1.Arenobufagin is a novel isoform-specific probe for sensing human sulfotransferase 2A1.
Xiangge TIAN ; Chao WANG ; Peipei DONG ; Yue AN ; Xinyu ZHAO ; Weiru JIANG ; Gang WANG ; Jie HOU ; Lei FENG ; Yan WANG ; Guangbo GE ; Xiaokui HUO ; Jing NING ; Xiaochi MA
Acta Pharmaceutica Sinica B 2018;8(5):784-794
Human cytosolic sulfotransferase 2A1 (SULT2A1) is an important phase II metabolic enzyme. The detection of SULT2A1 is helpful for the functional characterization of SULT2A1 and diagnosis of its related diseases. However, due to the overlapping substrate specificity among members of the sulfotransferase family, it is difficult to develop a probe substrate for selective detection of SULT2A1. In the present study, through characterization of the sulfation of series of bufadienolides, arenobufagin (AB) was proved as a potential probe substrate for SULT2A1 with high sensitivity and specificity. Subsequently, the sulfation of AB was characterized by experimental and molecular docking studies. The sulfate-conjugated metabolite was identified as AB-3-sulfate. The sulfation of AB displayed a high selectivity for SULT2A1 which was confirmed by reaction phenotyping assays. The sulfation of AB by human liver cytosols and recombinant SULT2A1 both obeyed Michaelis-Menten kinetics, with similar kinetic parameters. Molecular docking was performed to understand the interaction between AB and SULT2A1, in which the lack of interaction with Met-137 and Tyr-238 of SULT2A1 made it possible to eliminate substrate inhibition of AB sulfation. Finally, the probe was successfully used to determine the activity of SULT2A1 and its isoenzymes in tissue preparations of human and laboratory animals.
2.Comprehensive clinical assessment on imaging diagnosis of low back pain
Youzhuan XIE ; Xiaokui HOU ; Zhenan ZHU
Orthopedic Journal of China 2006;0(01):-
Objective To pursue the appropriate selection of radiographic examinations of low back pain and to reduce the expenses. Method In this study,61 cases of imaging tests were studied,including 38 cases of lumbar disc herniation,10 cases of lumbar spondylolisthesis,4 lumbar stenosis,4 lumbar infection and 5 lumbar neoplasm. Result Correct diagnosis was based on the clinical signs and symptoms.Imaging findings had its value only when basing on them.Conventional roentgenogram offered a cheap evaluation and high spatial resolution of bony structures.And it could give a clue to take further imaging studies.Low back disorders could be inferred from the contrast column defect of dura sac and nerve root sheath in the myelography.In the diagnosis of disc herniation,its limitations existed at far lacteral and L5S1 disc herniation.Computed tomography,a cross-sectional imaging modality,allows direct visualization of the bony structure of the spine.But routine imaging could only cover a limited number of levers.Another limitation was the difficulty in distinguishing the soft tissue changes in the spinal canal with the similar CT values.CT-myelography was performed to delineate more clearly the bony and soft tissue anatomy,and could determine whether lesions were medullary,intradual or extradual in location.MR imaging provided the larger anatomic region in multiple planes with high spatial resolution of soft tissue.One limitation of MR imaging was relatively low spatial resolution of bony structure. Conclusion Of all imaging studies conventional roentgenogram is the most important and the most fundamental.It could not be substituted by myelography,CT,CT-myelography,and MR imaging,which have their designated own roles respectively in the clinical decision-making process and in general were more complementary than competitive.
3.In vitro culture of bone marrow stromal cells immobilized in calcium alginate beads
Wenhui LI ; Xiaokui HOU ; Tingting TANG ; You WANG
Chinese Journal of Tissue Engineering Research 2005;9(30):260-262
BACKGROUND:Carrier material provides an essential part of microenvironment for seed cell culture. There are some investigations analyzing the long-term in vitro culture of cartilaginous cell in calcium alginate gel matrix.OBJECTIVE: This study was designed to investigate the activity, morphology and microstructure of cultured in vitro bone marrow stromal cells (BMSCs) immobilized in calcium alginate beads.DESIGN: Single sample studySETTING: It was conducted at the Orthopaedic Institute of Zhengzhou University and Orthopaedic Laboratory of Shanghai Second Medical University.MATERIALS: The experiment was conducted at the Orthopaedic Laboratory of Shanghai Second Medical University from September 2001 to March 2002. Eight New Zealand white rabbits, with an age ranging from 4 to 6weeks, were used for collecting BMSCs.METHODS: Eight rabbit were anaesthetized and 1.0-2.0 mL bone marrow was aspirated from the humerus of femur using a 16-gange biopsy needle pretreated with heparin for primary and passage culture of BMSCs.Sodium alginate solution was prepared. The sodium alginate-BMSCs composite formed calcium alginate beads, with a cell density of 1×109 L-1 and tology characteristics of BMSCs were examined using toluidine blue staintron microscope (TEM).scope, many aggregate cells with round shape, in all sizes, occasionally characteristics of BMSCs: Cells and matrix materials were well-mixed, with no materials stained. The cells were equably stained, in all sizes, with large nuclei and little plasma, and occasionally having two or more nuclei and cell division. These cells had an intact structure, well-defined border, and tures of BMSCs: The cells had normal structures, with no mitochondrion swelled and no degranulation in ribosomes. They had a continuous nuclear membrane, with babyhood morphology, round shapes,oval shapes or irregular shapes, and occasionally having two nuclei.CONCLUSION: BMSCs-calcium alginate beads exhibit advantages including a good hydrophilicity, easiness for nutritive matter to penetrate and proliferating actively. It is feasible to amplify BMSCs through in vitro cullage repair by tissue engineering (TE) method.
4.Experimental study of calcium alginate compound in the construction of tissue engineered bone and cartilage
Wenhui LI ; Xiaokui HOU ; Tingting TANG ; You WANG
Chinese Journal of Tissue Engineering Research 2005;9(26):237-239
BACKGROUND: Seed cells and supporting materials are two key problems for cartilaginous tissue engineering. Enough functional cells in supporting materials is the foundation in the formation of bone and cartilaginous tissue in vivo.OBJECTIVE:To perform a morphological study on the tissue engineered bone and cartilage constructed with calcium alginate, in vitro cultured bone marrow stem cells(BMSCs) and growth factors.DESIGN:Morphological comparative study with the results observed for 12 weeksSETTING:Orthopaedic Institute of Zhengzhou University and Orthopedic Laboratory of the Second Affiliated Hospital of Shanghai Medical University.MATERIALS:This experiment was completed from September 2001 to June 2002 in the Orthopedic Laboratory of the Second Affiliated Hospital of Shanghai Medical University. Nine New Zealand male rabbits at the age of 3 months were selected, their mean body mass was 3.2 kg.METHODS: Rabbits received dorsal subcutaneous injection of five kinds of the following combinations at two points for each combination and in the Ⅰ+TGF-β. Totally ten points were marked. Three rabbits were put to death at the 4th, 8th, 12th weeks after injection respectively and dorsal subcutaneous hyperplasia tissues were collected for observation by using hematoxylin and eosin (HE), toluidine blue, Masson, tomato red-O staining techniques.MAIN OUTCOME MEASURES: Histological observation of dorsal subcutaneous hyperplasia tissues.RESULTS: At the fourth week, cartilaginous like tissues island could be observed in calcium alginate+ BMSCs + IGF-Ⅰ group, calcium alginate+BMSCs +TGF-β group and calcium alginate + BMSCs + IGF-Ⅰ +TGF-βgroup, with chondrocblasts surrounded by alkaline matrix; A great deal of cartilaginous-like tissue appeared at the eighth week with part of it transforming into bone, and cells secrete much matrix, part of the chondroblast clustered. At the 12th week, approximately all of the cartilaginous tissues turned into bone in calcium alginate+ BMSCs + IGF- Ⅰ +TGF-β group, and began to absorb.CONCLUSION: The compound composed of calcium alginate, BMSCs and growth factor might form bone and cartilaginous tissue. Calcium alginate is suitable for the growth of BMSCs and is believed to be an optimal bone and cartilaginous engineering carrier.
5.Minimally-invasive management of injuries of knee joint
Chinese Journal of Orthopaedic Trauma 2004;0(06):-
The arthroscopic technique was first used for minimally invasive treatment and diagnosis of injuries inside the knee joint. It can make the diagnosis of acute cartilage injury more accurate. Since it can also correctly detect the location, type and scope of the meniscus injury, surgeons can reasonably decided whether the injured meniscus should be sutured or partially resected so as to maintain as most part of meniscus as possible. It can also be applied to repair or reconstruct the anterior and posterior cruciate ligaments. The combination of arthroscopic technique and minimally invasive internal fixation has often been used to treat fractures of tibial plateau, patella, and femoral condyles, as well as to assist the treatment of fractures of distal femur with retrograde interlocking nails. The internal fixation by percutaneous minimally invasive anatomical plates, e.g. Liss Invasive Stabilization System(LISS), has proved to be effective in the clinical treatment of fractures near tibial and femoral metaphyses.
6.Progress in treatment of intra-articular injury and arthroscopic techniques in China
Chinese Journal of Orthopaedic Trauma 2002;0(01):-
Intra-articular injury refers to acute and chronic trauma o f such joint structures as bone, cartilage and intra-articular soft tissues: li gament,cartilage disc and synovium. The limitations of traditional diagnosis and treatment usually result in joint instability and traumatic arthritis. Arthrosc opic surgery is a typical method of minimally invasive technique, and has been a pplied in our country since 1980s. With rapid progress and popularization of t he arthroscopic technology as well as great improvement in arthroscopic instrume nts and equipment (lens systems, fiberoptic systems and accessory operative inst ruments), the advanced arthroscopic operative method can be widely applied in di agnosis and treatment of acute joint traumas, intra-articular bone and cartilag e fractures, intra-articular ligament injuries, meniscus injuries and shoulder joint ailments. The article reviews the progress made in the past decade in the clinical application of arthroscopic technology.
7.Prospect of the basic fibroblast growth factor (bFGF) application in orthopaedics
Chinese Journal of Orthopaedic Trauma 2002;0(04):-
Basic fibroblast growth factor is one of the most important grow th factors with broad distribution and powerful bioactivities. Since it has positi vely inductive or stimulative effects on tissues, such as bone, cartilage, muscl e, vessel, and nerve, it will have extensive applications in orthopaedics in the future. The current review introduces new advances in the applications of bFGF in orthopaedics.
8.Seed cells and its culture methods in cartilage tissue engineering
Wenhui LI ; Xiaokui HOU ;
Chinese Journal of Orthopaedic Trauma 2002;0(03):-
Acute cartilage injury is one of the most common disorders in orthopaedic clinics.Articular cartilage is known to have a limited capacity to repair or regenerate itself. The disorder is difficult to be dealt with, usually affects the function of joint and may induce the onset of osteoarthrosis.This will result in great economic burdens on the patients and their family and greatly degrade the patient s quality of life. The recent progress in researches on seed cells, culture methods, biological material has provided a new method to repair the acute injury of articular cartilage and thus may solve many problems. The article introduces new advances in seed cells and their culture methods in cartilage tissue engineering.
9.True scapular roentgenographic diagnosis of subacromial impingement syndrome
Yan GU ; Kerong DAI ; Xiaokui HOU
Chinese Journal of Orthopaedics 2001;0(03):-
Objective To evaluate the acromion morphology of scapular roentgenography in the diagnosis of subacromial impingement syndrome. Methods Sixty four patients 75 shoulders with chief complaint of shoulder pain were examined roentgenographically by true scapular A-P and lateral views. The Neer impingement sign of all these patients were positive. Results In this series, there were 10 shoulders of type Ⅰ acromion, 13.3%; typeⅡ 17 shoulders, 22.7%; type Ⅲ 48 shoulders, 64.0%. Fifty nine cases 69 shoulders accepted Lidocaine test. The positive rate was: type Ⅰ, 4 shoulders, 44.4%; type Ⅱ, 8 shoulders, 53.3%; type Ⅲ, 38 shoulders, 84.4%. Among these three groups, the result of Chi test for Lidocaine tests: P0.05 (typeⅠ to type Ⅱ). Conclusion True scapular X ray film is a useful and effective diagnostic method for subacromial impingement syndrome. The incidence of subacromial impingement syndrome of the patients with type Ⅲ acromion is significantly higher than the other two types.
10.Arthroscopic synovectomy for rheumatoid arthritis in early and middle stages
Jun ZHANG ; Xiaokui HOU ; You WANG
Chinese Journal of Minimally Invasive Surgery 2001;0(05):-
Objective To compare the efficacy of arthroscopic synovectomy between early and middle stage rheumatoid arthritis. Methods A total of 34 cases of rheumatoid arthritis (42 knee joints) either in early stage (22 cases, 24 joints) or in meddle stage (12 cases, 18 joints) underwent arthroscopic synovectomy. The bipolar radiofrequency was used for the synovial debridement and hemostasis in 32 knee joints. Postoperatively, routine anti-rheumatoid drugs were administrated. Follow-up observations in the two groups included the evaluation of knee function, the erythrocyte sedimentation rate (ESR), the C-reaction protein (CRP), and the rheumatoid factor (RF). Results The rate of excellent or good results was 91.7% in early stage (22/24) and 66.7% in middle stage (12/18), without statistical significance between the two groups ( ? 2=2.705, P =0.100). Conclusions Arthroscopic synovectomy is effective for the treatment of rheumatoid arthritis in both early and middle stage. Bipolar radiofrequency is helpful for the complete elimination of the synovium, the prevention of joint hematoma, and the rehabilitation of joint function.

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