0.05)in the mild group compared with at the first day, but an obvious rise(P0.05), while continued to increase in the severe group(P

Objective To investigate the expression change of P2X3 receptor in spinal dorsal horn and dorsal root ganglion(DRG) in rats with experimental inflammatory pain.Methods Totally 30 SD rats received a subcutaneous injection of carrageenan in the right paw to establish inflammatory pain model,and then divided randomly and equally into 5 groups,that is 6,12,24,72 and 168 h after injection.Another 6 rats undergoing a subcutaneous injection of normal saline in the right paw served as control.Pain threshold was assayed by thermal radiation and von Frey filament in 2,4,6,12,24,72 and 168 h after injection.Expression of P2X3 receptor in spinal dorsal horn and DRG of every group was measured by immunohistochemical staining(IHC).Results After injection of carrageenan,thermal hyperalgesia and mechanical allodynia appeared in the rats.The thermal withdrawl latency(TWL) and 50% paw withdrawl threshold(PWT) were decreased obviously from 2 h,descended to the lowest at 12 h,then returned to the normal level after 72 h.The expression of P2X3 receptor in spinal dorsal horn was increased obviously from 24 h,while that at DRG was increased obviously from 12 h.Both of the expressions lasted for 72 h,and returned to the normal level at 168 h after injection.Conclusion Thermal hyperalgesia and mechanical allodynia appear in the rats after carrageenan injection,and the P2X3 receptor is activated and up-regulated in the DRG and spinal cord,suggesting that P2X3 receptor may play an important role in the incidence and development of inflammatory pain.

Objective To investigate the localization of Smad2, Smad3, Smad4, and Smad7 proteins and their expression changes in the 5/6 subtotally nephrectomized rat kidney. Methods The rat model of chronic renal failure was established by performing 5/6 subtotally nephrectomy (SNx) and rats in the control group underwent sham-operation. The rats were sacrificed at 4, 8, and 12 week after operation. The sites and levels of expressions of Smad2, Smad3, Smad4, and Smad7 proteins were examined by immunohistochemical staining. Renal fibrosis was assessed by measuring tissue hydroxyproline. Results Immunohistochemical staining indicated that Smad2, Smad3, and Smad4 proteins were mainly expressed in glomeruli and renal tubular cells, while Smad protein 7 was expressed in glomeruli, but rarely in proximal renal tubular cells. Expressions of Smad2, Smad3, and Smad4 proteins in glomeruli were significantly increased during 4-12 weeks after 5/6 nephrectomy, but the expression level of Smad protein 7 was significantly decreased, but accompanied increase of hydroxyproline content in the renal tissues. Conclusion These results indicate that TGF-?/Smad signaling is involved in the progress of chronic glomerulosclerosis. The high level expressions of Smad2, Smad3, and Smad4 proteins and the down-regulation of Smad7 protein may be the major cause of the glomerulosclerosis in this model.

Objective To investigate the regulating effect of heat reinforcing acupuncture manipulation on body energy metabolism enzymes in rheumatoid arthritis (RA) and preliminarily explain the mechanism of heat-producing action of heat reinforcing acupuncture manipulation. Method Forty chinchilla rabbits were randomized into normal, model, equal reinforcement and reduction, twirling reinforcement, and heat reinforcing acupuncture manipulation groups. A model of cold syndrome-type RA was made by ovalbumin induction and exposure to low temperature in the other four groups not including the normal group. From two days after successful model making, the normal and model groups were grabbed and fastened (bound) by the same way as for the acupuncture groups, 30 min once daily. The equal reinforcement and reduction group received even reinforcing-reducing method;the twirling reinforcement group, twirling reinforcement method;the heat reinforcing group, heat reinforcing acupuncture manipulation. The needle was manipulated for 1 min and retained for 30 min once daily, for a total of seven days. The RA rabbit knee joint circumference was measured and the inflammation score was recorded according to synovial histopathological sections before and after treatment. After the completion of intervention, the rabbits were sacrificed and the articular synovium was rapidly separated for frozen sections. Articular synovium lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and cytochrome oxidase (CCO) activities were measured by histochemical staining. Result After acupuncture intervention, the RA rabbit knee joint circumference was shortened in all the equal reinforcement and reduction, twirling reinforcement, and heat reinforcing acupuncture manipulation groups, but the shortening effect on the RA rabbit knee joint circumference was better in the heat reinforcing acupuncture manipulation group than in the equal reinforcement and reduction and twirling reinforcement method groups (P<0.05);the inflammation score recorded according to the RA rabbit synovial histopathological sections was also decreased more in the heat reinforcing acupuncture manipulation group than in the equal reinforcement and reduction and twirling reinforcement method groups (P<0.05). Synovial LDH integral optical density, total positive area and area percentage were significantly higher in the model group than in the normal group (P<0.05), and significantly lower in the equal reinforcement and reduction, twirling reinforcement, and heat reinforcing acupuncture manipulation groups than in the model group(P<0.05) and also in the heat reinforcing acupuncture manipulation group than in the equal reinforcement and reduction and twirling reinforcement method groups (P<0.05). Rabbit synovial SDH and CCO activities were significantly higher in the model group than in the normal group (P<0.05). Rabbit synovial SDH and CCO integral optical density, total positive area and area percentage were significantly higher in the equal reinforcement and reduction, twirling reinforcement, and heat reinforcing acupuncture manipulation groups than in the model group (P<0.05) and also in the heat reinforcing acupuncture manipulation group than in the equal reinforcement and reduction and twirling reinforcement method groups (P<0.05). Conclusion Heat reinforcing acupuncture manipulation has a definite therapeutic effect on RA and can increase SDH and CCO activities to enhance aerobic metabolism and produce more local energy in a rabbit model of RA. That may be the mechanism by which heat reinforcing acupuncture manipulation produces heat.

OBJECTIVE:To study the effects of different drying methods on the quality of North Qinglongyi,thus optimize the best drying way of North Qinglongyi. METHODS:North Qinglongyi was processed using five kinds of drying methods includ-ing drying in the shade,drying in the sun,oven drying,microwave drying,freeze drying and oven drying under different tempera-ture (40,50,60 ℃). The indicators were investigated,such as drying time,recovery rate of water,water content,alcohol ex-tract,content of active ingredients walnut quinone and juglone,in vitro anti-tumor activity of methanol extract to human gastric ade-nocarcinoma BGC823 cells and human lung cancer A549 cells. The effects of different drying methods on the quality of North Qinglongyi were analyzed comparatively. RESULTS:Among 5 kinds of drying method,microwave drying time was the shortest, and the time of drying in the shade was the longest;water content of sample with freeze drying was the highest,and that with mi-crowave drying was the lowest;the content of alcohol extract has little difference;the content of walnut quinone of sample with freeze drying was the highest,and that with microwave drying was the lowest;the content of juglone of sample dried at 40 ℃ was the highest,and that of freeze drying was the lowest;anti-tumor experiment in vitro showed that inhibition rate of sample with dif-ferent drying methods on the growth of 2 kinds of cells was in descending order of freeze drying>40 ℃ drying>drying in the shade,and the inhibition rate decreased with the increase of drying temperature. CONCLUSIONS:Different drying methods have obvious influence on the quality of North Qinglongyi. 40 ℃ drying method is the best for North Qinglongyi drying from the com-prehensive analysis including cost,the contents of effective composition,anti-tumor activity and practice.

AIM:To observe the effect of altitude hypoxia on ?-aminobutyric (GABA) content and prepro-somatostatin mRNA (PPS-mRNA) in the rat hypothalamus. METHODS: Using altitude hypoxia model,in situ hybridization and amino acid analyzer, the number of PPS-mRNA and GABA content in rat hypothalamus was determined. RESULTS:After altitude hypoxia, the contents of GABA in hypothalamus and the number of PPS-mRNA neurons in periventricular nucleus (PeVN), paraventricular nucleus (PaVN) and arcuate nucleus (ArcN) increased significiantly. Bicuculline, a GABA receptor antagonits, could enhance PPS-mRNA expression evoked by altitude hypoxia, but had no effect on GABA content. CONCLUSION: Altitude hypoxia can induce neurotransmitter imbalance of hypothalamus.

AIM: To observe the effect of altitude hypoxia on glutamate, aspartate and nitric oxide synthase (NOS) in the rat hypothalamus. METHODS: Using altitude hypoxia model, amino acid analysis system and the NADPH-d histochemistry, we determined the content of glutamate, aspartate and the number of NADPH-d neurons in the rat hypothalamus. RESULTS: After altitude hypoxia, the contents of glutamate, aspartate in the hypothalamus of rats were increased significantly, densely and deeply stained NADPH-d neurons were seen in hypothalamic paraventricular nucleus (PVN)and supraoptic nucleus(SON). If rat were pretreated with the NMDA receptor blockers Ketamine (ip,40 mg/Kg)or AP-V(i.c.v, 10 ?g) , the number of NADPH-d neurons in the rat hypothalamic PVN and SON was markedly less than that in corresponding altitude hypoxia group. CONCLUSION: NMDA receptor may take part in the expression of hypothalamic NOS induced by altitude hypoxia.

Objective To examine the expression of Noggin in CNS of the developing rat. Methods In situ hybridization histochemistry(ISHH) was performed using digoxigenin-labeled cRNA as probes. Results It was revealed that densely and deeply stained noggin positive cells were detected in cortex,hippocampus,cerebellum,and nucleus of hypothalamus and thalamus in embryonic day(E)16 rats.The number of noggin positive cells was increased in the thalamus and medulla oblongata at postnatal day(P)1-2,whereas decreased in the hippocampus and cortex.The number of noggin positive cells was decreased significantly in brain at 1 week postnatal(P1W),and began to increase at P2W,especially in the cortex and hippocamps.Strong positive signal can be detected in the frontal cortex,parietal cortex,cingulated cortex,hippocampus,olfactory and cerebellum at 1 month postnatal(P1M).The expression of noggin begins to decline at P3M,only sparse noggin positive cells can be seen in CNS at P18M.Furthermore,there is no noggin positive cells seen in the spinal cord of rats during development.Conclusion Our results indicated that noggin could play an important role in CNS development of rats.

Objective To examine the expression of BMP4 in CNS of the developing rat. Methods In situ hybridization histochemistry(ISHH) was carried out on tissue sections using specific digoxigenin\|labeled oligonucleotide probe. Results It showed that BMP4 mRNA positive cells were located mainly in cerebellum and olfactory at E16.Strong positive signal was seen in hypoglossal nucleus,and moderate signal also seen in spinocerebellar tract and spinal lemniscus at P1\|2.The number of BMP4 mRNA positive cells was increased in the frontal cortex,parietal cortex,and hippocampus subiculum at P1W.The peak of BMP4 expression was in cortex and periamydaloid cortex.Widely distributed BMP4 mRNA positive cells were detected in cortex and hippocampus of rats at P1M,strong positive signal was observed in temporal CNS at P3M,strong positive signal was observed in hippocampus,temporal corex and periamydaloid cortex,lateral nucleus of thalamus and paraventricular nucleus of hypothalamus.BMP4 mRNA positive cells were also found in corex,hippocampus,hypothalamus and thalamus at P18M.Conclusion\ These results indicated that BMP4 could play an important role in CNS development of rats.

Objective To clarify the effective location of propofol in central nervous system (CNS). Methods Forty-two Wistar rats were ramdomized into control group,50 mg/kg propofol,100 mg/kg propofol,150 mg/kg propofol,tail shearing,propofol followed by tail shearing (n=7 in each group). The NOS expressions in the CNS were recorded by NADPH-d histochemistry after anesthesia by intraperitoneal injection of propofol. Results Rather widely stained NOS positive neurons were observed in the control group. In propofol groups,the NOS expressions were decreased significantly as compared with the control group,mainly located in ACB,LS,Pe,VLG,Den,SO,SCh,AVPO,Sol,SuM,BL,PV,LHb and Icj,showing a negative dose-effect relation with propofol. Conclusion Propofol has the determined sites of action in CNS and the decrease of NO synthesis by the inhibition of NOS may play a role in propofol-induced general anesthesia.