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.

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

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

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.

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.

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 expression and the early diagnostic significance of heat shock protein 70 in acute allograft rejection of liver-transplamed rats. Methods The model of rat orthotopic liver transplantation was made by using a modified "two-cuff technique". The rats were randomly divided into 3 groups. For each group, donors and receptors all included 15 rats respectively. The control group: Wistar to Wistar liver transplantation; The untreated group: SD to Wistar liver transplantation, not receiving any immunosuppressant after liver transplantation; The treatment group: SD to Wistar liver transplantation, receiving intramuscular injection of tacrolimus (FKS06, 2mg kg-1. day-1) after operation. Five rats were executed randomly in every group on the post-transplantation day 3, 5 and 7 and the graft samples were obtained for optical microscopic observation. The expression of HSPT0 in grafts was detected by using immunohistochemical method and RT-PCR. The correlation between acute rejection following liver transplantation and the expression of HSP70 in grafted liver was studied. Results There was no acute rejection examined in the control group. The untreated group showed typical allograft rejection and the rejection activity index (RIA) went up gradually after the operation (P<0.01). The treatment group showed no rejection or borderline allograft rejection. The level of HSP70 was increased transiently after operation, then reduced in the control group (P<0.05). The level of HSP70 in the untreated group was higher than in the control groupand gradually increased with the prolongation of time after transplantation (P<0.01). A significant correlation was found between HSP70 and pathological score in the untreated group (P<0.01). The treatment group showed low levels of HSP70 of all the time. Conclusions The expression of HSP70 in grafts is closely related to the occurrence and development of the acute rejection and can be useful for early diagnosis of acute allograft rejection following liver transplantation.

Objective To identify the risk factors for early postoperative cognitive dysfunction (POCD) in the elderly patients undergoing spinal surgery.Methods One hundred and fifty ASA Ⅱ or Ⅲ patients,aged ≥65 yr,undergoing elective spinal surgery under general anesthesia,were studied.Venous blood samples were taken at 1 day before operation and 7 days after operation to determine the concentration of serum S-100B protein.Cognitive function was assessed by Mini-Mental State Examination (MMSE) at 1 day before operation and 7 days after operation.The patients were diagnosed as having POCD if MMSEpre-MMSEpost ≥ 3.The patients were divided into POCD group and non-POCD group.Age,body weight,sex,education,type of operations,complications,preoperative TCM syndrome type,MMSE score at 1 day before operation and 7 days after operation,duration of operation,emergence time,and intraoperative blood loss,cardiovascular events,and amount of fluid infused per hour were recorded.The risk factors for POCD were analyzed using multivariate logistic regression analysis.Results Thirty-eight patients developed early POCD (25.3％).The resuhs of logistic regression analysis showed that age ≥ 70 yr,elementary education or below,duration of operation ≥5 b,and the number of hypotension occurred during operation≥3 were the risk factors for early POCD.Conclusion Age≥70 yr,elementary education or below,duration of operation ≥ 5 h,and the number of hypotension occurred during operation ≥ 3 are the risk factors for early POCD in the elderly patients undergoing spinal surgery.