Three-dimensional visualization on the relationship between tubules after macula densa and afferent arterioles in mouse kidney / 解剖学报

Objective To establish the spatial course of distal tubule and afferent arterioles after macula densa, and to locate and detect the proteins in the adjacent parts by using three-dimensional visualization technology of microstructure. Methods C57 BL/6J mice were fixed by perfusion and embedded in epon 812. Tissue blocks were cut perpendicular to the longitudinal axis of the kidney. And a total of 720, 2. 5 μm-thick consecutive sections were obtained from the renal capsule to the outer stripe of the renal outer medulla. After aligning the digital microscopic images through computer registration procedures, the tubules and vessels were traced by 3D reconstruction program edited by C Language. Selecting the tissue sections of the contact site and applying the improved immunoperoxidase staining method to detect H

Morphometry of the ureteric bud branching in the developing mouse kidney / 解剖学报

[Abstract] Objective To investigate the branching pattern of the ureteric bud and the number of the nephron induced by each ureteric bud tip, through the three-dimensional tracing of the ureteric tree, combined with the morphological analysis and measurement of the ureteric tree. Methods The kidneys were obtained from three mice at various developing time points and prepared for paraffin and epoxy sections. Then the microscopic images were digitized and aligned from these sections. Based on the computer-assisted tracing and visualization of ureteric tree, the number of branches and the nephron induced by each ureteric bud tip were obtained by counting. In addition, paraffin sections were stained with HE staining for morphological observation of nephrogenic zone and ureteric bud, while in order to reflect the density of the ureteric bud tips at nephrogenic zone, the distance between two neighboring ureteric bud tips was measured aided with the Claudin-7 immunohistochemical staining. Results The ureteric bud branching tree revealed that the initial bifid iterative branching formed the framework of renal medulla, the branching became complicated and dense in cortex and nephrogenic zone, while the distance between ureteric bud tips were also decreasing. The number of the nephron induced by each ureteric bud tip increased from one (E14. 5) to two (E17. 5), and occasionally to three. Conclusion Threedimeasional Visualization of ureteric bud branching tree reveals regional complication, suggesting molecules in different regions drive different branching patterns; While the density of the ureteric bud tips at nephrogenic zone increases corresponding to decreasing of thickness of the nephrogenic zone, and the disappearance of the ureteric bud tips after birth is also consistent with the gradual consumption of nephron progenitor cells.

Morphometry of the capillary loop stage glomerulus in developing mice / 解剖学报

Objective To describe quantitatively the development of the capillary loop stage glomerulus (capG) with respect to the volume density of capillaries in the glomerulus based on the morphogenesis of the kidney. Methods The kidneys were obtained from mice at various developing time points and prepared for paraffin sections. The volume density of CD34 positive endothelial cells and surrounded capillary lumen in glomeruli was measured using a combination of immunohistochemical staining and the stereological grid system. Results The capG was divided into early, middle, and late phases, and middle phase capG was subdivided into early-middle and late-middle phases, according to the morphology of developing glomeruli and the arrangement of podocytes. As result, the volume density of capillary loops in early phase capG could not be measured due to the complex "glomerular" shape. The volume density of capillary loops increased from (35.95±6.45)% in the early-middle phase capG, to (58.36±6. 30) % in the late-middle phase capG, and to (79.89± 5.21) % in the late phase capG, compared to (93.61 ±1.96) % in the mature glomerulus. Furthermore, the volume density of capillary loops remained constant at same stage even though at different developmental time points. Conclusion This study demonstrated a significantly increased volume density of capillary loops with the kidney development. In addition, the results provide a descriptive and reliable parameter for the evaluation of glomerular development.

Three-dimensional visualization of the renal proximal tubules in developing mice / 解剖学报

Objective Adult proximal tubule ( PT ) is not only the segment most frequently involved in acute renal tubule injury, but also the easiest to repair. It may be consistent with the rapid growth and differentiation mechanism of this segment during the development of the kidney, while the developing information is insufficient. Therefore, we three- dimensional visualized the developing PT to analysis its spatiotemporal morphogenesis. Methods The kidneys were obtained from mice at various developing time point, embryonic day ( E ), postnatal day ( P ). The volume density of Claudin-2 positive PT in the cortex was measured using a stereological method in paraffin sections. After image recording and alignment of the serial sections, the spatial courses of the developing PT were traced and visualized in three dimensions using computer-assisted program. The length of the developing PT was calculated at the same time. Results The volume density of PT in the cortex of PI mice was significantly higher than that in the embryonic stage. Then it experienced a decline ( P3, P5 ), an increase ( start at P7 ) to a stable adult level ( P28 ). The tubular tracing showed that the lengths of developing PT and the number of convolutions of their convoluted part increased with the maturation, but lower than that of adultin E14. 5, E17. 5 and P5 PT in E14. 5 and E17. 5 mice were similar to that of adult with respect to general spatial courses. They were, however, significantly different from adult in the initial direction of PT and the arrangement of the straight part of PT in the medullary rays. While, it was in P5 that the spatial pattern of some PT was gradually approaching to the adult model. Conclusion This study demonstrated that the development of PT was consistent with the kidney development in terms of its volume density in cortex, length and spatial course. It started at the S-shaped body, kept throughout the embryonic period and continued to postnatal, ended at kidney maturation ( P28 ).

Comparative study of differentiation potential of mesenchymal stem cells derived from orofacial system into vascular endothelial cells / 北京大学学报(医学版)

OBJECTIVE@#To compare the proliferation and capacity of differentiation to vascular endothelial cells and angiogenesis induction among stem cells from human exfoliated deciduous teeth (SHED), dental pulp stem cells (DPSC) and human bone marrow mesenchymal stem cells (BMSC) from orofacial bone.@*METHODS@#SHED and DPSC were isolated from pulp tissue of the patients. BMSC were isolated from orthognathic or alveolar surgical sites. The surface markers of the cells were detected by flowcytometry. Cell counting kit-8 (CCK-8) assays were conducted to detect the proliferation ability of the cells. The cells were induced into endothelial cells with conditional medium and then the induced cells were cultured in Matrigel medium. The expression of angiogenesis-related genes such as platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2) and von Willebrand Factor (vWF) were quantified by real-time PCR. The cells were cultured in chick embryo chorioallantoic membrane (CAM) and the vessels were counted after 5 days.@*RESULTS@#The cell surface markers CD73, CD90, CD105 and CD146 of all the stem cells were positive, CD34 and CD45 were negative. The CD146 positive rate of SHED and DPSC was higher than that of BMSC. SHED had a higher proliferation rate than DPSC and BMSC. After angiogenic induction for 14 d, 3 kinds of cells emanated pseudopodia formed grid structure long vasculature in Matrigel media. The total length of tube formation of induced BMSC (7 759.7 μm) and SHED (7 734.3 μm) was higher than DPSC (5 541.0 μm). The meshes number of induced SHED (70.7) was higher than DPSC (60) and BMSC (53.7) in Matrigel medium. The expression of CD31, VEGFR2 and vWF genes of SHED were higher than those of BMSC and DPSC. VEGFR1 gene expression of BMSC was higher than that of the other groups, and SHED was higher than DPSC. The expression of VEGF showed no difference among the cells. No deference was showed between the effect of the stem cells and negative control on new formed vessels in CAM. The total length of vessels of SHED (30.4 mm) was higher than that of the negative control (20.9 mm) and BMSC (28.0 mm).@*CONCLUSION@#SHED, DPSC and BMSC can differentiate into vascular endothelial cells. SHED showed a stronger angiogenesis differentiation and proliferation potential compared with DPSC and BMSC.

Association of electronic devices usage and visual fatigue in Chinese college students / 眼科新进展

Objective To investigate the prevalence of visual fatigue among Chinese college students with the usage of electronic products.Methods A total of 6000 college students were recruited from 127 universities in 29 cities (except Taiwan,Qinghai,Tibet,Ningxia and Xinjiang).The questionnaire involved questions pertaining to the dependence of electronic products,use of electronic products in dormitory,home,vehicles,and prevalence of visual fatigue.Logistic regression was performed to analyze the potential risk factors for visual fatigue.Results A total of 4848 questionnaires were valid,involving 2259 male and 2589 female students.The results revealed that the incidence of visual fatigue in Chinese college students was 53.5％,a higher rate in females than in males.And 33.8％ of whole group admitted that they had become relied on electronic products.The proportion of electronic products used at the table,on the bed,transport equipment reached 91.3％,87.6％ and 74.8％,respectively.In terms of the usage duration,38.3％ of surveyed students use mobile phone,Tablet PC over 4 hours per day,with 29.7％ using electronic products over 1 hour on the bed and 49.1％ in vehicles over 0.5 hour.Univariate logistic regression analysis showed that the prevalence of visual fatigue were related to gender,dependence on electronic products,the use of electronic products in dormitory,home,vehicles.Conclusion The prevalence of visual fatigue is relatively high among Chinese college students,which may be associated with the following several factors,including gender,dependence on electronic products and the use of electronic products in dormitory,home and vehicles.

Preparation of high immunity yolk antibody against Vibrio parahemolyticus and comparison of effectiveness between different extraction methods / 吉林大学学报(医学版)

Objective:To compare the effectiveness between three methods for purifying the immunoglobulin of egg yolk(IgY) which are polyethylene glycol (PEG) method, chloroform extraction method and chloroform / PEG method, and to provide basis for obtaining the batch of IgY.Methods:The inactivated vaccine of Vibrio parahemolyticus (V. parahemolyticus) was prepared and the hens were immunized by multi-point intramuscular injection.The eggs were collected and the IgY was purified by PEG method, chloroform extraction method and chloroform/PEG method.The protein extraction rate, the IgY titer and the purity of the antibody which purified by different methods were detected.Furthermore, the operation process, cost and safety of the three methods were analyzed.Results:The protein contents of the extraction belonging three methods from high to low in turn were chloroform extraction method, chloroform/PEG method, and PEG method.There was no significant difference in the antibody titer between three methods, and the tiler of chloroform extraction method was slightly high.The purities of purified antibody from high to low in turn were PEG method, chloroform/PEG method and chloroform method.The PEG method had better security but relatively lower extraction efficiency and higher cost.The chloroform/PEG method had high extraction efficiency and good antibody purity.Conclusion:The PEG method is suitable for a small amount of extraction in the laboratory.The chloroform/PEG method is appropriate for extracting the high quality IgY in a batch as it has high extraction efficiency and good antibody purity.

Value of Serum ProGRP,NSE and CEA Detection in Diagnosis of Small Cell Lung Cancer / 现代检验医学杂志

Objective To investigate the diagnostic value of combined detection of serum gastrin-releasing peptide precursor (ProGRP),neuron specific enolization enzyme(NSE)and carcinoembryonicantigen(CEA)in small cell lung cancer(SCLC). Methods 471 patients with lung tumor from department of respiratory medicine and thoracic surgery and 162 healthy people from medical examination center were studied.Serum levels of ProGRP,NSE and CEA were detected by using electrochemi-cal luminescence method.ROC curves were drawn and the area under the curve (AUC)was calculated.Results The levels of ProGRP and NSE were significantly higher in patients with SCLC than those in NSCLC,lung benign disease group and normal control group (P <0.01).The levels of CEA were significantly higher in SCLC than those in patients with lung be-nign disease group and normal control group (P <0.05).The AUC of ProGRP,NSE and CEA in the diagnosis of SCLC were 0.933,0.777 and 0.554,respectively.The sensitivity of ProGRP,NSE and CEA in the diagnosis of SCLC were 82.6%,60.4%,41.6% and the specificity were 95.2%,83.3% and 71.7% respectively.The sensitivity of combined detec-tion of ProGRP,NSE and CEA was 91.3% and the specificity was 65.3%.Conclusion The serum ProGRP detection has a higher diagnostic value for SCLC.The combined detection of ProGRP,NSE and CEA is useful in the early diagnosis of SCLC.

Subtractive SELEX using agar beads for screening DNA aptamers with specific affinity to HIV gp41 antigen / 南方医科大学学报

<p><b>OBJECTIVE</b>To obtain DNA aptamers with a highly specific affinity to HIV gp41 antigen using SELEX screening for detection of HIV.</p><p><b>METHODS</b>The specific DNA aptamers of HIV gp41 antigen were screened from the double-stranded DNA derived from the single-stranded DNA (ssDNA) library with agarose beads as the supportive medium and HIV gp41 antigen as the target molecule using SELEX technique and real-time quantitative PCR.</p><p><b>RESULTS</b>The secondary ssDNA library obtained after 6 rounds of screening was amplified by PCR to obtain dsDNA. The dsDNA was linked with pMD18-T vector, cloned and sequenced to obtain 4 aptamers of HIV gp41 antigen. The affinities of the 4 aptamers (K) all reached the nanomolar level. Among the 4 aptamers, the No.15 aptamer showed the strongest affinity. Specificity analysis of the aptamers revealed that all these 4 aptamers had specific affinity to HIV gp41 antigen with no affinity to other non-specific proteins.</p><p><b>CONCLUSION</b>We successfully obtained DNA aptamers with highly specific affinity to the HIV gp41 antigen from random single-stranded oligonucleotide library, and the obtained aptamers have the ability to antagonize HIV gp41 antigen.</p>

Oligodendrocyte and spinal cord injury / 生物医学工程学杂志

Spinal cord injury (SCI) is frequently companied by necrosis and apoptosis of oligodendrocytes (OLs), which contributes to demyelination of myelinated nerve fibers and their electrophysiological defects. This pathological demyelination often results in sensory or motor deficits. Here, we first focus on the microenvironment changes after SCI that cause OLs' death, then discuss the major mechanism of endogenous oligodendrocytogenesis and axonal remyelination, and finally summarize current therapies targeting OLs protection and replacement.