Objective:To explore the distribution and clearance of 99mTc labeled diethylenetriamine pentaacetic acid(99mTc-DTPA)in different brain regions of adult rats after administration through brain extracellular space(ECS)pathway.Methods:After the injection of a volume of 2 μL and radioactive activity of about 3.7 MBq(100 μCi)of 99mTc-DTPA into the caudate nucleus and thalamus of SD rats through stereotactic positioning of rat brain,the single photon emission computed tomography/computed tomography(SPECT/CT)for small animals was used for imaging at different time points,and the dyna-mic distribution and clearance of the tracer in the whole body were observed continuously.The SD rats were injected with 99mTc-DTPA into thalamus and caudate nucleus respectively for biological distribution in vivo.They were put to death 4 h later.Their blood and urine were collected.The brain,cerebellum,heart,liver,spleen,lung,and kidney were taken and weighed by γ counter to measure its radioactivity.Results:SPECT/CT imaging results showed that after 99mTc-DTPA was administered through brain ECS,the radioactivity was concentrated in the brain,kidney and bladder.The tracer administered to the left caudate nucleus was preferentially drained to the right cerebellum,while the tracer administered to the right caudate nucleus was preferentially drained to the left cerebellum.There was a phenomenon of"con-tralateral cerebellar dominant drainage"in the caudate nucleus.The thalamic area preferentially drained to the ipsilateral cerebellum after administration.Four hours after administration via ECS,high radioac-tive uptake appeared in urine,cerebellum and brain,followed by blood and kidney.The radioactive up-take values of heart,liver,spleen and lung were low,which were mainly excreted through urinary sys-tem.Conclusion:Intracerebral ECS administration is a promising method of administration,but there are significant differences in distribution and clearance in different brain regions.This study further ex-pands the content and significance of"ECS regions",and also provides an important theoretical founda-tion for the treatment of encephalopathy and the research of new drugs through brain ECS in the future.

Objective:To explore the distribution and clearance of 99mTc labeled diethylenetriamine pentaacetic acid(99mTc-DTPA)in different brain regions of adult rats after administration through brain extracellular space(ECS)pathway.Methods:After the injection of a volume of 2 μL and radioactive activity of about 3.7 MBq(100 μCi)of 99mTc-DTPA into the caudate nucleus and thalamus of SD rats through stereotactic positioning of rat brain,the single photon emission computed tomography/computed tomography(SPECT/CT)for small animals was used for imaging at different time points,and the dyna-mic distribution and clearance of the tracer in the whole body were observed continuously.The SD rats were injected with 99mTc-DTPA into thalamus and caudate nucleus respectively for biological distribution in vivo.They were put to death 4 h later.Their blood and urine were collected.The brain,cerebellum,heart,liver,spleen,lung,and kidney were taken and weighed by γ counter to measure its radioactivity.Results:SPECT/CT imaging results showed that after 99mTc-DTPA was administered through brain ECS,the radioactivity was concentrated in the brain,kidney and bladder.The tracer administered to the left caudate nucleus was preferentially drained to the right cerebellum,while the tracer administered to the right caudate nucleus was preferentially drained to the left cerebellum.There was a phenomenon of"con-tralateral cerebellar dominant drainage"in the caudate nucleus.The thalamic area preferentially drained to the ipsilateral cerebellum after administration.Four hours after administration via ECS,high radioac-tive uptake appeared in urine,cerebellum and brain,followed by blood and kidney.The radioactive up-take values of heart,liver,spleen and lung were low,which were mainly excreted through urinary sys-tem.Conclusion:Intracerebral ECS administration is a promising method of administration,but there are significant differences in distribution and clearance in different brain regions.This study further ex-pands the content and significance of"ECS regions",and also provides an important theoretical founda-tion for the treatment of encephalopathy and the research of new drugs through brain ECS in the future.

RBM46 is a germ cell-specific RNA-binding protein required for gametogenesis, but the targets and molecular functions of RBM46 remain unknown. Here, we demonstrate that RBM46 binds at specific motifs in the 3'UTRs of mRNAs encoding multiple meiotic cohesin subunits and show that RBM46 is required for normal synaptonemal complex formation during meiosis initiation. Using a recently reported, high-resolution technique known as LACE-seq and working with low-input cells, we profiled the targets of RBM46 at single-nucleotide resolution in leptotene and zygotene stage gametes. We found that RBM46 preferentially binds target mRNAs containing GCCUAU/GUUCGA motifs in their 3'UTRs regions. In Rbm46 knockout mice, the RBM46-target cohesin subunits displayed unaltered mRNA levels but had reduced translation, resulting in the failed assembly of axial elements, synapsis disruption, and meiotic arrest. Our study thus provides mechanistic insights into the molecular functions of RBM46 in gametogenesis and illustrates the power of LACE-seq for investigations of RNA-binding protein functions when working with low-abundance input materials.
Animals ; Mice ; 3' Untranslated Regions/genetics* ; Cell Cycle Proteins/metabolism* ; Gametogenesis/genetics* ; Meiosis/genetics* ; Nuclear Proteins/genetics* ; RNA-Binding Proteins/genetics*

Objective To explore the anti-inflammatory and anti-angiogenic effect of miR-15a in diabetic retinopathy(DR).Methods High glucose model(HG)was established by treatment with 25 mmol/L glucose in human retinal pigment epithelial cell line HARPE-19,and HARPE-19 cells treated with 5 mmol/L glucose served as control(LG).HG group and LG group were divided into 4 subgroups:miR-15a mimic group,NC-mimic group,miR-15a inhibitor group and NC-inhibitor group.Cell proliferation ability was detected by MTT proliferation assay.Cell migration ability was detected by cell scratch assay.The relationship between miR-15a and ASM was identified by 3′UTR luciferase reporter assay.miR-15a,ASM,VEGF,IL-1β,IL-6,TNF-α mRNA expression was detected by RT-PCR method.Results In NC-mimic cells and NC-inhibitor cells,the expression of miR-15a in HG group was significantly lower than that in LG group(all P<0.05).The expression level of ASM mRNA in miR-15a mimic group was significantly lower than that in NC-mimic group (P<0.05),the expression level of ASM mRNA in miR-15a inhibitor group was significantly higher than that in NC-inhibitor group (P<0.05).The cell scratch width in miR-15a mimic group was significantly higher than that in NC-mimic group(P<0.05).The cell scratch width in miR-15a inhibitor group was significantly lower than that in NC-inhibitor group(P<0.05).The absorbance value in miR-15a mimic group was significantly higher than that in NC-mimic group(P<0.05),the absorbance value in miR-15a inhibitor group was significantly lower than that in NC-inhibitor group(P<0.05).In NC-mimic cells and NC-inhibitor cells,the mRNA expression levels of VEGF,IL-1β,IL-6 and TNF-α in HG group was significantly higher than those in LG group(all P<0.05).Conclusion miR-15a downexpression induces inflammatory reaction and angiogenesis in retinal cells.miR-15a is involved in the pathogenesis of DR by dual regulation of inflammation and angiogenesis.

Objective To study the protective effects of tBHQ on type 2 diabetic rats retina and its related mechanism.Methods Sixty SD rats were divided into normal control group (NC group),model group (diabetes mellitus,DM group) and tBHQ group.After feeding with high fat and high sugar diets for 4 weeks,the rats in model group were induced by intraperitoneal injection of STZ for the model of type 2 diabetes mellitus.1％ tBHQ were added into the high fat and high sugar feed 1 week later after successfully modeled in the tBHQ group.Fasting plasma glucose (FPG) and fasting serum insulin (FINs) were detected at 4 and 12 weeks after modeled.Immunohistochemical method and real-time fluorescent quantitative PCR (qRT-PCR) were respectively used to detect the distributions and relative expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2),heme oxygenase 1 (HO-1),B-cell lymPhoma 2 (Bcl-2) and vascular endothelial growth factor (VEGF) in retinas of the rats.Results FPG levels totally comparative differences were statistically significant between each group (F =78.531,P =0.000).The level of FPG in DM and tBHQ group were obviously higher than that in NC group,the 12 weeks was higher than 4 weeks in DM group,and the 12 weeks was lower than 4 weeks in tBHQ group (all P ＜ 0.05).Totally comparative differences in the level of FINs were statistically significant (F=22.480,P =0.000),NC group was lower than DM and tBHQ group,12 weeks was higher than 4 weeks (all P ＜ 0.05).Immunohistochemical detection showed that each factor in each group were expressed in the retina of rat,and the relative expressions of Nrf2,HO-1,Bcl-2 and VEGF protein in retina have totally statistically differences in different time points after modeled (all P ＜0.05).The expressions of each factor in DM group were more than those in the NC group.The Nrf2,HO-1 and Bcl-2 in tBHQ group were more than those in the DM group,but the VEGF was lower.At 12 weeks,the expressions of VEGF and Nrf2 in DM group were more than those at 4 weeks,but the HO-1 was lower;the Nrf2,HO-1 and Bcl-2 in tBHQ group were more than those at 4 weeks (all P ＜ 0.05).PCR tests revealed that the relative expressions of Nrf2,HO-1,Bcl-2 and VEGF mRNA in retina have totally statistically differences in different time points after modeled(all P ＜ 0.05).The expressions of each factor in DM group were more than those in the NC group.The Nrf2,HO-1 and Bcl-2 mRNA in tBHQ group were more than those in the DM group,but the VEGF mRNA was lower.At 12 weeks,VEGF mRNA in DM group and Nrf2,HO-1,Bcl-2 in tBHQ group were more than those at 4 weeks (all P ＜ 0.05).Conclusion tBHQ maybe have protection for islet function on diabetic rat,and can induce the expressions of Nrf2,HO-1,Bcl-2 in retina of diabetic rats and reduce the expression of VEGF,inhibit the oxidative stress of retinal tissue damage,reduce cell apoptosis and inhibit the proliferation of retinal blood vessels.tBHQ may protect the retina of diabetic rats through the Nrf2/HO-1/VEGF and Nrf2/Bcl-2 way.

Diabetic retinopathy (DR) is the most serious and common blinding ocular complication in all microangiopathy of diabetes mellitus.The occurrence of DR associated with a variety of factom,the pathogenesis has not yet been completely clear,and there are several theories.In recent years,the relationship between the occurrence of DR and sex hormone levels become a hot ophthalmic research,research shows that the prevalence of DR significantly increased in postmenopausal women,in which estrogen change plays the main role.Learning connection between estrogen and DR will contribute to the clinical diagnosis and treatment of DR.This article reviews recent research advances in the relationship between estrogen and DR.

Objective:To retrospectively analyze the CT features among gastric cancer, gastric lymphoma and gastric malignant stromal tumors, evaluated the application value of MDCT in gastric malignant tumor.Methods: Sixty two patients with gastric malignancies tumor confirmed by pathology underwent MDCT plain scanning and triphasic contrasted scanning before operation. Referring to the pathological results, to observe the CT signs, include: scope of tumors and involvement, the density of tumors and enhanced characteristics, mucosal membrane, the change of the stomach softness, peripheral lymph node enlargement, the involvement of other organs, and to make statistical analysis for imaging features of these tumors byx2 testing.Results: Sixty two cases of gastric malignant tumor included 42 cases of gastric carcinoma, 12 cases of gastric lymphoma and 8 cases of gastric malignant stromal tumor, were confirmed by postoperative pathology. The CT manifestations were as follows: the stomach mucous membrane surface involvement in all the gastric cancer, the positive rate was significantly higher than gastric lymphoma and stromal tumor. The area involved by lymphoma were more than two partitions, significantly higher than that of gastric cancer and stromal tumor (x2=36.643,x2=20.00;P<0.05). 39 cases (92.8%) of gastric cancer and 8 cases (100%) of malignant stromal tumors were significantly enhanced, but all gastric lymphoma were not obvious enhancement. The positive rate of enlarged lymph nodes in gastric cancer(61.9%) and lymphoma(83.3%) were higher than stromal tumor(12.5%)(x2=9.731,x2=6.603;P<0.05).Conclusion: The CT manifestations of the gastric cancer, lymphoma, malignant stromal tumors had different characteristics. Multislice CT for the diagnosis and differential diagnosis of gastric malignant tumor has important value.

BACKGROUND:Propagation phase contrast tomography can greatly improve the spatial resolution of chondrocytes and microvessels depending on the high colimation and high coherence performance of hard X-ray. OBJECTIVE: To detect the alteration of angioarchitecture after spinal cord injury in rats using propagation phase contrast tomography. METHODS: Eight male Sprague-Dawley rats were divided into two groups: in experimental group, an acute spinal cord injury model was induced in rats by the modified Alen’s method; in sham control group, rats were subjected to laminectomy. At 1 day after operation, normal and injured spinal cord segments were taken and treated with formaldehyde-methyl salicylate sequentialy for 48 hours. The segments were scanned by propagation phase contrast tomography in BL13W1 beamline experimental station of Shanghai Synchrotron Radiation Facility, China. Harvested data were analyzed by VGStudio Max 2.1 software for three-dimensional reconstruction and microvasculature quantitative analysis. RESULTS AND CONCLUSION:The propagation phase contrast tomography successfuly simulated the morphology of angioarchitecture folowing spinal cord injury. After acute spinal cord contusion, the destruction of nerve tissues was accompanied by severe microvasculature damage. Intramedulary tissue damage and loss of blood supply was spread from the central zone to the ends. Three-dimensional microvascular quantitative data showed that after spinal cord contusion, the number of microvessels and vascular perfusion volume drasticaly reduced (P < 0.01). These findings indicate that the propagation phase contrast tomography without angiography has potential as a new ultra high-resolution visualization technique for three-dimensional microvessel imaging and quantitative analysis.

Background Studies showed that vascular endothelial growth factor (VEGF) plays an important role in the development and progress of diabetic retinopathy (DR),and the association between VEGF-2578C/A polymorphism(SNPs) and risk for DR is a hotspot.Objective This Meta analysis aimed to investigate the comprehensive and reliable conclusion in the association of VEGF-2578C/A SNPs and risk for DR in different races.Methods A systematic search of electronic databases including PubMed,Cochrane Library,EMbase,VIP,Wanfang technological,CNKI and reference lists of relevant articles was carried out until April,2014.Case-control studies on the relationship between VEGF-2578C/A SNPs and DR were selected based on inclusion and exclusion criteria,and the relevance of VEGF-2578C allele to DR,the relevance of VEGF-2578C/A SNPs to DR and the relevance of VEGF-2578A allele to Caucasian DR were quantitatively analyzed.Begger funnel plot of publication biases on the relationships of VEGF SNPs with the risk of DR under the allele and dominant models was drown.RevMan 5.0 software was used for the statistical analysis.The pooled odds ratio (OR) and corresponding 95％ confidence interval (CI) were used to assess the strength of the association.Results A total of 1 228 DR cases and 1 224 diabetes controls without retinopathy(DWR) were included from 8 independent studies (9 groups of data).A significant relationships between VEGF-2578A allelic gene and VEGF-2578AA gene type with DR were found in all samples,and the A allelic gene and AA gene type were the risk genes of DR (A versus C:OR=1.39,95％ CI=1.08-1.80,Z=2.52,P=0.01;AA versus CC+C/A:OR=1.53,95％ CI=1.05-2.24,Z=2.20,P=0.03;CC versus AA+C/A:OR=0.70,95％ CI=0.50-0.98,Z =2.10,P =0.04).When the other two studies which did not meet the HardyWeinberg Equilibrium were incorporated in a sensitivity analysis,the results were not materially altered.VEGF-2578 A allelic gene was the risk gene to Europeans with DR (OR =1.50,95％ CI=1.02-2.21,Z =2.07,P =0.04),but not among Asians in subgroup analysis (P＞0.05).No significant publication bias was found.Conclusions The Meta analysis demonstrates that VEGF-2578C/A is associated with DR in Europeans but not in Asians.Further case-control studies based on larger sample size are still needed,especially in Asians.

Background Thyroid-associated ophthalmopathy (TAO) is a kind of clinically common and incurable ocular disease,and its incidence is at top place.The etiology and pathologic mechanism of TAO are still unknown because of shortness of replicative animal models and difficulty to acquire the ocular tissues in the early stage of the disease.To better understand the pathogenesis of TAO and investigate effective treatable measures, an appropriate animal model should be developed.Objective This study was to immunize female BALB/c mice with the recombinant plasmid of human thyroid-stimulating hormone receptor (TSHR) extracellular domain in cationic liposomes for the establishement of TAO models.Methods Thirty-two 6-to 8-week-old female BALB/c mice were randomly assigned to four groups according to computer random allocation.pcDNA3.1 +/hTSHR289 of 100 μg in an adjuvant cationic liposomes was injected via anterior tibialis muscle and peritoneal cavity separately in the recombinant plasmid injection group in 0, 3,6 weeks, and pcDNA3.1 or cationic liposomes was injected in the liposomes injection group or the blank plasmid group in the same way, respectively, and normal saline solution was injected in the blank control group.Body weight of the mice was measued before and 1 month,2,3 and 4 months after initial injection.The manifestations were observed after modeling.The mice were sacrificed 17 weeks after initial injection,and the histopathology examination was carried out on the thyroid gland and orbital tissue.The heart blood was collected from the mice,and serum contents of total thyroxin 4 (TT4) and thyroid-stimulating hormone (TSH)were assayed by ELISA.Results Protrusion, eyelid swell and keratitis occurred in 12 eyes of 6 mice in the recombinant plasmid injection group after immunization.A significant difference in the body weight of the mice was found among the blank control group, blank plasmid group, liposomes injection group and recombinant plasmid injection group (Fgroup =3.425, P =0.028), and the body weight was considerably reduced in the recombinant plasmid injection group in comparison with the blank control group, blank plasmid group,liposomes injection group (Ftime =0.838 ,P=0.023).The serum levels of TT4 were (7.75±1.00), (7.96±0.76), (6.76±1.10) and (4.43±2.88) μg/dlin the blank control group, liposomes injection group, blank plasmid group, and recombinant plasmid injection group, and those of TSH were (6.36±2.58),(4.83±3.96),(6.63±1.71) and (1.60 ±1.76) ng/ml, showing significant differences among the groups (F =7.150, P＜0.001;F =5.521, P＜0.01) , and the serum levels of TT4 and TSH were remarkably lower in the recombinant plasmid injection group than those of the blank control group,liposomes injection group and blank plasmid group (all at P ＜ 0.05).Histopathology revealed the lymphocyte infiltration of thyroid gland in 6 mice and proliferation of orbital adipose tissue, infiltration of lymphocytes and mastocytes,deposition of hyaluronic acid as well as swell, breakage and inflammatory cell infiltration of extraocular muscle in 15 eyes of the recombinant plasmid injection group.Conclusions A murine model of TAO can be successfully induced by immunization with recombination plasmid pcDNA3.1 +/hTSHR289 and cationic liposomes.The histopathology characteristics and ocular findings of the animal models are similar to human TAO.