Aim To investigate the changes in the expression of protein kinase C gamma (PKC?), phosphorylated cAMP response element binding protein(pCREB)and immediate-early gene(c-fos and c-jun) in the spinal cord in formalin-induced inflammatory pain and study the effect of agmatine on the changes of PKC? activation, phosphorylation of CREB and expression of c-fos and c-jun.Methods Rats were decapitated at 10, 20 min or 2 h after intraplantar injection of 50 ?l 5% formalin and L_4, 5 spinal cords were dissected. Immunohistochemistry and western blotting analyses were used to observe the expression of PKC?, pCREB, c-fos and c-jun in the spinal dorsal horn and the effect of agmatine on the changes of their expression. Results Unilateral peripheral inflammation induced PKC? activation and CREB phosphorylation bilaterally while c-fos and c-jun expression ipsilaterally in rat spinal cord. PKC activity increased in membrane fractions with unchanged levels in the cytosolic fractions. Pretreatment intraperitoneally with 160 mg?kg-1 agmatine 15 min before inflammation significantly inhibited the activation of PKC? in the membrane fraction, suppressed the phosphorylation of CREB and the expression of c-fos and c-jun. Conclusion The mechanism of the analgesic effect of agmatine may be associated with inhibiting PKC? activation in the plasma membrane, CREB phosphorylation, c-fos and c-jun up-regulation which play roles in the hyperalgesia with peripheral inflammation.

Objective To investigate the clinical features of and gene mutations in a Chinese Han pedigree with piebaldism. Methods Clinical data were collected with informed consent from a pedigree with piebaldism, processed and documented. A clinical genetic analysis was conducted and pedigree chart was drawn. Genomic DNA was extracted from the peripheral blood of 14 patients and 40 unaffected individuals in the family as well as 50 unrelated human controls, and subjected to the amplification of 21 exons and flanking sequences of the KIT gene by PCR. Sequence analysis was performed by Mutation SurveyorTM. Results There were 73 members in the family, and of them, 14 were diagnosed with piebaldism according to typical clinical features. Piebaldism was inherited in an autosomal dominant pattern in this family. A heterozygous 4-base insertion mutation 1900insATGA in exon 13 of KIT gene was identified in all the 14 affected family members, which resulted in a frame-shift mutation at codon 634 and produced a premature translation termination codon. This mutation was undetected in either the unaffected family members or unrelated controls. Up to the time of this writing, this mutation had not been previously reported. Conclusion The novel mutation 1900insATGA in the KIT gene may be the cause of clinical phenotype of piebaldism in the family.

The noninvasive measurement of liver stiffness (LS) was evaluated by transient elastography (FibroScan) and the possible influencing factors from the patients' clinical situations including age, gender, liver inflammation represented by alanine transaminase (ALT) and total billirubin (TBIL) level, HBV replication (HBV DNA loads), portal vein pressure (portal vessel diameter, PVD), splenic thickness (SPT) and body mass index (BMI) were analyzed in patients with chronic hepatitis B (CHB). A total of 466 patients including 31 patients with acute-on-chronic liver failure (ACLF), and 435 patients with chronic hepatitis B (CHB) among which 82 patients were diagnosed with liver cirrhosis (LC) by clinical manifestations and liver B-type ultrasonic inspection were enrolled at Tongji Hospital from April to December 2009. LS was measured by a FibroScan device (EchoSens, France). Simultaneously, ALT and TBIL levels, HBV DNA loads, PVD, SPT and BMI in all patients were also tested. Forty-one healthy volunteers served as controls. The values of LS were correlated positively with ages of CHB patients and significantly higher in males than in females. In patients with BMI>28 kg/m(2) (obesity) and abnormal levels of ALT and TBIL, LS values were significantly increased as compared with those having normal levels of ALT and TBIL. The patients with ACLF had the highest LS value. Furthermore, LS values in the patients with LC were significantly higher than those in patients without LC. It is concluded that noninvasive measurement of liver fibrosis by FibroScan provides an alternative method to evaluate liver fibrosis of patients with CHB. In order to properly illustrate the stiffness value taken by transient elastography, patients' gender should be taken into consideration and it is also suggested to avoid possible influencing factors including liver inflammation (high levels of ALT and TBIL) and obesity (high BMI).

OBJECTIVETo evaluate the therapeutic effect on pain in the dressing change after perianal abscess surgery treated with warm suspending moxibustion at Changqiang (GV 1).

METHODSOne hundred and sixty patients were randomized into a moxibustion group and a far-infrared therapy group, 80 cases in each one. In the moxibustion group, the warm suspending moxibustion at Changqiang (GV 1) was applied. In the far-infrared therapy group, the infrared ray was radiated at Changqiang (GV 1). The treatment started in 10 min after dressing change in the two groups. Totally, 8 treatments were required. The time requirement being equal to or less than 2 score in the assessment face scale (AFS), the case number of pain relief in 10 min after treatment and the intention of the two therapies after everyday dressing change were observed in the patients of the two groups.

RESULTSIn the moxibustion group, the time of pain relief was shorter than that of the far-infrared therapy group [(11.5 +/- 3.9) min vs (17.8 +/- 3.8) min on the 1st day; (9.4 +/- 3.6) min vs (15.2 +/- 4.3) min on the 2nd day; (7.8 +/- 2.7) min vs (14.3 +/- 3.2) min on the 3rd day; (6.4 +/- 3.3) min vs (10.5 +/- 2.9) min on the 4th day; (5.9 +/- 2.9) min vs (11.2 +/- 1.9) min on the 5th day; (5.3 +/- 2.2) min vs (8.8 +/- 2.3) min on the 6th day; (5.4 +/- 1.8) min vs (9.2 +/- 2.1) min on the 7th day; (5.1 +/- 1.4) min vs (9.5 +/- 2.6) min on the 8th day, all P < 0.05]. The case numbers of pain relief in 10 min after treatment were 74 cases, 75 cases, 77 cases and 78 cases from the 1st to the 4th day separately in the moxibustion group, which were much more than 63 cases, 65 cases, 68 cases and 69 cases in the far-infrared therapy group (all P < 0.05). The treatment intention scores in the moxibustion group were better than those in the far-infrared therapy group (all P < 0.05).

CONCLUSIONThe warm suspending moxibustion at Changqiang (GV 1) relieves pain rapidly in the dressing change of much more patients after perianal abscess surgery in much shorter time as compared with the far-infrared therapy. Moreover, the patients have more expectation to be treated with moxibustion therapy.

Abscess ; surgery ; Adolescent ; Adult ; Aged ; Anus Diseases ; surgery ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Pain, Postoperative ; therapy ; Young Adult

Objective To explore the clinical characteristics and outcome of correction with Ilizarov fixator for the knee deformities.Methods From May 2003 to April 2010,21 patients (22 knees) underwent knee deformity correction with Ilizarov fixator,including 12 males and 9 females with an average age of 20.3 years (range,8-48).Causes of the deformities included poliomyelitis in 4,burn in 2,osteomyelitis in 2,trauma in 9,Blount diseases in 2,and multiple osteochondromatosis in 2.Five patients had fixed flexion contracture due to soft tissue,they were corrected through a combination of Ilizarov's frame crossover the joint with a pair of hinges by gradual posterior distraction.Eight patients (9 limbs) had one way bony deformities and 7 patients had complex deformations.The frame with 4 hinge-posts was used for correction by restoring the alignment firstly,and then gradual lengthening to correct bone shortening.Additionally,an overlay frame of the above mentioned combinations was applied for correction of bony deformity combined with soft tissue contracture for 1 patient.Results The average time in frame was 22.3 weeks (range,12-36).At the time of removing frame,satisfactory alignment was achieved in all of the affected knees,and solid bony healing was obtained in osteotomy or bone lengthening area in 16 patients (17 limbs) with bony correction.All patients were followed up for an average of 32.1 months (range,6-86).The range of motion was improved from 102.14°±49.36° preoperatively to 126.90°±24.31° at the final follow-up.Additionally,the Japanese Orthopaedic Association knee score was also increased from 50.24±23.64 before operation to 85.71±10.52 at the final follow-up.All of them were able to walk without crutches,deal with daily life independently.Only 2 patients with the range of motion of the knee less than 90° were not able to squat.Conclusion Ilizarov fixator has advantages of minimal intervention to local tissue in operation and nimble adjustment at any time,and disadvantages related to a longer time in frame.

CD4(+)CD25(+)CD127(dim/-) regulatory T cells (Tregs) have been implicated in suppressing T cell immune responses to hepatitis B virus (HBV), but the inhibition mechanism has not being clear yet. This study investigated the effects of soluble FGL2 (sFGL2) secreted by Tregs on immune suppression in chronic HBV-infected patients. We verified that sFGL2 protein and mRNA were highly expressed in Tregs. The separated Tregs by using magnetic beads from peripheral blood mononuclear cells (PBMCs) in 20 patients with chronic hepatitis B were co-cultured with PBMCs at a ratio of 1:3 with anti-CD3 stimulating antibody or FGL2 blocking antibody. The proliferation index of CD8(+)T cells after blocking FGL2 was higher than that in blank group (3.58±0.18 vs. 3.28±0.17, P=0.034) in 18 of 20 samples, and lower than that in CD3 stimulation group (3.82±0.19, P=0.026) in 16 of 20 samples. The IFN-γ secreted in the mixed culture in the absence of Tregs was higher than that in the culture in the presence of Tregs, but it could be abolished by FGL2 blocking antibody. These results suggest that sFGL2 protein secreted by Tregs suppresses the proliferation and function of CD8(+) T cells in chronic hepatitis B.

Recently, the Th17 cells and IL-17 have been shown to play a critical role in the immune-mediated liver injury in hepatitis B, while their functions in acute liver failure have not been well elucidated yet. In this study, we primarily investigated the role of IL-17 in the development of mouse hepatitis virus strain 3 (MHV-3)-induced acute liver failure. IL-17 mRNA levels in liver tissue were quantified by using quantitative real-time polymerase chain reaction, and cytokine IL-17 levels in liver tissue and serum were determined by using ELISA in MHV-3-induced murine fulminant hepatitis model. The IL-17 expression levels on CD4(+)T and CD8(+)T cells were determined by using flow cytometry. The correlation between IL-17 level and liver injury was studied. Th17 associated cytokines were also investigated by intracellular staining. Our results showed that the IL-17 expression was significantly elevated in the liver and serum of BALB/cJ mice infected with MHV-3. Moreover, a time course study showed that the percentage of both IL-17-producing CD4(+)T cells and IL-17-producing CD8(+)T cells was increased remarkably in the liver starting from 48 h and peaked at 72 h post-infection. There was a close correlation between hepatic or serum IL-17 concentration and the severity of liver injury defined by ALT level, respectively. Th17 associated cytokines, IL-6, IL-21 and IL-22, were also increased significantly at 72 h post-infection. It was concluded that IL-17 may contribute to the pathogenesis of MHV-3-induced acute liver failure.

Aim To evaluate the analgesic effect of agmatine on inflammatory pain and its influence on the analgesic effect of morphine. To investigate whether the mechanism of analgesic effect of agmatine is related to activation of imidazoline receptor or to affect the release of endogenous glutamate and gamma-aminobutyric acid (GABA) from rat spinal cord slices. Methods The formalin test in rats was used as a long-lasting inflammatory pain model. Effects of agmatine on basal and K+ evoked release of endogenous glutamate and GABA from rat spinal cord slices were determined by high performance liquid chromatography (HPLC). Results Pretreatment with agmatine (ip or sc) inhibited the second phase of the nociceptive response of rats and potentiated the analgesic effect of morphine in phase 2, but not in phase 1. Idazoxan did not attenuate the analgesic effect of agmatine. Agmatine (1, 10, 100, 1000 ?mol?L -1 ) had no effect on the basal release of glutamate and GABA from spinal cord slices, nor did it affect the K+ (50 mmol?L -1 ) evoked release of glutamate and GABA contents. Conclusions Agmatine has an analgesic effect and enhances morphine analgesia in the second but not the first phase of formalin-induced nociception. Its analgesic effect does not likely involve imidazoline receptor. The mechanism of the analgesic effect of agmatine may not be associated with inhibiting glutamate release nor increasing the GABA content.

BACKGROUND: Previous studies showed that the prevalence and extent of carotid artery stenosis increased with thedevelopment of coronary artery disease. There was a higher incidence of intracranial small-vessel disease, but lower of carotid artery disease in the Chinese stroke patients as compared with the white.OBJ ECTIVE: To observe the distribution of carotid and intracranial artery stenosis in patients with 3-vessel coronary artery disease.DESIGN: An observational study.SETTING: Department of Neurology; Heart Center, Beijing Chaoyang Hospital affiliated to Capital Medical University.PARTICIPANTS: From August 2003 to August 2004, The coronary angiography was performed in the outpatients and inpatients suspected to be coronary arteriosclerotic cardiopathy in the Department of Neurology, Beijing Chaoyang Hospital affiliated to Capital Medical University, and 126 patients of them with 3-vessel diseases were examined with carotid arteriography, including 56 males and 70 females, 47-76 years of age. Informed contents were obtained from all the participants.METHODS: Digital substraction angiography (DSA) was performed immediately after coronary angiography in the 126patients. All catheterizations were performed through a transfemoral approach using the Seldinger technique, and thenan appropriate amount of nonionic Ominipaque was injected. The angiography of bilateral carotid arteries, subclavian artery, or vertebral artery was taken from different angles. The percentage of stenosis was calculated directly from DSAmachine. Evaluative standards: Based on the stenosis degree from carotid angiography results, the patients were divided into 5 categories as normal, mild stenosis, moderate stenosis, severe stenosis and complete occlusion.MAIN OUTCOME MEASURES: Severity of carotid stenosis.RESULTS: All the 126 patients were involved in the final analysis of results. There were 13 (10.32%), 18 (14.29%), 12(9.5%), or 10 (7.9%) patients found to have mild, moderate, severe carotid stenosis, or complete occlusion, and the incidences of these changes were fairly similar. However, the incidence of angiographic carotid stenosis coupled with 3-vessel carotid artery disease was 42.06%.CONCLUSION: The prevalence of carotid stenosis in patients with 3-vessel carotid artery disease was as high in the Chinese population as that in Westem countries. In patients with 3-vessel disease, the prevalence of carotid stenosis was higher than that of intracranial artery stenosis, thus they may require both coronary and carotid interventions.

Objective In order to evaluate that whether Pluronic P85 coated poly(butylcyanoacrylate) nanoparti-cles was able to deliver antiepileptic drug phenytoin into the brain va bypassing mesial temporal lobe epilepsy (MTLE)-induced Pgp in a rat model of MTLE. Methods The rat model of MTLE, induced by li-pilocarpine, was divided in-to two groups (6 for nanoparticle drug group and 7 for PHT drug group). Immunohistochemistry assay was performed to detect Pgp expression at the hippocampus. Nanoparticles were prepared by interfacial polymerization method. Dialysate samples of brain were collected at 30, 60, 120, 180, 240 and 300 min after drug administration by microdialysis tech-nology. Samples were analyzed by high performance liquid chromatography (HPLC). Results The area under the curve (AUC) ratio of brain/plasma in Nanoparticle drug group was 0.370.10 which was significantly higher compared with 0.190.06 in conventional PHT drug group (P<0.05). The Pgp immunopositive area, as assessed by analysis of labeled surface area, was higher in the DG, CA3 and CA1 sector in the hippocampus of MTLE rats when compared to the normal rats. Conclusions Pluronic P85 coated PBCA nanoparticles can significantly deliver PHT into brain via bypassing MTLE-induced Pgp in a rat model of MTLE.