Background The early disorder of diabetic retinopathy (DR) is the damage of retinal neural cells induced by high glucose and lack of oxygen.Previous studies show that bushenhuoxue serum can enhance the activity of glutamine synthetase (GS) in Müller cells under hypoxia,and glutamate-mediated retinal excitotoxicity also can be reduced by bushenhuoxue serum intervention.However,whether the concentration of glycine can be increased by bushenhuoxue serum is not clear.Objective This study was to investigate the protective effects of bushenhuoxue serum on retinal ganglion cells (RGCs) under hypoxia.Methods The Sprague Dawley (SD) rat serum containing bushenhuoxue was prepared.The RGCs of newborn SD rats were purified and identified by a twostep immunopanning procedure.After 72 hours,all RGCs were cultured in 96-well plates and divided into four groups:normal control group (cultured in adult SD rats normal serum),bushenhuoxue group (cultured in bushenhuoxue serum),hypoxia group (cultured in 1 mmol/L sodium dithionite); hypoxia + bushenhuoxue intervention group (cultured in bushenhuoxue serum+sodium dithionite).Glutamate and glycine contents in the extracellular fluid were detected by L-8800 automatic amino acid analyzer,and the content of lactate dehydrogenase (LDH) was assayed using LDH kits in 24,48 and 72 hours after culture.Results Cultured cells showed the green fluorescence under the immnofluorescence microscope.The contents of glutamate,glycine and LDH in the extracellular fluid were (0.0805±0.0010)mg/L,(0.0554±0.001 5)mg/L and (1 626.03 ±122.10)μmol/(min · L) in the normal control group in 24 hours after culture,and those in the hypoxia group were (0.022 5±0.001 1) mg/L,(0.014 6±0.001 1)mg/L and (1 458.68±94.23)μμmol/(min · L),showing significant reducing in the hypoxia group (q =-3.53,P =0.00 ; q =-2.45,P =0.00 ; q =-2.98,P =0.02).Compared with the normal control group,LDH and glycine contents in the extracellular fluid were significant raised in the hypoxia group 48 hours after culture (q =2.55,P =0.01 ;q =4.48,P =0.00).Seventy two hours after culture,the glutamate and glycine contents in the hypoxia group were higher than those of the normal control group (q =2.45,P =0.00 ;q =3.72,P =0.00).In 48 and 72 hours of culture,the contents of glycine were (0.017 4±0.001 5) and (0.019 2±0.001 2) mg/L in the hypoxia+bushenhuoxue intervention group,which were significantly higher than (0.016 0±0.001 2) and (0.018 0±0.000 8) mg/L in the hypoxiagroup (q=2.28,P=0.04;q=2.33,P=0.03),but the LDH level were (1 632.94±264.31) and (1 875.00±137.45)μmol/(min · L) in the hypoxia+ bushenhuoxue intervention group,which were lower than (1 688.49 ± 112.86) and (2 267.86 ± 175.21) μmol/(min · L) of the hypoxia group (q =-2.95,P =0.02 ; q =-2.35,P=0.00).No significant differences were seen in the glutamate content 24,48 and 72 hours after culture (P=0.55,0.28,0.46).A positive correlation was seen between the glutamate content and glycine content in the extracellular fluid (Kendall coefficient =0.519,Spearman coefficient =0.696,both at P =0.000).Conclusions The release levels of glutamate and glycine increase in the hypoxia RGCs,which probably is a compensatory response of RGCs.Bushenhuoxue serum can protect RGCs against injury by increasing the release of glycine and decreasing the LDH leakage from RGCs.

Objective To study the relationship between GSR and firing distance, and to discuss whether the relationship can be used to estimate the firing distance. MethodsShotting porkets with "5.4" hand gun at the firing distance of 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 120cm respectively, then observing the distributing and element of GSR with SEM/EDX and finally building the equation of regression. ResultsThe relationship between the quantity of GSR and firing distance is linear and the equation is built. When the firing distance is from 10 to 90cm, the result is preferably. ConclusionWe can estimate the "5.4"hand gun firing distance through testing GSR around gunshot wounds.

Objective To investigate the effects of normobaric hyperoxia intervention on learning and memory abilities of valproic acid(VPA) autism model rats and the morphology of pyramidal cells in hippocampus CA1 area.Methods Animal model groups of autism were obtained in male offspring of the Wistar rats that received intraperitoneal injection of 600 mg/kg VPA at the 12.5 day after pregnancy.According to the eye opening time,behavior,weaning weight and the learning and memory abilities which were evaluated by the Y electricity maze test at the 28th day after birth,40 male VPA autism model rats were randomly selected 20 only and divided into normobaric hyperoxia model group (group A,n =10),atmospheric air model group (group B,n =10).Normal control groups were obtained in male offspring of Wistar rats that received intraperitoneal injection of equivalent physiological saline at the same period pregnancy.(group C,n =10).Rats in group A were treated with oxygen for 1 h per day and lasted 1 week;group B and C were treated with normal air.The learning and memory abilities of three groups were assessed at the 35th day after birth.The immunohistochemistry methods and image analysis were used to observe the pyramidal cells of autism model rats in hippocampal CA1 region.The effect of normobaric hyperoxia therapy on pyramidal cell of autism model rats in hippocampal CA1 region were evaluated by HE staining technique.Results The trying times of group A after treatment were less than those before treatment (31.15 ± 0.99 vs 31.54 ± 0.97,t =2.739,P =0.018).The memory times were more than those before treatment (3.00± 0.58 vs 2.69 ± 0.48,t =-2.309,P =0.040).The trying times of group A after treatment were less than those in group B after treatment (P =0.016).The memory times of group A were not different from that in group B after treatment(P=0.810).The morphology of pyramidal cells in hippocampal CA1 region showed that the pyramidal cells of the autism model rats had apoptosised.The number of apoptotic cells reduced and the number of normal form cells increased after the normobaric hyperoxia intervention compared with the autism model rats.Conclusion Normobaric hyperoxia intervention can improve the learning and memory abilities of the autism model rats.The apoptosis of the pyramidal neurons in hippocampal CA1 might be reduced after the normobaric hyperoxia intervention.

Objective To observe the cerebral activation pattern in patients with cervical dystonia (CD) during finger movements and the changes caused by botulinum toxin injection by functional MRI.The possible etiological mechanism of CD and effect of peripheral botulinum toxin treatment on the level of central nerves system are investigated.Methods A designed functional MRI block with complex finger movements was applied and 11 patients with CD as well as 11 age and gender matched controls were scanned.Compare the activation pattern of CD pre/post treatment groups versus health controls.Evaluate and compare the symtom severity with Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS).Make a correlation analysis between activation pattern alteration and TWSTRS change in CD pre/post treatment groups.Results An reduced extent of activation in patients with CD was demonstrated compared to healthy controls in ipsilateral putamen,prefrontal cortex and contralateral somatosensory cortex to the direction of the head deviation,while an elevated extent of activation in ipsilateral precuneus and fusiform with statistic significance.At the time point of 4 weeks after botulinum toxin treatment patients showed no evident difference with healthy controls except for the decreased activation in contralateral precuneus to the direction of the head deviation.TWSTRS of patients with CD decreased from (20.02 ± 5.52) to (4.11 ± 4.34) with statistic significance (t =11.71,P =0.000) after botulinum toxin treatment.There was a positive correlation between cerebral activation pattern change in some cerebral areas (ipsilateral primary somatosensory cortex,premotor cortex,supplementary motor cortex,insula,fusiform,hippocampus with parahippocampa as well as contralateral middle temporal cortex and hippocampus with parahippocampa to the direction of the head deviation) and TWSTRS score decrease.Conclusions There are widespread abnormalities in cortical and subcortical activation pattern in patients with CD,which might due to dysfunction of sensory-motor integration.We speculate a basic pre-dystonic state is present in affected body parts prior to clinical symptoms appear.Botulinum toxin fulfills its subsequent central effect by reorganizing and normalizing the cerebral cortex in patients with CD.

to common antifungal drugs.

Objective To report a case of Majocchi's granuloma induced by var. raubitschekii of Trichophyton rubrum, and to evaluate the relationship between deep and superficial fungal infection with genotyping technique. Methods The patient underwent physical, pathological and mycologic examination,which included microscopic observation, fungal culture, and urease reaction. The sequence of intertranscribed spacer (ITS) of rDNA was analysed by PCR and sequencing. Isolates from affected toes and tissue as well as one reference strain and six clinical strains of T. rubrum, were subjected to analysis of the tandem repeat subelement(TRS-1) in nontranscribed spacer(NTS)of rDNA by PCR. Results A 48-year-old female patient presented with a 2-month history of red papules and nodular lesions on the back, buttock and thigh,as well as a 3-year history of onychomycosis which had become more severe after a liver transplantation 9 months before. Pathological and mycologic examinations confirmed the diagnosis of Majocchi's granuloma. The pathogen was identified as var. raubitschekii of T. rubrum by microscopic examination, fungal culture, positive urease reaction and the sequence of ITS. As shown by the amplications of TRS-1 of NTS, the genotypes of strains from affected nails and tissue were identical, but differed from those of other clinical strains of T.rubrum. Conclusions There is a polymorphism in TRS-1 of rDNA NTS of T. rubrum, with the genotypes of isolates from affected nails and tissue being identical, which suggests they are of the same origin.

Arteriovenous Fistula ; therapy ; Child ; Embolization, Therapeutic ; Humans ; Male ; Pulmonary Artery ; abnormalities ; Pulmonary Veins ; abnormalities

OBJECTIVETo observe the effect differences on post-stroke dysphagia among acupoint injection combined with neural electrical stimulation, acupoint injection, neural electrical stimulation, and swallowing training respectively, so eligible intervention can be applied to this kind of disease.

METHODSOne hundred and eight-three patients of post-stroke dysphagia were randomized into a comprehensive treatment group (42 cases), an acupoint injection group (44 cases), a neural electrical stimulation group (49 cases) and a swallow training group (48 cases) and were treated with the comprehensive therapy of acupoint injection and neural electrical stimulation, acupoint injection, neural electrical stimulation and swallowing training separately. The treatments for 10 days made one session. There were 3 days at the interval among treatment sessions and 3 sessions were required totally. The cases in those treatment groups were blankly controlled with the other 47 patients of post-stroke dysphagia. All the patients received basic rehabilitation treatment. The modified water swallowing test was conducted to assess the efficacy before treatment, 10 days after treatment and 30 days after treatment in each group separately. The clinical efficacy, score of water swallowing test and improvement in water swallow test were compared among the groups.

RESULTSAfter 10-day treatment, the differences in efficacy and score of water swallow test were not significant in each group (all P > 0.05). After 30-day treatment, the effective rate (94.29%, 33/35) in the comprehensive treatment group was apparently better than 68.75% (22/32) in the acupoint injection group, 80.00% (32/40) in the neural electrical stimulation group, 67.50% (27/40) in the swallowing training group and 42.86% (12/28) in the blank group separately. The score in water swallow test in the comprehensive treatment group was lower than that in each of the other groups (1.37 ± 0.60 vs 2.03 ± 1.00, 1.90 ± 0.90, 2.20 ± 0.72, 2.71 ± 0.90, all P < 0.05). The differences in the effective rate and score in water swallow test were not significant among the acupoint injection group, neural electrical stimulation group and swallowing training group (all P > 0.05), which indicated that the improvement in swallowing function in the comprehensive treatment group was significantly superior to the other groups (all P < 0.05).

CONCLUSIONThe comprehensive therapy of acupoint injection and neural electrical stimulation achieves the much better efficacy on post-stroke dysphagia.

Acupuncture Points ; Adult ; Aged ; Combined Modality Therapy ; Deglutition ; Deglutition Disorders ; drug therapy ; etiology ; physiopathology ; therapy ; Electric Stimulation Therapy ; Female ; Humans ; Male ; Middle Aged ; Stroke ; complications ; Treatment Outcome ; Vitamin B 12 ; administration & dosage ; Young Adult

Objective To study the immune response gene and the mechanism of membranous glomerulonephritis in the mouse. Methods Reproduced and identitied the animal model of MGN in mice ,extracted the total RNA of pathology group and the control group,amplified and verified I-Aβ1 gene through RT-PCR. Then sequenced and analyzed the I-AβI gene from the PCR production. Results The mutation rate of the I-Aβ1 gene was 2. 578‰ in the pathology group, and 0. 286‰ in the control group. It was obviously higher in the pathology group than in the control group(P ＜0.01). Conclusion I-Aβ1 gene rose in mice,may be related to membranous glomerulonephritis.

Objective:To investigate the efficiency of Agrobacterium tumefaciensmediated transfor-mation of Aspergillus fumigatus by using pyrG as a recessive selectable marker.Methods: FAP1 and SHO1 genes target sequences,composed of a selectable marker pyrG and the flanking sequences of the FAP1 and the SHO1 genes,were cloned into a binary plasmid pDHt/sk,respectively.The produced plasmids were transformed into A.tumefaciens.The A.tumefaciens and uracil auxotroph A.fumigatus were cocultured in induction medium without uricil and uridine at 24 ℃ for 48 h.To inhibit growth of A.tumefaciens and to select transformants,the cultures were transferred to 37 ℃ and incubated for another 48 h.Results: In this study,A.tumefaciens-medidated transformation of A.fumigatus produced high homologous recombination rates,which was 44%(7 of 16) for FAP1 and 35%(7 of 20) for SHO1.Conclusion: Our study showed that A.tumefaciens-medidated transformation by using pyrG as a recessive selectable marker is an efficient tool for target gene deletion of A.fumigatus.