Simultaneous 3D Visualization of the Microvascular and Neural Network in Mouse Spinal Cord Using Synchrotron Radiation Micro-Computed Tomography / 神经科学通报·英文版

Effective methods for visualizing neurovascular morphology are essential for understanding the normal spinal cord and the morphological alterations associated with diseases. However, ideal techniques for simultaneously imaging neurovascular structure in a broad region of a specimen are still lacking. In this study, we combined Golgi staining with angiography and synchrotron radiation micro-computed tomography (SRμCT) to visualize the 3D neurovascular network in the mouse spinal cord. Using our method, the 3D neurons, nerve fibers, and vasculature in a broad region could be visualized in the same image at cellular resolution without destructive sectioning. Besides, we found that the 3D morphology of neurons, nerve fiber tracts, and vasculature visualized by SRμCT were highly consistent with that visualized using the histological method. Moreover, the 3D neurovascular structure could be quantitatively evaluated by the combined methodology. The method shown here will be useful in fundamental neuroscience studies.

Simultaneous 3D Visualization of the Microvascular and Neural Network in Mouse Spinal Cord Using Synchrotron Radiation Micro-Computed Tomography / 神经科学通报·英文版

Effective methods for visualizing neurovascular morphology are essential for understanding the normal spinal cord and the morphological alterations associated with diseases. However, ideal techniques for simultaneously imaging neurovascular structure in a broad region of a specimen are still lacking. In this study, we combined Golgi staining with angiography and synchrotron radiation micro-computed tomography (SRμCT) to visualize the 3D neurovascular network in the mouse spinal cord. Using our method, the 3D neurons, nerve fibers, and vasculature in a broad region could be visualized in the same image at cellular resolution without destructive sectioning. Besides, we found that the 3D morphology of neurons, nerve fiber tracts, and vasculature visualized by SRμCT were highly consistent with that visualized using the histological method. Moreover, the 3D neurovascular structure could be quantitatively evaluated by the combined methodology. The method shown here will be useful in fundamental neuroscience studies.

Arthroscopic debridement and meniscectomy in treatment of meniscus injury combined with osteoarthritis / 中国内镜杂志

Objective To investigate the effect of arthroscopic debridement and meniscectomy in treatment of moderate or severe meniscus injury combined with knee osteoarthritis in early or middle stage. Methods 156 cases diagnosed with moderate or severe meniscus injury combined with knee osteoarthritis in early or middle stage were collected from October 2011 to October 2014. Lysholm knee score and preoperative examinations such as anteroposterior, lateral, axial radiographs, the standing full leg length X-ray film and MRI scan of the knee were recommended to definitively understand the osteoarthritis staging and meniscus injury grading. All patients were treated with arthroscopic debridement and meniscectomy. After operation, physical rehabilitation exercises and regular clinical follow-up were carried out as planned. The Lysholm knee score data from preoperation and terminal follow-up was statistical analyzed. Results No patient experienced any perioperative and postoperative complications. Statistical analysis showed that the Lysholm knee score of postoperation was significantly higher than that of preoperation [(87.3 ± 7.9) vs (67.5 ± 4.9), P < 0.05). Conclusion Arthroscopic debridement and meniscectomy in treatment of moderate or severe meniscus injury combined with knee osteoarthritis in early or middle stage, gains beneficial effects for its minimal invasion and quick recovery.

Application of simulation technology to designing aerosol sprayers / 军事医学

Objective To develop a portable aerosol sprayer .Methods With computer aided design(CAD)technology,a three-dimensional model of the aerosol sprayer was built , and the simulation analysis of nozzles was conducted using the computational fluid dynamics ( CFD) technology .Results A model of portable aerosol sprayers was designed .According to simulation analysis , the speed and pressure of aerosol in different locations were obtained and were proved to change with time, providing a theoretical basis for determination of working parameters and structural design parameters of the aerosol sprayer nozzle so that design efficiency and quality could be improved; time and cost could be reduced .Conclusion Through CAD technology,an effective method for rational design and structure optimization of the aerosol sprayer is esta -bished.Hydromechanic problems that used to be solved with physical analogue experiments can be resolved by means of CFD simulation analysis .

Histomorphological analyse of accelerating the fibrocartilage layer repair of patella-patellar tendon junction in rabbits by low intensity pulsed ultrasound stimulation / 中南大学学报(医学版)

OBJECTIVE@#To analyse the effect of low intensity pulsed ultrasound stimulation (LIPUS) on accelerating the fibrocartilage layer repair of patella-patellar tendon junction.@*METHODS@#A total of 60 mature female New Zealand white rabbits undergoing standard partial patellectomy were divided into 2 groups randomly. The control group was given comfort treatment and the treatment group was given LIPUS treatment starting from day 3 to the end of week 6 postoperatively. The scheduled time points of animal euthanization would be at week 6, week 12 and week 18 postoperatively. The patella-patellar tendon (PPT) complex would be harvested and cut into sections after decalcification for H&E staining, Safranine o/fast green staining. The thickness and gray value of fibrocartilage layer were analyzed by SANO Microscope Partner image analyzer.@*RESULTS@#At week 6, week 12 and week 18 postoperatively, the fibrocartilage layer in the treatment group was significantly thicker than that in the control group (P<0.01), and the gray value of fibrocartilage layer was significantly smaller than that in the control group (P<0.01).@*CONCLUSION@#LIPUS helps to accelerate the fibrocartilage layer repair of patella-patellar tendon junction in rabbit models.

In vivo study on three-dimensional structure of lumbar facet joints based on computerassisted medical image processing method / 医用生物力学

Objective To study three-dimensional feature of the thickness of subchondral cortical bone, the surface area and space width of the facet joints as well as its morphological characteristics and variation, and to compare the differences of these parameters in patients with low back pains and healthy people, based on three dimensional reconstruction images of the facet joints. Methods CT scanning from L1/2 to L5/S1 at supine position was performed on 90 subjects to obtain two-dimensional images of the facet joints and three-dimensional point-cloud data of the subchondral cortical bone, the surface area and space width of the facet joints after reconstruction. Distributions of these parameters between the superior and inferior facet joints and at different regions of the articular process were investigated, and differences of these parameters in lumbar segments, gender, age and symptoms were also compared. Results (1)At superior facet, the largest thickness was located at the cranial region, while at inferior facet, it was located at the caudal region. The thickness of subchondral cortical bone was significantly smaller at central region than at the other regions. A more obvious difference could be observed at the lower lumbar levels（L4/5, L5/S1）. (2)The average surface area of the facet joints was (173.2±3.6) mm2, and it was increased with the lumbar level and age increasing, especially at the L4/5 level and at the age of more than 40. The surface area of superior facet joints was significantly larger than that of inferior facet joints for patients with low back pains (except at L5/S1 level), who had an obviously larger surface area of the facet joints than healthy people. (3)The average thickness of space width of the facet joints was (1.46±0.08) mm, and it was decreased with age decreasing, especially at the age of more than 40. The female had significantly larger space width than the male, while patients with low back pains had significantly smaller space width. At each facet joint, the space at cranial region was significantly larger than that at caudal region, and it was also significantly larger at central region than at surrounding region. These differences were more obvious at the lower three lumbar levels. Conclusions Computer-assistant image processing and reconstruction analysis techniques can accurately measure the relative parameters of lumbar facet joints with complicated structure in three dimensional space. The feature of the facet joints was correlated with shape of the facet joint surface，stress on the joint and degeneration of the joint, so it was a reflection of adaptive changes in human body structure due to the long-term loading on the facet joint.

Effect of tetramethylpyrazine on the expression of macrophage migration inhibitory factor in acute spinal cord injury in rats / 中南大学学报(医学版)

OBJECTIVE@#To determine the effect of tetramethylpyrazine (TMP) on the expression of migration inhibitory factor (MIF) in acute spinal cord injury (ASCI) in rats.@*METHODS@#Allen's weight-drop method was used to establish a rat model of ASCI at T10. A total of 110 adult SD rats were divided into a sham operation group (group S, n=10), a control group (group C, n=50), and a TMP group (group T, n=50). Spinal cord functionality was measured by a modified Rivilin loxotic plate degree, BBB score, and combined behavioral score (CBS) at 1, 3, 5, 7, 14 and 21 d postoperatively. The injured spinal cord tissue samples were harvested at 1, 3, 6, 12 h and 1, 3, 5, 7, 14, 21 d postoperatively (n=5 at each time point) and used to prepare continuous histological sections, in which the expression of MIF was analyzed by immunohistochemistry.@*RESULTS@#The degree in group T measured by modified Rivlin loxotic plate test after the ASCI was significantly higher than that in group C at 7, 14, and 21 d (P<0.05). BBB score in group T was significantly higher than that in group C at 5, 7, 14, and 21 d after the ASCI (P<0.05). CBS score in group C was significantly higher than that in group T at 5, 7, 14, and 21 d after the ASCI (P<0.05). The significantly low number of MIF positive cells was shown in group T when compared with that in group C at 12 h and 1, 3, 5, 7 d after the ASCI (P<0.05). As time passed, there was negative correlation between modified Rivlin loxotic plate degree and MIF expression and also between BBB score and MIF, and there was positive correlation between CBB score and MIF expression.@*CONCLUSION@#TMP has protective effect after the ASCI, and may promote the repair of injured spinal cord tissues. TMP may decrease the MIF expression in cells after the ASCI.

Stability of the hydrogenase from Tetraselmis subcordiformis and its preliminary purification / 生物工程学报

Tetraselmis subcordiformis, a marine green alga, can produce hydrogen by photobiologically hydrolyzing seawater with hydrogenase. In this study, the preliminary purification of the enzyme was explored by ammonium sulfate precipitation, and the impact of sodium dithionite, beta-mercaptoethanol and glycerol on the enzyme stability during the process was investigated. The experimental results illustrated that sodium dithionite provided significant protection on the hydrogenase by depleting oxygen, while glycerol, a protectant against the structure instability of the enzyme, also presented protection. Crude enzyme with specific activity of 0.557 U/mg protein was extracted using 60%-70% saturated ammonium sulfate solution supplemented with 200 mmol/L sodium dithionite and 5% glycerol, and the hydrogenase recovery yield was about 30%.

Bone regeneration and maturation in a rabbit tibia lengthening model under low-intensity purse ultrasound stimulation / 中国组织工程研究

BACKGROUND: Experiments has demonstrated that low-intensity pulse ultrasound stimulations(LIPUS) can accelerate bone healing. However, Its affect mechanisms on maturation of regenerated bone remain unclear.OBJECTIVE: The hypothesis that LIPUS has positive effect on maturation of regenerate bone was put forward creatively, and wants to vedfy the hypothesis by compare the difference between LIPUS intervention and no intervention.METHODS: Thlrty-six adult, healthy, New Zealand White rabbits were randomly assigned to LIPUS treatment group and control.group, with 18 animals in each group.All animals were undergone mid-diaphyeaal tibia ostactomy and were immobilized in Orthofix M103 Mini lengtheners. After a eaven-day latency period, gradual distraction of 0.5 mm per twelve hours for 10 days to produce 10 mm distraction. A 4-weeks course of LIPUS treatment was applled over the distraction site for twenty minutes daily starting immediately after the completion of distraction. At weeks 4, 8, 12 after distraction, the bone healing was evaluated by X-ray film, and total mineralized areas were measured by Image J software. The newly formed bone In the lerigthening field was harvested and stained with haematoxylin-eosin, Masson's tdchroma end VG. Thearaas of newly formed bone were measured to calculate a percentage of the total callus area.RESULTS AND CONCLUSION: Thirty-six rabbits ware included in the final analysis, At week 4 after distraction, the maturation of newly formed bone in LIPUS treatment group appeared eadier than that in the control groups. The bona callus image presented in the marginal and center region of experimental defect, increased greatly and had higher density until filled of the lengthening fields. Many bone-like masses appeared in the newly formed tissues and the tissues surrounding the bone granules partially changed Into bone-like tissues, and the rate of callus production was increased (P ＜ 0.05). There was no significant difference between two groups at weeks 8 and 12 after distraction. Histological results showed that that new bone formation of the LIPUS group was earlier than the control group at weeks 4, 8, and 12 after distraction. The percentage of new bona on total callus area of the LIPUS group was greater than that of the control group at weak 4, but there was no obvious difference at weeks 8 and 12 after distraction. LIPUS accelerates new bone formation, increases the size of the distraction callus, exhibits highly effective in achieving maturation of bone in the animal modal with distraction osteogenesis.

Effect of low-intensity pulsed ultrasound stimulation on maturation of regenerate bone / 中南大学学报(医学版)

Objective To explore the effect of low intensity pulsed ultrasound stimulation (LI-PUS) on the maturation of regenerate bone in a rabbit limb lengthening model. Methods Sixty skeletal mature female New Zealand white rabbits were randomly divided into an LIPUS treatment group and a control group. All rabbits were underwent mid-diaphyseal tibial osteotomy and immobilized in an Orthofix M103 Mini lengther. Gradual distraction at 0. 5 mm every 12 h for 10 d was performed at day 7 postoperatively. A 4-week course of LIPUS treatment group was applied over the distraction site for 20 min daily starting immediately after the completion of the distraction only for the treatment group. Rabbits were euthanized and the mid-diaphyseal tibia was harvested for evaluation at 4, 8, and 12 wk after the completion of the bone lengthening protocol. Radiographic analysis was performed to study the formation of bone callus using the ImageJ software at 12 wk after the completion of the bone lengthening protocol. Bone mineral density (BMD) of regenerate bone was measured by Dual energy X-ray absorptiometry (DEXA) . Torsional testing to failure was performed on the tibia specimens at 8 and 12 wk after the completion of the bone lengthening protocol. Results Radio-graphic measurement showed higher relative gray scale of bone callus in the LIPUS group than that in the control group at 12 wk (P ＜ 0. 05) . BMD in the LIPUS group was significantly higher than that in control group at 8 and 12 wk (P ＜ 0. 05). Biomechanical testing showed that the ultimate torque, ultimate torsional stiffness, and energy absorption at failure of regenerated bone at 8 and 12 wk in the LIPUS treatment group were better than those in the control group (P ＜ 0. 05) Conclusion LIPUS as a biophysical stimulation may accelerate the formation and maturation of regenerate bone in rabbit tibia lengthening model.

Mitochondrial DNA deletion mutations in articular chondrocytes of cartilage affected by osteoarthritis / 中南大学学报(医学版)

Objective To detect the changes of mitochondrion DNA (mtDNA) sequence in articular chondrocytes of cartilage affected by osteoarthritis and to clarified the pathogenetic mechanism of osteoarthritis. Methods We analyzed the mtDNA 4 977 bp deletion mutations of articular chondrocytes in 10 patients with osteoarthritis and 3 normal cartilages using the gap-PCR amplification method. We designed a two round PCR detection method, in which total DNA was isolated from articular chondrocytes as the template of the first round PCR reaction and products from the first round were the template in the second round reaction. Results The results of the first rounds of PCR reaction showed the mtDNA 524 bp amplified products in the osteoarthritis group and in the corresponding peripheral blood samples were not detected, but the 533 bp products were detected. However, the results of the second round reaction revealed that the 524 bp zones were detected in 2 of the 10 osteoarthritis patients and the corresponding peripheral blood samples were not detected. The 533 bp products were detected in all specimens. The mtDNA 524 bp amplified products in all the normal articular chondrocytes and the corresponding peripheral white blood cells contrast were not detected in both rounds PCR. Conclusion This was the first study to evaluate the mtDNA 4 799 bp large fragment deletion mutational accumulation between nt8 470～nt13 447 of articular chondrocytes in osteoarthritic cartilage. Osteoarthritis may be related to mtDNA mutation of articular chondrocytes.

Histopathological analysis of choroidal melanoma / 中华眼底病杂志

Objective To investigate the histopathological characteristics of choroidal melanoma. Methods The histopathological data from 64 patients with choroidal melanoma were analyzed retrospectively. The tumor size and the cytological types were observed and detected. The locations of the tumor were classified according to the involved part invaded by the anterior margine of the tumor, and the degrees of the development of the tumor were graded according to the extent of the outward infiltration of the tumor cells. Results In 64 patients with choroidal melanoma, There were large, medium, and small tumors in 25(39.1%), 31(48.4%), and 8(12.5%) respectively. The spindle cell type was found in 42 patients (65.6%) including spindle cell A and B type in 15(23.4%) and 27(42.3%) respectively; epithelioid and mixed cell type was found in 7(10.9%) and 10(15.6%) respectively; the other types were found in 5(7.8%). Twenty-five cases(39.1%)had no invasion with sclera, 22(34.4%)had but limited to sclera, 12(18.8%)penetrated through sclera and 5(7.8%)had intra-orbit infiltration. Conclusion The histopathological characteristics of choroidal melanoma are multiple, and spindle cell type is the most common one. The choroidal melanoma can easily invade the sclera.

The Effect of Fragmin in Treatment of Unstable Angina / 医药导报

Objective:To study the effect of Fragmin in treatment of unstable angina (UA).Methods:100 patients with UA were randomized into treatment group and control group.In the control group,the subjects were given routine drugs such as sorbitrate,nifedipine and aspirin,while those in the treatment group given fragmine in addition to the routine drugs.Results:The response rates in the treatment and control groups were 82.1% and 54.5%,respectively (P＜0.01).The total effective rates in the treatment and control groups were 92.9% and 63.6%,respectively (P＜0.01).The total effective rate of EKG improvement in the treatment and control groups were 80.4% and 50.0%,respectively (P＜0.01).Conclusion:Fragmin is effective in the treatment of UA.

Ultrastructure observation of biological living skin equivalent / 实用口腔医学杂志

Objective:To observe the ultrastructure of biological living skin equivalent in vitro.Methods:Keratinocytes and fibroblasts were sourced from the back skin of legally aborted human foetus.Fibroblasts were seeded into bovine type I collagen gel and cultured for 3 days.Keratinocytes were seeded on the surface of collagen gel and cultured for another 2 days,then the equivalent skin was exposed to air-liquid interface to form a protective cornified layer.Histological structure and ultrastructure of the equivalent skin were observed by light microscope and transmission electron microscope.Results:The morphogy of equivalent skin of full thickness consisted of epithlium and dermis.Epithelium was made up of stratum basale,stratum spinosum,stratum granulosum and stratum corneum.lntercellular bridge existed within the cells of different layers.Some ultrastructures in epithelium such as desmosome,Lamelle,tonofibrils and lipid droplet were found in the equivalent skin.Conclusion:Tissue-engineered skin may have the features of natural skin.