Objective To observe the distribution features of phosphorylated p 44/42 extracellular signal-regulated kinase(p-ERK) induced by affective pain in the anterior cingulate cortex (ACC) of the rat.Methods Twelve male SD rats were randomly divided into the control group (n=6) and spinal nerve ligation (SNL) group (n=6).The rats of the SNL group were exposed to unilateral L5 spinal nerve ligation (SNL) surgery.The behavior was examined by mechanical pain thresholds (MPTs) and the pain affect was measured using open-field test, elevated zero maze.The expressions of p-ERK in ACC (AP 3.2,2.7,2.2mm from Bregma) were detected with immunofluorescence test .Results In comparison with the control group , the unilateral mechanical pain threshold in the SNL group decreased significantly after operation . The operation also resulted in pronounced anxiety-like behavior .The p-ERK labelled cells in different levels of ACC ( AP 3.2,2.7,2.2mm from Bregma) of SNL rats were (11.89 ±2.57),(32 ±4.67 ) and (17.56 ±2.04 ), respectively, while the control group rats were (12.44 ±2.16),(10 ±0.87 ) and (10.11 ±1.36).These data indicated that compared with the control group, the expressions of p-ERK in ACC(AP 2.7,2.2mm from Bregma) increased significantly in the SNL group after operation(P<0.01), besides for the level of ACC(AP 3.2mm from Bregma) (P>0.05).Conclusion The results suggest that the neuropathic pain can result in anxiety-like behavior and increased expressions of p-ERK in ACC of the rat.This change may be closely related to the expressions of p-ERK in ACC(AP 2.7,2.2mm from Bregma), but not to the level of ACC(AP 3.2mm from Bregma).

Objective To explore the relationship between mechanical withdrawal threshold ( MWTs) and local cutaneous blood perfusion ( BP ) in rats with acute inflammation induced by carrageenan.Methods Twenty male Sprague-Dawley ( SD) rats were randomly divided into control and model groups.The acute inflammatory rat model was established by subcutaneously injecting with carrageenan into the left hindpaw.MWTs were measured by Dynamic Plantar Aesthesiometer 37450 before injection ( as base) of carrageenan and at 4 h, 24 h, 48 h, 72 h after carrageenan injection. The local cutaneous BP was detected by Pericam Perfusion Speckle Imager ( Pericam PSI) at the time after measuring of MWTs.The above two behaviors were compared and the relationship between them was analyzed.Results MWTs of the model group rats were decreased while BP significantly increased than that in the control group ( P<0.01 ) .The MWTs and BP in the model rats showed a negative correlation, especially the correlation index showed significant differences at 4 h and 72 h after carrageenan injection ( P<0.01) .Conclusions Carrageenan-induced acute inflammation in rats causes significant changes in mechanical pain threshold, which has a negative correlation with local cutaneous blood perfusion.

Trigerninal neuralgia is a common refractory disease in clinic. Professor FANG Jianqiao has rich experience through diagnosing and treating the disease for many years. In the first diagnosis, professor FANG underlines the position of damaged neuron and syndrome differentiation. He considers acupuncture should be implemented by stages,namely according to whether the patients are in the period of pain attack, different acupuocture prescriptions are made. Acupuncture manipulation and needle-retention time should be adjusted according to the condition of disease. And the appropriate application of electroacupuncture and transcutaneous electrical acupoint stimulation can strengthen the effect.
Acupuncture Points ; Acupuncture Therapy ; history ; China ; History, 20th Century ; History, 21st Century ; Humans ; Trigeminal Neuralgia ; diagnosis ; therapy

Objective By simulating a high-glucose condition of peritoneal dialysis (PD)fluid,to explore the effect of high glucose on the expression of leptin and its relationship with peritoneal angiogenesis.Methods Adipocytes differentiated from 3T3-L1 were divided into high glucose group (139 mmol/l glucose) and high mannitol group.Leptin levels in supernatant collected at 0 h,12 h,24 h and 48 h were measured by ELISA.Endothelial cells (ECs) were respectively cultured with normal glouse,high glucose,high mannitol condition,supernatants of adipocyte induced by normal glouse,high glucose and high mannitol,high glucose supernatants+leptin antibody,and high mannitol supernatants + leptin antibody.Tubular structure formation and migration of ECs were detected.Results Adipocytes exposed to high glucose for 48 h produced more leptin as compared with control group,high mannitol group,12 h-high glucose group and 24 h-high glucose group (all P ＜ 0.05).Compared with ECs in normal group,ECs in high glucose had less tubular structure formation and increased migration (all P ＜ 0.01).Compared with those of ECs in high glucose,the tubular structure formation and the migration of ECs in adipocyte supernatants induced by high glucose had increased (all P ＜ 0.01),and these effects were reduced by leptin antibody (all P ＜ 0.01).Conclusion There is an up-regulation of leptin in adipocytes exposed to high glucose,which may be an alternative way to prevent peritoneal angiogenesis.

Objective To study the chemcial constituents in the root and stem of Piper laetispicum. Methods Compounds were separated and purified by silica gel column. Their structures were identified on the basis of spectral methods. Results Six compounds were isolated and identified from the root and stem of P. laetispicum: N-isobutyl-(3, 4-methylendioxyphenyl)-2E, 7E-nonadienamide (Ⅰ), pipercallosidine (Ⅱ), N-isobutyl-7-(3, 4-methylendioxyphenyl)-2E, 4E-heptadienamide (Ⅲ), 4, 5-dihydropiperlonguminine (Ⅳ), 1-cinnamoylpyrrolidine (Ⅴ), piperitol (Ⅵ). Conclusion Compound Ⅰ is a novel one, named as laetispiamide B and compound Ⅱ is isolated from P. laetispicum for the first time.

Objective To analyze the balance function of injured lower lim b by dynam ic posturography. Methods Using the dynam ic posturography instrum ent, the postural evoked responses of sixty-two norm al people and two hundred and fifty-eight people with injured lower lim b bones and joints were detected. The test was included sensory organization test(SO T) and adaption test(ADT).The results of two groups were com pared by t test. Results Com pared with the norm al people, the im paired people had significant statistical differences in balance scores of SO T3-SO T6 and proportion score of dynam ic proprioception (P<0.05). There was no obvious decrease in ADT. Conclusion The balance function of injured lower lim b significantly decreases.

ObjectiveTo study the effect of testing amino acids levels in brain microdialysate by method of high performance liquid chromatography (HPLC) with α-aminobutyric acid (AABA) as the internal standard.MethodsReversed phase high performance liquid chromatography (RP-HPLC), orthophthaladehyde (OPA) precolumn derivatization method and fluorescence detector were used. The separation was performed using an ODS-C18 column. AABA, an isomeric compound of GABA, served as the internal standard, phosphate buffer solution and methanol as the mobile phase, gradient elution lasted in 35 min.Results7 kinds of amino acids and internal standard were separated completely. The mean recovery of amino acid was 88.2%—102.3%. The mean correlation coeffeicient of the linear relationship was 0.998±0.0015.Limits of detection for amino acids were 1.0—8.6 ppm respectively.ConclusionThe method mentioned above is simple and sensitive for operation, and can be used for determined levels of amino acids in microdialysate.

AIM: To investigate whether a pathological micro-environmental culture system consisting of cholestatic sera induces embryonic stem cells (ESC) to differentiate into hepatocyte-like cells in vitro, and select hepatic stem cells from differentiating embryonic stem cells. METHODS: Mouse ESC, E14 cell line, were cultured in Dulbecco's modified Eagle's medium containing 106 U/L recombinant mouse leukemia inhibitory factor (rmLIF) and 10% FCS. After embryonic bodies formed by the hanging drop culture method, they were exposed to fibroblast growth factor-4 (FGF-4) and hepatocyte growth factor (HGF) for one week, and then placed to a pathological micro-environmental culture system consisting of 5% cholestatic sera and cultured for 2 weeks. Morphological examination, immunocytochemical staining of albumin and CK8/18 were carried out, and mRNA level of albumin and transthyretin were detected by RT-PCR. Glycogen storage and urea synthesis of the cells were tested with PAS staining and colorimetric assay, respectively. RESULTS: The proliferation of cells was inhibited at the early stage when cultured in a pathological micro-environmental culture system consisting of 5% cholestatic sera, but 2 weeks later, a large number of epithelial-like cell colonies were observed, which exhibited hepatocellular phenotype, expressing albumin and CK8/18, transcribing mRNA of albumin and transthyretin and synthesizing glycogen and urea. CONCLUSION: A pathological micro- environmental culture system consisting of 5% cholestatic sera could not only induce embryonic stem cells to differentiate into hepatocyte-like cells, but select hepatic stem cells from differentiating embryonic stem cells initially induced by FGF-4 and HGF in vitro as well.

Objective To Study on CXCR1/CXCR2 antagonist G31P anti-inflammatory reaction mediated by neutrophils.Methods Detect whether G31P can block chemotaxis of neutrophils induced by human IL-8 and inhibit the release of IL-8 by epithelia of segmental bronchus;establish HEK293 cell line transfected by pcDNA3.0-CXCR1 ,2,4 and detect the chemotaxis of IL-8 for HEK293 ;establish the experi-mental model of pneumonia induced by the P.aeruginosa,take count of the nucleated cells in the bronchoal-veolar lavage fluid(BALF),analyze myeloperoxidase(MPO) of lung tissue and observe the histopathology changing of it.Results G31P can inhibit the chemotaxis for neutrophils and transfected HEK293 cell line,inhibit the A549 releasing of inflammatory mediators;the proportion of neutrophils declines in G31P treat-ment group,pathology examination appears clear discrepancy.Conclusion G31P can block the chemotaxis of chemotactic factor with ELR+ CXC to neutrophils,block the combination of chemotactic factor with its re-ceptor CXCR2,block the CXCR2 on the surface of alveolar epithelia and vascular endothelial cells.Accordingly,neutrophils recruiting to topoinflammation can be prevented.

Purpose To analyze EGFR exon 18 ~21 gene mutations in lung cancer, and compare xTAG liquidchip technology and Sanger sequencing technology in clinical practice. Methods 1 139 tumor tissue samples from phaseⅠtoⅣlung cancer patients were randomly collected. DNA was extracted from the samples. EGFR gene mutation status in exon 18~21 was detected by xTAG liquidchip technology and Sanger sequencing technology respectively. Results The mutation status of EGFR was obtained by xTAG liquidchip technology in 1 134 patients, and 1 105 by Sanger sequencing technology, detection success rate was 99. 56% and 97. 01% respective-ly. The sensitivity and specificity of xTAG liquidchip technology Comparing with Sanger sequencing was 99. 59% and 94. 54%. Sever-al cases of multiple mutations were detected by both methods. All mutation types detected by two methods are fully consistent. Conclu-sions Comparing with Sanger sequencing technology, xTAG liquidchip technology, which is able to detect mutations of exon 18~21 simultaneously, is more convenient and efficient for EGFR gene mutation detection in lung cancer.