had isolate CSM operatively treated at the same time. Results 1)The average age of the patients in the group of CSM secondary to athetoid cerebral palsy was 37.94 years,and in the group of isolate CSM was 52.48 years, with significant statistic difference(P

[Objective]To study the effect of intra-operative three-dimensional fluoroscopy-based spinal navigation guidance for vertebroplasty to treat osteoporotic vertebral compression fracture(OVCF).[Method]A clinical study was performed in which 41 patients suffering painful osteoporotic vertebral compression fractures underwent vertebroplasty procedure.During this procedure,cannulation of the pedicle and vertebral body was performed with the aid of isocentric 3D fluoroscopy-based spinal navigation(group A) and biplanar fluoroscopy(group B).RDQ and VAS of each group was compared between preoperation and postoperation.Total operating time,intra-operative fluoroscopy time and pedicle puncturation accuracy were compared between group A and group B.Possible complications such as cement extravasations were evaluated with X-ray fluoroscopy.[Result]Forty-six vertebrae were successfully injected with polymethyl methacrylate.The two groups had statistically significant difference in RDQ and VAS between preoperation and postoperation,but no statistically significant difference between the two groups.Mean operating time and intra-operative fluoroscopy times of group A were shorter than those in group B.There was no case of broken pedicle in group A.One vertebra was found to have bone cement leakage to soft tissue in group B,but the vertebral canal and pedicle intact.The author followed up for an average of 21 months(range 8～38 months) and found no severe complication or collapse of vertebra.[Conclusion]With the aid of intra-operative three-dimensional fluoroscopy-based spinal navigation,vertebroplasty can be performed more accuractly and securely.The shorter total operating time,intra-operative fluoroscopy times are convincing and less possible complication can be expected.

[Objective] To evaluate the early effect of posterior dynamic lumbar stabilization in lumbar degenerative disease.[Methods]The clinical outcomes of 31 patients with lumbar degenerative disease treated by posterior decompression with Wallis posterior dynamic lumbar stabilization implant or combined with posterior lumbar fusion were retrospectively studied,and assessed with visual analogue scale(VAS)and spinal operative standard of Chinese Medical Association.The early effect and complications associated with Wallis posterior dynamic lumbar stabilization were recorded.[Results]The operative procedure of Wallis posterior dynamic lumbar stabilization implant was easy and less invasive.The VAS scores were 7.9?2.0,2.6?1.2 and 1.7?0.8 at one day preoperatively,two week postoperatively and final follow-up,respectively.The good to excellent result was 94.4% at the lastest follow-up.No compliction related with Wallis posterior dynamic lumbar stabilization was found.[Conclusion]It is easy and safe to use Wallis posterior dynamic lumbar stabilization in treatment of degenerative lumbar disease,and the early effect is good.The Wallis system provides an alternative for treatment of lumbar degenerative disease.

OBJECTIVE:To analyze bacteria distribution and drug resistance of pediatric severe sepsis in our hospital,and to provide reference for clinical rational use of antimicrobial agents. METHODS:57 pediatric severe sepsis patients were collected from pediatric intensive care unit of our hospital during Jan. 2014 to May 2015. The results of pathogen culture and drug sensitivity tests were analyzed retrospectively. RESULTS:Of 57 children,pathogen were detected in 18 cases(31.58%). A total of 91 pathogen were detected,of which there were 24 strains of Gram-positive(G+)bacteria(26.37%)mainly including Staphylococcus and Entero-coccus,60 strains of Gram-negative (G-) bacteria (65.93%) mainly including Klebsiella pneumoniae and Acinetobacter calco-acetcus-A. baumannii complex and 7 strains of fungus (7.69%) as Candida. 4 strains of methicillin-resistant Staphylococcus,22 strains of carbapenems-resistant K. pneumoniae,21 strains of multi-drug resistant K. pneumoniae and 7 strains of multi-drug resistant A. calcoacetcus-A. baumannii complex were all detected. Methicillin-resistant Staphylococcus,Staphylococcus aureus and Streptococ-cus pneumoniae were sensitive to vancomycin and linezolid,with resistant rate of 0. K. pneumoniae was completely resistant to ampi-cillin sodium and sulbactam sodium,piperacillin sodium and tazobactam sodium,imipenem and cephalosporin,with resistant rate of 100%. Resistant rate of A. calcoacetcus-A. baumannii complex to major common antimicrobial agents was higher than 50%. Esche-richia coli was resistant to cefotaxime,and resistant rates of other antimicrobial agents were lower than 40%. CONCLUSIONS:Main pathogen of pediatric severe sepsis is G- bacteria in our hospital,and carbapenems-resistant K. pneumoniae is detected,to which should be pay attention. The multiple drug-resistant treatment should be adopted for pediatric severe sepsis caused by multiple drug-re-sistant bacteria. Antimicrobial agents should be selected rationally according to pathogen type and the results of drug sensitivity test.

Objective To explore the effect of enhanced physical therapy with integrated traditional and western medicine on muscularweakness after selective posterior rhizotomy. Methods 44 children with muscular weakness after selective posterior rhizotomy were dividedinto the treatment group (n=28) and the control group (n=16). Exercise therapy, physical therapy, electroacupuncture and Chinese massagewere applied to the treatment group, while exercise therapy was applied to the control group only. The muscle strength of the leg of childrenwere compared right after the surgery and 2 weeks after the surgery. Results The muscle strength of the leg of children in treatment group recoveredbetter than the control group 2 weeks after the surgery (P<0.05), and it was almost at the same level with that before surgery (P<0.05). Conclusion The enhanced physical therapy with integrated traditional and western medicine method could rapidly recover the musclestrength of the leg of children after SPR.

Objective To explore the expression profiles and their clinical significance of microRNA (miRNA) molecules in different stages of chronic hepatitis B virus (HBV) infection.Methods The miRNA molecule expressions of 12 patients with chronic hepatitis B,12 chronic HBV carriers,12 inactive hepatitis B surface antigen (HBsAg) carriers,and 9 healthy controls were screened using miRNA microarray.The miRNA profiles were validated by the real time fluorescence quantitative polymerase chain reaction (PCR).The t-test was used for comparison between twogroups,whereas one-way ANOVA and SNK-q tests were used for multiple comparisons.Mann-Whitney and Kruskal Wallis H tests were used for comparison of two or more groups of data with skewed distribution.The receiver operation characteristic (ROC) curve was constructed to evaluate the diagnostic significance of miRNA molecules.Results Compared with the healthy controls,significant differences in the expression profiles of miRNA molecules were found in peripheral blood mononuclear cells of chronic HBV carriers (2 molecules up-regulated,and 18 down regulated) and chronic hepatitis B patients (33 molecules up-regulated and 19 down-regulated).No significant difference was found between inactive HBsAg carriers (2 molecules up regulated,and 3 down-regulated) and controls.The results of six miRNA molecules detected by real-time fluorescence quantitative PCR were basically consistent with the results detected by microarray.The area under ROC curve of the six miRNA molecules of hsa miR-4711-3p,hsa-miR-3191 5p,hsa-miR-5704-5p,hsa-miR 548ah-5p,hsa-miR-146a-5p and hsa-miR-29b-3p in distinguishing immune tolerance and clearance of chronic HBV infection were 0.994,0.984,0.967,1.000,1.000 and 0.996,respectively.Conclusions The different expression profiles of miRNA molecules could be used to distinguish the different stages of chronic HBV infection,and are closely related with the immune tolerance and activation in chronic HBV infection.

Objective miRNA-122 levels may correlate with liver cancer prognosis,and therefore understanding its expression is crucial for future treatment.This study investigates the effect of DNA methylation on the expression of liver specific miRNA-122 and its effects on proliferation and apoptosis of hepatocellular carcinoma cells.Methods Methylation sequencing detected the methylation of the miRNA-122 promoter region,and the level of miRNA-122 expression was measured by using real-time quantitative PCR.The proliferation and apoptosis of hepatocellular cell lines were detected by flow cytometry and CCK8.Results Compared with human primary hepatocytes [(21.9 ± 11.4)％],the level of miRNA-122 promoter methylation in Huh7,HepG2,and QSG-7701 cell lines were (87.6±9.3) ％,(89.0 ± 14.3)％,and (69.5 ±11.5)％,respectively.This represents a significant increase (P=0.000),especially in Huh7 and HepG2 cell lines.Compared with human primary hepatocytes (2.83× 104 ±3746),the levels of miR-122 expression in the above three cell lines were significantly decreased,especially in Huh7 and HepG2 cell lines (P=0.007).After treatment with 5-Aza-dc,the degree of methylation in Huh7 and HepG2 cell lines were significantly lower than that of the blank group (P=0.038,P=0.025),and the levels of miRNA-122 expression were significantly elevated (P=0.008,P=0.003).Also,compared with the control groups,the apoptosis of Huh7 cells and HepG2 cells were significantly increased (P=0.001,0.027).Conclusion The expression of miRNA-122 is regulated by DNA methylation and correlated with the apoptosis of liver cancer cells.Therfore,the methylation regulation of miRNA-122 expression might be involved in the occurrence and development of hepatocellular carcinoma.

BACKGROUND: Mesenchymal stem cells are few in human bone marrow, and their number will decrease with aging or body weakening, so a large amount of amplification is necessary. However, the biological characteristics of human mesenchymal stem cells of each passage remain poorly understood.OBJECTIVE: To analyze and compare the biological characteristics of each passage of bone marrow-derived mesenchymal stem cells (MSCs) so as to provide a basis for clinical demands of tissue engineering.DESIGN,TIME AND SETTING: Cytological observation in vitro. The experiment was performed at the Department of Orthopedics and Central Laboratory, Dongzhimen Hospital, Beijing University of Chinese Medicine from March to October 2008.MATERIALS: From bone marrow of patients with non-hematopoietic disease, MSCs were provided by Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine.METHODS: Bone marrow was collected form posterior superior iliac spine of patient, MSCs were isolated and cultured by Percoll method. When the cells were confluent at 90%, they were trypsinized and observed by inverted miscroscopy. The second passage of cells were collected for index detection.MAIN OUTCOME MEASURES: Cell morphological characteristics and immunophenotype; cell activity was detected by MTT; cell division and apoptosis in the proportion of necrosis were analyzed by flow cytometry analysis.RESULTS: The passaged MSCs exhibited uniform appearance in fusiform shape, and their growth was slowed down after 9 passages, exhibiting cytoplasm vacuolization and body enlarging. The second passage of MSCs was positive for CD44, CD106,and CD105, but negative for CD34 and CD45. MTT values peaked at passage 9, and gradually decreased since passage 10. At passage 11, the number of MSCs at division stage was increased, but from the sixth passage, the number of apoptotic cells increased significantly, reaching more than 60% at passage 8.CONCLUSION: According to biological characteristics analysis of MSCs at each passage, the third to the sixth passage cells are recommend for clinical therapy.