Objective To investigate abnormal myocardial contractile responses induced by dobutamine in patients with idiopathic dilated cardiomyopathy (DCM).Methods Eighteen DCM patients underwent low dosage dobutamine stress echocardiography (5,10,20 ?g?kg~ -1?min~ -1).Transthoracic echocardiogram was recorded with the use of a HP Sonos 5500 color echocardiographic diagnostic system.Wall motions in 16 myocardial segments were assessed using a four-point scale recommended by the American Society of Echocardiography.During dobutamine infusion,abnormally contracting segments were classified into four different patterns of contractile response: improved,unchanged,worsened and biphasic.Unchanged,worsened and biphasic segments were defined as abnormal contractile responses.Worsened and biphasic segments were judged to be ischemia-like responses.Results All patients showed abnormal myocardial contractile responses of wall motions (100%),and thirteen were ischemia-like responses ( 72.2%).In total 225 segments,126 segments showed abnormal contractile responses ( 56.0%).Among them,97 segments were unchanged segments ( 43.1%),29 segments were ischemia-like responses ( 12.9%),in which 16 segments were worsened ( 7.1%),and 13 segments were biphasic ( 5.8%).The statistical analysis showed that incidence of improved segments was highest in two point segments( 62.1%) compared with three point and four point segments; incidence of ischemia-like responses was highest in three point segments( 24.6%); incidence of unchanged segments was highest in four point segments( 68.4%).Conclusions DCM has the abnormal myocardial contractile responses induced by dobutamine stress echocardiography.It indicates there is the ischemia-like energy mismatch between demand and supply in DCM,which would be related to etiology and progress of DCM.

Objective To investigate whether dobutamine could induce or worsen electromechanical activity synchronism in the patients with idiopathic dilated cardiomyopathy(DCM). Methods Eighteen patients with DCM[mean age ( 45.8 ? 13.9 ) years, NYHA class Ⅱ-Ⅲ, left ventricular ejection fraction(LVEF)

BACKGROUND:Intervertebral disc degeneration is the pathological basis of degenerative spinal diseases. Studies on the influentialfactors of intervertebral disc degeneration contribute to the prevention and treatment of degenerative spinal disease.
OBJECTIVE:To observe the growth and proliferation of nucleus pulposus cels isolated by trypsin plus type II colagenase digestion in complete medium with different glucose concentrations, exploring the optimal glucose concentration for growth of nucleus pulposus cels.
METHODS:Nucleus pulposus cels isolated and cultured by trypsin plus type II colagenase digestion method were observed under an inverted microscope, and thecelnumber was counted. Morphology of nucleus pulposus cels was observed afterhematoxylin-eosinstaining and toluidine blue staining. Colagen type II immunoreactivity was detected by immunohistochemical staining combined with immunofluorescent staining.Nucleus pulposuscels were incubated in complete medium containing various glucose concentrations (0, 6.25, 12.5, 17.5, and 25 mmol/L) for 24 hours, and then cel proliferation and apoptosis were determined.
RESULTS AND CONCLUSION:The stained nucleus pulposus cels showed polygonal and short spindle, with one or two nuclei. Celularpseudopod appeared gradualy and then became slim with increased passage numbers. The isolated and cultured nucleus pulposus cels positively expressed colagen type II and aggrecanProliferative activity of nucleus pulposus cels cultured in medium with 17.5 mmol/L glucose was significantly higher than that in medium with 0 and 25 mmol/L glucose (P< 0.05 orP< 0.01). There wasno significant differencein cel apoptosis between these groups except for 0 mmol/L glucose (P<0.05). These results confirm that a large number of nucleus pulposus celscan beharvested by trypsin plustype II colagenase digestion and the optimal glucose concentration is 17.5 mmol/L.

Objective To prepare highly expressed, purified and refolded SCR15-18 of human soluble complement receptor type 1 (sCR1-SCR15-18) protein. Methods The expression of recombinant pET32-sCR1-SCR15-18 in E.coli.. BL21 was induced by IPTG of different concentrations for different time period under different temperatures and the bacteria were split by sonication. The sCR1-SCR15-18 protein was purified by Ni2+-NTA resin affinity chromatography. The purified protein was refolded under different conditions. Then the bioactivity of the protein was analyzed. Results The sCR1-SCR15-18 protein of high expression, purity and bioactivity was attained. Conclusion The parameters of expression, purification and refolding of sCR1-SCR15-18 protein were optimized, which may pave a way for further studies.

AIM: To set up population pharmacokinetics/pharmacodynamics (PPK/PD) model of valproate (VPA) in children with epilepsy in China, and promote reasonable use of antiepileptic drugs(AEDs)in clinical practice. METHODS: Sparse data of VPA serum concentrations from 246 pediatric children were collected. These patients were divided into three groups: PPK-Model group ([WTBX]n=146), to calculate PPK parameter values of VPA and set up a PPK model; PPK-Valid group ([WTBX]n=100), to valid the PPK model; and PPD group ([WTBX]n=69), to set up PPK/PD model. Based on the data of PPK-Model group and PPK-Valid group, a PPK model of VPA in children with epilepsy in China was successfully set up by using NONMEM software by ourselves. Now, using the data of 69 patients in PPD group who were on VPA monotherapy and this PPK model, we set up PPK/PD model by NONMEM software. Efficacy of epilepsy treatment was divided into 5 grades according to the percentage of seizure frequency decreased (PSFD%): grade 1: PSFD% was 100%; grade 2: PSFD% was 75%-100%; grade 3: PSFD% was 50%-75%; grade 4: PSFD% was 25%-50%; grade 5: PSFD% was less than 25%. The quantitive relationship between the VPA serum concentrations and the probability for its efficacy score was characterized by Logistic regression analysis with NONMEM. RESULTS: Logistic regression analysis showed that, VPA serum concentrations and the probability for its efficacy grades 5, 4, 3, 2, and 1 were (23 ?g?ml -1, 5, 50%), (30 ?g?ml -1, 4, 32.3%), (50 ?g?ml -1, 3, 26.3%), (65 ?g?ml -1, 2, 36.5%), (78 ?g?ml -1, 1, 50%), and (100 ?g?ml -1, 1, 84.2%)respectively. CONCLUSION: A PPK/PD model of VPA in children with epilepsy in China is successfully established by using NONMEM software, and the probability of efficacy grade for any concentration can be calculated. It will be valuable to facilitate individualized dosage regimen.

[Objective]To study the pathogenesis,clinical manifestation and surgical treatment of the posterior margin separation of lumbar vertebral epiphysis.[Method]Sixteen patients suffering from the posterior margin separation of lumbar vertebral epiphysis were followed up.The clinical manifestation and radiologic examination were analyzed and results of surgical treatment were evaluated.[Result]The patients were usually young and manifested with the sign and symptom of lumbar disc herniation and/or lumbar stenosis.CT was helpful for the accurate diagnosis of this disease.The different surgical measures were taken for the treatment according to the type and range of protrusion.[Conclusion]The posterior margin separation of lumbar vertebral epiphysis were divided into three types:end plate separation and moving into posterior margin,Schmorl node and avulsion fracture.The good results can be obtained with surgery.

Objective To study the biological effects of pSNAV2-hTGF?1 and pSNAV2-hTGF?3 on the reversion of rabbit disc degeneration. Methods Rabbit nucleus pulpous and annulus fibrosus cells were isolated and cultured. The fluorescence labled pSNAV2 were used to detect the transfect rates of rabbit disc cells at first. Then, the pSNAV2-hTGF?1 and pSNAV2-hTGF?3 were transfected into the degenerated rabbit disc cells respectively. The biological effects of hTGF?1 and hTGF?3 on degenerated rabbit disc cells were detected with Western-bloting and 35S detection to analyze and compare the matrix synthesis of the tranfected cells. Results pSNAV2 could transfect degenerated disc cells effectively in the early stages. Both the pSNAV2-hTGF?1 and pSNAV2-hTGF?3 could stimulate the synthesis of collagen Ⅱ and proteoglycan of the rabbit disc cells. For the early stage of degenerated disc cells, the synthesis of collagen Ⅱ and proteoglycan were greater transfected with pSNAV2-hTGF?1 than transfected with pSNAV2-hTGF?3. The pSNAV2-hTGF?1 could promote the degenerated rabbit annulus fibrosus cells to synthesize collagen Ⅰ and pSNAV2-hTGF?3 could promote the degenerated nucleus pulpous cells of later stage to synthesize the collagen Ⅱ. Conclusion Both pSNAV2-hTGF?1 and pSNAV2-hTGF?3 can promote the degenerated rabbit disc cells of early stage to synthesize the matrix. pSNAV2-hTGF?3 can efficently promote the seriously degenerated nucleus pulpous cells to synthesize the collagen Ⅱ.

Objective To evaluate the reversion possibility of hVEGF165 and TGF?1 to intervertebral disc degeneration by gene method. Methods The hVEGF165cDNA obtained from plasmid pcDNA3(+)-hVEGF165 was subcloned into the packaging plasmid pSNAV of AAV by molecular clone ways. The recombinant plasmid pSNAV-hVEGF165 was identified by restriction enzymes analysis and sequencing analysis, and then transfered to the HEK293 cell and VEC by lipofectamine mediated gene transfer method. The protein hVEGF165 was detected by immunofluorescence for immunocytochemistry and explored the influence to the proliferation of vascular endothelial cell by MTT. Whereafter the AAV-hVEGF165 was packaged by Benyuan Zhengyang Company. AAV-hVEGF165 and AAV-TGF?1 were cotransfected into annulus fibrosus cell of intervertebral disc, then the expression of hVEGF165 and TGF?1, and the change of collagen Ⅰin annulus fibrosus cell were detected by Western blot. Results The recombinant pSNAV-hVEGF165 was completely constructed and confirmed by restriction enzymes analysis and sequencing analysis. The protein hVEGF165 was detected by immunofluorescence for immunocytochemistry in experimental group, and hVEGF165 could promote the proliferation of vascular endothelial cell. The bioactive AAV-hVEGF165 was successfully constructed. The expression of AAV-hVEGF165 and AAV-TGF?1 were manifested in degenerative annulus fibrosus cell by Western blot, and the expression of collagen Ⅰin annulus fibrosus cell cotransfected by AAV-hVEGF165 and AAV-TGF?1 was markedly more than that of the monogenic transfected cell. Conclusion hVEGF165 could cooperate with TGF?1 to promote the expression of collagen Ⅰ.

Objective To study the method of isolating and culturing stem cells from rat adipose tissue and to determine if adipose-derived stem cells(ADSCs)harvest from rat could differentiate into osteogenic and chondrogenic in vitro.Methods ADSCs were isolated from rat inguinal fat pads after extensive washing with phosphate-derived saline and digesting with collagenase.After primary culture in control medium and expanded to two passages,the cells were incubated in either an osteogenic medium or an chondrogenic medium for 2-4 weeks to induce osteogenesis and chondrogenesis,respectively.Evidences of Osteogenic differentiation,were detected by a ALP solution,and while chondrogenic differentiation was confirmed using the histologic Alcian blue staining at acidic pH.Expressions of osteocyte and chondrocyte specific genes were confirmed by RT-PCR.Results ADSCs can be isolated from rat adipose tissue and,expanded rapidly.It exhibited a heterogeneous population of fibroblast like cells morphologically.ADSCs induced to osteogenesis were stained positively for alkaline phosphatase activity after 2 weeks and formed mineralized nodular structures,as conformed by Von kossa staining.Expression of osteocyte specific genes,such as ALP,osteopontin,osteocalcin,was detected.ALP and osteopontin,were expressed constitutively in osteogenic medium after 2 and 4 weeks of culture.Expression of osteocalcin,was induced by osteogenic growth factors at 4 weeks.Induced to chondrogenesis cells were positive of Alcian blue staining under acidic conditions and expression of aggrecan and typeⅡ/Ⅹ collagen genes.Aggrecan and typeⅡcollagen genes were abundant after 2 weeks in chondrogenic medium.TypeⅩ collagen gene was detected at 4 weeks.Conclusion Adipose-derived stem cells can be isolated from rat adipose tissue.Their biological characteristics are similar with mesenchymal stem cells(MSCs),and have the potential to differentiate into osteogenic and chondrogenic lineage.It may be an idea source of ADSCs for tissue engineering.

[Objective]To evaluate the accuracy and influenceing factors of somatosensory evoked potential in spinal cord monitoring during cervical and thoracic spinal surgery and intraoperative nerve root monitoring in lumbar surgery.[Method]The somatosensory evoked potential(SEP) were used during arvical and thoracic spinal surgery and evaluated the accuracy of SEP according to the record of different stages and spinal cord function after surgery.The EMG were used to monitor the nerve root function in lumber operation to estimate whether nerve root being stimulated or tensioned.In addition,affected fators of SEP and EMG during operation were observed.[Result]Of 128 cases of cervical and thoracic surgery,116 cases did not reach the warning standards(amplitude decreasing 50% or diappearing) and showed no postoperative enhancement of symptom of nerve roots injury.12 cases reached the warning standards intraoperatively and the surgeon were warned to take some steps to finish the operations,only in one case incompletely transient paralysis occurred due to the time of amplitude decreasing of intraoperative SEP more than 10 minutes.Effect of other factors such as anaesthesia and low blood pressure did not reach the warning standards.There were 3 artifical negative cases.Only 1 was artifical positive case.of 40 cases of lumbar surgery,12 cases were found myoelectic responses,which warning the surgeon at any time to avoid nerve roots injury,no nerve roots injury were found after operation.[Conclusion]During cervical and thoracic spinal operation,the somatosensory evoked potential can reflect the physiological and pathological conditions of spinal cord after ruling out the interfering factors.Intraoperative spontaneous electromyography can reflect the nerve roots function promptly and accurately and assure the safety of lumbar surgery.