BACKGROUND: The assessment of chronic nerve root injury is mainly depend on indirect evidences of imaging results.OBJECTIVE: This study is designed to find out the relationship between the degrees of the pathological changes of nerve roots and motor evoked potential (MEP) changes due to chronic compression.DESIGN: A self-controlled trial with animals as subjects.SETTING: Spine Surgery Department of the Second Clinical Hospital of Jinan University.MATERIALS: The trial was completed in the Spinal Surgery Department of the Second Clinical Hospital of Jinan University from October 2001 to May 2003. The subjects were 30 healthy cats of either gender weighing 3 to 5 kg.INTERVENTIONS: The compression model was established by wedging a Ⅴ-shape bone flap into the right intervertebral foramens between C7, C8 and T1 vertebrae. The left foramens served as controls. The cats underwent CT examination on the 2nd, 4th, 8th and 12th postoperative week to show the foramen changes. Six randomly selected cats underwent magnetic stimulation MEP each time on the 2nd, 4th, 8th, 12th and 24th week. The injuries to the compressed nerve roots were assessed according to Mackinnon Standard and they were compared with the MEP wave.MAIN OUTCOME MEASURES: Whether the superficial magnetic stimulated MEP monitoring result was in correspondence with the pathological changes of the chronically compressed nerve roots.RESULTS: The MEP did not show any abnormal changes for Grade Ⅰ injury prolonged to 9.6- 10. 2 ms. There was almost no evoke potential at all stimulation points for grade Ⅴ nerve root injury.CONCLUSION: That the injured nerve root presents dysesthesia and normal MEP results suggests grade Ⅰ injury. Prolonged latency including prolonged F wave suggests grade Ⅱ injury. And additional amplitude decrease together with widened and unclassifiable waves implies the injury be grade Ⅲ. The prominent features of grade Ⅳ nerve injury is much more decreased amplitudes and more prolonged latency. For grade Ⅴ injury there is always disappearance of whole MEP or some potential components. The superficial magnetic stimulated MEP monitoring results are in correspondence with the pathological changes of the nerve roots due to chronic compression, which means MEP monitoring results could serve as the quantitative indication to pathological changes of nerve injury.

BACKGROUND: Many clinical cases have proved that the satisfactory reposition, fusion and internal fixation in the internal fixation for inferior lumbar spondylolisthesis are not consistent with the postoperative symptoms and physical signs, and functional restoration.OBJECTIVE: To investigate the application of somatosensory evoked potential(SEP) in the monitoring during internal fixation for inferior lumbar spondylolisthesis, and the effects of SEP monitoring on the improvement of postoperative symptoms and the spinal functions as well.DESIGN: A randomized controlled trial.SETTING: Inpatient department of spinal surgery, an affiliated hospital of a university. PARTICIPANTS: Fifty-two patients with inferior lumbar spomlylolisthesis including 23 male and 29 female cases aged between 18 and 68 years old were admitted by the Department of Spinal Surgery, Second Affiliated Hospital(Shenzhen People's Hospital) of Jinan University, from June 2000 to December 2003. All cases were randomly divided into control group(n = 20) and monitor group ( n = 32).METHODS: SEP induced by segmental stimulation in cutaneous nerve was used in the control group for preoperative and intraoperative monitoring, and postoperative follow up. The intraoperative potential changes in patients with excellent and good improvement in postoperative functions had been retrospectively investigated to confirm the effective indices for intraoperative monitoring, which thereby provided references for intraoperative monitoring in patients of monitor group.MAIN OUTCOME MEASURES: ① Evaluation of postoperative spinal function; ② SEP latency and amplitude.RESULTS: SEP values after postoperative anesthesia were set as basis.The manifestations of intraoperative potential alterations: ① If the latency reduced 10% -15% or the amplitude increases more than 40% after fixation, it suggested favorable prognosis; ② The potential indices were stable during monitoring, or the reduction of latency was less than 10%,or the increase of amplitude was less than 30%, the fixation could be continued, and partial functions of nerve root and symptoms could be improved after surgery; ③ During the surgery, if potential indices had transient lluctuation, which could be recovered to basic potential within 15 to 20 minutes, fixation should be stopped during the fluctuation. Still partial functions of nerve root and symptoms could be improved after surgery; ④If the intraoperative latency prolonged more than 5%, or amplitude reduced more than 10%, or part of the components disappeared, or the wave shape dispersed, it might suggest postoperative aggravation of pain and dyskinesia. So intraoperative adjustment was necessary. The coincident rate of the improvement of the indices of intraoperative monitoring and the improvement of postoperative spinal function reached 93.75%.CONCLUSION: SEP induced by segrmental stimulation in cutaneous nerve is an objective and effective method in the monitoring and instruction of decompression, reposition, fixation of internal fixation for inferior lumbar spondylolisthesis, which has important merits in the improvement of the function of nerve root and the restoration of spinal function.

Objective To explore the effects and the underlying mechanisms of inhibiting three kinds of DNA methyltransferases(DNMT1,DNMT3a and DNMT3b) on the re-expression of cancer/testis antigen(CTA) in hepatic cells.Methods Transient transfection of HepG2 cells with siRNA was targeted against DNMT1,DNMT3a and DNMT3b(DNMT1+3a+3b),DNMT1 and DNMT3a(DNMT1+3a),DNMT1 and DNMT3b(DNMT1+3b),DNMT3a and DNMT3b(DNMT3a+3b),respectively.The other batch of cells was treated with 5'-aza-deoxycytidine(5-aza-dC) as positive control.Real-time PCR and Western blotting were used to detect the expressions of DNMTs and CTAsm,and the promoter methylation of partial CTA genes was detected with methylation specific PCR(MSP).Results Expressions of DNMT1,DNMT3a and DNMT3b declined significantly after siRNA interference in HepG2 cells.Two CTAs,CT10 and SSX1,re-expressed in the cells transfected by DNMT1+3a and DNMT1+3b.MAGE1 and MAGE3 were equally expressed in all cells despite been transfected or not.Demethylation occurred in the promoter region of CT10,while the promoter region of MAGE1 was in a state of unmethylation.Conclusions In HepG2 cells,CTA promoter can be demethylated via interference of DNMTs,which may lead to the re-expression of CTA that was originally unexpressed due to methylation.

For all the time the book Symptomatic Figures of Lip and Tongue(Chunshe Zhenghou Tu)was described as written by Li Jun of Qing dynasty. We made studies on historical records on Li Jun and the features and contents of this book, and proved that Symptomatic Figures of Lip and Tongue was not written by him, but a Chinese medical book written by Japanese.

objective To investigate the clinical signiifcance of the changes of angiotensionⅡ(AngⅡ) in children with IgA nephropathy (IgAN). Methods Thirty children diagnosed as primary IgA nephropathy by renal biopsy (IgAN group) and 30 healthy children (control group) were recruited from May 2008 to December 2012. The serum and urine AngⅡwere measured by ELISA and compared between IgAN group and control group. The AngⅡexpression in the renal tissue of IgAN group was detected by immuno-histochemical method, and was correlated with other clinical data.. Results Urine AngⅡwas signiifcantly higher in the primary IgAN group than that of control group (P<0.05);AngⅡexpression in the urine is positively correlated with proteinuria (r=0.37, P=0.046), and is associated with the severity of clinical presentation; AngⅡexpression in kidney tissue increased with the severity of the renal histopathologic grading (r=0.69, 0.79, P=0.000), while AngⅡin blood and proteinuria, AngⅡexpression in kidney tissue were not signiifcantly correlated with the number of crescents. Conclusions Urine AngⅡin children with IgAN is signiifcantly correlated with the severity of the pathologic stage and the level of proteinuria. Urine AngⅡdetection may be useful to assess the progress and prognosis of chronic kidney disease.

To study the cartilage differentiation of mouse mesenchymal stem cells (MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 (0, 10, 20, 50 and 100 ng/mL). After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen II mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen II mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesenchymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.
Bone Marrow Cells/*cytology ; Bone Morphogenetic Proteins/*pharmacology ; Cell Differentiation/*drug effects ; Cells, Cultured ; Chondrocytes/*cytology ; Chondrogenesis/drug effects ; Chondrogenesis/physiology ; Mesenchymal Stem Cells/*cytology ; Recombinant Proteins/pharmacology

Objective To find the optimal perfusion temperature of focal hypothermia in rats with traumatic brain injury(TBI).Methods Modified Feeney's free-falling model was used in the study.Forty-nine male Sprague-Dawley(SD)rats were randomly divided into seven groups:TBI group,systemic hypothermia group,focal hypothermia groups(including 0℃,10℃,20℃ or 25℃ subgroups,respectively)and control group.The contents of total sodium,potassium and water of traumatic brain were measured,and pathological changes were examined in the seven groups.Results The damaged neurons were significantly fewer in focal hypothermia groups(including 10℃,20℃ or 25 ℃ subgroups,respectively)than those in systemic hypothermia group at 72 hours after TBI(P

Objective To compare the three kinds of methods for in vitro primary culturing of rheumatoid arthritis synovial fibroblast-like cells (RASFs), in order to get fast and effective culture methods. Methods Synovial tissue from RA synovial arthroscopic resection were treated by collagenase digestion method, modified tissue culture method, double enzyme digestion method respectively. By using an inverted phase contrast microscope, cell morphology and growth characteristics were observed and identified with vimentin staining. Trypan blue was used to count the number of living cells after culturing for 14d. Results The three primary methods could successfully isolate and culture RASFs, and RASFs met the morphological characteristics of vimentin-positive cells>95%, namely, the proportion of RASFs cell confluence was 70% after 16-20days by the collagenase digestion method,whose cell confluence proportion reached 95%after 4 weeks;and the cell confluence proportion was above 70%after 10-14days by modified tissue culture method,and the cell confluence proportion reached 85%after 4 weeks by the double enzyme digestion method. The comparison of the viable cells number cultured same number of synovial tissue by the three methods show the viable cells number cultured by the modified tissue culture method were (1.60±0.08) ×106, those by the collagenase digestion method were (1.41±0.08) ×106, those by the double enzyme digestion method were (1.19 ±0.05) ×106, which were with significant difference among them (P<0.05) .The comparison of incubation time of RASFs primary cells showed it took (267.50±16.58) mins by the collagenase digestion method, (183.75 ±11.08) mins by the double enzyme digestion method, and 149.10 ±13.71mins by the modified tissue culture method, with significant differences (P<0.05) .Conclusion Modified tissue culture for RASFs is an efficient and fast culture method, the number and purity of RASFs can meet the requirements for biology experiments.

Objective To investigate the role of Rho/ROCK signaling pathway in the protective effects of hydrogen gas (H2) on acute lung injury (ALI) in a mouse model of sepsis.Methods Eighty male C57BL/6 mice were randomly divided into four groups (n =20 per group):sham surgery group,H2 control group (sham + H2 inhalation),sepsis model group and H2 treatment group (sepsis + H2 inhalation).The mouse model of sepsis was created by cecal ligation puncture (CLP),and the mice in sham surgery group didn't undergo cecal ligation and puncture.The mice in the H2 inhalation groups received inhalation of 2％ H2 for 1 hour at 1 hour and 6 hours after CLP or sham surgery,respectively.Ten mice in each group were selected and subjected to Evans blue (EB) test to evaluate the pulmonary endothelial permeability at 24 hours after CLP operation.The rest of 10 mice in each group were sacrificed at 24 hours after CLP operation,the bronchoalveolar lavage fluid (BALF) was collected for the measurement of protein concentration,tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1 β) content,and polymorphonuclear neutrophils (PMN)counts.The lung tissues were obtained to determine the content of malonaldehyde (MDA) and the activity of superoxide dismutase (SOD),the wet/dry lung weight ratio (W/D) was calculated,the lung pathological changes in hematoxylin and eosin (HE) stained sections were evaluated under a light microscope,the activity of Rho/ROCK signaling pathway and expression of zonula occluden 1 (ZO-1) were detected by Western Blot,and the distribution and expression of ZO-1 were also examined by immunofluorescence staining.Results There was no statistical difference in the above indexes between the sham surgery group and the H2 control group.Compared with the sham surgery group,the sepsis group demonstrated significant increases in the concentrations of protein,TNF-α,IL-1 β and PMN counts in BALF,the lung EB and MDA content,W/D ratio,the ratio of Rho-GTP/total Rho,the expressions of ROCK1 and ROCK2,the ratio of phosphorylated-myosine phosphatae targeting subunit 1 (p-MYPT1)/MYPT1,and significant decreases in the lung SOD activity and ZO-1 expression.Compared with the sepsis group,the H2 treatment group showed statistically significant decreases in the concentrations of protein,TNF-α,IL-1 β,PMN counts in BALF [protein (g/L):3.12 ± 0.33 vs.6.37±0.56,TNF-o(ng/L):128.45± 17.33 vs.563.83±61.72,IL-1β (ng/L):75.76± 14.35 vs.245.52±30.56,PMN counts (× 105/L):7.46± 1.34 vs.18.55± 5.73],and permeability of lung [EB concentration (μg/g):73.33±6.98vs.144.83± 12.38],the lung MDA content (mmol/g:3.66±0.53 vs.6.04± 1.13),the lung W/D ratio (5.02± 0.34 vs.7.26 ±0.56),the ratio of Rho-GTP/total Rho,the expressions of ROCK1 and ROCK2,the ratio of p-MYPT1/MYPT1 [Rho-GTP/total Rho:(43.12 ± 4.69)％ vs.(68.82± 5.44)％,ROCK1 (gray value):2.42 ± 0.42 vs.6.03 ± 0.64,ROCK2(gray value):2.56± 0.52 vs.4.85 ± 0.53,p-MYPT1/MYPT1:(57.83 ± 8.67)％ vs.(112.50± 13.43)％],and statistically significant increases in the lung SOD activity (kU/g:18.58± 1.68 vs.13.31±2.20) as well as the expression of ZO-1 (gray value:0.61 ± 0.07 vs.0.32 ± 0.06,fluorescence intensity:0.77 ± 0.06 vs.0.54 ± 0.05;all P ＜ 0.05).Moreover,lung HE staining showed that there were obvious lung injuries in the sepsis group which were alleviated in the H2 treatment group.Conclusion H2 could improve endothelial permeability and suppress inflammation and oxidative stress to alleviate ALI in septic mice through inhibition of Rho/ROCK signaling pathway.

Objective To simulate the chemical microenvironment of traumatic brain injury (TBI) under mild hypothermia, and investigate the effect of such microenvironment on umbilical cord mesenchymal stem cells (UCMSCs) in vitro.Methods Eighteen SD rats were allocated to shamoperated group, TBI group and mild hypothermia group according to the random number table, with 6 rats per group.Rat models of TBI were made by electric cortical contusion impactor.After systemic mild hypothermia (33℃) for 4 h, brain tissue homogenate extracts were harvested.Polyacrylamide gels mimicking the elastic modulus of brain were manufactured.Human UCMSCs were isolated and cultured on the gels, added with brain tissue extracts from each group.After 24 h, the apoptosis level of UCMSCs was checked, and the medium was changed with normal one.Cell growth and morphological changes in each group were given dynamic observation.Seven days later, cell immunofluorescence was implemented, with the differentiation level of each group estimated.Results Apoptotic rate in TBI group was 73.47％,significantly higher than 10.42％ in sham-operated group (P ＜0.01).While the apoptotic rate was 28.57％ in mild hypothermia group, indicating mild hypothermia significantly reversed the apoptosis of cells in TBI group (P ＜ 0.01).Cell immunofluorescence demonstrated rate of neuronal differentiation of UCMSCs in sham-operated group, TBI group and mild hypothermia group was 16.48％, 2.59％ and 11.83％ respectively.Mild hypothermia resulted in significantly improved neuronal differentiation of UCMSCs after TBI (P ＜ 0.05).Conclusions More apoptosis and lower neuronal differentiation ability are observed in UCMSCs in the chemical microenvironment after TBI.However, mild hypothermia significantly reverses the elevation of apoptosis and restores the neuronal differentiation capacity of UCMSCs after TBI.