Effects of echinomycin on endothelin-2 expression and ovulation in immature rats primed with gonadotropins

Echinomycin is a small-molecule inhibitor of hypoxia-inducible factor-1 DNA-binding activity, which plays a crucial role in ovarian ovulation in mammalians. The present study was designed to test the hypothesis that hypoxia-inducible factor (HIF)-1alpha-mediated endothelin (ET)-2 expressions contributed to ovarian ovulation in response to human chorionic gonadotropin (hCG) during gonadotropin-induced superuvulation. By real-time RT-PCR analysis, ET-2 mRNA level was found to significantly decrease in the ovaries after echinomycin treatment, while HIF-1alpha mRNA and protein expression was not obviously changed. Further analysis also showed that these changes of ET-2 mRNA were consistent with HIF-1 activity in the ovaires, which is similar with HIF-1alpha and ET-2 expression in the granulosa cells with gonadotropin and echinomycin treatments. The results of HIF-1alpha and ET-2 expression in the granulosa cells transfected with cis-element oligodeoxynucleotide (dsODN) under gonadotropin treatment further indicated HIF-1alpha directly mediated the transcriptional activation of ET-2 during gonadotropin-induced superuvulation. Taken together, these results demonstrated that HIF-1alpha-mediated ET-2 transcriptional activation is one of the important mechanisms regulating gonadotropin-induced mammalian ovulatory precess in vivo.

A magnetic resonance imaging study on the morphological variation of the lateral ventricles in Alzheimer Disease patients / 中华行为医学与脑科学杂志

ObjectiveTo investigate the variation of the lateral ventricles volume and the angle of lateral ventricles in AD patients.Methods20 AD patients and 20 healthy persons as the control were enrolled in this study and performed with craniocerebral scanning using 3.0T MRI,respectively.Quantitative measurements were made with stereology,including the volume of lateral ventricles,the relation of the variation of the angle and illness degree.ResultsThere were significant differences between normal controls and patients with AD in the whole volume of lateral ventricles,the left whole volume of lateral ventricles ( AD (71.53 ± 3.25 ) cm3,healthy person ( 65.39± 4.59) cm3,t =- 4.88,P ＜ 0.00),the left whole volume of lateral ventricles ( AD ( 36.14 ± 1.75 ) cm3,healthy person(31.88 ±3.24)cm3,t =- 5.16,P ＜ 0.00),the right whole volume of lateral ventricles( AD(35.42 ±2.89 ) cm3,healthy person ( 33.22 ± 2.48 ) cm3,t =- 2.58,P ＜ 0.01 ).There were significant differences between normal controls and patients with AD in left anterior horn and right anterior horn of the lateral ventricles.There were significant differences between normal controls and patients with AD in width of the left temporal horn and the right temporal horn of the lateral ventricles( AD the width of the left temporal horn(5.28 ± 1.72)mm,healthy person(2.13 ±0.97)mm,t=8.74,P＜0.05,AD the width of right temporal horn(5.13 ± 1.83) mm,healthy person (2.08 ± 1.01 )mm,t =8.01,P ＜ 0.05 ).There was significant correlation between the width of the left temporal horn,the right temporal horn and the scores of MMSE and FOM.ConclusionThe changes of the lateral ventricular volume and point of presence of patients with Alzheimer's disease were characteristically which may be reference in evaluating of Alzheimer's disease.

Cloning and prokaryotic expression of a glutathione peroxidase from tick Hemaphysalis longicornis / 中国人兽共患病学报

Ticks transmit various diseases to livestock and human beings. The researches on ticks are of great importance in medical and veterinary sciences. To express a glutathione peroxidase gene (HlGPx) from Hemaphysalis longicornis in Escherichia coli (E. coli) so as to provide basis for further studies, the gene was amplified from cDNAs of H. longicornis adult ticks by PCR prompted by information from EST library. The TGA codon encoding selenocysteine in the gene was mutated into the universal genetic code for cysteine,TGC, by site-directed mutagenesis. E. coli was transformed with the recombinant expression vector pGEX-4T-1/HlGPx'(the mutated HlGPx) and induced to express recombinant HlGPx', which was then analyzed by sodium dodecylsulphate-polyacrylamide gel electrophoresis and Western blotting. The result showed that the complete coding sequence of HlGPx was obtained successfully, the deduced polypeptide was 223 aa, with a calculated molecular mass of 25.0 kDa; the recombinant fusion protein was approximately 51 kDa, correspondent to the calculated. The antibody against glutathione S-transferase (GST) recognized the recombinant protein fused with GST in a Western blotting assay, which confirmed the result above-mentioned. In conclusion, the mutated HlGPx was expressed in E. coli successfully and it could be used for further preparation of immune serum and functional analysis.

Research progress of ovarian tissue cryopreservation / 生物医学工程学杂志

Ovarian cryopreservation can save a large number of germ cells during the cryogenic process, and can restore ovarian endocrine function and ovulation potential after thawing and transplantation, which is an ideal way to retain fertility for patients with cancer. Many factors influence the effect of ovarian cryopreservation, like cryoprotectant (CPA), frozen carrier, cortical block size and freezing procedure. An efficient and standard transplantation procedure is needed to develop for further clinical application and scientific research. In order to optimize this technology, we analyzed different factors to improve the recovery of ovarian function after freezing during ovarian cryopreservation.

Morphometric and functional alterations of amygdale and hippocampus in patients with depression: a MRI study / 中华放射学杂志

Objective To explore the morphometric and functional alterations of amygdale and hippocampus in patients with depression by anatomical and functional MRI, and try to reveal the pattern and pathogenesis of the changes in depression. Methods Sixty patients (divided equally into mild, moderate and major groups according to patient′s scores of HAMD) and 20 healthy control groups were scanned using T1WI and fMRI. The outlines of hippocampus and amygdale were drawn manually by observer and the volumes were calculated and normalized subsequently. Functional MRI was processed using SPM5 and individual activation map was got subsequently. Dunnett-t test and Pearson correlation analysis were separately used to analyze the morphometric and functional changes and the correlations between cerebral changes and clinical severity. Results The hippocampal volumes of control groups were 2296±202 left for left side and 2283±199 for right side. The hippocampal volumes of depressive patients were smaller than those of control groups, especially for the major group (left 1978±176,Dunnett-t=-10.0,P0.05,right 2210±191,Dunnett-t=-1.6,P>0.05). The amygdale′s volumes of control groups was 1762±185,the right was 1749±182, while those in patient group reduced along with the patient′s condition, i.e., the mild groups (left 1992±200,Dunnett-t=4.8,P<0.01,right 1989±191, Dunnett-t=5.0,P<0.001), the moderate groups (left 1889±192, Dunnett-t=2.8,P<0.05,right 1896±195,Dunnett-t=2.8,P<0.05), and the major groups (left 1539±178,Dunnett-t=-6.8,P<0.01,right 1543±180,Dunnett-t=-7.0,P<0.01).For fMRI study, patient group demonstrated more activation of the amygdale and hippocampus under the stimulations of negative images than controls. Furthermore, the strengthens of activation decreased along with the patient′s condition, i.e., the major ones showed the weakest activation among the patients, though it was higher than that of control group. In patient group, both the volumes and activations of hippocampus and amygdale showed significant negative correlations with HAMD scores(r=-0.80--0.90,P＜0.05). Conclusion The hippocampal volumes of depressive patients reduced, which may be due to the change of the amygdale, and the amygdale′s volumes were changed along with the patient′s condition. There were more activation in the amygdale and hippocampus of depressive patients under the stimulations of negative images.

MRI enhancement scanning features and pathology of the orthotropic transplantation nude mouse model with human pancreatic cancer / 中华胰腺病杂志

Objective To investigate the MRI imaging features, and pathologic basis of the orthotropic transplantation nude mouse model with human pancreatic cancer. Methods Adopting Siemens Magnetom Trio Tim 3.0 Tesla superconductive MRI and breast coil was used to examine 30 orthotropic transplantation nude mouse models of the human pancreatic cancer, these mouse were sampled to acquire TSE-T1 -weighted and T2-weighted transverse axial images. Intraperitoneal injection of Gd DTP A was used to perform continuous dynamic enhancement scanning. Signal intensities of tumors were measured in plain scanning and each phase' s enhancement scanning images, respectively. Intensification rates of tumors were calculated. Pathologic examination of tumors was performed to be compared with the findings of MRI scanning. Results The successful rate of inoculation of 30 nude mice was 100%. The histological findings were comparable with poorly differentiated adenocarcinoma. Compared with signal of adjacent tissues, the MRI findings of the tumors were uniformly slightly hypointensity (90% , 27/30) , or unevenly (10% , 3/30) on TSE-T1WI; uniformly (20% , 6/30) or unevenly (80% , 24/30) hyperintensity with equal or more hyper signal spots on TSE-T2WI. Signal intensities on plain scanning was 228.35 ±11.71, and 1.5,3,6,9, 12 min after enhancement scanning, thesignal intensities were 258.20 ± 11.17, 301.75 ± 17.09, 358.65 ±25.13, 480.05 ± 19.01, 558.35 ± 40.49, which were significantly higher than those in plain scanning (P <0.01). The intensification rate of every phase was 0.13 ±0.04, 0.35 ±0.11, 0.56 ±0.10, 1.10 ±0.10, 1.45 ±0.18, and the difference among these phases was statistically significant (P <0.01). The significantly intensified area was the area where the tumor cells grew actively with rich capillaries; the central area without intensification was the area of necrotic tissue and/or densely packed tumor cells and few capillaries. Conclusions High resolution MRI imaging of implanted tumors can be obtained by intraperitoneal injection of contrast, and it is consistent with pathologic examinations.

Dynamic research about the change of the behavior and hippocampal volume of rat depressive model / 中华行为医学与脑科学杂志

Objective To explore the dynamic change and correlation of the behavior and hippocampal volumes by MRI of rat depressive model. Methods Based on the evaluation of ethology, thirty SD rats were randomly and equally divided into control group and model group which was exposed to chronic unpredictable mild stress for 8 weeks. The body weight, fluid consumption test and Morris water maze test were done at the beginning and per week while the bilateral hippocampal volumes of every rat were measured by high field MRI at the beginning and per two weeks. Results Based on the behavioral data,the rat depressive model was established successfully,and changed dynamically. On 4th week,the left hippocampal volumes of depressive rat were significantly reduced (model group(37.13 ±2.40)mm3 ,control group (39.05 ±2.05)mm3, t=2.36, P＜0.05) while the right ones did not. On 6th week, the right ones began to reduce significantly ( model group ( 37.85 ± 2.11 ) mm3, control group ( 39.44 ± 2.10 ) mm3, t = 2.07, P ＜ 0.05 ) while the left one still reduced, but the extent of the left's reduction was bigger than the right( left:15% ,right:8% ). And there was apparent correlation between the behavior and the reduction of hippocampal volumes. Conclusion The behavior and hippocampal volumes of rat depressive model changes dynamically,and there was a regularity of them.

Carotid Atherosclerotic Plaque:MR Imaing at 3.0T Magneton / 实用放射学杂志

Objective To study the capability in detection and qualitative analysis of carotid atherosclerotic plaque with 3.0T MR scanner.Methods Bilateral carotid arteriae in thirty-two patients with arterosclerosis which detected by Doppler ultrasound were examined with a 3.0T MR scanner.The pulse sequences included 3D time of flight,FLASH-3D,TSE-DIR T_1WI,TSE-DIR PDWI-FS,DIR TSE-T_2WI-FS and TSE-T_2WI.According to AHA modified criteria of atherosclerosis typing,the signal characteristics of plaques were analysed.Results Carotid plaques were homogeneous signal intensity on all sequences in 30/49,heterogeneous signal intensity on at least one sequence in 15/49,among them,7 with lipid core,3/49 plaques contained calcification ,2/49 appeared as thrombosis and resulting in vessel occulusion,28/49 plaques presented the character of thick fibrous cap,13/49 plaques showed character of thin fibrous cap,the surfaces of 5/49 plaques were rough.Conclusion High resolution multiple sequences imaging with 3.0T MR scanner can be used for detection and qualitative analysis of the carotid atherosclerotic plaque in a certain degree.

Establishment and magnetic resonance imaging monitoring of the orthotopic transplantation nude mouse model with human pancreatic cancer cell line PANC1 / 中华胰腺病杂志

Objective To establish a stable orthotopic transplantation nude mouse model of the human pancreatic cancer and to explore the role of monitoring tumor growth with noninvesive MRI.Methods The tumors cells suspension made by the subcutaneous injection of human pancreatic cancer cell lines PANC1 were used as the source of tissue for orthotopic implantation of tissue.and transplanted into the pancreas of 20 BALB/C-nu nude mice.After implantation,the successful rate,tumor formation time,tumor growth speed,tumor shape and the change of signal of the tumor were monitored and recorded noninvasively by MRI.At the end of the 7th week,all the specimens were examined by pathological methods.Results Thirty-five percent (7/20)mice with implantation of primary human PANC1 adenocarcinoma cells were detected to have orthotopic implanted tumors by MRI after 15 days,and all the 20 nude mice developed pancreatic tumor within 27 days after operations,and the successful rate was 100%.Compared with adjacent normal tissue,the T1 WI imaging of 90%(18/20)of all the tumors showed uniformly hypo-intense signal,10%(2/20)showed iso-intense signal,the T2W 75%(15/20)showed uniformly hyper-intense signal.The tumor size 2,3,4,5,6,7 weeks after implantation was(912.6±2.4)mm3,(94.3±11.2)mm3,(175.9±82.5)mm3,(395.8±126.6)mm3,(1290.2±167.2)mm3,(1583.4±87.4)mm3,respectively.Pathologic examination confirmed poody differentiated pancreatic adenocatcinoma and it remained the primary tumor's biolosic features.Conclusions The orthotopic transplantation nude mouse model was in accordance with the feature of human pancreatic cancer and was easy for noninvasive monitoring,which provided an effective and stable in vivo experimental system.

Effect of the chemically assisted enucleation on the enucleation of sheep oocytes and the development of their reconstructed embryos / 生物工程学报

In order to enhance the efficiency of sheep somatic cell nuclear transfer, we used a chemically assisted enucleation with colchicine to study the effects of the concentration of colchicine, the incubation time of oocytes in colchicine and the maturation time of oocytes on the enucleation rates and the development of reconstructed embryos. The results showed that 1) there were no significant differences in the rates of cytoplast protrusion and enucleation between oocytes that were incubated in colchicine (0.4 microg/mL) for 0.5 h and oocytes that were incubated in colchicine (0.4 microg/mL) for 1 h, and the rate of cytoplast protrusion can be 85.4% while the rate of cytoplast enucleation is 100%. 2) There was no significant difference in oocyte enucleation between oocytes treated with medium containing 0.2 microg/mL colchicine for 0.5 h and oocytes treated with medium containing 0.4 microg/mL colchicine for 0.5 h. 3) A maturation time of 18-23 h did not affect the rates of cytoplast protrusion and enucleation by chemically assisted enucleation, whereas the rate of enucleation of oocytes by blind enucleation was found to decrease with a prolonged incubation time. 4) The development rates of reconstructed embryos could not be influenced by these two enucleation methods, increased from oocytes matured for 21-23 h. These results demonstrate that sheep oocytes can be enucleated fast and effectively by optimized colcholine chemically assisted enucleation, which can enhance the enucleation rate of sheep oocytes and the early development of reconstructed embryos in vitro.

Identification methods of embryonic stem cells / 中国组织工程研究

OBJECTIVE: Embryonic Stem Cell (ES) are characterized by totipotency and normal karyotype and provide the theoretical base in the following fields: embryonic developmeut of mammalian, cell differentiation, expres sion of exogenous genes and utilization of Ess to develop animal model for human inherited diseases. The identification of ES therefore is very important to research and utilize ES.DATA SOURCES: The relevant articles to embryonic stem cell between January 1980 and December 2003 were computer searched for in Medline with the key words "embryonic stem cell, embryo, Alkaline phosphatase,Oct-4"in English. Similarly, the relevant articles to embryonic stem cell between January 1980 and December 2003 were computer searched for in China Journal Full-text Database (CJFD) with the key word "embryonic stem cell" in Chinese.STUDY SELECTION: The articles were browsed firstly. The inclusion criteria were for those articles about the identification of embryonic stem cells. The exclusion criteria were for those about repetitive studies, reviews and other similar articles.DATA EXTRACTION: 16 articles about the identification of embryonic stem cells were collected. Then, the full-texts of the articles were looked through.DATA SYNTHESIS: The selected data were summarized in the following order: ①Preliminary identification of Ess based on morphology and growth;②Immunological valuation; ③Chromosome related identification; ④Identi fication of totipotency and pluripotency.CONCLUSION: The identification of embryonic stem cells is not the result of only one identifying method, but a process of identification. During this process of comprehensive identification, it is recommend to conduct AKP test firstly, karyotype analysis secondly, then examination of surface markers and finally identification of Ess totipotency when Ess are sufficient, takingcare to repeat every identification.