Objective To evaluate the changes in the expression of DJ-1 protein during myocardial ischemia-reperfusion (I/R) in diabetic rats.Methods Fifty male Sprague-Dawley rats,weighing 220-280 g,were used in this study.Type 1 diabetes mellitus was induced by intraperitoneal streptozotocin 65 mg/kg and confirmed by fasting blood glucose ＞ 16.7 mmol/L.Forty animals with type 1 diabetes mellitus were randomly divided into 3 groups:diabetes group (group D,n =10),diabetic sham operation group (group DS,n =15) and diabetic I/R group (group DIR,n =15).Another 10 non-diabetic rats in which citrate buffer 6 ml/kg was injected intraperitoneally were served as control group (group C).Myocardial I/R was produced by occlusion of the anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in group I/R.At 120 min of reperfusion,5 rats were sacrificed and myocardial specimens were c(on)tained for determination of infarct size in groups DS and DIR,and 10 rats were sacrificed and myocardial specimens were obtained for microscopic examination and for determination of cell apoptosis,malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and expression of DJ-1 and phosphatase and tensin homologue (PTEN) protein.Apoptotic index (AI) was calculated.Linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein was analyzed.Results Compared with group DS,the myocardial infract size was significantly increased in group DIR (P ＜ 0.05).Compared with group C,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in groups D,DS and DIR (P ＜ 0.05).Compared with groups D and DS,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in group DIR (P ＜ 0.05).There was no significant difference in the parameters mentioned above between groups D and DS (P ＞ 0.05).There was linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein and the correlation coefficients (r) were-0.734,0.593,-0.818,and-0.812 in turn.Conclusion Down-regulation of DJ-1 protein expression is involved in myocardial I/R injury in diabetic rats via decreasing anti-oxidative stress responses and upregulating PTEN protein expression.

Objective To investigate the effects of Dihuangyinzi(DHYZ) on behaviors and RAGE/p38 pathway in APP/PS1 mice.MethodTwenty APP/PS1 dementia mice were randomly divided into model group(n=10) and Chinese medicine group(n=10).The blank group was C57 BL/6 J normal mouse(n=10).The mice in Chinese medicine group were intragastric administration with DHYZ (9.75 g·kg-1·d-1).The mice in model group and blank group were treated with distilled water.After 30 days,the abilities of learning and memory of mice were detected by Morris water maze.The expression of amyloid-beta1-42(Aβ1-42) in the hippocampus and cortex was detected by immunohistochemistry.Reactive oxygen species of brain tissue were detected by DCFH-DA Methods in the brain of APP/PS1 mice.Gene expression level of receptor for advanced glycation end products(RAGE) was measured by real-time polymerase chain reaction (RT-PCR) in the cortex and hippocampus of APP/PS1 mice.The expression of phospho-mitogen-activated protein kinases (p38) was analyzed with Western blot and immunofluorescence analysis in the cortex and hippocampus of APP/PS1 mice.Results Behavioral Results showed that DHYZ significantly increased the distance((23.088±7.083)cm) and residence time((1.961±1.230)s)of effective area in Morris water maze on the fifth day(P<0.05,P<0.01)and remarkably increased the number of effective area crossings((1.607±0.405) times) and plats((0.893±0.283) times) in Morris water maze on the fifth day(P<0.01,P<0.05).DHYZ also significantly reduced the intracelluar ROS level(122.611±7.630) in the brain(P<0.01),and DHYZ could depress the expression of RAGE(1.467±0.081,7.983±0.136) and phosphorylation of p38 (0.376±0.026,0.538±0.016)in the cortex and hippocampus of APP/PS1 mice(P<0.01,P<0.05).Conclusions The Results demonstrate that DHYZ can partly improve memory impairment of APP/PS1 mice by the inhibition of RAGE/p38 pathway.

Objective To investigate the effects of Dihuang Yinzi (DY) on the receptor for advanced glycation end-products(RAGE)/reactive oxygen species(ROS)/apoptosis pathway in SH-SY5Y cells induced by amyloid-beta1-42 (Aβ1-42) oligomer. Methods Firstly, we adopted methyl thiazolyl tetrazolium(MTT) method to detect the cell vitality in fetal bovine serum (FBS) group, blank serum group, and low-, middle- and high- dose DY-containing serum groups, so as to confirm the optimal concentration and treatment time of DY-containing serum. Secondly, we applied MTT method to detect cell vitality and applied Annexin V/propidium iodide (PI) staining method to observe the apoptosis of SH-SY5Y cells treated with 0~20 μmol/L Aβ1-42 for 24 and 48 h, so as toconfirm the optimal concentration and treatment time of Aβ1-42 for establishing Alzheimer's disease (AD) model in vitro. Thirdly, MTT method was used for the detection of cell vitality, and Annexin V/PI staining method was used for detection of the apoptosis of SH-SY5Y cells in blank serum group, model group, western medicine control group and low-, middle-and high-dose DY-containing serum groups, and Dihydroethidium (DHE) method was used for the assay of ROS contents, so as to observe the effect of DY on the recovery of injured SH-SY5Y cells induced by Aβ1-42. Finally, we applied Western blot method to detect the expression level of RAGE in SH-SY5Y cells of blank group, model group and DY-containing serum group; after Aβ1-42-induced SH-SY5Y cells were transfected with RAGE gene, we adopted DHE staining method and Annexin V/PI staining method to detect ROS content and cell apoptotic rate in all of the above groups, so as to observe the effect of DY on SH-SY5Y cell apoptosis and RAGE expression. Results The cell vitalities were increased in low- and middle-dose DY-containing serum groups at 24 h (P < 0.05 or P < 0.01 compared with that in the blank serum group). The conditions for the establishment of AD model in vitro were as follows: the optimal concentration of Aβ1-42 was 5μmol/L, and the treatment time was 24 h. The cell vitalities were significantly enhanced, the cell apoptotic rate and ROS content were significantly lowered in Aβ1-42-induced SH-SY5Y cells of the medication groups(P <0.05 or P < 0.01 compared with those in the model group) , and the cell vitality was the highest and the cell apoptotic rate was the lowest in the middle-dose DY-containing serum group. The RAGE expression level was decreased in Aβ1-42-induced SH-SY5Y cells of the middle-dose DY-containing serum group(P < 0.05 compared with that in the model group) . ROS content and cell apoptotic rate were decreased in Aβ1-42-induced SH-SY5Y cells transfected with RAGE gene in the middle-dose DY-containing serum group (P<0.01). Conclusion DY may play an anti-oxidative role through inhibiting the production of ROS and cell apoptosis, thus to suppress RAGE protein and to achieve the preventive and therapeutic effect for AD.

OBJECTIVETo explore the influence of chromosome polymorphisms on the outcome of in vitro fertilization embryo transfer (IVF-ET).

METHODSPatients who completed the first cycle of in vitro fertilization fresh embryo transfer were retrospective studied. Patients with the chromosome polymorphisms were classified as to the study group (200 treatment cycles), all patients with normal chromosomes at the same period were classified as the control group (4777 treatment cycles).

RESULTSNo significant difference was found between the chromosome polymorphisms and the control groups in terms of clinical pregnancy rate (44.50% vs. 39.85%, P=0.750), early abortion rate (15.73% vs. 10.79%, P=0.163) and live birth rate per cycle (34.5% vs. 30.73%, P=0.437) except for fertilization rate (60.94% vs. 64.08%, P=0.001), cleavage rate (95.01% vs. 97.09%, P=0.000) and good quality embryo rate (53.8% vs. 58.2%, P=0.001).

CONCLUSIONChromosomal polymorphisms appeared to have no adverse influence on the outcome of IVF-ET treatment.

Adult ; Chromosome Aberrations ; Chromosomes ; genetics ; Embryo Transfer ; methods ; Female ; Fertilization in Vitro ; methods ; Humans ; Male ; Polymorphism, Genetic ; genetics ; Pregnancy ; Pregnancy Outcome

To investigate the changes of endocrine, cyclic nucleotide and immune systems in subjects of yang deficiency constitution, and to explore the relationship among characteristics and causes of yang deficiency constitution, the physiological and biochemical parameters.

Objective To investigate the effect of schisandrin (SCH) treatment on learning and memory abilities and their mechanism in APP/PS1 dual transgenic dementia mice,and explore the effect of Chinese medicine on Alzheimer's disease (AD).Methods Thirty-five APP/PS1 dementia mouse models were randomly assigned into APP/PS1 model group (n=17) and APP/PS1+SCH group (n=18);another 10 male C57BL/6J mice were chosen as blank control group.The mice in the APP/PS1+SCH group were given intragastric administration of SCH at 2.6 mg/(kg· d) for 30 d;the mice in the APP/PS1 model group and blank control group were treated with distilled water for 30 d.The learning and memory abilities of these APP/PS1 mice (n=7) were detected by Morris water maze.Mice from the three groups were sacrificed;Nissl staining was used to observe Nissl bodies of neurons in brain tissues;real-time fluorescence quantitative PCR (qPCR) was used to detect the mRNA content of terminal glycosylationend products receptor (RAGE) in brain tissues;Western blotting was used to detect the expressions of RAGE and phosphorylated P38 mitogen-activated protein kinase (p-p38) in brain tissues.Results (1) The results of water maze space exploration experiment showed that the times of crossing the platform area in the three groups were statistically significant (P＜0.05);as compared with the APP/PS1 modelgroup,the times of crossing the platform area in the APP/PS1+SCH group were significantly increased (P＜0.05).(2) Nissl staining results showed that the contents of Nissl bodies in the hippocampal CA1 area and cortical neurons of the APP/PS 1 model group were significantly reduced,with light staining and cell body atrophy;the lesions in mice of the APP/PS1+SCH group were less severe than those of APP/PS1 model group,some neurons were atrophic,and the content of the neuronal nileite bodies in the hippocampal CA1 region was relatively abundant.(3) The qPCR results showed that there were statistically significant differences in RAGE mRNA expression levels in the cortex and hippocampus of the three groups (P＜0.05);as compared with the APP/PS1 model group,the APP/PS1+SCH group had significantly reduced RAGE mRNA expression in the hippocampal area (P＜0.05).(4) Western blotting results showed that RAGE and p-p38 protein expression levels in two parts of mice of APP/PS1+SCH group were significantly reduced as compared with those in the APP/PS1 model group (P＜0.05).Conclusion SCH may improve the functional status of hippocampal and cortical neurons and improve the spatial exploratory memory ability of APP/PS1 mice by down regulating the RAGE and P38 expressions.

Objective:To evaluate effect of diammonium glycyrrhizinate(DG)on lipopolysaccharide(LPS)-induced TLR4/NF-κB inflammatory signaling pathway in BV2 microglial cells and to explore its potential regulatory mechanisms on ameliorating neu-roinflammation.Methods:BV2 microglia injury model was induced by LPS and treated with 20 and 60 μmol/L DG.MTS was used to determine vitality of cells.NO level secreted by cells was detected by Griess.Inflammatory factors TNF-α,IL-6,and IL-1β levels in cell supernatant were measured by ELISA.TNF-α,IL-6,IL-1β,TLR4 and MyD88 mRNA levels were detected by RT-qPCR.Western blot was used to determine expressions of TLR4,MyD88,NF-κB,p-NF-κB,IκB-α and p-IκB-α proteins of cells.Results:Compared with control group,LPS-treated BV2 microglia had significantly lower viability(P<0.05),significantly higher NO secretion(P<0.05),significantly up-regulation levels of inflammatory factors TNF-α,IL-6 and IL-1β(P<0.05),and significantly higher mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),TLR4,MyD88,NF-κB,p-NF-κB,IκB-α,and p-IκB-α protein expressions were significantly increased(P<0.05).Compared with LPS group,BV2 microglia intervened with different concentrations of DG had significantly higher viability(P<0.05),significantly lower NO secretion(P<0.05),significantly lower TNF-α,IL-6 and IL-1β inflammatory factors levels(P<0.05),significantly lower mRNA levels of TNF-α,IL-6,IL-1β,TLR4 and MyD88(P<0.05),and TLR4,MyD88,p-NF-κB and p-IκB-α protein expressions were significantly reduced(P<0.05).Conclusion:DG can inhibit LPS-induced neuroinflammatory responses in microglia,whose underlying mechanism is suppression of TLR4/NF-κB signaling pathway.

Objective To increase the awareness of ectopic ACTH syndrome in patients with thymus carcinoid. Methods Clinical manifestations, especially atypical presentation of ectopic ACTH syndrome, laboratory findings and imaging data (chest X-ray and CT, etc.) in 6 patients were analysed. Results In all 6 patients, the thymus tumors removed were pathologically proved to be thymus carcinoid. Conclusion Patients having typical manifestations of Cushing′s syndrome with hypokalemia, edema, proteinuria and equivocal result of dexamethasone inhibition test, should be considered as ACTH syndrome. And thymus carcinoid seems to be the cause of the disease.

OBJECTIVE: To determine the sperm-zona pellucida (ZP) binding and ZP-induced acrosome reaction in patients with unexplained infertility, and to discuss the relationship between ZP-induced acrosome reaction and fertilization rate. METHODS: We compared the fertilization rate and good embryo rate in patients with unexplained infertility after fertilization in 2 ways. Based on the causes of infertility, patients were divided into an unexplained infertility group (Group A) and a pure female tubal factor group (Group B). Oocytes which were obtained by super ovulation from 25 patients with unexplained infertility were randomly divided into 2 groups with conventional in vitro fertilization (IVF) (Group A1) and intracytoplasmic sperm injection (ICSI) fertilization (Group A2). The pure female tubal factor group (Group B) had conventional IVF. We conducted sperm-ZP binding and ZP-induced acrosome reaction experiments with 2 groups of men's sperms separately. We compared the number of sperm-egg binding and ZP-induced acrosome reaction rate and discussed the relationship between the ZP-induced acrosome reaction and fertilization rate, and also the fertilization rate, good embryo rate and pregnancy rate in patients with unexplained infertility after fertilization in 2 ways. RESULTS: The average number of sperm-egg binding (78.29 ± 16.31) and the ZP-induced acrosome reaction rate (55.87 ± 27.69) % in Group A were lower than those of Group B [94.63 ± 6.72, (82.53 ± 17.99)%]. The difference between the average number of sperm-egg binding and the ZP-induced acrosome reaction was significant (P <0.01). The fertilization rate of Group A1 was significantly lower than that of Group B and Group A2 (P <0.01). But there was no significant difference in the good embryo rate among the 3 groups. There was no significant difference between Group A2 and B in fertilization rate and good embryo rate (P <0.05). There was no significant difference in pregnancy rate between Group A and B (P <0.05). Fertilization rate and the rate of acrosome reaction had marked positive correlation with statistical significance (r =0.932, P <0.01). CONCLUSION ZP binding and ZP-induced acrosome reaction are very important experiments in sperm function test for patients with unexplained infertility. It can not only effectively avoid no embryo transferring due to complete failure of fertilization but also get a desirable outcome of pregnancy using half-ICSI fertilization in patients with unexplained infertility.
Acrosome Reaction ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility ; etiology ; therapy ; Male ; Oocytes ; physiology ; Ovulation Induction ; Sperm Injections, Intracytoplasmic ; Sperm-Ovum Interactions ; physiology ; Treatment Outcome ; Zona Pellucida ; physiology

Objective To investigate the effect ofDihuangyinzi (DHYZ)-medicated serum on receptor for advanced glycation end product (RAGE)/p38 miotgen-activated protein kinase (MAPK) /nuclear factor (NF)-κB pathway in SH-SY5Y cells induced by Aβ1-42.Methods Male SD rats were randomly divided into normal control group and experimental group (n=20);natural sera medium and DHYZ sera medium were prepared.(1) SH-SY5Y cells were divided into control group,model group and DHYZ treatment group;natural sera medium,natural sera medium+Aβ1-42 oligomer,and DHYZ sera medium+Aβ1-42 oligomer were given to the cells,respectively.Westem blotting was used to detect the protein expressions of NF-κB p65,p38 and phosphorylate (p)-p38.(2) SH-SY5Y cells were given DHYZ sera medium+Aβ1-42 oligomer treatment,and at different time points of Aβ1-42 oligomer treatment (15 min,30 min,60 min,12 h,24 h,48 h and 72 h),Western blotting was used to detect the protein expressions of p38 and p-p38.(3) SH-SY5Y cells were divided into 6 groups:mock-transfected RAGE blank group,transfected RAGE blank group,mock-transfected RAGE model group,transfected RAGE model group,mock-transfected RAGE herb group and transfected RAGE herb group;herb groups were given DHYZ-medicated serum;inflammatory factors,interleukin (IL)-1β,IL-6,and tumor necrosis factor (TNF)-a,were measured by ELISA and cytometric bead array.Results (1) As compared with model group,DHYZ treatment group had significantly decreased NF-κB p65 and p-p38/p38 protein expression.(2) The p-p38 protein expression began to increase 30 min after Aβ1-42 treatment,reached to its peak level 24 h after Aβ1-42 treatment,and began to decrease 48 h after Aβ1-42 treatment.(3) The IL-1β,IL-6 and TNF-α levels were increased significantly in the transfected RAGE model group as compared with those in the mock-transfected RAGE model group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were increased significantly in the transfected RAGE herb group as compared with those in the mock-transfected RAGE herb group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were decreased significantly in the mock-transfected RAGE herb group as compared with those in the mock-transfected RAGE model group (P＜0.05);the IL-1β,IL-6 and TNF-α levels were decreased significantly in the transfected RAGE herb group as compared with those in the transfected RAGE model group (P＜0.05).Conclusion DHYZ-medicated serum could inhibit the RAGE-p38 pathway and improve the inflammatory reaction in Aβ1-42-induced SH-SY5Ycells transfected with RAGE gene to protect the SH-SY5Y cells.