This paper reported a new method of Armillaria mellea isolation-Gastrodia elata tissue isolating. Compared with normal isolating method-rhizomorph isolating method, it showed that the success rate of new method (78% ) was higher than the rhizomorph isolating method (16% ) , besides this, the new method was easier, and growth characteristic of obtained strain was superior to that obtained from rhizomorph isolating method.

Objective:To investigate the changes and the adjusting effects of brain-gut peptides (BGP) on immunologic function in the rats before and after relief of biliary obstruction.Methods:One hundred Wistar rats were randomly divided into five groups,including sham common bile duct ligation group (group A),common bile duct ligation group (group B),sham relief of biliary obstruction + normal saline group (group C) ,relief of biliary obstruction + normal saline group (group D) and relief of biliary obstruction + Sandostatin (group E).At 1d,7d after the operation of each group,the changes of plasma vasoactive intestional peptide (VIP),substance P(SP),serum interleukin-2(IL-2) and serum T-lymphocytic subsets (CD4 +,CD8 +)were determinded by radioimmunoassay and flowcytometry .Results:Plasma VIP and SP were increased in group B compared with group A(P

Avellino corneal dystrophy(ACD) is an autosomal dominant eye disorder caused by mutation of R124H in the transforming growth factor-beta induced gene (TGFBI) on chromosome 5,which was responsible for accumulating of abnormal TGFBI.Although the underlying mechanism by which mutations cause abnormal TGFBI deposition is not yet clear,but we have a better understanding of the etiology and possible pathogenesis of corneal dystrophy with the rapid development of human genetics and molecular biology,and summarizes the current achievement of this disease and understand the roles of TGFBI and its interaction with Periostin,which may contribute to further research in ACD.

Objective To evaluate the relationship between the amount of grafted cells and the incidence of acute graft versus host disease (aGVHD) after haploid hematopoietic stem cell transplantation (haplo-HSCT). Methods Data of 68 patients who underwent haplo-HSCT from Jan 2009 to Dec 2013 were analyzed retrospectively. Influences of different factors on the incidence of Ⅲ-Ⅳ degree of aGVHD after HSCT were evaluated. Results 68 patients including 42 males and 26 females were 5/10-9/10 HLA match with 19 father donors, 24 mother donors, 16 sibling donors and 9 children donors. 51 patients not suffered Ⅲ-Ⅳdegree of aGVHD included 32 males and 19 females with the mean age of 20 years old (5-55 years old). 17 patients sufferedⅢ-Ⅳdegree of aGVHD including 10 males and 7 females with the mean age of 23 years old (5-54 years old). There were no significant differences in the amount of the grafted mononuclear cells (MNC) and CD34+cells, and the white blood cell counts (WBC) and platelet count (Plt) recovered time between two groups (P>0.05). However, MNC number was related to CD34+cell number (P<0.05) and WBC recover time (P<0.05), and the CD34+cells number was related to WBC and Plt recover time (P< 0.05). Conclusion The incidence of Ⅲ-Ⅳ degree of aGVHD is unrelated to the amount of grafted MNC, and CD34+cells.

Objective To determine the incidence and risk factors of iatrogenic retinal breaks in eyes undergoing pars plana vit- rectomy for idiopathic macular pucker.Design Retrospective case series.Participant 88 consecutive vitrectomies performed on eyes with idiopathic macular pucker.Method Consecutive vitrectomies performed on eyes with idiopathic macular pucker at Beijing Tongren Eye Center between 2002 and 2006 were retrospectively reviewed.Cases with iatrogenic retinal breaks were recorded and analyzed. Main Outcome Measure Number and location of retinal breaks,and anatomic outcome after surgical managements.Result A total of 88 consecutive vitrectomies were included in the study.Of the 88 eyes,8 eyes had 14 iatrogenic retinal breaks detected,with an aver- age incidence of 9.1%.Peripheral retinal breaks(8.0%)were more common than posterior retinal breaks(1.1%).All peripheral retinal breaks occurred around the selerotomy sites(100%)and the quadrant of predominant hand was involved most commonly(62%).Most of the breaks(88%)were detected during the surgery.All eyes with iatrogenic retinal breaks obtained anatomic retinal reattachment (100%).Conclusion Despite improvements in instrumentation and surgical techniques,iatrogenic retinal break continues to be an im- portant complication of pars plana vitrectomy in eyes with idiopathic macular pucker.This complication tends to occur more commonly at peripheral retina and is mainly selerotomy-related.

Objective To investigate the influence of citalopram on the fast response action potential,slow response action potential,in vitro electrocardiogram(ECG) and in vivo ECG of cardiac myocytes,and explore its mechanism of adverse cardiac effects. Methods Conventional microelectrode technique was employed to record the fast and slow response action potentials of the isolated papillary muscles of guinea pigs.In vivo and in vitro ECG were recorded from anesthetized animals and Langendorff-perfused hearts,respectively. Results Citalopram could prolong the RR interval and QRS duration of in vivo ECG.The premature ventricular contraction and atrial ventricular block were induced by 12.5?10-6 mol/L citalopram.The maximum ascending velocity of 0 phase(Vmax),action potential amplitude(APA) and action potential duration(APD50 and APD90) were dose-dependently decreased by citalopram in the fast and slow response action potentials of guinea pigs,respectively. Conclusion Citalopram can inhibit sodium and calcium channels effectively,which may be the ionic mechanism that citalopram induces arrhythmia in the clinical practice.

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 investigate the changes of reactive oxygen species(ROS) level and mitochondria fission-fusion-balance in cortical neurons of rats with chronic fluorosis and reveal the correlation between these two factors. Methods One hundred and twenty rats were randomly divided into 3 groups(control group, low-dose fluorosis group, high-dose fluorosis group) and 40 rats were in each group according to body weight and the experiments were carried out for 3 months or 6 months. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose group were fed with water containing NaF 10.0,50.0 mg/L, respectively. The level of ROS and the morphology in mitochondria fission-fusion balance in neurons of the cortex of rat brains prepared with cortical frozen sections were detected with ROS fluorescent probe and MitoTracker RED probe, respectively. Results Significant differences of the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were found between 3 groups at the experiment period of 3 month and 6 month(F= 3.07,3.06,3.05,3.07, all P < 0.05). As compared with control group(10.43 ± 5.98,4.12 ± 3.86) at the experiment period of 3 month, the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were obviously increased in high-dose fluorosis group(25.48 ± 6.09,20.47 ± 6.09, all P < 0.05), whereas no significant changes were found in low-dose fluorosis group(11.67 ± 3.49,6.68 ± 3.48, all P> 0.05). Furthermore, the increases in both ROS level and abnormal numbers of mitochondria were significant observed in the cortical neurons of low-dose fluorosis group (63.02 ± 8.15, 49.33 ± 8.61) and high-dose fluorosis group(65.60 ± 7.40,53.10 ± 6.95) as compared with the control group (25.26 ± 6.41,20.26 ± 6.41) at the experimental period of 6 month (all P < 0.05). The abnormal numbers of mitochondria correlated with ROS level(r = 0.93,0.81, all P < 0.05). Conclusions Taking excessive amount of fluoride results in high level of oxidative stress and impaired the balance of mitochondrial fission-fusion,which is dependent on the feeding times and doses of fluoride. The mechanism of the mitochondrial abnormalities might be associated with the high level of oxidative stress induced by chronic fluorosis.

Background Deficiency of neurotrophic factor is associated with the damage of optic nerve in glaucoma.Reaserches showed that ectopically applied neurotrophic factor has a transient neuroprotective effect in glaucoma model,and the viral expression of adeno-associated neurotrophie factor may provide long-term supplementation of neurotrophic factor and neuroprotection in tissues.Objective The present study Was to investigate the neuroproteetive effect of adeno-associated viral(AAV)-mediated brain-derived neurotrophic factor (BDNF)on retinal ganglion cells(RGCs)in DBA/2J mice with experimental glaucoma. Methods 10 clean DBA/2J mice were administered intravitreal injection with 1 microliter of AAV-BDNF-GFP in the left eyes at the age of 6 months,and the right eyes were injected with the same volume of saline solution as control.Intraocular pressure (IOP)was measured with Tonolab in the mice every month.Retinas were obtained after 3 months for the investigation of GFP expression in RGCs using fluorescence microscopy.Immunohistochemistry Was performed by applying TUJ1 and Cy3 antibodies to identify surviving RGCs. Results The IOP of DBA/2J mice were 11.90 mmHg and 11.40 mmHg in the right eyes and left eyes,respectively,at 4 months.The IOP of DBA/2J mice began to rise at 5 months and reached its peak in 8 month-old mice.There was no statistically significant difference in IOPs between the right eyes and the left eyes from 4 month-through 9 month-old mice(t=-1.78-0.61,P=0.11-0.90).Three months after intrlavitreal injection of AAV-BDNF-GFP,GFP was positively expressed in RGCs of retinas with the expression rate of 46.33%±8.08%.The over-expression of BDNF led to more RGCs survival than the control eyes (3168.13±1319.33/mm2 vs 2024.81±796.38/mm2,t=2.75,P=0.02). Conclusion These data suggest that BDNF can exert a protective effect in DBA/2J glaucoma mice.

BACKGROUND:Alogeneic hematopoietic stem cel transplantation (alo-HSCT) is an effective mean to cure severe aplastic anemia, and especialy haplotype transplantation is regarded as a transplantation system with Chinese characteristics, and rank at the international leading level. OBJECTIVE:To explore the patterns of haplotype alo-HSCT as a new immune tolerance method for severe aplastic anemia and to solve the transplantation rejection and graft-versus-host disease. METHODS:Twelve patients with severe aplastic anemia who underwent haplotype alo-HSCT at the Department of Hematology, General Hospital of Beijing Military Area, China from April 2013 to May 2014 were enroled. Al these patients received the new regimen of inducing immune tolerance through the application of high-dose cyclophosphamide (400 mg/m2, consecutively 3 days before transplantation; 50 mg/kg, consecutively 3 days after haplotype transplantation). RESULTS AND CONCLUSION:The median time of neutrophil recovery was 17 (13-21) days, and the median time of platelet recovery was 21 (15-31) days. After transplantation, there were one case of degree II acute graft-versus-host disease and one case of chronic graft-versus-host disease, both of which were controled. The folow-up time was 6 months at least, and the median time was 11 months. During the folow-up, one case died of rejection reaction and one case died of severe lung infection. These findings indicate that the new method of inducing immune tolerance with high-dose cyclophosphamide after transplantation for severe aplastic anemia has significant effects in reducing graft-versus-host disease and transplantation-related mortality rate.