Acute Disease ; Case-Control Studies ; Child, Preschool ; Echocardiography, Doppler ; Female ; Heart ; physiopathology ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; diagnostic imaging ; physiopathology

Background Establising the culture model of Müller cells for obtaining the highly putified target cells is essential for the study about the physiology and pathology of retinal Müller cells. The exsiting purifing method for culturing Müller cells is dissatisfactory. Objective This study was to establish a method to obtain high purifing Müller cells. Methods The retina from 5 clean newborn SD rats were isolated and digested by 0. 01% trypsin and cultured in DMEM containing 10% fetal bovine serum. The cellular suspension was then prepared,and the target cells were screened using flow cytometry based on the size and the quantity of cells. Cultured and passaged cells were identified by transmission electron microscope and light microscope. Immunocytochemistry was used to detecte the expression of GFAP in cultured cells for the determination of type and purity of the cells. Results The cells showed the similar shape to retinal Müller cells after primarily culture with the large volume, and some small other types of cells could been seen. The growth of cells was quickly 3 weeks later. The fibroblasts were removed using sticking-wall by steps,and neurons were eliminated following passage. Aboundent of cellular organs were seen under the transmission electron microscope. The positive response rate of the cells for CFAP was 100%. Conclution Flow cytometry offer a rapid and feasible approach for purifying Muller cell and it builds the foundation for further study about Müller cells.

Due to the low effectiveness of traditional assisted reproductive technology (ART), new technological possibilities are constantly explored. Lots of studies have demonstrated the potential of microfluidics to revolutionize the fundamental processes of in vitro fertilization (IVF). With the advantages of high efficiency, short time, harmless collection, real-time observation of separation, similar microenvironment, and automation, the application of microfluidics in sperm isolation and IVF has shown an evident superiority over the conventional approaches and provided a new platform for ART. This review highlights the application of various microfluidic techniques in sperm motility assessment and isolation, sperm chemotaxis assay, IVF, sperm concentration, and sperm separation and enrichment in recent years. It also briefly introduces the basic principles, structural design, and operation processes of the microfluidic platform, focusing on the advantages and disadvantages of each method and the potential of their clinical application. Obviously, there are still some challenges to the application of microfluidics in ART. However, it is believed that the development of this new technology would be toward a highly integrated application of several steps in one single device, known as IVF-lab-on-a-chip.
Fertilization in Vitro ; methods ; Humans ; In Vitro Techniques ; Male ; Microfluidics ; methods ; Reproductive Techniques, Assisted ; Sperm Motility ; Spermatozoa

The aim of this study is to establish a method to culture and purify cerebral astrocyte of tree shrew (Tupaia belangeri), a kind of new laboratorial animal which is a relative of primates. Newborn tree shrews were used in this experiment. The cortex of cerebrum was isolated and placed in 4°C for 20 min to injure neurons. The cortical tissue was disaggregated by trypsin digestion. Differential attachment method was used to remove fibroblasts. The mixed culture was rinsed by trypsin (0.005%) solution to remove neurons. Upon reaching 70% confluence, the culture was subjected to static trypsin digestion until a white slice film exfoliated from the bottom of culture bottle. This film, i.e. astrocyte layer, was taken out and cultured, and the third passage was identified by immunocytochemical staining and immunofluorescence with anti-glial fibrillary acidic protein (GFAP) antibody. The result showed the purity of tree shrew astrocytes was more than 98%. Thus the method to culture highly purified astrocyte of tree shrew was successfully established, which would contribute to further study in central nervous system physiology and diseases in this new laboratorial animal.
Animals ; Astrocytes ; cytology ; Brain ; cytology ; Cell Separation ; methods ; Primary Cell Culture ; methods ; Tupaiidae

Objective The stentless full root aortic bioprosthesis has superior hemodynamics.Clinical data of Bio-Bentall procedure using stentless full root bioprosthesis of our center was retrospectively analyzed in this perspective for validation.Methods From November 2001 to March 2009,317 adult patients ( 196 male and 121 female) underwent modified Bio-Bentall procedure using the Medtronic Freestyle xenograft as a full root replacement.Two hundred and three patients received an isolated root replacement or a root and ascending aortic replacement (ARR).In 114 patients a variety of concomitant procedures including coronary artery bypass grafting ( n =32 ),mitral valve repair ( n =11 ) and aortic arch replacement ( n =36 ) were performed.(ARR + ).Results Mean patient age was (70.3 ± 10.2) years (range 17 -94 years),97 patients were 75 and older at time of procedure.Mean operative time for the ARR was ( 190 ± 57 ) min with a clamp time of 88 - 27 min.Mean operative time for ARR + group was (282 ±93) min with an average clamp time of (110 ±32) min.Overall operative mortality was 7.9％ (25/317),for ARR it was 5.4％ (11/203 ).Mean ICU stay was (4.9 ± 8.1 ) days,mean hospital stay being (9.8 ± 8.1 ) days.Necessity for bailout bypass surgery among patients with ARR was low at 1.5％ (3/203) comparable to stented xenograft implantations.Echocardiography demonstrated excellent clinical results with low transvalvular gradients especially when a single suture inflow anastomosis technique was used.Conclusion Full root stentless valve implantation preserving porcine root integrity is a valuable option in aortic valve/ascending aorta surgery.Though technically a more challenging operation,it does not lead to increased perioperative morbidity and mortality and can be beneficial mainly for elderly patients with small aortic roots with or without aortic root pathology.

Objective To evaluate the safety of interventional radiotherapy with X-ray angiography. Methods To compare X-ray radiation levels with and without radioprotect with X-ray protective monitor by simulating operators' body's parts. Results The difference is significant. Conclusion Interventional radiotherapy with angiography with effective protect is safe.

Chinese medicine pair (CMP) was frequently applied in traditional Chinese medicine (TCM) clinic, and its significance was shown in long-term clinical practices and many accumulated experiences. It is the unique combination of two relatively fixed Chinese medicines in TCM clinic with the basic feature and principle of TCM compatibility, is the most fundamental and the simplest form of TCM formulae with certain theory basis and combinatory reason, which is proven effective. And the unique combination is frequently used for achieving mutual reinforcement or detoxication. CMP is an intermediate point between single herb and many TCM formulae, reflecting the regularity of TCM formulae compatibility and connotation of differential treatment. This paper analyzed and summarized the basic characteristics, development process and research significance of CMP, which aims to lead the modern basic and applied research on compatibility theory of CMP.
Drug Interactions ; History, 16th Century ; History, 17th Century ; History, 18th Century ; History, 19th Century ; History, 20th Century ; History, 21st Century ; History, Ancient ; Medicine, Chinese Traditional ; history ; methods

1H NMR metabonomics approach was used to reveal the chemical difference of urine between patients with Xiao-Chaihu Tang syndrome (XCHTS) and healthy participants (HP). The partial least square method was used to establish a model to distinguish the patients with Xiao-Chaihu-Tang syndrome from the healthy controls. Thirty-four endogenous metabolites were identified in the 1H NMR spectrum, and orthogonal partial least squares discriminant analysis showed the urine of patients with Xiao-Chaihu Tang syndrome and healthy participants could be separated clearly. It is indicated that the metabolic profiling of patients with Xiao-Chaihu Tang syndrome was changed obviously. Fifteen metabolites were found by S-pot of OPLS-DA and VIP value. The contents of leucine, formic acid, glycine, hippuric acid and uracil increased in the urine of patients, while threonine, 2-hydroxyisobutyrate, acetamide, 2-oxoglutarate, citric acid, dimethylamine, malonic acid, betaine, trimethylamine oxide, phenylacetyl glycine, and uridine decreased. These metabolites involve the intestinal microbial balance, energy metabolism and amino acid metabolism pathways, which is related with the major symptom of Xiao-Chaihu Tang syndrome. The patients with Xiao-Chaihu Tang syndrome could be identified and predicted correctly using the established partial least squares model. This study could be served as the basis for the accurate diagnostic and reasonable administration of Xiao-Chaihu-Tang syndrome.
Humans ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Syndrome ; Urinalysis

AIMThe changes of tissue inhibitor of metalloproteinase-4 (TIMP-4) expression in mouse ovary during pregnant and postpartum period were studied to investigate the role of TIMP-4 in corpus luteum (CL).

METHODSRT-PCR was used to deter mine the change of TIMP-4 mRNA and indirect immunofluorescence was used to observe the change of TIMP-4 protein. The expression of TIMP-4 mRNA was observed in various periods throughout the stage of pregnancy and postpartum day 1.

RESULTSThe expression of TIMP-4 was gradually enhanced from day 1 to day 8, reached a maximal expression at day 8, while decreased at day 11 and to the lowest level at postpartum day 1. Indirect immunofluorescence results further indicated that TIMP-4 protein was localized to CL and theca-intera cells in various periods throughout the pregnancy and postpartum day 1. In addition, the change pattern of TIMP-4 protein agreed with that of the TIMP-4 mRNA in pregnancy CL.

CONCLUSIONThe expression of TIMP-4 in mouse ovary during pregnancy and postpartum is in spatio-temporal pattern and it may be involved in the formation and function maintain of CL during pregnancy in mice.

Animals ; Female ; Mice ; Mice, Inbred Strains ; Ovary ; metabolism ; Postpartum Period ; Pregnancy ; RNA, Messenger ; genetics ; Tissue Inhibitor of Metalloproteinases ; metabolism

Objective To investigate the correlation of clinicopathological parameters and prognosis with serum and pancreatic cancer tissue klotho. Methods Immunohistochemistry EnVision two step method was used to assess klotho protein expression of a tissue microarray ( TMA) of 79 pairs of pancreatic tissue and normal surrounding tissue. The serum klotho levels in 39 pancreatic cancer patients and 39 healthy controls who had matched clinical data were measured by ELISA. The relationships between the expression of klotho and the clinicopathological features and survival were analyzed. Results Klotho expression positivity in pancreatic cancer tissue was significantly higher than that in adjacent normal tissues (59. 5％ vs 96. 3％);serum level of klotho was markedly higher in pancreatic cancer patients than that in control group [(670. 30 ± 82. 24)pg/ml vs (310.35 ± 34.65) pg/ml], and both the difference was statistically significant (P<0.001). Klotho expression was negatively associated with tumor clinical stage and lymph node metastasis (P<0. 05), and the expression of klotho did not correlate with patients' gender, age, tumor size, location, local invasion depth and the like. The median survival time in pancreatic cancer patients with positive klotho expression were longer than that in in pancreatic cancer patients with negative klotho expression [(48. 31 ± 6. 94) months vs (19. 50 ±6. 78)months], and the difference was statistically significant (P<0. 01). ROC analysis on serum klotho gave a cutoff value of 376. 51 pg/ml to diagnosis pancreatic cancer with a sensitivity of 84. 6％ and specificity of 87. 2％. Conclusions Klotho level in serum and tissue of pancreatic cancer patients was closely correlated with clinicopathological parameters and prognosis, which may be a potential biomarker for pancreatic cancer.