Objective To understand the mechanism of how cardiomyocytes exit from the cell cycle,we examined the expression of polo-like kinase 1(plk1) in the postnatal developmental process of cardiac myocytes. Methods Mitotic Index(MI) of cardiomyocytes was examined in the neonatal,2-week-old,4-week-old,and adult rat hearts(five cases per groups) by double immunofluorescence stained with H3P and ?-sarcomeric actin antibodies.plk1 mRNA and protein expression during the postnatal developmental process of cardiac myocytes were detected by RT-PCR and Western blot analysis in rat hearts. Results The MI of cardiomyocytes in 0-day-old hearts(0.905?0.087%) was approximately 2.4 times over that in 2-week-old hearts(0.372?0.094%)(P

Objective To study the diagnostic value of multi-direction adjusted multiplanar reconstruction (MPR)by 64-slice CT myelography (CTM)in diagnosing cervical nerve injury,and the possibility of the MPR to replace conventional myelography and CT direct-scanning axial images.Methods Twenty-six patients with cervical nerve root injury were examined by conventional myelograpby and 64-slice CT using isotropic parameters.Then multi-direction MPR were performed to display nerve roots on coronal and sagittal planes besides axial images.Twenty-six patients were performed surgical operations and diagnosis were obtained.The coincident diagnosing rate with surgical operations results were compared statistically among multi-direction MPR,direct-scanning axial CT images,conventional myelography.The numbers of images were also compared between axial MPR and direct scanning axial CT images.Results Direct sign of nerve root avulsion was the loss of normal nerve root defect seen in the Isovist filled thecal sac in 6d-slice CT,which was found in 31 nerve roots.Indirect signs included:(1)Traumatic pseudomeningocele: 29 nerve roots showed the leak of Isovist into nerve root sheath,and extended into foramina;(2)Arachnoid cyst: 26 nerve roots clearly displays cystic distension in nerve root,which has low- density fine clew form septation from subarachnoid cavity and no nerve root in the cyst ;(3)Deformity of the subarachnoid space : deformity of thecal sac,partially lack of Isovist into arachnoid space,which was found in 17 nerve roots.The coincident diagnosing rate of cervical nerve root injury by multi-direction adjusted coronal MPR imaging was 92.6% (50/54),which was higher than by axial CT (77.8%,42/54)and conventional myelography (68.5%,37/54),There was significant difference between the conventional myelography, direct-scanning axial CT,multi-direction MPR images (Kappa = 0.686,0.772,0.920, respectively,P

BACKGROUND: The study of cell transplantation to repair injured cardiac muscle and improve cardiac function of dilated cardiomyopathy (DCM) has become a hotspot in recent years. However, the effect of cardiac electrophysiology following transplantation is still unknown.OBJECTIVE: To explore the influence of ailogenic implanted mesenchymal stem cells (MSCs) on cardiac function, structure and electrophysiology of rabbits with DCM.DESIGN, TIME AND SETTING: Randomized controlled animal trial was performed at Electrophysiology Laboratory of Institute of Pediatrics, Chongqing Medical University between January 2004 and May 2006.MATERIALS: Forty-three New Zealand whiterabbits, weighing 3.0-3.5 kg, irrespective of gender, were selected. Adriamycin was produced by Haizheng Medical Products Company of Zhejiang Province, China, No. 050307. The Ultrasonograph SSD-5000 came from Aloka, Japan, and RM6240 multiplying channel electrophysiolograph of Chengdu Instrument Company was applied.METHODS: All animals were randomized into normal group (n=12), cell transplantation group (n=13), and DCM model group (n=13). Except the normal group, adriamycin was applied to create rabbit DCM model, I mg/kg, twice a week for successive 8 weeks. Bone marrow solution was harvested from the remaining 5 rabbits, and MSCs were isolated, amplified and purified using attachment method. Three weeks after modeling, the MSCs were transplanted into left ventricle anterior wall at four sites in cell transplantation group, model group was injected with matching DMEM/FI2 medium, while the normal group was not given any treatment.MAIN OUTCOME MEASURES: At four weeks postoperatively, left ventricular end-diastolic dimension, end systolic dimension,left ventricular ejection fraction, and shortening fraction were monitored by ultrasonograph; the value for monophasic action potential amplitude (MAPA) and the maximum velocity in 0 phase (V,,~), the value for 50% monophasic action potential durations (MAPDs0) and 90% monophasic action potential durations (MAPDg0) were detected by electrophysiolograph. In addition, the cardiac tissues harvested from implanted region were observed by light microscope and electron microscope, and also the expression of cardiac troponin T (cTnT) and connexin 43 (Cx43) was detected through immunohistochemistry.RESULTS: Of 43 rabbits, 9 rabbits each of the transplantation group died of the acute or chronic toxic effects of Adriamycin, finally, 25 rabbits were included in final analysis. Compared with model group, the end systolic dimension was diminished, and the left ventricular ejection fraction and shortening fraction in cell transplantation group were significantly increased (P ＜ 0.05). The MAPDs0 and MAPDgo in cell transplantation group prolonged significantly compared with model group (P ＜ 0.05). In model group,cardiac myocytes appeared mitochondria swelling and hyperplasia, and their sarcomere had different lengths, arranged unevenly,accompanied by abnormal contraction bands. In addition, myolysis was found in parts of myocardium under electron microscope.However, the structural abnormalities in the cell implantation group were less than the DCM model group, and the implanted MSCs could express cTnT and Cx43.CONCLUSION: Allogenic MSCs transplantation can improve cardiac function, lessen structural abnormalities and may inhibit the progression of electrophysiology derangement.

Objective:To determine the derivative chromosome 9 by the method of detecting the ASS gene,and to explore the relationship between the deletion of derivative chromosome 9 and the efficacy and prognosis of the chronic myeloid leukemia (CML)patients. Methods:The materials of 34 CML patients with positive BCR-ABL fusion gene whose ASS genes were detected were retrospectively analyzed. All patients were treated with Extra-signal (ES)probe to detect the derivative chromosome 9.All patients were divided into two groups according to whether they carried the derivative chromosome 9.The blastic phase or the accelerated phase rates in two groups were compared by using Fisher exact probability. Results:All patients were detected by FISH (BCR-ABL ES probe),and all the BCR-ABL fusion signals were positive.6 of 34 patients were found the deletion of ASS gene, among them 1 case blonged to chronic phase,and 5 cases developed into blastic phase or accelerated phase. In the patients without ASS gene deletion,there were 22 cases in chronic phase,and 6 cases in plastic phase or accelerate phase,there was significant difference of blastic phase rate/accelated phase rate between them (P<0.05).A total of 26 patients were treated with tyrosine kinase inhibitors (TKI).5 of 26 patients belonged to the ASS gene deletion group,1 of 5 patients treated with TKI got molecular remission,4 of 5 patients developed into blastic phase or accelerated phase.21 of 26 patients belonged to the group without ASS gene deletion,and among them,19 cases got molecular remission,2 cases developed into plastic phase or accelerated phase after treatment of TKI,there was significant difference between them (P < 0.05 ). 6 patients were treated with traditional chemotherapy (hydroxyurea,interferon);1 of 6 patients belonged to the ASS gene deletion group,finally developed into the blastic phase or accelerated phase;5 of 6 patients belonged to the group without ASS gene deletion,2 cases got the hematological remission,and 3 patients developed into blastic phase or accelerated phase after treatment,and there was no significant difference of blastic phase rate/ accelerated phase rate between them (P > 0.05 ). Conclusion:The CML patients with derivative chromosome 9 (ASS gene deletion)prone to get disease progression, and have a higher proportion in the blastic phase or accelerated phase patients.Derivative chromosome 9 is related to the bad treatment efficacy of TKI and the poor prognosis of CML.

Objective To compare the characterization and myocardial differentiation capacity of amniotic fluid-derived mesenchymal stromal cells (AF MSCs) and umbilical cord Wharton's Jelly-derived mesenchymal stromal cells (WJ MSCs). Methods The human AF MSCs were cultured from amniotic fluid samples obtained by amniocentesis. The umbilical cord WJ MSCs were obtained from Wharton's Jelly of umbilical cords of infants delivered full-term by normal labor. The morphology, growth curves, and analyses by flow cytometry of cell surface markers were compared between the two types of cells. Myocardial genes (GATA-4, c-TnT, α-actin, and Cx43) were detected by real-time PCR and the corresponding protein expressions were detected by Western blot analysis after myocardial induced in AF MSCs and WJ MSCs. Results Our findings revealed AF MSCs and WJ MSCs shared similar morphological characteristics of the fibroblastoid shape. The AF MSCs were easily obtained than the WJ MSCs and had a shorter time to reach adherence of 2.7 ± 1.6 days to WJ MSCs of 6.5 ± 1.8 days. The growth curves by MTT cytotoxic assay showed the AF MSCs had a similar proliferative capacity at passage 5 and passage 10. However, the proliferative capacities of WJ MSCs were decreased at 5 passage relative to 10 passage. Both AF stem cells and WJ stem cells had the characteristics of mesenchymal stromal cells with some characteristics of embryonic stem cells. They express CD29 and CD105, but not CD34. They were positive for Class I major histocompatibility (MHC I) antigens (HLA-ABC), and were negative, or mildly positive, for MHC Class II (HLA-DR) antigen. Oct-4 was positive in all the two cells types. Both AF MSCs and WJ MSCs could differentiate along myocardium. The differentiation capacities were detected by the expression of GATA-4, c-TnT, α-actin, Cx43 after myocardial induction. Conclusions Both AF MSCs and WJ MSCs have the potential clinical application for myogenesis in cardiac regenerative therapy.

Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion (I/R) injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation (H/R).Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo (24.4％ ± 9.4％ vs.2.2％ ± 1.9％,P ＜ 0.01,n =5) and in vitro (14.12％ ±0.92％ vs.2.22％ ± 0.08％).The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.

Objective To preliminarily explore the effects and brain protective mechanism of intermittent hypoxia preconditioning ( IHP) on rats with seizures induced by lithium-pilocarpine ( Li-pilo) .Methods A total of 96 8-week old male Sprague Dawley rats ( clean grade ) were randomly divided into control group , seizure group and four IHP-seizure groups.The animal model of epilepsy was established by intraperitoneal injection of Li-pilo in the seizure group and four IHP-seizure groups (Li-pilo was injected at 1, 3, 7, or 14 days after a 5-day regimen of IHP).Subsequent seizure behavior , the latency period and percentage of generalized seizures were quantitatively evaluated for 240 min and the cognitive function was tested by Morris water maze task , and followed by the detection of hippocampus neuron apoptosis and related protein (BCL-2, Bax, and cleaved-caspase-3) by TUNEL labeling and Western blot, respectively.Results The induced seizure peaked on an average between 50-150 min after Li-pilo administration , scored using a modified Racine scale.The average scores of modified Racine scale in the IHP-3d seizure group was significantly lower than that in the other groups.The latency period and percentage of generalized seizures in the IHP-3d seizure group rats were significantly different from the parameters in the seizure group rats (P<0.05).IHP-3d seizure rats showed lower escape latency, neuronal apoptosis counts and higher percentage of time in the probe quadrant compare with the seizure group and the other three IHP-seizure groups ( P <0.05 ) .Compared with the control group , the parameters of water maze and apoptosis detection in the IHP-3d seizure group showed no significant changes (P>0.05).Conclusions The results indicate that IHP treatment may help to decrease the susceptibility to epilepsy by reducing abnormal apoptosis , and has a brain protective effect on the seizure rats .

Doctor WANG Zhi-zhong in the Southern Song Dynasty proposed the acupoint view of "location of disease", which explained the connotation of acupoints from the angle of clinic. Its meaning included two levels, one level meant pathological change on the body surface, that was the location of acupuncture diagnosis-treatment, and the other one indicated that the body surface which was the reflecting point of pathological change on the distal area or inside the body was the location of acupuncture diagnosis-treatment. The specific connotations of clinical acupoints were: location of pathogenic factors or reflection of pathogenic factors, regularity between acupoints un- der disease and specific organ, morphological differences and positioning variability after acupoints under disease, and acupoints examination, diagnosis and treatment.
Acupuncture Points ; Acupuncture Therapy ; history ; China ; History, Ancient ; Humans ; Medicine in Literature ; Meridians

Objective To isolate, culture and identify mouse umbilical cord mesenchymal stem cells( mUCMSCs) and to study whether they can be induced to differentiate into adipocytes, chondrocytes and osteoblasts.Methods The mUCMSCs were isolated and expanded by adherent culture from fresh mouse umbilical cord.The morphological characteris-tics of the resulting cells were observed under inverted phase contrast microscope, and their expression of mesenchymal sur-face markers was identified and analyzed by flow cytometry.Then mUCMSCs were induced to differentiate into chondro-cytes, adipocytes and osteoblasts in vitro.Results Fibroblast-like cells could be isolated from the fresh umbilical cord by adherent culture.These adherent cells highly expressed mesenchymal markers including CD29, CD90 and CD105 while low expression of CD34.The cells were successfully induced to differentiate into chondrocytes, adipocytes and osteoblasts. Conclusions The mUCMSCs isolated from fresh mouse umbilical cord by adherent culture have potential of differentiation into chondrocytes, adipocytes and osteoblasts.

Objective To investigate that butyrate-producing probiotic clostridium butyricum improves the intestinal epithelial barrier function in food allergic mice by regulating TWIK-related potassium channel-1 (Trek1) expression in intestinal epithelial cells.Methods An intestinal allergy mouse model was created,then the model construction effect was verified by detecting the related indicators by ELISA,flow cytometer.The change of small intestinal tissue permeability was detected by the Ussing chambers.The Trek1 expressions in mouse jejunum tissue in control group and allergy group were detected by Western blot and immunofluorescent method;in the Transwell system,T84 cells were used to establish epithelial cellular monolayer for exposing to the allergic mediators,the Trek1 mRNA and protein expression were detected by qRT-PCR and Western blot.Then the allergic mice were grouped and treated by different methods including normal saline,SIT,SIT/SB,SB,SIT/CB,CB,SIT/CB/Spadin,the expression of mice intestinal Trek1,intestinal barrier function and allergic reaction indicators were detected.Results Compared with the control group,the small intestinal Trek1 protein and tissue expression level in the food allergic mice were significantly decreased,the intestinal mucosal permeability was significantly increased,the differences were statistically significant(P＜0.05);after T84 cells exposing to the allergic mediators,Trek1 mRNA and protein expression were significantly decreased(P＜0.05),but adding p38 inhibitor in advance could antagonize this change;the single use of SIT,clostridium butyricum and sodium butyrate could increase the intestinal Trek1 expression in food allergic mice,and alleviated the allergic reaction(P＜0.05),whereas the combined use of SIT and clostridium butyricum or sodium butyrate had more significant effect(compared with the SIT group,P＜0.05),moreover could significantly decrease small intestinal mucosal permeability and improved the intestinal barrier function(P＜0.05).Conclusion Sodium butyrate or butyrate-producing probiotic clostridium butyricum can restore the intestinal barrier function,alleviates the allergic reaction and strengthens the SIT curative effect in allergic mice by increasing Trek1 expression.