1.Correlation between serum osteocalcin and type 2 diabetes mellitus in elderly patients
Jie DENG ; Yikun ZHU ; Shuhong SHI ; Xing LI ; Baozhen ZHAO
Chinese Journal of Geriatrics 2013;(5):510-512
Objective To investigate the correlation between serum osteocalcin and type 2 diabetes mellitus (T2DM) in elderly patients.Methods A total of 55 T2DM patients (diabetic group) and 50 non-diabetic subjects (control group) aged ≥60 years were enrolled in this study.The levels of fasting blood glucose (FBG),glycated hemoglobin (HbAlc),insulin resistance index (HOMA IR),osteocalcin (OC),body mass index (BMI) and bone mineral density (BMD) in groups were compared.The correlations between serum osteocalcin and the above indicators were analyzed.Results The levels of OC and BMD were higher in control group than in diabetic group [(11.2±3.2)μg/Lvs.(4.1±3.0)μg/L,(-1.3±0.3) vs.(-2.6±0.5),respectively,both P<0.05].The levels of FBG,HbAlc,FINS,HOMA-IR were lower in control group than in diabetic group [(4.7±2.0) mmol/L vs.(9.4±2.1) mmol/L],[(4.8±1.5) % vs.(7.6±1.6)%,(7.4±3.2) U/L vs.(23.7±3.0) U/L,(1.5±0.7) vs.(9.9±1.2),respectively,all P<0.05].Serum osteocalcin concentration was negatively correlated with the levels of FBG,HbAlc,FINS and HOMA-IR in elderly patients with T2DM (r=-0.739,-0.713,-0.613,-0.092,all P<0.01).Conclusions Serum osteocalcin concentration is correlated with the levels of blood sugar and insulin resistance index in elderly patients with T2DM.A further study on the correlation between osteocalcin and T2DM may provide a new target for the treatment of type 2 diabetes mellitus.
2.Binocular integration of binocular neurons in striate cortex of Kitten
Fang, XIE ; Xue-feng, SHI ; Li-min, XU ; Teng-yue, ZHANG ; Jia-xing, WANG ; Yu-xian, NING ; Kan-xing, ZHAO
Chinese Journal of Experimental Ophthalmology 2011;29(6):485-488
Background The integration of segregated pathways from the two eyes first appears in V1 neurons,where it not only plays a critical role in the generation of a three-dimensional visual representation.Abnormal visual experiences in critical period usually lead to amblyopia and binocular integration defects.Objective Present study was to investigate how neurons of kitten coordinate their activity patterns in response to synchronous dichoptic stimulus inputs in striate cortex.Methods Spike rate and local field potential(LFP) gamma band(20-90Hz) power of three kitten(1-1.2Kg,8-10 weeks old) to monocular and synchronous dichoptic presented gratings were assessed for 28 binocular neurons in V1 of kitten by in vivo extracellular record method under anaesthesia and paralysis.Ocular dominance index(ODI) and binocular integration index(BII) were assessed and the correlation between these two indexes were analyzed.Results In 28 cells with binocular characteristic,the absolute value of spike-ODI was significant larger than that of LFP-ODI(t=2.606,P=0.021).A positive linear correlation between the ocular preferences of spike and LFP was found(R2=0.513,F=27.423,P=0.003).In dichotic trails,binocular facilitation with BII for spike was 2.348±0.996,showing a significant reduce in comparison with BII for LFP(3.678±1.974)(t=2.671,P=0.019).Binocular integration index for two signals were greater when monocular responses of both eyes were similar(P=0.035 and P=0.124,respectively).Conclusion Both spike rate and gamma band power of LFP exhibited binocular facilitation to synchronous presented dichotic stimuli with significant facilitation induced by balanced monocular responses.Spiking activity and LFP reflect neural activities of different spatial scales and source components.
3.Cross-linking and immunoprecipitation assay and its development
Lingyue XING ; Yan ZHANG ; Jing HUANG ; Ping LI ; Yang WANG ; Minglei SHI ; Zhihu ZHAO
Military Medical Sciences 2013;(11):867-869
It′s reported that RNA-binding proteins ( RBP) play key roles in post-transcriptional regulation of eukaryotic genes.The aberrations of RBP are associated with a large number of human disorders , particularly autoimmune and neuro-logic diseases .The interaction between RNA and proteins has been widely explored since the development of the method known as RNA immunoprecipitation with differential display or microarray analysis (RIP-ChIP) around the year of 2000. Since then, diverse derivatives of the RIP-ChIP, such as ultraviolet crosslinking and immunoprecipitation ( CLIP), high-throughput sequencing of CLIP cDNA library (HITS-CLIP), photoactivatable -ribonucleoside-enhanced crosslinking and immunoprecipitation ( PAR-CLIP) , and individual nucleotide resolution CLIP ( iCLIP ) have been developed .All these methods have some advantages over the original RIP-ChIP and greatly facilitate the study of RBP-RNA interactions .Addi-tionally , aided by the next-generation sequencing , transcriptome-wide identification of RBP target sites has become possible and the RNA-binding site resolution of RBP has also improved to some degree .We introduced the basic principles and processes of the interactions between proteins and RNA , focusing on the advantages , disadvantages and prospect of the present genome-wide version of CLIP .
4.Effects of different experimental conditions on Kunming and BALB/C mice in forced swimming test
Hui ZHANG ; Junfeng WANG ; Bo XING ; Yan ZHAO ; Fei LIU ; Jianguo SHI ; Yonghui DANG
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(4):306-308
Objective To investigate the sensitivity and variability factors that were assessed on the forced swimming test (FST) using BALB/C and Kunming mice. Methods The immobility time of FST was compared using Kunming and BALB/C mice in different experimental conditions including circadian rhythm ( day and night) ,gender and water temperature ( 12,22 and 32℃ ) . Results (①) The immobility time of BALB/C during the daytime( ( 142.42 ± 33.58) s) was significantly increased than that at night ( ( 104.89 ± 34.33 ) s). (② The immobility time of Kunming mice( (91.95 ± 40.32) s) was significantly decreased than that of BALB/C mice ( ( 142.42 ± 33.58 ) s). (③)The immobility time under the water temperature of 22 C ( ( 92.24 ± 25.81 ) s) was significant longer than that under the water temperature of 32C ( (60.72 ± 11.11 ) s). Conclusion BALB/C stain,male mice,daytime and water temperature of 22℃ should be chosen in the FST.
5.The protection of 3-AB, a PARP Inhibitor, on injures of adenohypophysis in rats with severe acute pancreatitis
Wenhong DENG ; Kailiang ZHAO ; Bo YANG ; Qiao SHI ; Xing ZHOU ; Weixing WANG
Chinese Journal of General Surgery 2013;28(10):787-791
Objective To evaluate the protection of 3-aminobenzamide (3-AB),an inhibitor of Poly (ADP-ribose) polymerase (PARP),on severe acute pancreatitis associated adenohypophysis injury in rats.Method Forty Wistar rats were randomly divided into 4 groups:sham operation group (SO group,n=8),SAP group (n=12),3-AB pretreatment group (n =12),drug control group (n =8).The bilepancreatic duct was cannulated through the duodenum and SAP model was induced by a standardized pressure-controlled retrograde infusion of 5% sodium taurocholate (0.1 ml/100 g) into the bile-pancreatic duct.In 3-AB group,3-AB (20 mg/kg) was administered via femoral vein 30 min prior to the operation;other procedures were identical to SAP group.In SO group,pancreas was flipped several times only.In drug control group,3-AB (20 mg/kg) was administered via femoral vein 30 min prior to the operation.Serum amylase,lipase were measured.Pancreas and pituitary tissue were taken for pathological examination under light microscope.PARP and NF-κB antibodies for adenohypophysis immunohistochemical stains.Adenohypophysis cell was observed under electronic microscope.Result Serum amylase,lipase and pancreas pathological scores were significantly higher in 3-AB group compared with SO group (P < 0.05),but lower than that in SAP group (P < 0.05).Adenohypophysis pathological injury was less severe in 3-AB group.Expressions of PARP and NF-κB in adenohypophysis cells were significantly higher in 3-AB group compared with SO group,but lower than that in SAP group (P < 0.05).Ultrastructural change of thyrotroph cell was relieved in 3-AB group.No significant difference was observed between SO group and drug control group in PARP and NF-κB expression nor adenohypophysis pathological injury.Conclusions 3-AB exerts the protective effect against acute pancreatitis associated adenohypophysis injury by inhibition of PARP and NF-κB.
6.Changes of PARP and NF-κB in adenohypophysis of rat model of severe acute pancreatitis
Wenhong DENG ; Kailiang ZHAO ; Bo YANG ; Qiao SHI ; Xing ZHOU ; Weixing WANG
Chinese Journal of Emergency Medicine 2013;22(10):1090-1095
Objective To investigate the changes of poly-ADP-ribose polymerase (PARP) and NF kappa B (NF-κB) in adenohypophysis in rat model of severe acute pancreatitis (SAP),and their role in the mechanism of adenohypophysis injury in SAP.Methods Forty Wistar rats were randomly (random number) divided into 5 groups:the sham operation group (SO group,n =8),SAP 1 h,3 h,6 h and 12 h groups (n =8 in each group).SAP model was induced by retrograde injection of 5% sodium taurocholate into the biliopancreatic duct.Serum levels of amylase,lipase and ascites were measured.After sacrifice of experiment rats,pancreas and adenohypophysis tissues were taken for pathological examination under light microscope.Adenohypophysis cells were observed under electronic microscopy as well.PARP and NF-κB expressions in adenohypophysis cell was studied by using immunohistochemisty assay.Results After modelling,serum levels of amylase,lipase and ascites in SAP group increased gradually,which were higher than those in SO group (P < 0.05).Adenohypophysis cell swelling and partial necrosis were observed under light microscope.As the time prolonged,their nuclei became dark and pyknotic more and more,and the endoplasmic reticulum and mitochondrial swelling in adenohypophysis cells were observed under electronic microscopy.The expressions of PARP and NF-κB in SAP group increased gradually,which were higher than those in SO group.Conclusions Significant pathological and ultrastructural injuries were observed in adenohypophysis cells in severe acute pancreatitis.These changes might correlate with PARP and NF-κB signaling pathway.
7.Influence of peroxisome proliferator activated receptor γ2 endogenous ligands on mRNA expression of bone metabolism related genes in osteoblastic cells
Yikun ZHU ; Liting LI ; Guangxia XI ; Shuhong SHI ; Xing LI ; Baozhen ZHAO
Chinese Journal of Endocrinology and Metabolism 2012;28(3):221-225
Objective To observe the effect of oxidized low-density lipoproteins (Ox-LDL),15-Deoxy-△ 12,14-prostaglandin J2 ( 15d-PGJ2 ),leukotrienes B4 ( LTB4 ) on mRNA expressions of peroxisome proliferator activated receptor γ2 ( PPARγ2 ),receptor activator of NF-κB ligand (RANKL),alkaline phosphatase ( ALP),and osteoprotegerin(OPG) in osteoblastic cells of rats; and to investigate the influence of these PPARγ2 endogenous ligands on bone metabolism.Methods Rat osteoblastic cells were cultured in vitro for 24 h in medium with different PPARγ2 endogenous ligands at various concentrations ( the final concentrations of Ox-LDL were 0,12.5,25,50μg/ml; the final concentrations of 15 d-PGJ2 were 0,10,20,30 μmol/L; the final concentrations of LTB4 were 0,0.1,1.0,10 μ mol/L).RT-PCR was performed to determine the mRNA expressions of PPARγ2,RANKL,ALP,and OPG in osteoblastic cells.Results RT-PCR analysis showed that Ox-LDL,15d-PGJ2,and LTB4 all down-regulated the mRNA expressions of RANKL,ALP,and OPG,while up-regulated the mRNA expressions of PPARγ2 in osteoblastic cells in a dose-dependent manner.Significant differences were found in interclass comparisons( P<0.05 or P< 0.01 ).Conclusions These findings suggest that Ox-LDL,15d-PGJ2,and LTB4 suppress the expressions of osteogenic genes through activating the transcription activity of PPARγ2,and this may be a plausible mechanism of senile osteoporosis.
8.Establishment of a radioresistant human lung cancer cell subline and its mechanism of radioresistance
Wei ZHAO ; Qing WANG ; Li LIU ; Xing SHI ; Gan DING ; Gang WU
Chinese Journal of Radiological Medicine and Protection 2008;28(5):508-511
Objective To establish a radioresistant cell subline from a human A549 lung cancer cell line and investigate the mechanism of radioresistance. Methods Two proposals were applied for the non-small cell lung cancer A549 cells irradiated with X-rays:A group of A549 cell line was irradiated five times, the fractionated dose was 600 cGy, and the other group was exposed 15 times, the fractionated dose was 200 cGy. After the completion of irradiation, two monoclones were obtained from the survival of cells and named the subline A549-S1 and A549-S2. The radiosensitivity and cell cycle distribution of these two clones,together with its parental A549 cells were measured by clone formation assay and flow cytometry.The mRNA and protein levels of Notch1 in A549 cell line and the sublines were determined by RT-PCR and Western-blots. Results Compared with the parental A549 cells, A549-S1 cells showed significant resistance to radiation with D0, Dq and N values increased, and a broader initial shoulder as well as 1.38-fold increased value of SF2. The A549-S1 subline also showed higher percentage of cells in S phase and G2/M phase, but lower percentages in G0/G1 phase (P<0.05). The expression of Notch1 in A549-S1 was enhanced obviously than in A549 cells. But for A549-S2 the radioseasitivity was slightly increased compared with the parental cells with D0, Dq and N values decreased and a curve initial shoulder. The ratio of cells in S and G0/G1 phase ratio was lower than that in parental A549 cells, but that in G2/M phase ratio was higher significantly (P<0.05) .The expression of Notch1 had no marked change compared to A549 cell. Conclusions The radioresistance of the A549 cell subline is correlated with the irradiation program. The cell subline shows a different cell cycle distribution from their parental line. The cell cycle distribution has a close correlation with the expression of Notch1.
9.Effects of maternal chronic aluminum exposure on memorial behaviour and hippocampal intracellular Ca~(2+) concentration on their offspring
Huihui LIU ; Jie SUN ; Wei XING ; Kui CAI ; Yuan CAI ; Zhuo ZHAO ; Qiushi TANG ; Lide SHI
Chinese Pharmacological Bulletin 2003;0(07):-
Aim To investigate the effects of maternal chronic aluminum exposure on memorial behaviour and hippocampal intracellular Ca2+ concentration ([Ca2+]i)on their offspring after the induction of LTP(long-term potentiation). Methods Adult Wistar rats (150~200 g) were exposed to aluminum by drinking distilled water, the concentration of AlCl3 is 0.015 mol?L-1(2 g?L-1) and 0.03 mol?L-1(4 g?L-1) aluminum chloride (AlCl3) solution, respectively, for 30 days prior to mating and during the whole gestation and suckling period. Their offspring were distributed into three experimental groups: control group; two exposed groups (represented by 0.2%-Al and 0.4%-Al ) administrated aluminum exposure ended at postnatal day 21. The brain tissue and blood aluminum levels were measured by Atomic absorption spectrometry (AAS). Memorial ability of the offspring was tested by Step down test.[Ca2+]i was measured by the technique of Fura-2/AM calcium ions fluorescence indicator. Results The mean aluminum content in blood and brain tissue was significantly higher than the control group(P0.05), but was significantly decreased in 0.4%-Al exposed group(P
10.Development of spatiotemporal frequency turning in rat lateral geniculate neuron
Xu-hong, ZUO ; Xue-feng, SHI ; Fang, XIE ; Li-min, XU ; Kan-xing, ZHAO
Chinese Journal of Experimental Ophthalmology 2012;30(5):388-391
BackgroundRecent researches suggested that properties of neurons in the lateral geniculate neuron (LGN) may represent an important neural limitation on the development of basic spatial and temporal vision,and even binocular rivalry.However,previous studies on the properties of spatiotemporal frequency tuning of LGN were rather concentrated on a monkey or cat,whereas little is known about rat.ObjectiveThis study was to examine the development of spatiotemporal frequency tuning in rats LGN.MethodsTwenty Wistar rats were collected and divided into 14-16 day,20-22 day,27-30 day and 60 day groups according to the different ages after their eyes opened and 5 rats were assigned for each group.Extracellular single neuron recording was carried out in the rats to study the spatio-temporal receptive field properties of neurons in LGN by sinusoidal gratings visual stimuli.Dynamic changes of the spatio-temporal receptive field properties of neurons in LGN with the development of Wistar rats were evaluated.ResultsThe differences between band-pass and low-pass distribution of temporal frequency or spatial frequency of rat LGN were not statistically significant (x2 =0.68,0.47,P>0.05 ).The optimal temporal frequency of receptive field in rat LGN went up to the maximum value until 60 day in Wistar rats.The mean optimal temporal frequencies of neurons in the four different age groups were ( 2.5 ± 1.3 ),( 2.6± 1.2),(2.6± 1.1 ) and ( 3.6± 1.1 ) Hz with significant differences among the 4 groups (F=4.53,P<0.05 ),and those in the 14-16 day group,20-22 day group,27- 30 day group were significantly lower than in the 60 days group ( q =4.43,4.10,4.03,P < 0.05 ).No significant differences were seen among the 14-16 day group,20-22 day group and 27-30 day group ( P>0.05 ).The optimal spatial frequency values in the four groups were ( 0.04 ± 0.04 ),( 0.04 ± 0.03 ),( 0.05 ± 0.03 ),( 0.05 ± 0.04 ) cpd,respectively without statistical difference among them ( F =0.58,P > 0.05 ).The temporal and spatial bandwidth values in the various age groups were not statistically significant among the four groups ( F =0.37,1.22,P>0.05). Conclusions The development of temporal and spatial frequency characteristics of the rat LGN receptive field may be related to its functional visual pathway.