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.

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

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 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.

This study was aimed to probe into the mechanism of moxibustion in the treatment of Alzheimer’s disease (AD). A total of 40 male 15-month-old SD rats were randomly divided into the normal group, model group, moxibustion group and sham-operation group. Stereotactic injection of agglutinated Aβ25-35 into rat’s bilateral hippocampus was used to prepare AD models. Equal amount of normal saline was injected to rat’s bilateral hippocampus in the sham-operation group. Model rats were treated by moxibustion at the distance of 2-3 cm above points of‘BL23-Shenshu’, ‘ST36-Zusanli’ and ‘GV20-Baihui’. No intervention was given to rats in the normal group. The learning and memory ability was detected by Morris water maze test. Changes on expressions of Bcl-2, Bax and Caspase-3 of hippocampus zone were detected by immunohistochemical method. The results showed that in the model group, the average escape latency of five days and the last three days were significantly lengthened. And the times across the platform position were significantly reduced. Compared with the normal group and the sham-operation group, expressions of Bax and Caspase-3 in the hippocampus were significantly increased, and the expression of Bcl-2 was obviously decreased (P < 0.01). In the moxibustion group, the average escape latency of five days and the last three days were shortened obviously, and the times across the platform position were significantly increased. Compared with the model group, expressions of Bax and Caspase-3 were significantly reduced, and the expression of Bcl-2 was significantly increased (P < 0.01). It was concluded that the action mechanism of AD treatment with moxibustion may be through the reducing of proapoptotic protein Bax and Caspase-3 releasing, promoting the releasing of anti-apoptotic protein Bcl-2, so as to improve the learning and memory impairment caused by Aβ.

Objective To observe the effects of moxibustion on learning and memory ability and nerve cell ultrastructure in CA1 region of hippocampus of rats with Alzheimer's disease (AD);To discuss its mechanism of action to AD.Methods A total of 40 male SD mice was randomly divided into normal group, model group, moxibustion group and sham-operation group. Stereotactic injection agglutinated Aβ25-35 into bilateral hippocampus of rats in model group and moxibustion group to prepare AD models. Rats in moxibustion group were treated by moxibustion at the points which were located above 2-3 cm away from Shenshu, Zusanli and Baihui. Rats in normal group received no treatment. The learning and memory ability was detected by Morris water maze test. Morphological changes of rat nerve cell ultra structure in hippocampal CA1 region were observed by transmission electron microscope.Results In model group, the average escape latency of five days and the last three days was significantly lengthened, and the times across the platform position significantly decreased. Transmission electron microscope indicated nerve cell structures were damaged and displayed unclearly;mitochondrion swelled, even mitochondrial crista was abnormal and deformed, endoplasmic reticulum of inter organelle expanded and the deposition of lipofuscin increased. After the treatment by moxibustion, the average escape latency of five days and the last three days was shortened obviously, and the times across the platform position significantly increased. In moxibustion group, the edema of nerve cells significantly decreased;dilation of endoplasmic reticulum and mitochondria swelling significantly improved;golgi bodies, mitochondria and ribosomes obviously increased in comparison with the model group. Conclusion Moxibustion can improve the learning and memory impairment caused by Aβ by promoting repairing of nerve cells in brain.

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 damage of rat articular cartilage induced by different doses of T-2 toxin, and to explore the relationship between mini-dose T-2 toxin and articular cartilage damage. Methods A total of 120 Wistar rats, weighing 50 - 70 g, were randomly divided into four groups according to their body weights: T-2 toxin group 0(control), 100, 200, 300 μg/kg, 30 rats in each group. Animals in the control group were fed standard rat chow, and animals in the three T-2 toxin groups were fed T-2-toxin-contaminated chow (the dose was 100, 200, 300 μg/kg, respectively). After 6 months, rats were euthanized by ether asphyxiation. The bilateral knee joints were collected and section prepared. The articular cartilage was examined by light and electronic microscope. Results Light microscope showed, the rat articular chondrocytes were clear and arranged orderliness in the control group. The rat articular chondrocytes were disarranged in 100 μg/kg T-2 toxin group.Degeneration and necrosis were found in 200 μg/kg group. Chondrocytes were shrunken with hypereosinophilia cytoplasm and fragmented pyknotic nuclei, extensive areas of chondrocyte loss and chondrocyte clones were visible in 300 μg/kg group. Scanning electronic micrograph(SEM) showed, the rat articular chondrocytes were clear, well formed and arranged tidy in the control group. The surface of articular cartilage was rough in 100 μg/kg group.Collagen fasciculi ruptured and stacked up in 200 μg/kg group. Presented a typical articular dryness phenomenon,the cartilage surface collapsed and many pits appeared in 300 μg/kg group. Transmission electronic microscope (TEM) showed that chondrocytes were abundant with cytoplasm, well-developed rough endoplasmic reticulum in the control group; agglomerate chromatin scattered along the karyotheca, nuclear membrane was thickening, with vacuolar degeneration of the endoplasmic reticulum in the 100 μg/kg group; endoplasmic reticulum expended, with protein retention and organelles breaks in the 200 μg/kg group. A large number of chondrocytes lost organdles, the membrane structures disrupted and the cartilage matrix stromatolyzed in the 300 μg/kg group. Conclusions Within the range of 100 - 300 μg/kg, T-2 toxin induces dose-related articular cartilage injury, the greater the dose, the more serious damage.

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.