0.05). When MS and NID groups compared with NNID group, the difference was extremely significant(all P

Objective:To investigate the therapeutic efficacy of immunotherapy with dendritic cells-based vaccine against intracranial gliomas in rats.Methods:C6 glioma cells were injected into the brain of Wistar rat under stereotactic monitor to establish an animal model of glioma.The precursors of dendritic cells were isolated from bone marrow of rats,stimulated in vitro with recombinant rat granulocyte-macrophage colony-stimulating factor(rrGM-CSF)and interleukin-4(rrIL-4).Cultured cell populations were confirmed to be functional DCs.These DCs were then pulsed ex vivo with C6 tumor-lysates prepared by three cycles of freezing to -80℃ and thawing to 0℃ and subsequently injected subcutaneously into rats harboring intracranial C6 tumor.Rats from different group were treated with five weekly subcutaneous injections of either control media,unpulsed DCs,or DCs pulsed with tumor-lysates.The animals were followed for survival,the percentage of CD8 +T cells in peripheral blood and cytotoxicity assay in vitro were determined by FACS.The level of cytokine IFN-? and IL-10 were detected by ELISA.Results:The results indicated that C6 glioma model rats treated with tumor-lysate-pulsed DCs led to prolonged survival time,increased the percentage of CD8 + T lymphocyte in peripheral blood in comparing with control group.Cytotoxicity assays suggest that vaccination with these tumor-lysate-pulsed DCs can induce specific cytotoxic T lymphocytes against C6 tumor cells compared with control group.Furthermore,significantly enhanced IFN-? and reduced IL-10 (even undetectable)were observed in rats treated with pulsed-DCs.Conclusion:Data supported the therapeutic efficacy of systemic vaccination with DCs pulsed with tumor-lysates against intracranial glioma.

Objective To investigate whether or not extracorporeal shock wave (ESW) treatment is safe for treating low back pain caused by lumbar facet joint degeneration by observing its effects on the lumbar segment of the spinal cord.Methods Five adult dogs were used for this study.ESWs were focused on their right L_(4～5) facet joints. The energy output was set at 40% of the maximum output,and a total of 1000 shocks were delivered at a frequency of 120/min.Before and 3 days after the treatment,the somatosensory evoked potentials (SEPs) induced by stimulating the tibial nerve were recorded from the animals' scalps,and the motor function of their hind limbs was evaluated with Tarlov's scale.The animals were then sacrificed and the L_(4～5) segments of their spinal cords were taken for histopatho- logical observation,with the T_(12) segments taken as controls.Results Three days after ESW treatment,the left and the right hind limbs scored 5 and 4 on the Tarlov scale.The amplitude of SEPs decreased and their latency increased. Histopathological observation showed mild swelling of the L_(4～5) segment of the spinal cord with congested veins on the surface.Erythrocyte and leukocyte exudation as well as myelin sheath swelling in the spinal cord were also observed un- der the microscope,but with no demyelination.Conclusion ESW had some adverse effects on the spinal cord when focused on the facet joint.Further studies of the safety range of ESW energy and distance are needed.

Hearing Loss ; diagnosis ; therapy ; Humans

Objective To construct an expression system to produce the virus-like particles containing a part of the sequence of PSA mRNA, which are ribonuclease-resistant due to the encapsulation of the mRNA by bacteriophage MS2 coat proteins. Methods The PCR products of PSA cDNA fragments were cloned to TA vector pBS-T, then the targeted segments could be obtained when the pBS-T-PSA were digested by restriction endonuclease Hind Ⅲ and cloned to prokaryocytic expression vector pNCCL1. The recombinant plasmids named PNCCL1-PSA were transfected into E. Coli BL21-DE3 and induced to express with IPTG. Results The recombinant plasmids were successfully constructed. The bacteriophage MS2 coat protein which expressed in BL21 can self- assemble to form ribonuclease resistant virus-like particles and the PSA mRNA was encapsulated into virus-like particles. Conclusions The virus-like particle containing PSA mRNA can be expressed in prokaryocyte and it can be used as standard and control in detecting PSA mRNA. It provides a new, stable and ribonuclease-resistant RNA standard in RNA detection.

ObjectiveTo evaluate the diagnostic performance of CTCA in predicting myocardial perfusion defects through comparative analysis between MPI defects and severity of coronary stenosis on CTCA.MethodsFour hundred and seventy-eight patients who underwent CTCA and 99Tcm-MIBI MPI in the same period were analyzed retrospectively.According to the severity of coronary stenosis judged by visual evaluation of the vessel diameter,the patients were divided into five groups:no stenosis,mild stenosis,moderate stenosis,severe stenosis and total occlusion.MPI results were classified as negative or positive for perfusion defects,and the prevalence of perfusion defects in every group was calculated per-patient and per-vessel basis.The cut-off of stenotic severity for predicting myocardial perfusion defects was designated as 50％ or 75％,with MPI as standard reference.True positive,true negative,false positive and false negative statistics were thus determined separately on patient and vessel basis.The diagnostic performance for CTCA were calculated and compared.Pearson Chi-square and its partition tests or Fisher exact test were used to compare ordinal variables.ResultsFifty-eight patients showed myocardial perfusion defects.Either by patientbased or vessel-based analysis,the prevalence of myocardial perfusion defects showed an increasing trend with greater coronary artery stenosis in each group,and there were statistical differences among them (x2 =116.62 and 483.83,both P ＜ 0.05).On patient-based analysis,sensitivity ( SN),specificity ( SP),positive predictive value( PPV),negative predictive value(NPV) and accuracy (AC) for CTCA predicting myocardial perfusion defects were 62.1 ％ ( 36/58 ) and 34.5％ ( 20/58 ) (x2 =8.84,P ＜ 0.05 ),84.5％(355/420) and 97.1％ (408/420) (x2 =40.16,P ＜0.05),35.6％ (30/101) and 62.5％ (20/32) (x2 =7.19,P＜0.05),94.2％ (355/377) and 91.5％ (408/446) (x2 =2.18,P ＞0.05),81.8％ (391/478)and 89.5 ％ (428/478) (x2 =11.66,P ＜ 0.05 ) when the cutoff was set to 50％ and 75％,respectively.On vessel-based analysis,the SN,SP,PPV,NPV and AC for CTCA predicting myocardial perfusion defects were 58.8％ (40/68) and 30.9％ (21/68) (x2 =10.73,P ＜ 0.05),95.9％ (1768/1844) and 99.0％ (1826/1844) (x2 =36.72,P ＜ 0.05 ),34.5％ (40/116) and 53.8％ (21/39) (x2 =4.59,P ＜0.05 ),98.4％ (1768/1796) and 97.5％ ( 1826/1873 ) (x2 =4.14,P ＜ 0.05 ),94.6％ ( 1808/1912 ) and 96.6％ ( 1847/1912 ) (x2 =10.31,P ＜ 0.05 ),respectively.ConclusionsThe prevalence of myocardial perfusion defects correlates positively with the severity of coronary stenosis seen on CTCA.CTCA may predict perfusion defects with high SP and NPV.However,the PPV of CTCA in predicting myocardial perfusion defects is poor when the stenosis cut-off is set at 50％.It is significantly improved when the cutoff value is set at 75 ％.

Objective To explore the value of color doppler ultrasonography by bed side in the early diagnosis of HIE in full term neonates.Methods The changes of cerebral parenchymal and cerebral arterial blood stream parameter on 35 cases of neonates clinically diagnosed HIE of mild and moderate degree and 40 cases of normal newborns on the 24,48 and 72 hours after birth were observed by color doppler ultrasonography by bed side.Results 1.The cerebral parenchyma was even echo in normal newborns,but it was maldistributed and reinforced in mild asphyxia neonates and it was more serious in moderate degree.The echo of cerebral parenchyma in mild degree was near normal in 48 hours after birth,while the echo of cerebral parenchyma in moderate degree was still maldistributed and reinforced in 48 and 72 hours after birth.2.There was obvious changes in the cerebral arterial blood stream parameter and hemodynamics of the asphyxia newborns compared with normals.The systolic peak velocity(Vs)and end diastolic velocity(Vd)of the cerebral arteries in mild and moderate degree were obviously lower than that of control group in 24,48 hours after birth(Pa0.05).3.Resistance index(RI)of the cerebral arteries in mild and moderate degree were higher than that of control group in 24,48 hours after birth(Pa0.05).Conclusion Color doppler ultrasonography by bed side is a convenient,noninvasive method for diagnosing HIE.

In temporomandibular disorders (TMD), pain takes place when neuropeptides stimulate synovial tissue to produce several cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, which activate neurons and glia of synovial membrane at the bilaminar regions of temporomandibular joint (TMJ). It has been reported that, after neurogenic differentiation, the synovial mesenchymal stem cells (SMSCs), deriving from TMJ, possess the same cytological features as the neuronal cells. This study examined the ability of substance P (SP) and calcitonin gene-related peptide (CGRP) to stimulate SMSCs and neurogenic SMSCs secreting inflammatory cytokines during TMD, evaluated the mutual effects of inflammatory cytokines and neuropeptides and tested the analgesic effect of hyaluronic acid (HA). The levels of IL-1β, IL-6 and TNF-α in SMSCs and neurogenic SMSCs in the presence of neuropeptides were measured by ELISA. SP and CGRP produced by SMSCs and neurogenic SMSCs were determined by RT-PCR and Western blotting. The results showed that the expression of SP and CGRP was significantly enhanced in the neurogenic SMSCs in response to IL-1β, IL-6 and TNF-α, and the effect was remarkably inhibited by HA. IL-1β, IL-6 and TNF-α, in return, could be enhanced in the neurogenic SMSCs upon stimulation by SP and CGRP. Neuropeptides and inflammatory cytokines might work mutually on the TMD pain. The HA-mediated analgesic effect may be implicated in the inhibition of SP and CGRP expression in neurogenic SMSCs.

Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Diagnosis, Differential ; Histiocytoma, Benign Fibrous ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neprilysin ; metabolism ; Receptors, Cell Surface ; metabolism ; Skin Neoplasms ; metabolism ; pathology ; surgery ; Thigh ; Vimentin ; metabolism

The hom gene encoding for homoserine dehydrogenase was amplified from the genomic DNA of Corynebacterium glutamicum ATCC 13032.After the kanamycin-resistant gene(Km)cassette from plasmid pET28a was inserted into the center of hom,the hom::Km cassette was then electroporated into the competent cell of C.glutamicum ATCC 13032.And kanamycin-resistant clones were obtained.PCR was performed to confirm whether the Km gene was integrated into the hom gene of these clones and the recombinant strains of hom-disrupted were screened out.Fermentation results showed that the lysine yield of the hom-disrupted strain C.g-hom::Km-8 reached 4.7 g/L,which was 6.7 times that of C.glutamicum ATCC 13032.