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ObjectiveThe mechanism of histone phosphorylation modification in oocyte meiosis is less studied. This study is designed to investigate the pattern of histone H3 phophorylation and regulation of maturation process in the porcine oocytes.MethodsThe histone H3Ser10 (H3S10) phosphorylation expression was examined on the porcine oocyte meiotic process. The porcine cumulus oocyte complexes (COCs) were divided into four groups, one group was cultured as control group, and the other 3 groups were supplemented with 5, 10, and 30 μmol/L ZM447439, and cultured in vitro for 27 h, respectively, 5, 10, and 30 μmol/L ZM4474349 treatment group. The proportion of each meiotic stage was counted. The phosphorylation pattern of histone H3S10 and the expression level of protein kinase Aurora B were detected at the porcine oocytes.ResultsCompared with histone H3S10 phosphorylation level of oocyte GVBD phase, the MI and AI phases were significantly increased (P<0.05), and H3S10 phosphorylation level of AI phase was remarkedly higher than that of MII phase (P<0.05). Compared with the control group, the proportion of oocytes at the GVBD phase in the 10 and 30 μmol/L ZM4447439 treatment group [(32.14±0.51)%, (95.34±0.59)%]was higher than that of the control group [(2.56±0.03)%, P<0.05], the proportion of oocytes at the MI phase [(66.88±0.13)%, (4.66±0.04)%] significantly decreased than that of the control group [(87.42±0.14)%, P<0.05], and the proportion of oocytes at the AI stage [(1.01±0.03)%, (0.000±0.00)%] significantly decreased compared with the control group[(10.02 ± 0.21)%, P<0.05]. Compared with the control group (0), oocytes H3S10 dephosphorylation modification ratio in the 10 μmol/L and 30 μmol/L ZM4474349 treatment group [(35.2±0.39)%, (95.4±0.65)%]significantly increased (P<0.05). Compared with the control group, the relative expression level of Aurora B in the 10 and 30 μmol/L ZM4447439 treatment group was significantly reduced (P<0.05).ConclusionHstone H3S10 phosphorylation plays arolein the maturation of mammalian oocytes. AuroraB kinase inhibitors (ZM447439) treatment can reduce H3S10 phosphorylation and Aurora B expression level and lead to oocytesmaturation disorder.
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Objective@#To investigate the role of pH2AX in the reversibility of mouse testicular reproductive function impaired by single heat stress.@*METHODS@#Twenty-four C57 male mice were randomly divided into heat stress and control groups and immersed in water at 43℃ and 25℃, respectively, for 15 minutes. At 1, 7, and 14 days of heat exposure, all the mice were sacrificed and their testis tissues collected for determining the apoptosis of the germ cells by TUNEL and measuring the expression level of the pH2AX protein by immunohistochemistry and Western blot.@*RESULTS@#The highest percentage of apoptotic cells were found in the seminiferous tubules of the mice in the heat stress group on the 1st day of the exposure and almost no apoptosis was observed at 7 and 14 days. The pH2AX protein was expressed in the nuclei of the basement membrane of adjacent seminiferous tubules. Compared with the control group, the expression of pH2AX was significantly increased on the 1st day of exposure (0.47 ± 0.02 vs 1.61 ± 0.04, P <0.01), then decreased at 7 days (0.85 ± 0.03) in comparison with that on the 1st day (P <0.01), and again elevated at 14 days (1.72 ± 0.02) as compared with either those at 1 and 7 days (P <0.01) or that of the control (P <0.01).@*CONCLUSIONS@#Heat stress causes dynamic changes of the pH2AX expression in the testis of the mouse, which are associated with heat stress-induced proliferation and division of the testicular spermatogenic cells.
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Animals , Male , Mice , Apoptosis , Blotting, Western , Heat Stress Disorders , Histones , Metabolism , Hot Temperature , Immunohistochemistry , In Situ Nick-End Labeling , Mice, Inbred C57BL , Random Allocation , Seminiferous Tubules , Cell Biology , Spermatozoa , Cell Biology , Metabolism , Testis , Time FactorsABSTRACT
Objective:To investigate the effects of the recent upper respiratory tract infections (URI) on the incidence of perioperative respiratory adverse events in children scheduled to undergo general anesthesia and elective surgery.Methods:In the study,232 children undergoing general anesthesia with laryngeal mask airway (LMA) for elective ophthalmic surgeries at Peking University First Hospital,Beijing,China,from Nov.1,2015 to May 10,2016 were enrolled.On the day of the surgery,the parents of the children were preoperatively asked to fill out a questionnaire regarding the baseline characteristics and medical history of the children,including gender,age,height,weight,history of URI within the last 2 weeks before anesthesia,history of premature,long-term passive smoking exposure,habitual sleep snoring,and history of asthma.In addition,all adverse respiratory events throughout the perioperative periods (oxygen desaturation,cough,copious secretions,laryngospasm and bronchospasm) as well as perioperative variables (number of attempts to insert the LMA successfully,anesthesia duration and so on) were recorded.Multivariate Logistic regression analysis was applied to identify independent risk factors of perioperative respiratory adverse events.Results:Among the 232 children included in the study,28.0％ (65/232) presented with a history of a recent URI within the last 2 weeks before anesthesia.The presence of the recent URI increased the incidence of oxygen desaturation (23.1％ vs.12.0％,P =0.034),copious secretions (15.4％ vs.6.6％,P =0.036) and any of all the adverse respiratory events (32.3％ vs.18.6％,P =0.024).Multivariate Logistic regression analysis identified two independent risk factors of perioperative adverse respiratory events:a history of URI within the last 2 weeks before general anesthesia (OR =2.021,95％ CI:1.023-3.994,P =0.043) and habitual sleep snoring (OR =3.660,95％ CI:1.517-8.832,P =0.004).Conclusion:A history of a recent URI within 2 weeks before general anesthesia was associated with a higher incidence of oxygen desaturation,copious secretions and the overall respiratory adverse events.For the children with recent URI,we recommend the general anesthesia and elective surgery should be postponed for at least 2 weeks after the URI.
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Objective To analyze the treatment outcome of MDR -TB (Multidrug -resistant Tuberculosis)patients and to provide suggestions for standard management of MDR -TB patients.Methods Information of MDR -TB patients enrolled in Global Fund Multi -drug Resistant Tuberculosis Project between July 2009 and June 201 0 were collected and analyzed. Results Among the 68 treated patients,44 patients were cured and 1 patient finished the course.The cure rate was 66.1 8%.While 5 patients died,5 patients quitted the treatment as a result of side effects.Significant differences on the cure rate were observed between patients who had relapsed and failure of treatment (P <0.05).Conclusion The cure rate of MDR -TB needs to be improved.It is necessary to promote standard treatment and management and to improve the patient compliance.
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Objective To evaluate the prevalence of depression,anxiety and suicide behavior in patients suffering from tuberculosis in Hangzhou and to explore their relationship with medication adherence. Methods Demographic characteristics,self-rating anxiety scale (SAS),the center for epidemiological studies -depression (CESD),social support rating scale (SSRS),suicide behavior information and the morisky medication adherence scale (MMAS)were investigated in 973 tuberculosis patients who were selected by systematic random sampling.Results The means of SAS and CESD were 39.71 ±8.30 and 14.16 ±10.77 respectively,which were both higher than the norms(P<0.01).Totally 102 (10.48%)patients had anxiety and 333 (34.22%)were depressed.Out of 973 patients,60 (6.17%)reported suicide ideation after tuberculosis diagnosis.The prevalence of non -adherence was 20.55%,which was defined with MMAS score above one and more.The non -adherence group had higher anxiety,depression and suicide ideation prevalence than the adherence group (15.50%vs.9.18%,46.50%vs.30.66%,11.00%vs.4.92%respectively,P<0.01).The mean score of SSRS,subjective support,objective support and utilization of support in the non-adherence group were 30.71 ±5.15,4.61 ±2.07,19.74 ±4.55 and 6.34 ±1.93 respectively,which were 34.06 ±7.39,6.62 ± 2.27,20.67 ±5.27 and 6.77 ±2.23 in the adherence group respectively.SSRS and its three dimension scores were significantly lower in the non-adherence group than that in the adherence group (P<0.01).Conclusion These findings show a quite serious situation of psychological problems of tuberculosis patients in Hangzhou and suggest psychological intervention should be included in adherence intervention.
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BackgroundOur previous study determined that tetrandrine (Tet) has an inhibitory effect on the proliferation of human Tenon capsule fibroblasts ( TCFs ) in vitro,but its mechanism is poorly understood.ObjectiveThis study was to investigate the effect mechanism of Tet on human TCFs.MethodsHuman TCFs were isolated and cultured from scleral tissue of donor using explant technique.The cells were identified by vimentin antibody staining and morphology.The third generation of cells were seeded in the culture plate at the density of 1 × 105 cells/ml.Twenty-four hours after inoculation,the Tet of 1 × 10-5 mol/L was added in the well of culture plate,and the cells cultured only in 1640 medium served as the control group.The apoptosis of the cells was assessed by TUNEL,and the expressions of bax,bel-2,transforming growth factor-β2 (TGF-β2 ) in TCFs were detected using immunochemistry.Results The cultured cells showed the features of the fibroblasts in shape with the positive response for vimentin.A number of TUNEL positive cells were seen in Tet group and no TUNEL positive response was found in control group.The expression levels (A value) of bax,bcl-2 and TGF-~ protein in TCFs were 0.577 ± 0.009,0.430±0.012 and 0.341 ±0.017 in Tet group,and those in control group were 0.320±0.015,0.819±0.021 and 0.624±0.014 respectively,showing statistically significant differences between two groups( t =33.277,-35.356,-28.093,P＜0.01 ).Conclusions Tet suppresses the proliferation of human TCFs through up-regulating the expression of bax and down-regulating the expressions of bcl-2 and TGF-β2 in vitro.
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Objective: To investigate dihydroartemisinin(DHA)-induced apoptosis of human prostate cancer cells and the underlying molecular mechanism. Methods: Nude mice were implanted with human prostate cancer PC-3 cells to establish tumor-bearing mouse model. Twenty models mice were evenly randomized into four groups: control group, solvent group (DMSO), large dose DHA (200 μ/kg) group and low dose DHA (100 μ/kg) group. The implanted tumors were observed on day 13 after drug administration. Morphological changes of PC-3 cells were observed by transmission electron microscope(TEM). The protein expression of apoptosis associated gene Bcl-2 and Bax were examined by immunohistochemical method. Apoptosis was detected by TUNEL assay. Results: TEM examination revealed scattered apoptotic cells and apoptotic bodies in tumor tissues of mice in the DHA groups; immunohistochemical examination revealed that the protein expression of apoptosis-associated gene Bcl-2 was decreased and that of Bax was increased in DHA groups (P < 0. 05). TUNEL staining revealed that the rate of cell apoptosis increased significantly in DHA groups(P < 0. 05). Conclusion: It is demonstrated that DHA has strong effect in inducing apoptosis in human prostate cancer cell line PC-3 in vivo, which might be related to the down-regulation of Bcl-2 and up-regulation of Bax.
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Objective To study the prevalence and risk factors of human immunodeficiency virus (HIV) among men who have sex with men (MSM) in Beijing. Methods A sample of 550 MSM was recruited between March and June in 2008 in Beijing. Interviewer-administered interviews were conducted to collect information on demographics and behaviors. Blood samples were collected to test for HIV,syphilis,HBsAg and HCV infections. Chi-square test and logistic regression model were used to identify risk factors associated with HIV infection. Results Of the 550 eligible MSM surveyed,HIV prevalence appeared to be 4.5% (25/550). Data from multivariate logistic regression analysis demonstrated that factors as reporting unprotected anal intercourse with causal male sexual partners in the last month (X~2=11.381,P=0.001),rectal douching after homosexual anal intercourse in the past three months (X~2=5.326,P=0.021),feeling sad in the last month (X~2=8.809,P=0.003) were significantly associated with HIV infection. Conclusion Health education on safer sex behavior and mental health care should be taken to hinder the speed of HIV transmission among MSM.
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Ghrelin, an endogenous ligand for the growth hormone secretagogue (GHS) receptor, stimulates feeding and increases body weight. The primary action site of ghrelin has been reported to be the neuropeptide Y (NPY)/agouti-related peptide (AgRP) neurons in the hypothalamic arcuate nucleus (ARC). In addition to the hypothalamus, the caudal brainstem also appears to be an important mediator for the orexigenic activity of ghrelin. However, it is not clear whether ghrelin applied directly to the caudal brainstem activates forebrain structures. The aim of this study was to determine whether recruitment of forebrain structures was required for hyperphagic responses stimulated by ghrelin delivery within the caudal brainstem. In our experiment, all rats were surgically implanted with indwelling cannulas in the dorsal vagal complex (DVC), and ghrelin (20 pmol in 0.5 μL) was delivered to the DVC. After the injection, the orexigenic response to ghrelin was recorded by Feeding and Activity Analyser, and NPY/AgRP mRNA expressions in rat hypothalamus were detected by real-time PCR. In addition, the NPY immunoreactive neurons in the ARC were assayed by immunohistochemistry. The results showed that ghrelin significantly increased cumulative food intake at 1, 2 and 3 h after ghrelin injection, maximal response occurring at 2 h after injection. NPY/AgRP mRNA levels in ARC treated with ghrelin increased significantly compared with those in control group (injected with saline). The highest levels of NPY and AgRP mRNA were detected at 2 h after injection. The total number and mean optical density of NPY-positive neurons increased in ghrelin treated rats compared with those in control group. Consistently, ghrelin's effect was most pronounced at 2 h after injection. Taken together, we conclude that the activation of NPY/AgRP neurons in the ARC is involved in the mediation of the hyperphagic response to brainstem ghrelin administration in neurologically intact rats.
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Animals , Male , Rats , Agouti-Related Protein , Genetics , Metabolism , Arcuate Nucleus of Hypothalamus , Metabolism , Physiology , Brain Stem , Metabolism , Physiology , Feeding Behavior , Ghrelin , Pharmacology , Hyperphagia , Hypothalamus , Metabolism , Physiology , Neurons , Metabolism , Physiology , Neuropeptide Y , Genetics , Metabolism , Peptide Fragments , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To investigate the risk factors related to outcome of chronic severe hepatitis B.</p><p><b>METHODS</b>A total of 336 consecutive patients with chronic severe hepatitis B (CSHB) were analysed retrospectively. According to the outcome, objects were divided into survival group (n = 137) and death group(n = 199), then to observe the differences between them in respect to age, sex, family history, prothrombin activity (PTA), complications including ascites, infection, electrolyte disturbance, upper gastrointestinal bleeding, hepatic encephalopathy, hepatorenal syndrome and the corresponding quantity of complications in each individual, antivirus therapy, artificial liver support system (ALSS) therapy, and alprostadil therapy. Finally, risk factors related to prognosis were selected by stepwise Logistic regression analyse.</p><p><b>RESULTS</b>In univariate analyse, significant differences between the two groups were found related to age, PTA, complications and its quantity (P < 0.01 for all), and antivirus therapy (P < 0.05) rather than sex, family history and treatment of ALSS or alprostadil. Logistic regression revealed that risk factors comprised of PTA and quantity of complications, antivirus therapy was the only protective factor.</p><p><b>CONCLUSION</b>A numbers of factors including age, PTA, complications and its quantity, and antivirus therapy affect the prognosis of CSHB, among which, antivirus therapy can reduce the death rate.</p>
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Adult , Female , Humans , Male , Hepatitis B, Chronic , Diagnosis , Logistic Models , Prognosis , Retrospective Studies , Risk Factors , Treatment OutcomeABSTRACT
<p><b>AIM</b>To investigate the mechanism through which insulin affect the learning and memory abilities of the Alzheimer's disease-like rats.</p><p><b>METHODS</b>Okadaic acid (OA) was injected into the CA1 region of the rat hippocampus and the insulin was injected into the lateral cerebral ventricle of the rats. The learning and memory abilities of the rats were assessed through Morriswater maze behavioral test, and the expressions of nicotinic acetylcholine receptors and GFAP were observed by Westem blotting and immunohistochemistry, respectively.</p><p><b>RESULTS</b>Compared with the control rats, the abilities of learning and memory were lowered significantly (P < 0.01) and the expressions of the nicotinic acetylcholine receptors were decreased and the GFAP positive astrocytes were increased greatly in the model rats (P < 0.05). In the rats injected with insulin, it was found that their learning and memory abilities were improved significantly (P < 0.01) and that the expression of the nicotinic acetylcholine receptors were increased and GFAP positive astrocytes were decreased obviously (P < 0.05), as compared with the model rats.</p><p><b>CONCLUSION</b>Insulin is able to enhance the learning and memory abilities of the Alzheimer's disease-like rats, possibly by improving the function of the acetylcholine system and decreasing the astrocytes proliferation in the brain.</p>
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Animals , Male , Rats , Alzheimer Disease , Glial Fibrillary Acidic Protein , Metabolism , Insulin , Pharmacology , Learning , Memory , Random Allocation , Rats, Wistar , Receptors, Nicotinic , MetabolismABSTRACT
The reform and practice on teaching approaches of microbiology integrated case-based study,heuristic teaching,interaction teaching,comparison expression teaching and inductive learning was explored,which was contraposed the education aims of independent college and student's trait,and combined to the fact of University of Electronic Science and Technology of China Zhongshan Institute.The result of practice showed that student enthusiasm for study was mobilized,and better results in teaching were got by applica-tion of diversiform teaching approaches.
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Aim To investgate the mechnism through which ginkgolides affect learning and memory capabilities of the Alzheimers disease-like rats. Methods Okadaic acid(OA)was injected into the CA1 region of the rat hippocampus and the rats were gavaged with ginkgolides. The learning and memory abilities of the rats were assessed through Morris water maze behavioral test, and the expressions of nicotinic acetylcholine receptors and ChAT were observed by Western blotting and immunohistochemistry, respectively.Results Compared with the control rats, the capabilities of learning and memory were lowered significantly(P