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 establish docetaxel (Doc) resistant MDA‐MB‐231/Doc model and epirubicin (Epi) resistant MDA‐MB‐231/Epi mode from triple negative breast cancer cell line MDA‐MB‐231 and to explore their biological characteristics .Methods The MDA‐MB‐231/Doc and MDA‐MB‐231/Epi drug‐resistant cell lines were respectively established by gradually increasing Doc or Epi concentrations induction method in 12 months .The biological characteristics of the cell lines were compared by the cell mor‐phological observation ,MTT and flow cytometry ;the real‐time fluorescent quantitative PCR was used to detect multi‐drug resist‐ance gene (MDR1) mRNA expression;the expression of P glycoprotein(P‐gp) ,estrogen receptor (ER) ,progesterone receptor (PR) and human epidermal growth factor receptor 2 (Her‐2) was detected by Western Blot .Results After the 12‐month induction ,the established MDA‐MB‐231/Doc could grow stably in the medium containing 12 nmol/L Doc ,and MDA‐MB‐231/Epi could grow stably in the medium containing 800 nmol/L Epi;in the same drug concentration ,the growth proliferation rate of the drug resistant cell line was significantly higher than that of the parental generation cells ,their drug resistance indexes were 8 .32 times and 64 .93 rimes of parental generation sensitive cells ,moreover which showed the mutual cross drug resistant status .Compared to the parental generation cells ,the cells of stage G1 and G2 in two cell lines were increased and the cells of stage S were decreased ,with the prolon‐gation of drug withdrawal time ,the cell proliferation speed was accelerated .The expression level of MDR1 gene was increased in the two drug‐resistant cell lines ,which were 4 .05 times and 5 .96 times of parental generation cells respectively ,P‐gp protein expression was positive .Compared with the MCF‐7 cell line ,ER ,PR and Her2 expression in the MDA‐MB‐231 cell line was negative and typi‐cal triple negative breast cancer cell line .Conclusion The drug resistance cell lines of MDA‐MB‐231/Doc and MDA‐MB‐231/Epi are successfully established with stable growth and drug resistance .

Objective To detect the changes of visceral sensitivity in rats presenting intestinal dysbacteriosis and the expressions of tight junction protein (ZO-1)and Toll-like receptor 4 (TLR4)so as to explore the effect of intestinal dysbacteriosis on visceral sensitivity and the possible mechanisms.Methods We randomly divided 30 male SD rats of SPF grade into normal control group (n = 12 )and dysbacteriosis group (n = 18 ).Rats in dysbacteriosis group were administered with lincomycin hydrochloride (300 mg/mL),1 mL each time per rat once a day for 7 consecutive days;those in normal control group were fed with the same amount of saline.On the eighth day,six rats were randomly selected from normal control group and dysbacteriosis group respectively to detect whether the model was successful.After the model was successfully constructed,the remaining 12 dysbacteriosis rats were randomly divided into the negative control group and the probiotics intervention group with 6 in each.Rats in the intervention group were given probiotic bifidobacterium triple viable capsules (Bifico)orally,one capsule with 1/3 mL of saline,1 mL each time per rat once a day for 7 consecutive days;those in the negative control group received the same amount of saline.On the eighth day,fresh feces was cultured for flora to detect visceral sensitivity by abdominal withdrawal reflex (AWR),the mRNA and protein expressions of ZO-1 and TLR4 in the colon,and the expression of serum inflammatory cytokines IL-10 and TNFα.Results The expression of ZO-1 in the colon was significantly lower in the rats of dysbacteriosis group than those in the control group,and the expression of TLR4 was also significantly increased.Correspondingly,the expression of pro-inflammatory factor TNFα in the serum of the rats in dysbacteriosis group was significantly increased,while that of anti-inflammatory factor IL-10 was significantly lower than in the control group (P <0.05).Furthermore,compared with dysbacteriosis group,the expression of ZO-1 was increased significantly and TLR4 was decreased in probiotics group in varying degrees. Similarly,the expression of TNFα was obviously lower while that of IL-10 in the serum was higher (P < 0.05 ). Conclusion Inhibiting the expression of ZO-1 and increasing the expression of TLR4,thus leading to chronic low-grade inflammation, may be one mechanism of visceral hypersensitivity caused by intestinal dysbacteriosis. Probiotics may restore the dysbacteriosis and thus improve visceral hypersensitivity.

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 observe the clinical efficacy of pricking anti-Ashi points with fire needles in treating acute lumbar sprain.Method Seventy-six patients with acute lumbar sprain were randomized into a treatment group and a control group, 38 cases in each group. The treatment group was intervened by pricking anti-Ashi points with fire needles, while the control group was by warm needling. The Visual Analogue Scale (VAS) and Roland-Morris Dysfunction Questionnaire (RMDQ) were adopted to evaluate lumbar pain and dysfunction after 1-day and 5-day treatment, and the clinical efficacies were compared between the two groups.Result Respectively after the first treatment session and 5 treatment sessions, the VAS and RMDQ scores were significantly changed in both groups (P<0.01). Respectively after the first treatment session and 5 treatment sessions, the VAS and RMDQ scores in the treatment group were significantly different from that in the control group (P<0.05). The total effective rate and recovery rate were respectively 94.7% and 63.2% in the treatment group, versus 86.8% and 42.1% in the control group, and the differences were statistically significant (P<0.01).Conclusion Pricking anti-Ashi points with fire needles is an effective method in treating acute lumbar sprain, and it can produce a significant real-time analgesic effect.