Objective To observe the influence of ATP protection after brachial plexus injuries. Methods A total of 80 female Wistar rats,weighting 280～300 g,were randomly divided into ATP and con- trol groups.The right C_5～T_1 nerve roots were transected and then the intraperitoneal injection of 4m[ of ATP or normal saline was given immediately and once daily to the rats,respectively.The rats were sacrificed on postoperative days 14,28 and 42 respectively.The C_5-T_1 segments of the spinal cord were harvested.NT-3 activity was measured by enzymo-histochemistry method.Four weeks and 6 weeks postoperatively,ultrastruc- ture of the denervated skeletal muscles was observed.Results Compared to the control group,the expres- sions of NT-3 was increased in the treated groups with ATP injection (P

This study tested the effects of the gastrointestinal pulse train electrical stimulation with different parameters and at different locations on the neuronal activities of the lateral hypothalamus area (LHA) in obese rats in order to find the optimal stimulation parameter and location. Eight gastric electrical stimulations (GES) with different parameters were performed and the neuronal activities of gastric-distension responsive (GD-R) neurons in LHA were observed. The effects of stimulations with 8 parameters were compared to find the optimal parameter. Then the optimal parameter was used to perform electrical stimulation at duodenum and ileum, and the effects of the duodenal and ileac stimulation on the GD-R neurons in LHA were compared with the gastric stimulation of optimal parameter. The results showed that GES with the lowest energy parameter (0.3 ms, 3 mA, 20 Hz, 2 s on, 3 s off) activated the least neurons. The effects of GES with other parameters whose pulse width was 0.3 ms were not significantly different from those of the lowest energy parameter. Most gastric stimulations whose pulse width was 3 ms activated more LHA neurons than the smallest energy parameter stimulation, and the effects of those 3 ms gastric stimulations were similar. Accordingly, the lowest energy parameter was recognized as the optimal parameter. The effects of stimulations with the optimal parameter at stomach, duodenum and ileum on the LHA neuronal activities were not different. Collectively, gastrointestinal electrical stimulation (GIES) with relatively large pulse width might have stronger effects to the neuronal activities of GD-R neurons in LHA of obese rats. The effects of the GIES at different locations (stomach, duodenum and ileum) on those neurons are similar, and GES is preferential because of its easy clinical performance and safety.

Objective To investigate the proliferative characteristics of fibroblast-like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immunosuppressive effect of total glucusides of paeony(TGP).Methods FLS of OA and non-inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of TGP.After incubation,the survival fraction(SF)of FLS was evaluated by MTI' and the TNF-?,IFN-?and bFGF level in cultured FLS supernatant was measured by ELISA.The expression of FLS c-los mRNA and cell cycle of OA-FLS was observed by RT-PCR and flow eytometry respectively at the same time.Results No statistical significant differences were noted between the OA and NS FLS in pro- liferating double time.High doses of TGP suppressed FLS-SF more evidently in OA patients than in NS(P0.05).Conclusion High dose TGP can inhibit OA-FLS proliferation,modulate cy- tokine secretion and c-fos expression in OA.This suggests that TGP has immunosuppressive effect on OA syn- ovitis,probably by preventing the synovial hypertrophy in OA.

Objective To investigate the proliferation characteristics of fibroblast like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immnnosuppressive effect of T-614 [N-(3-formy- lamino-4-oxo-6-phenoxy-4H-chromen-7-yl)methanesulfonamide ] on them.Methods FLS of OA and non- inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of T-614.After incu- bation,the survival fraction(SF)of FLS was evaluated by MTT,cell cycle was observed using fluorescence - activated cell sorting(FCS)method and the expression of c-fos and COX-2 mRNA was examined by RT- PCR in FLS of OA patients.Results No statistically significant difference was noted between the OA and NS FLS in proliferation ability and cell cycle.High dose T-614 suppressed FLS SF obviously in OA and NS sta- tistically(P＜0.05),whereas the inhibition degree was not different between the two kinds of FLS.The agent induced cell apoptosis and reduced the accumulation of c-fos mRNA in OA-FLS at dose 1000 ml/L,prolonged G_1 term and shortened S term at dose 200 ml/L.The expression of COX-2 mRNA in OA FLS was suppressed obviously by T-614 at dose 1000 ml/L.Conclusion OA FLS do not display a distinct activated unlimited viability compared with NS cells,without stimulated by proinflammatory cytokine in vitro.High dose T-614 moderately inhibits the proliferation and differentiation of FLS,directly affects gene of the c-fos and COX-2 expression in OA,which may contribute to its immunosuppressive effect on OA'synovitis.

OBJECTIVETo explore the distribution laws of TCM syndrome types and to analyze the distribution of dynamic blood pressure curve, atherosclerosis, and age in senile hypertension patients.

METHODSTotally 1 131 senile hypertension patients were recruited from 7 provinces and municipal cities. Features of TCM syndromes, classification and distribution curves, and syndrome distribution laws were observed. The distribution curves of dynamic blood pressure, carotid atherosclerosis, and age were compared in each TCM syndrome types.

RESULTSThere were four main syndrome types in 736 cases (56.15%), i.e., excessive accumulation of phlegm-dampness syndrome (210 cases, 16.02%), yin deficiency and hyperactivity of yang syndrome (177 cases, 13.50%), Gan-Shen yin deficiency syndrome (79 cases, 6.03%), and deficiency of qi and yin syndrome (252 cases, 19.22%). Besides, there were two more sub-types, i.e., collateral obstruction by blood stasis syndrome and collateral obstruction by phlegm and stasis. Circadian blood pressure monitor was completed in 211 cases. Of them, abnormal circadian blood pressure occurred in 152 cases (accounting for 72. 38%); yin deficiency and hyperactivity of yang syndrome, excessive accumulation of phlegm-dampness syndrome, deficiency of qi and yin syndrome plus collateral obstruction by blood stasis syndrome were most often seen. Color ultrasound of carotid artery was performed in 660 patients of main syndromes. The incidence was quite higher in those of excessive accumulation of phlegm-dampness syndrome (182 cases, 27. 58%), deficiency of qi and yin syndrome plus collateral obstruction by blood stasis syndrome or collateral obstruction by phlegm and stasis (322 cases, 48.79%). Yin deficiency and hyperactivity of yang syndrome was dominant in patients 60 -79 years old, while deficiency of qi and yin syndrome and Gan-Shen yin deficiency syndrome were dominant in patients older than 80 years.

CONCLUSIONSExcessive accumulation of phlegm-dampness syndrome, yin deficiency and hyperactivity of yang syndrome, Gan-Shen yin deficiency syndrome, and deficiency of qi and yin syndrome were main syndrome types in senile hypertension patients. There was statistical difference in the distribution curves of blood pressure, atherosclerosis, and age of various TCM syndrome types.

Aged ; Asian Continental Ancestry Group ; Atherosclerosis ; epidemiology ; Biomedical Research ; Blood Pressure ; Humans ; Hypertension ; epidemiology ; Medicine, Chinese Traditional ; Qi ; Research Design ; Risk Factors ; Yin Deficiency ; epidemiology

ObjectiveTo investigate the clinical effects and side-effect of mesenchymal stem cell(MSCs) transplantation on spinal cord injury(SCI),traumatic brain injury(TBI),multiple sclerosis(MS) or Parkinson's disease(PD).MethodsThe bone marrow(222～350 ml) of 11 patients with SCI(n=6),TBI(n=3),MS(n=1) or PD(n=1) were harvested from the patients' ilia and then MNCs were isolated.The MNCs were injected intravenously or into subarachnoid space by lumbar puncture.The neural function and side-effect were observed before and after MSCs transplantation and the patients were followed up.ResultsThe data demonstrated the improvement of sense and motor function in 5 patients with SCI,one had no improvement by 2 months following-up.These patients' sense and motor levels improved obviously.Their muscle strength of lower extremity increased,the muscular tone decreased and urinary bladder function improved.Changes in neurological deficits and improvements in function may appear within 2 days after transplantation,most of them within 2 weeks.There were significantly amelioration in 3 patients with TBI treated with MSCs transplantation,one of them could walk with cane independently after 3 months.One's PVS score elevated from 5 to 8 scales after transplantation.The tremor was alleviated after 1 week,and the muscular tone decreased,which lead to reduce the dose of Madopar after 3 months,in patient with PD.The patient with MS showed no improvement in short time.The side-effect included fever(7/11),headache(2/11) and abdominal dissension(1/11).1 patient feel numb in his legs while injection into subarachnoid,and appeared meningeal stimulation after injection.ConclusionThere were significantly clinical effects in treatment of SCI,TBI,MS,and PD with MSCs transplantation in short time,and with few side-effect. The long-term clinical effects need more observation with larger samples.

BACKGROUNDPodocyte has inflammatory role in the development of diabetic nephropathy (DN). Mycophenolate mofetil (MMF), an anti-inflammatory agent, can suppress macrophage infiltration and reduce renal injury in streptozotocin-induced diabetic rats. Angiotensin II receptor blocker (ARB), another renal protecting agent, can decrease podocyte loss in DN. In this study, we detected the expression levels of monocyte chemoattractant protein-1 (MCP-1) and nephrin to evaluate podocyte's role in inflammatory reaction in DN, observe and compare the effect of MMF alone and in combination with valsartan, on preventing podocyte loss in streptozotocin (STZ) induced diabetic rats.

METHODSDiabetic model was constructed in uninephrectomized male Wistar rats by single peritoneal injection of STZ (65 mg/kg). The successfully induced diabetic rats were randomly divided into four groups: diabetes without treatment group (DM), valsartan treated group (DMV), MMF treated group (DMM), and combined therapy group (DMVM). Normal rats of the same sibling were chosen as control (NC). At the end of the 8th week, serum biochemistry, 24-hour urinary protein (UP) and the ratio of kidney weight/body weight (RWK/B) were measured. The rats were sacrificed for the observation of renal histomorphology through light and electron microscope. Nephrin, desmin and MCP-1 levels were detected by semi-quantitative immunohistochemical assays. Real-time quantitative PCR was used to detect the mRNA levels of nephrin and MCP-1.

RESULTSCompared with group NC, serum glucose level, 24-hour UP and RWK/B in group DM were significantly higher (P < 0.01), and the nephrin mRNA level in DM group was significantly lower (P < 0.05). The nephrin mRNA expression levels in group DMV, DMM and DMVM were all higher than that of DM group (P < 0.05) and no significant differences were found among the three treatment groups (P > 0.05). Treatment with MMF, valsartan or their combination could significantly decrease the 24-hour UP and RWK/B, and suppress glomerulosclerosis and interstitial fibrotic lesions in diabetic rats. In diabetic rats, the high expressions of desmin and MCP-1 in kidney were suppressed by valsartan, MMF or their combination.

CONCLUSIONSPodocytes are involved in the inflammatory reaction of diabetic rats. MMF could suppress MCP-1 and desmin expression, enhance nephrin expression, and attenuate proteinuria in diabetic rats. The combined therapy of valsartan and MMF did not show any superiority over monotherapies on renal protection. MMF may have renoprotective effect in early stages of diabetic nephropathy through preventing podocytes loss and anti-inflammatory activity.

Animals ; Chemokine CCL2 ; analysis ; Desmin ; analysis ; Diabetic Nephropathies ; drug therapy ; pathology ; Drug Therapy, Combination ; Immunohistochemistry ; Male ; Membrane Proteins ; analysis ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; therapeutic use ; Podocytes ; drug effects ; pathology ; Rats ; Rats, Wistar ; Tetrazoles ; administration & dosage ; therapeutic use ; Valine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Valsartan

OBJECTIVETo observe the effect of Cornus Officinalis total glycosides (COTG) and Cornus polysaccharides (CP) on myocardial mitochondria and expression levels of glycogen synthase kinase-3β (GSK-3β) of acute myocardial infarction (AMI) rats.

METHODSThe AMI rat model was established by ligating the left anterior descending branch of coronary artery. Rats were divided into 5 groups according to random digit table, i.e., the sham-operation group, the model group, the COTG prevention group, the CP treatment group, the COTG treatment group, 12 in each group. Normal saline was administered to rats in the normal control group and the model group by gastrogavage. Corresponding medication was respectively administered to rats in the rest 3 groups by gastrogavage. The cardiac function was detected by echocardiography and hemodynamics. The infarct size was determined by Masson trichrome staining. The expression of mitochondrial biogenesis genes such as a subunit of peroxisome proliferators-activated receptor-γ coactivator-1 (PGC-1α), PGC-1β, nuclear respiratory factor-1 (NRF-1), and GSK-3P mRNA were detected by Real-time PCR.

RESULTSCompared with the sham-operation group, the myocardial infarction size increased, cardiac function decreased, the expression of PGC-1α, PGC-1β, and NRF-1 mRNA decreased, and the expression of GSK-3β mRNA increased (all P <0. 05). Compared with the model group, myocardial infarction sizes were reduced, cardiac function was improved, the expression of NRF-1 mRNA was elevated in the COTG prevention group, the CP treatment group, the COTG treatment group; the expression of the PGC-1α and PGC-1β mRNA was elevated in the COTG prevention group and the CP treatment group; the expression of GSK-3β mRNA was reduced in the CP treatment group (all P <0. 05). Compared with the CP prevention group, fractional shortening (FS) and aortic systolic blood pressure (SBP) increased in the CP treatment group; ejection fraction (EF) decreased in the CP treatment group; the expression of PGC-1α, PGC-1β, NRF-1 mRNA were reduced in the the CP treatment group and the COTG treatment group; the expression of GSK-3β mRNA decreased in the CP treatment group (all P <0. 05). Compared with the COTG treatment group, FS, EF, left ventricular end systolic pressure (LVESP), SBP, and the expression of GSK-3β mRNA were reduced in the CP treatment group (P <0. 05).

CONCLUSIONSCOTG and CP could improve cardiac function, reduce the myocardial infarction area, and promote biogenesis of myocardial mitochondria. Their protective effects on the mitochondria of cadiocytes might be achieved by GSK-3β signalina pathway.

Animals ; Cornus ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Glycogen Synthase Kinase 3 ; Glycogen Synthase Kinase 3 beta ; Glycosides ; Heat-Shock Proteins ; Mitochondria, Heart ; physiology ; Myocardial Infarction ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Polysaccharides ; Protective Agents ; pharmacology ; therapeutic use ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Transcription Factors

OBJECTIVETo explore the inflammatory cascade mechanism through Toll like receptor 2 (TLR2) pathway after cerebral ischemia/reperfusion, and to study molecular mechanisms of Guanmaitong (GMT) Tablet for protecting brain damage.

METHODSWe used bolt-line method to block/release the middle cerebral artery, causing cerebral ischemia/reperfusion (I/R) injury model. GMT Tablet was given by gastrogavage. Rats were then divided into the high dose GMT group (1200 mg/kg), the middle dose GMT group (600 mg/kg), the low dose GMT group (300 mg/kg), the positive control group (Tanakan, 20 mg/kg). Their right brain tissues were fixed in 10% neutral formalin. TLR2 expressions were detected by immunofluorescence staining. The total protein was extracted from right brain tissues by ultrasonica- tion. Expression levels of extracellular regulated protein kinases (ERK), phospho-extracellular regulated protein kinases (p-ERK), p38-mitogen activated protein kinases (p-ERK), phospho-p38-mitogen activated protein kinases [p-p38-MAPKs(p-p38)] were assessed by Western blot. Abdominal aortic blood was withdrawn. IL-6 and IL-1β levels were detected by ELISA in brain tissues and serum.

RESULTSCompared with the sham-oepration group, expression levels of TLR2, ERK, p-ERK, p38, p-p38 protein were up-regulated (P < 0.05, P < 0.01), and contents of IL-6 and IL-1β in brain tissues and serum were increased in the model group (P < 0.01). Expression levels of TLR2, ERK, p-ERK, p38, p-p38 were down-regulated (P < 0.05, P < 0.01), and contents of IL-6 and IL-1β were reduced in brain tissues and serum in middle and high dose GMT groups (P < 0.05, P < 0.01).

CONCLUSIONSTLR2 pathway was involved in cerebral I/R injury. GMT protected neurons by down-regulating protein expressions of TLR2, ERK, p-ERK, p38, p-p38 and contents of IL-1β and IL-6.

Animals ; Blotting, Western ; Brain Ischemia ; metabolism ; Cerebral Infarction ; Down-Regulation ; Drugs, Chinese Herbal ; therapeutic use ; Interleukin-1beta ; Interleukin-6 ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; Tablets ; Toll-Like Receptor 2 ; metabolism ; Up-Regulation ; p38 Mitogen-Activated Protein Kinases ; metabolism

AX15 is a mutein of naturally occurring human ciliary neurophic factor (hCNTF), with improved biological activity, stability and solubility. AX15 is susceptible to protease degradation when expressed in Pichia pastoris. Amino acid sequencing revealed the degradation was occurred behind position 12 and 13 amino acid residues, which constitute a dibasic site, RR. Based on the substrate specificity of KEX2, a KEX2 resistant mutein of AX15-AX15 (R13K) was constructed, in which RR was replaced by RK. It was demonstrated that the stability of AX15 (R13K) improved significantly, as no degradation was detected even after 120 hours of induction. AX15 (R13K) was purified to homogeneity by ultrafiltration and gel filtration. TF-1 cell survival bioassay showed AX15 (R13K) had equivalent specific activity to AX15. The protease resistant mutein of AX15 may have greater in vivo stability and thus have superior therapeutic potential.
Ciliary Neurotrophic Factor ; biosynthesis ; genetics ; Genetic Vectors ; Humans ; Mutant Proteins ; biosynthesis ; genetics ; Mutation ; Peptide Hydrolases ; chemistry ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification