Objective　To establish the method of 2-dimensional electrophoresis(2-DE) for proteins of human spermatozoa and to construct a protein map of human spermatozoa. Methods　The sperm pellet was prepared with simple Percoll layer protocol. We studied the effects of various sample preparation methods, loading quantities and isoelectric-focusing protocols on the quality of silver-stained 2-DE map, and constructed a primary protein map of human spermatozoa. Result　Up to 703 protein spots were acquired with sample preparation Method Ⅰwhile only 194～210 spots with Method Ⅱ.With immobilized pH gradients and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(IPG-DALT) we could acquire over 700 spots while only 280～300 with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(ISO-DALT). Conclusion　It is satisfactory to lyse sperm with sample preparation Method Ⅰ and to separate sperm proteins by IPG-DALT for establishing 2-D map of human sperm.

BACKGROUND:Spermatogonial stem cel s with abilities of differentiation, self renewal and proliferation are a kind of adult stem cel s that can transfer genetic information into offspring, which have great application prospects in medicine, genetics and zoology. OBJECTIVE:To review the source, biological characteristics, and application of spermatogonial stem cel s as wel as self-renewal and molecular regulation underlining these differentiations. METHODS:PubMed and CNKI databases were searched by the first author using key words of“spermatogonial stem cel , biological characteristics, self-renewal, differentiation”in English and in Chinese, respectively, to retrieve relevant articles published from 1990 to 2015. Literatures addressing spermatogonial stem cel s were included, and 46 articles were chosen for further analysis eventual y. RESULTS AND CONCLUSION:Spermatogonial stem cel s can be cultured in vitro, cryopreserved, and genetical y modified as wel as used for al ogeneic or xenogeneic transplantation, al of which contribute to understanding the mechanisms of spermatogenesis, thereby providing new means for treatment of male sterile disease and genetic diseases and providing new hopes for chemotherapy-induced germ cel damage in young cancer patients. Microenvironment and Plzf, GDNF, SCF/c-Kit signaling pathways can play an important role in the regulation of spermatogonial stem cel self-renewal and differentiation. As a cel model, spermatogonial stem cel s become an important tool for the researches on spermatogenesis mechanism, regeneration of spermatogenesis in sterile individuals and reproduction of transgenic animals.

Objective To analyze the correlation of traditional Chinese medical constitution types withcardiovascular risk factors in community hypertension patients. Methods We defined the constitution types of 1108 community hypertension patients, created personal health archives and provided routine examination for the subjects. After that, we input and analyzed the relevant data and results by computer. Results In 1108 cases of hypertension patients, 229 (20.67%) were defined as harmony physique, 879 (79.33%) as biased physique. In patients with biased physique, phlegm-dampness physique accounted for 25.54% (283 cases) , and then came yin deficiency physique and Qi deficiency physique, which accounted for 15.16% (168 cases) and 13.18% (146 cases) , respectively. The biased physique was dominated by single biased physique with a total of 590 cases ( 67.12%) , but the complex physique was also in large quantity, with a total of 289 cases ( 32.88%) . The complex physique consisting of two kinds of single physique was commonly seen in the patients with complex physique, accounting for 15.70%. The complex physique consisting of more kinds of single physique had less percentage. Phlegm-dampness physique was a kind of physique involving the most cardiovascular risk factors. Patients with phlegm-dampness physique had higher incidence of overweight, abnormal lipid level, family history of cardiovascular disease, lack of physical exercise and electrocardiographic abnormality, and the difference was statistically significant compared with other physique types ( P<0.05 or P<0.01) . Conclusion Phlegm-dampness, yin deficiency and Qi deficiency physique are the three commonly-seen constitution types in hypertension patients. Phlegm-dampness physique is the most commonly-seen biased physique and is also a kind of physique involving the most cardiovascular risk factors. Prevention and management in consideration with the constitution characteristics and cardiovascular risk factors will be a new prospective method for the treatment of hypertension patients in community.

Objective To investigate the effects of Annexin-A1 ( Anxa1 ) gene silencing induced by siRNA on the growth and migration of microglial BV-2 cells and its possible mechanisms.Methods A synthesized siRNA duplex targeting Anxa1 gene was transfected into BV-2 cells.The efficiency of siRNA-in-duced Anxa1 gene silencing was evaluated on both mRNA and protein levels by using reverse-transcription PCR and Western blot assay.MTT assay was performed to measure the proliferation of BV-2 cells with si-lenced expression of Anxa1 gene.Flow cytometry with Annexin V-FITC/PI double staining was used to de-tect the apoptosis rate of BV-2 cells.Transwell chambers were used to analyze the effects of siRNA-induced Anxa1 gene silencing on the migration of BV-2 cells.Western blot assay was performed to detect the expres-sion of signaling proteins related to cell cycle and migration.Results Compared with the siRNA negative control ( siRNA-NC) group, the inhibitory rates of siRNA-induced Anxa1 gene silencing on the proliferation of BV-2 cells were significantly increased at the time points of 24 h, 48 h and 72 h after intervention [(16.9 ±2.1)%, (23.1±3.6)%and (42.4±1.7)%vs (1.35±0.5)%, (2.06±0.7)% and (8.65±0.9)%, P<0.05 ].The apoptosis rate of BV-2 cells transfected with Anxa1 siRNA was (18.4±2.1)%, which was significantly elevated as compared with that of the siRNA-NC group (5.2±0.3)%and control group (4.3±0.2)%.Cell migration of the Anxa1 siRNA transfected BV-2 cells was inhibited remarkably at 48 h as com-pared with that of the siRNA-NC group (28.7±5.2 vs 173.4±11.4, P<0.01).Moreover, the suppressed expression of Cyclin D1 protein and activation of p38 and JNK signaling pathways were induced by silenced expression of Anxa1 gene in BV-2 cells.Conclusion The growth and migration of BV-2 cells were signifi-cantly inhibited by silencing the expression of Anxa1 gene with siRNA, the possible mechanisms might be associated with the suppressed expression of Cyclin D1protein and the activation of p38 and JNK signaling pathways.

A new dibenzocyclooctadiene lignan, renchangianin E (1) was isolated from the stems of Kadsura renchangiana. Its structure and stereochemistry were elucidated by spectroscopic methods, including 2D-NMR techniques.

Objective To investigate the induction of B-cell specific phenotype in classical Hodgkin lymphoma (cHL)upon all-trans retinoic acid(ATRA)incubation. Methods To construct B-cell specific promoter(CD19, CD79a,CD79b)driven reporter plasmid with NEO cassette to realize stable transfection and selection of cHL reporter cells. To verify the intact integration by amplification of the promoter and luciferase sequences,and to functionally validate the B-cell specific promoter by ABF1 interference and luciferase assay. Repoter cells were incubated with various doses of ATRA and luciferase activity was detected at 24,48 and 72 hours. Reporter cells were treated alone or in combination with 5-Aza and ATRA followed by luciferase assay. Endogenous B-cell specific genes(CD19, CD20, CD79a and CD79b) transcription and expression levels were detected by real-time PCR and immunoblot, respectively. The expression level of CD30 antigen on Hodgkin lymphoma cell membrane upon ATRA was assessed by flow cytometry. Results ATRA treatment stimulated B-cell specific signature in cHL cells including CD19,CD79a and CD79b while down-regulated their CD30 expression. Conclusions ATRA induces B-cell phenotype deficient cHL cells to regain their B-cell transcriptional program while abolishes their Hodgkin-specific machinery.

This paper was aimed to explore the features of single ventricle with dual source computed tomography (DSCT) and the value of the modality in diagnosis of the single ventricle disease. Dual-source computed tomography imaging of 14 cases of single ventricle compared with the results of surgery and cardiac catheter were retrospectively analyzed. Firstly, 14 cases were classified into three types according to Anderson classification method, including 7 cases(50. 0%) left ventricular type, 3 cases(21. 4%) right ventricular type and 4 cases (28. 6%) undecided ventricular type. Secondly, in accompanying malformation respect, pulmonary stenosis, bilateral superior vena cava, atrial septal defect (ASD) and common atrioventricular valve were presented frequently. Thirdly, affiliated cardiac chamber of left ventricular type were mostly in front of the major cardiac chamber (5 cases,71. 4%), while affiliated cardiac chamber of right ventricular type were behind of the major cardiac chamber totally. Fourthly, more than half of aortas and pulmonary arteries stemming from common cardiac chamber can be seen (8 cases, 57. 1%). Meanwhile the aorta valves were more on the right of pulmonary valve (9 cases, 64. 3%). The classification of 10 cases of single ventricle was compared with the results of surgery, and the coincidence rate is 100%. DSCT can diagnose the single ventricle disease accurately. The aorta, pulmonary artery and coronary artery can be displayed completely and simultaneously as well.
Adolescent ; Child ; Child, Preschool ; Female ; Heart Ventricles ; abnormalities ; diagnostic imaging ; Humans ; Infant ; Male ; Radiography, Dual-Energy Scanned Projection ; methods ; Retrospective Studies ; Tomography, X-Ray Computed ; methods

This paper is to compare dual-source computed tomography (DSCT) with trans-thoracic echocardiography (TTE) in the evaluation of anomalous pulmonary venous connection (APVC) to explore the advantages of DSCT. Thirty-one patients with APVC detected by DSCT were enrolled in this study. In this cohort, 16 patients underwent echocardiography followed by operative treatments; and among the remained 15 patients who did not received surgical treatment, echocardiography was performed in 10 patients. The numbers and drainage sites of anomalous pulmonary veins, and accompanied defects on DSCT and TTE were retrospectively analyzed. In terms of diagnosis of the numbers and the drainage sites of anomalous pulmonary veins, the accuracy of DSCT was 98% (49/50) and 93.8% (15/16), while the accuracy of echocardiography was 80% (40/50) and 56.3% (9/16), respectively. There were statistical differences between the two modalities (P < 0.05). Although the detection rates of accompanied defects were 89.1% (41/46) and 78.3% (36/46) for DSCT and echocardiography, respectively, no statistically significant difference was found between the two modalities (P > 0.05). The 15 patients without receiving surgical treatments were partial APVC confirmed by DSCT, but APVC was found in 40% (4/10) patients by echocardiography. In conclusion, DSCT could be superior to evaluating APVC in comparison with echocardiography, and should be recommended as an important procedure for preoperative assessment of the anomaly in patients with APVC.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Echocardiography ; Evaluation Studies as Topic ; Female ; Humans ; Infant ; Male ; Middle Aged ; Radiographic Image Interpretation, Computer-Assisted ; Retrospective Studies ; Scimitar Syndrome ; diagnostic imaging ; Tomography, X-Ray Computed ; methods ; Young Adult

The purpose of this study was to explore the clinical value of ECG-gated dual-source CT angiography (DSCTA) in evaluating coarctation of aorta (CoA). 23 patients suspected with CoA underwent DSCTA and transthoracic echocardiography (TTE). Surgical results were taken as reference standard. 23 patients were diagnosed with CoA, 12 cases had focal stenosis of aorta, while 11 had tubular stenosis. 19 cases were associated with other cardiovascular malformations, including 5 cases of hypoplastic aortic arch, 5 of patent ductus arteriosus, 10 of ventricular septal defect, 2 of bicuspid aortic valve, 4 of collateral arteries, 2 of aberrant left subclavian artery and 2 of persistent left superior vena cava, respectively. The mean diameter of stenotic segment of aorta was (7.0 +/- 3.7) mm. In 9 patients with tubular CoA, the diameter and length of stenotic segment of aorta were (6.2 +/- 7. 3) mm and (29.2 +/- 5.9) mm, respectively. The accuracies of DSCTA and TTE in the diagnosis of CoA were 100% and 91.3% (P > 0.05), respectively. It is well concluded that ECG-gated DSCTA could accurately evaluate CoA and associated cardiovascular malformations, and provide detailed anatomic information before surgery.
Adolescent ; Adult ; Angiography ; methods ; Aortic Coarctation ; diagnostic imaging ; surgery ; Cardiac-Gated Imaging Techniques ; methods ; Child ; Child, Preschool ; Contrast Media ; Echocardiography ; Electrocardiography ; methods ; Female ; Humans ; Infant ; Male ; Tomography, X-Ray Computed ; methods ; Young Adult

The purpose of this study was to determine the morphological and dynamic features of normal pulmonary valve by using dual-source CT. Thirty patients who underwent dual-source CT angiography with right ventricular outflow tract well-opacified were enrolled. Ten phases of images were reconstructed at 10% R-R interval, and pulmonary valve area and pulmonary annular diameter were measured on all 10 phases. As the pulmonary valve opened, its orifice appeared as oval or proximate triangle. As the pulmonary valve closed, it appeared as "Y-shape". Pulmonary valve opened from 10% to 30% phases, with maximum pulmonary valve area of (4.74 +/- 0.69) cm2 in 10% phase. Pulmonary valve closed from 40% to 90% phases. Pulmonary annular diameter varied with cardiac cycle, with maximum pulmonary annular diameter of (2.36 +/- 0.24) cm in 0% phase and minimum pulmonary annular diameter of (1.82 +/- 0.32) cm in 40% phase (P < 0.001). The proximal phase for assessing pulmonary valve was 70% phase. Dual-source CT may evaluate the morphological and dynamic features of normal pulmonary valve.
Adult ; Female ; Humans ; Iohexol ; analogs & derivatives ; Male ; Middle Aged ; Pulmonary Valve ; anatomy & histology ; diagnostic imaging ; physiology ; Tomography, Spiral Computed ; methods