Objective To retrospectively analyze clinical manifestations of autonomic nervous system (ANS) and skin sympathetic response (SSR) in Lambert-Eaton myasthenic syndrome (LEMS).Methods Fifty-three LEMS patients' medical records were reviewed and information regarding clinical symptoms and signs of ANS and SSR testing results were collected.Results ( 1 ) The most common initial symptom of LEMS was weakness of lower extremities ( n =41 ) and the most common symptom of ANS dysfunction was constipation ( n =25 ) and dry-mouth ( n =23),which could be occurred before the onset of the legs (n =7).(2) In symptoms of ANS,cardiovascular system dysfunction was found in 4 patients include one of ingone of bradycardia,one of postural hypotension and 2 of tachycardia- Secretory glands dysfunction was found in 34 patients:23 dry-mouth,6 dry-eyes,and 8 patients sweating dysfunctions.Twenty-eight patients complained of alimentary dysfunction including constipation and diarrhea.Bladder dysfunction was found in 2 patients,who complained of urinary incontinence.Seven male patients complained of sexual dysfunction.Abnormal skin scratch test was found in 17 patients.(3) SSR was performed in 33 patients and 18 found abnormal.Conclusions ANS manifestations are common and prominent in LEMS patients.SSR abnormality is also common in LEMS.More electrophysiology tests are needed in LEMS patients.

Objective:To investigate the protective role of heme oxygenase-1 and its reaction product,carbon monoxide against acute liver injury induced by carbon tetrachloride in rats.Methods: Thirty male Sprague-Dawley rats were randomly divided into six groups with five in each.The control group received a single dose of corn oil injection.Carbon tetrachloride was injected intraperitoneally(i.p) to establish acute liver injury models in rats.Hemin(50 ?mol/kg) was administered i.p.12 hours before CCl_4 treatment,with an aim to induce HO-1 protein expression in the liver of rats.Carbon monoxide was injected i.p.12 hours prior to CCl_4 injection,resulting in about 8%-12% carboxyhemoglobin concentration in vivo.The expression of HO-1 in the liver of hemin-treated rats was determined by western blot method at different time points.At 24 h after carbon tetrachloride administration,all rats were sacrificed to collect blood samples for the examination of ALT,AST levels and to remove liver tissues for analysis of MDA concentration,SOD activity and caspase-3 activity as well as TNF-a contents.In addition,histopathological changes were investigated and hepatocyte apoptosis was detected by TUNEL method.Results: The administration of carbon tetrachloride to rats caused a marked hepatic damage,characterized by significant elevation of serum ALT,AST levels(2 136.3?163.4 U,1 422.7?221.7 U) and liver MDA con-tent(5.28?0.93 ?mol/g),caspase-3 activitiy(optical density value(4.69)?1.02) and TNF-? level(256.3?27.3 ng/L) combined with a remarkable reduction in liver SOD activity(45.9?14.8 U/mg) as compared with the control rats.Histopathological observations revealed severe damage in the liver and prominent hepatocyte apoptosis took place in CCl_4treated rats.However,pretreatment with hemin could induce high expression of HO-1 protein and exert potent protective effects against liver injury,as demonstrated by a significant decrease in ALT,AST levels(287.1?24.3 U,246.2?21.7 U) and MDA concentration(3.27?1.34 ?mol/g),reduction in caspase-3 activity(optical density value 2.49?1.47) and TNF-? level(132.6?19.5 ng/L),as compared with the CCl_4-treated rats.Moreover,hepatocyte apoptosis and liver injury were both attenuated remarkably in the liver of rats pretreated with hemin.In contrast to hemin administration,single injection of exogenous CO produced the same protective effects,as indicated by the remarkable reduction of ALT,AST levels and caspase-3 activity and TNF-a levels.Conclusion: The above results suggest that HO-1/CO system has a potent protective effect on acute liver injury induced by carbon tetrachloride in rats.Induction of HO-1 expression and low concentration of CO can inhibit the progress of hepatic damage,which might be due to the alleviation of lipid peroxidation and reduction of caspase-3 activity or inhibition of TNF-? level.

BACKGROUND:The enamel matrix derivative has been used in the clinical treatment of severe periodontitis; however, the mechanism(s) by which enamel matrix derivative promotes periodontal regeneration is stil obscure. OBJECTIVE:To explore the effects of enamel matrix derivatives on the differentiation and proliferation of periodontal ligament stem cels. METHODS:Periodontal ligament stem cels were isolated and identified from human teeth. Cloning forming efficiency, surface antigen expression and pluripotency were detected and identified. Enamel matrix derivatives with different concentrations (20, 50, 100 mg/L) were used to culture periodontal ligament stem cels for 2 and 4 weeks. Colagen synthesis and mineralized nodule formation were detected using Trichrom staining and Von Kosa’s staining, respectively; real-time RT-PCR was employed to detect expressions of colagen type I, osteocalcin, and RUX2; MTT and cel growth rate assay were used to detect the proliferation of periodontal ligament stem cels. RESULTS AND CONCLUSION:Periodontal ligament stem cels were spindle-shaped and showed a higher colony forming efficiency than periodontal ligament cels. The expressions of surface antigens of periodontal ligament stem cels-CD105, CD29, CD45, CD44 were respectively 99.8%, 99.7%, 1.26%, 98.8%, indicating periodontal ligament stem cels have the multilineage differentiation potential. Enamel matrix derivatives improve the colagen synthesis and mineralization nodule formation of periodontal ligament stem cels in a time-dose dependent manner. They also can improve the expression of osteogensis-related genes colagen type I, osteocalcin, RUX2 and proliferation of periodontal ligament stem cels.

Objective To investigate dynamic changes of heme oxygenase-1 and carbon monoxide in acute liver injury induced by carbon tetrach loride(CCl_4)in rats.Method Male SD rats were randomly allocated to induce acute liver injury by CCl_4 injection.Hepatic HO activity was examined at different time point following CCl_4 treatment.Expression and location of HO-1 protein was determined by western blot and immunohistochemical methods.Serum ALT,AST levels and hepatic SOD,MDA concentrations were also analyzed.Results Administration of CCl_4 to rats caused a marked hepatic damage,characterized by significant elevation of serum ALT,AST levels and liver MDA content combined with a remarkable reduction in liver SOD activity.HO activity was elevated significanfly in a time-dependant manner after CCl_4 injection,while the expression of HO-1 protein increased remarkably from 6 to 36 hours.CO concentration in the liver homogenate of control rats remained very low but was elevated significantly after CCl_4 treatment,which was in accordance with changes of HO-1. Conclusions HO-1 activity and protein expression as well as CO production are higher in rats with acute liver injury induced by CCl_4 than in control group.HO-1/CO system plays an important role in the pathogenesis of acute hepatic damage and may have potent protective effect against liver injury.

Objectives To assess the value of external anal sphincter electromyography (EASEMG) in evaluating the related autonomic dysfunction in Parkinson's disease ( PD), parkinsonism dominant multiple system atrophy (MSA-P) and progressive supranuclear palsy (PSP). Methods From the records of EAS-EMG collected in our lab (total 562 cases), 60 PD (male 41, female 19), 68 MSA-P (male 35,female 33) and 13 PSP (male 10, female 3) were included in the analysis in this study. Mean duration,polyphasic ratio and satellite potential occurrence rate were comparable among the groups. Mean duration prolongation were graded as normal ( ＜ 10.0 ms), mild ( 10.0-11.9 ms), moderate ( 12.0-13.9 ms)and severe ( ≥ 14.0 ms). Results Among all related autonomic symptoms, the occurrence rate of constipation, urinary incontinence, urgency and frequency in patients with MSA-P(95.8% (23/24) ,94.6% (53/56) ,87.7% ( 50/57 ), 85.7% (42/49), 76.5% ( 39/51 ) ) were higher than that of PD ( 61.5%(16/26), 62.3% (33/53), 30.6% (15/49), 46.2% (24/52), 45.7% (21/46)) and PSP (75.0%(3/4) , 62.5% (5/8), 50.0% (4/8), 42.9% (3/7), 42.9% (3/7)). The abnormal rate of EAS-EMG in PD, MSA-P and PSP were 60.0%, 94.2% and 84.6%, accordingly. Mean duration ( PD ( 12.0 ± 1.6)ms, MSA-P (15.4±3.0) ms, PSP (13.8±1.8) ms), polyphasic ratio (PD 46.2% ±19.2%, MSA-P 63.9% ± 15.8%, PSP 51.5% ± 12.1% ) and satellite potential occurrence rate ( PD 9.5% ± 8.3%,MSA-P 26.5% ± 15.9%, PSP 19.2% ± 12.5% ) varied significantly different among the groups ( F =31.724, F = 17.412, x2 =45. 335, all P ＜0.01 ). Severe mean duration prolongation was overwhelming in MSA-P (66.2% ) , compared with mild 10.3% and moderate 23.5%. The predominant prolongation degree was moderate in PSP (61.5%, mild 7.7%, severe 30.8% ), and mild in PD (36.7%, moderate 36.7% ,severe 11.7%, normal 15.0% ). Conclusions EAS-EMG could play a role in evaluating the related autonomic dysfunctions in PD, MSA-P and PSP. The EAS-EMG impairment was severe and frequent in MSA-P, mild and infrequent in PD, moderate in PSP. The spectrum of mean duration prolongation suggested the possibility of Onuf's nucleus involvement in these diseases.

Objective To observe the prevalence and risk factors of chronic low back pain in puerperas after childbirth.Methods Eight hundred and eighty-one puerperas were selected,among whom 459 cases had uterine-incision delivery,and 422 cases had spontaneous delivery.The age,height and weight of pregnant women,birth weight of newborn,history of preoperative low back pain,parity and mode of delivery were recorded.The rate of chronic low back pain occurring within 1 month after childbirth and continuing for 3 months was recorded by telephone.The factors with P values less than 0.05 would enter the Logistic regression analysis to screen the risk factors of chronic low back pain.Results Two hundred and fifty-nine puerperas (259/881,29.4％) appeared chronic low back pain,of whom 157 puerperas (157/459,34.2％)delivered by uterine-incision and 102 puerperas (102/422,24.2％) delivered spontaneously,and the difference was statistically significant (P ＜ 0.01).Six hundred and fifty-eight puerperas had no history of preoperative low back pain,and 150 puerperas (150/658,22.8％) appeared newly developed chronic low back pain.Logistic regression analysis showed that mode of delivery,parity and history of preoperative low back pain were the risk factors of chronic low back pain.Condusions The rate of chronic low back pain in puerperas after childbirth is 29.4％,and the newly developed chronic low back pain is 22.8％.Uterineincision delivery,multiparity and history of preoperative low back pain are the risk factors of chronic low back pain for puerperas after childbirth.

Objective The aim of the study was to investigate the expression of P-glycoprotein (P-gp)and nuclear factor κB (NF-κB) in the cerebral cortex of rat with chronic fluorosis,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of fluoride.Methods Sixty SD rats were randomly divided into three groups.The rats in each group were given drinking water containing different levels of fluoride:control group less than 0.5 mg/L,small amount of fluoride exposure group 10.0 mg/L and large amount of fluoride exposure group 50.0 mg/L.The animals were examined at the sixth month after initiating the experiment.Protein levels of P-gp and NF-κB in brain tissues were detected by immunocytochemistry and Western blotting,and the P-gp protein and mRNA level by quantitative real time PCR method.Results As compared to the control group(28.21 ±6.13),the numbers of positive staining cells by P-gp antibody in the cortex of rat brains were significantly increased in both the small and the large amount of fluoride exposure groups[(48.46 ± 8.00),(53.72 ± 9.15),respectively,all P ＜ 0.05] ; the protein levels in the control group(100.00 ± 3.86)％ detected by Western blotting were significantly increased in the cortex of rat brains treated with fluoride in both the small and the large amount of fluoride exposure groups[(189.47 ± 3.14)％,(191.36 ± 11.09)％,respectively,all P ＜ 0.05].The significantly increased expression of NF-κB at the protein level was observed in the cortex of rat brains of the small and the large amount of fluoride exposure groups[(365.97 ± 6.04)％ and (417.15 ± 10.89)％,respectively] as compared with the control group[(100.00 ± 10.07)％,all P ＜ 0.05].The mRMA level of P-gp in the cortex of rat brains of the small and the large amount of fluoride exposure groups(2396 ± 427,3479 ± 371,respectively) were higher than that of the control group(260 ± 106,all P ＜ 0.05).Conclusion The increased expressions of P-gp and NF-κB in the cortex of rat brains are induced by chronic fluorosis,which might be connected with the mechanism of brain damages.

Objective To observe the expression of mitochondrial fission protein locus Fis1 and ultrastructural changes in the renal cells of rats with chronic fluorosis,and to reveal the mechanism in mitochondrial damage of the renal cells.Methods Sixty SD rats were randomly divided into 3 groups according to sex and body mass(20 in each group):control group,lower fluoride group and higher fluoride group.All the rats were fed with different doses of sodium fluoride in drinking water(0,10 and 50 mg/L,respectively).Six-month later,the expression of Fisl in renal cells was determined by real-time fluorenscence quantitative PCR and immunohistochemistry technology,the mitochondrial morphology of renal cells was observed under transmission electron microscopy (TEM).Results As compared with the control group(28.70 ± 12.41),Fis1 mRNA levels(91.48 + 34.83 and 582.09 ± 184.69) in renal cells of the lower fluoride and the higher fluoride groups were increased(all P ＜ 0.05).As compared with the control group(10.49 ± 7.66),Fisl protein levels(16.33 ± 10.26 and 21.50 ± 5.24) in renal cells of the lower fluoride and the higher fluoride groups showed a trend of increasing,the higher fluoride group was higher than that of the control group(P ＜ 0.05).By TEM,mitochondrial crest in renal cells of the lower fluoride and the higher fluoride groups was vague or disappeared,mitochondrial division section appeared.Conclusions Fluoride is a kind of toxicant that can cause damage to mitochondrion of renal cells,induce the expression of Fis1 in transcriptional and protein level,and lead to the obstacles of mitochondrial fusion-fission and ultrastructural abnormality of mitochondrion,which may play an important role in mechanism of mitochondrial damage in the renal cells of rats with chronic fluorosis.

BACKGROUND: Forest frog is a rare medicinal animal in China, but the skin of forest frog is waste after Oviductus Ranae production. The forest frog skin is rich of collagen, and is large in quantity without pollution and disease risk. So the forest frog skin has potential to be developed into collagen sponge; however, there is no research on collagen sponge preparation as yet.OBJECTIVE: To optimize the preparation of collagen sponge from forest frog skin, and to investigate the physical properties and in vitro cell compatibility.METHODS: Chitosan and glutaraldehyde were used to modify collagen sponge from forest frog skin. Chitosan/collagen (w/w) (1:1, 1:2, 1:4) and glutaraldehyde concentration (1%, 1.5% and 2%) were selected as the experimental factors.The significant water absorbency, mechanical properties and thermal denaturation temperature were chosen as the indexes. Using the orthogonal experimental design, we optimized collagen sponge preparation process. We also investigated the in vitro cell compatibility and surface morphology of the collagen sponge. The nine kinds of collagen sponges from forest frog skins were co-cultured with human foreskin fibroblasts to detect cell proliferation.RESULTS AND CONCLUSION: When the chitosan/collagen was 1:1 and the glutaraldehyde concentration was 1%, we could get the collagen sponge with ideal water absorbency (water absorption capacity=5.22), mechanical properties (elongation at break=10.96%) and thermal denaturation temperature (81.24 ℃). The aperture of the forest frog skin collagen sponge was 200-400 μm, and the pores were consistent in the size and arranged regularly. Except the chitosan/collagen of 1:4 and the glutaraldehyde concentration of 1% or 5%, all kinds of forest frog skin collagen sponges could promote the viability of human foreskin fibroblasts and exerted benefits to cell viability and growth. To conclude,the forest frog skin collagen sponge has good biocompatibility and apparent morphology, in aggreement with the requirements of biological materials.