1.Rotational animal brain injury models dynamically and quantitatively detected by diffusion tensor imaging correlated with β-amyloid precursor protein expression
Jingtong XIONG ; Jingwen ZHANG ; Jianlin WU ; Yuguo FANG ; Lin LIN
Chinese Journal of Radiology 2016;50(6):469-474
Objective To investigate dynamic regulation of quantifying fractional anisotropy (FA) and apparent diffusion coefficient (ADC) values in rotational brain injury models of rabbit using diffusion tensor imaging (DTI), and its correlation with β?amyloid precursor protein (β-APP). Methods Forty-two 6-month-old New Zealand rabbits were randomly divided into three groups, including control group (n=6), mild injured group (n=18) and severe injured group (n=18), and preformed on autonomous rapidly rotational brain injury device. The rotational angles of 45° for mild injured group and 90° for severe injured group were condemned, and MRI and pathology were conducted at 6 h, 12 h, 24 h, 48 h, 72 h and 1 week after injury (3 rabbits per subgroup). Routine sequences and DTI technique were performed on 3.0 T MRI. FA and ADC values in subcortical white matter, corpus callosum and brain stem were measured. Independent t?test was performed to evaluate the significance of the intergroup difference in FA and ADC values in mild and severe injured groups of verious brain regions by timing, one?way ANOVA was performed to evaluate its timing variation and its correlation with the number of the β-APP positive axons was analyzed by Pearson correlation analysis. Results FA and ADC values of the severe injured group were lower than that of the mild in most brain regions(P<0.05), and the difference in mild injured group was smaller than that in severe injured group. Both FA and ADC values in brain stem of the severe injured group were lower at 6 h after injury compared to mild injured group, which were sensitive to injury. Furthermore, FA and ADC values in each brain regions of mild and severe injured groups showed similar dynamic trends, namely gradually decreasing by time, and FA values were more sensitive to injury than ADC values. FA values in subcortical white matter and brain stem reduced in severe injured group at 6 h after injury compared with that before injury(P<0.05), and decreased in various brain area of both injured groups at 12 h after injury(P<0.05). Meanwhile, ADC values in all regions were declined in the severe injured group at 12 h after injury(P<0.05), and decreased in various regions in both injured groups at 24—48 h after injury(P<0.05)except for subcortical white matter in mild injured group. There were statistically negative correlations between FA and ADC values and the number of β-APP positive axons in 12—48 h after injury in most regions(P<0.05). Conclusions DTI can quantitatively detect and assess the pathological process in white matter and axons of TBI in earlier stage of the brain injury, and can be applied in evaluation and quantitative diagnose in these patients.
2.Inhibition of Proliferation of Megakaryoblastic Leukemic Cells by 1,25-Dihydroxyvitamin D_3 and Retinoic Acid
Tao CHENG ; Fang YAN ; Jinghua WAN ; Jianlin YIN
Academic Journal of Second Military Medical University 1982;0(01):-
The effect of inhibition of proliferation of a novel human megakaryoblastic leukemia cell line (HIMeg) by two physiological factors, 1.25-dihydroxyvitamin D3 [1,25(OH)2 D3] and 13-cis-retinoic acid (RA), was investigated. At the range of 10-9 - 10-6 mol/L, 1,25(OH)2 D3 and RA showed significant inhibition of proliferation of the megakaryoblastic leukemia cells,which was demonstrated by count of survival cells,incorporation of [3H]-TdR and [3H]-UR, and cloning efficiency. From above, it can further explain the m.echanism of differentiation-inducers and the effect of 1,25(OH)2D3 on myelofibrosis. It is possible for 1,25(OH)2D3 and RA to be used for a new -treatment.
3.Local intra-articular injection of sodium hyaluronate delays articular cartilage degeneration after traumatic osteoarthritis
Jianlin ZHOU ; Shuang DENG ; Hongsong FANG ; Hao PENG ; Bo QIU
Chinese Journal of Tissue Engineering Research 2015;(33):5295-5300
Abstract BACKGROUND: Sodium hyaluronate is an effective treatment for osteoarthritis, but the underlying mechanism remains unclear. There is evidence that abnormal expressions of matrix metaloproteinase (MMP)-1, -3 and -9 and tissue inhibitor of metaloproteinase (TIMP)-1 and -2 show great effects on osteoarthritis. OBJECTIVE: To assess the influence of intra-articular injection of sodium hyaluronate on expressions of MMPs-1, 3, 9 and tissue inhibitor of TIMPs-1, 2 in the rabbit cartilage after osteoarthritis. METHODS: Twenty-four mature New Zealand white rabbits were divided into normal control, model, and sodium hyaluronate groups. The model and sodium hyaluronate groups underwent unilateral anterior cruciate ligament transection, and rabbits in the sodium hyaluronate group received 0.3 mL of 1% sodium hyaluronate via intra-articular injection at 4 weeks after modeling, once a week for 5 weeks. At 11 weeks folowing surgery, the rabbits were kiled and the cartilage was harvested to extract total RNA. mRNA expressions of MMPs-1, 3, 9 and TIMPs-1, 2 in the cartilage were analyzed using real-time PCR for each group. RESULTS AND CONCLUSION:Compared with the model group, the range and extent of cartilage damage was reduced in the sodium hyaluronate group (P < 0.01), and Mankin scores were noticeably decreased (P < 0.05). In the cartilage, mRNA expressions of MMPs-1, 3, 9 were enhanced and mRNA expressions of TIMPs-1, 2 were down-regulated in the model group. However, the mRNA expression levels of MMPs-1, 3, 9 and TIMPs-1, 2 in the articular cartilage were not obviously changed in the sodium hyaluronate group. These results suggest that MMPs-1, 3, 9 and TIMPs-1, 2 are involved in the progression of osteoarthritis and the therapeutic mechanism of sodium hyaluronate is not realized through the down-regulation of their expressions during development of osteoarthritis. Sodium hyaluronate for treatment of osteoarthritis is a complex process and the underlying mechanisms require further investigation.
4.Correlation between venous blood phase values detected by enhanced gradient echo T2 star weighted angiography sequence and neurological severity scale scores in mild and severe traumatic brain injury models of rabbits
Jingtong XIONG ; Jingwen ZHANG ; Jianlin WU ; Yuguo FANG
Chinese Journal of Radiology 2016;(2):132-137
Objective To explore the change of venous blood phase values detected by enhanced gradient echo T2 star weighted angiography(ESWAN)sequence in mild and severe traumatic brain injury (TBI) models of rabbits in diverse phases and investigate their association with neurological severity scale (NSS) scores. Methods Fifty-one New Zealand rabbits, which were randomly divided into control group (n=3) ,mild injured group (n=24) , and severe injured group (n=24) by random digital table method, underwent routine MRI and ESWAN sequence at the time points of baseline, 1 h, 6 h, 12 h, 24 h, 48 h, 72 h, 1 week, and 2 weeks after injury(3 rabbits per subgroup)respectively. Blood phase values in veins of interest were recorded. Observation of behavior characteristics and abnormalities, followed by NSS, was executed post injury, and the correlation between venous blood phase values and NSS scores was statistically analyzed. Two independent-samples t-test was applied to compare venous blood phase values of diverse measured veins in each group separately at every time ponits. One-way ANOVA was used to analyze venous blood phase values varying over time of measured vessels in each injured group. Least significant difference t-test was applied to compare blood phase values within the subgroups with each other at each time point. NSS scores of mild and severe injured groups were compared by two independent-samples t-test. Correlations between venous blood phase values and NSS scores in each vein and group was analyzed with Spearman correlation analysis. Results Blood phase values in veins of interest presented an inclination of descending, which was more obvious in severe injured groups than in the mild. Change of venous blood phase values over time featured continuing reduction in earlier phases which reached to the minimum in 24-48h, and then increased gradually, especially in VMV, VLV, DSS, and MDVB (P<0.05). ICV and DCV also showed similar trend. Values of NSS scores in the two group were (15.5 ± 3.1) and (33.2 ± 6.5) respectively (t=3.543,P=0.001). Blood phase values in most of the measured veins correlated with NSS scores (P<0.05) after 6h post injury, especially significant during 24-72h among all the measured veins(P<0.05). Higher accuracy and sensitivity were presented in the supratentorial or superficial veins compared with subtentorial or profound veins. Conclusions It is feasible to evaluate oxygen saturation of veins after TBI by measuring venous blood phase values on ESWAN images. The method is effective in the assessment of the degree of injury and clinical status, indicating a favorable application prospect.
5.Osteochondral autograft and allograft and tissue engineering materials for repair of articular cartilage injury
Jianlin ZHOU ; Hongsong FANG ; Hao PENG ; Shuang DENG ; Jinqing WENG ; Feng LIU ; Sen CHEN ; Guanjin ZHOU
Chinese Journal of Tissue Engineering Research 2015;(34):5530-5535
BACKGROUND:In recent years, repair of articular cartilage injury has become an important field in basic medical research. Because injured articular cartilage is difficult to repair, the repair of articular cartilage injury has become a difficult hotspot.
6.Biological characteristics of different sources of mesenchymal stem cells
Hongsong FANG ; Jianlin ZHOU ; Hao PENG ; Shuang DENG ; Jinqing WENG ; Feng LIU ; Sen CHEN ; Guanjin ZHOU
Chinese Journal of Tissue Engineering Research 2015;(32):5243-5248
BACKGROUND:Mesenchymal stem cels, underin vivo orin vitro specific induction conditions, can differentiate into the cartilage, muscle, tendons and so on. Clinical trials concerning mesenchymal stem cels mainly include tissue repair (such as bone, cartilage and joint repair) and treatment of heart, liver, spinal cord injury and nervous system diseases.
OBJECTIVE:To compare the biological characteristics of mesenchymal stem cels from different sources.
METHODS: PubMed and CNKI databases were retrieved for articles related to sources of mesenchymal stem cels and biological characteristics of mesenchymal stem cels published from 1987 to 2015. The retrieved articles were summarized and analyzed in the folowing aspects: cel surface marker, proliferation, differentiation, migration, and function, so as to explore the merits and demerits of mesenchymal stem cels from different sources.
RESULTS AND CONCLUSION:A difference in the proliferation ability and surface markers is found between different sources of mesenchymal stem cels. Immunological competence of mesenchymal stem cels from different sources may be correlated with their activation status, species differences, tissue sources and culture conditions, resulting the immunological competence of mesenchymal stem cels from different sources is not exact the same. In-depth understanding of the factors and mechanisms by which influence the migration of mesenchymal stem cels from different sources can enhance the migration ability of different sources of mesenchymal stem cels, and increase their efficiency in wound healing, tissue repair and regeneration treatment.
7.Expression of SODD and P65 in ALL of children and its relationship with chemotherapeutic drugs.
Hongfang, TAO ; Qun, HU ; Jianlin, FANG ; Aiguo, LIU ; Shuangyou, LIU ; Liuqing, ZHANG ; Ying, HU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(3):326-9
The expression of silence of death domains (SODD) and its clinical significance and relationship with phospho-NF-kappaB-p65 proteins in bone marrow cells of childhood acute lymphoblastic leukaemia (ALL) were explored, and the expression of SODD and phospho-NF-kappaB-p65 in Jurkat cells treated with chemotherapeutic drugs was detected in order to find a new chemotherapeutic target. The expression of SODD and phospho-NF-kappaB-p65 proteins in bone marrow cells was detected by immunohistochemistry in 25 children with ALL. The apoptosis rate was measured by Annexin-V-Fluorescence/PI double-labeling flow cytometry and the expression of SODD and phospho-NF-kappaB-p65 proteins determined by Western blotting in the Jurkat cells. It was found that the expression of SODD and active P65 in ALL was significantly higher than that in normal control group (P<0.05). The expression of the SODD and phospho-NF-kappaB-p65 proteins in the high-risk (HR) group was significantly higher than that in the standard-risk (SR) group (P<0.05). The Pearson rank correlation analysis revealed that there was a positive correlation between SODD and phospho-NF-kappaB-p65 expression (P<0.01, r=0.69). VCR could effectively induce the apoptosis of Jurkat cells, and down-regulate the expression of SODD and phospho-NF-kappaB-p65 proteins in a time-dependent manner, but DNR could not down-regulate the expression of SODD effectively. It was concluded that SODD may be closely related to the clinical classification and prognosis of ALL in children. The expression of SODD and phospho-NF-kappaB-p65 had a definite synergistic relationship with the onset and development of ALL. VCR could down-regulate the expression of SODD and inhibit the NF-kappaB activation, which could recover the sensibility of apoptosis in leukemic cells.
8.Therapeutic effects of all trans-retinoitc acid combined with transarterial chemoembolization on Walker-256 hepatoma in rats.
Jianlin, FANG ; Chuansheng, ZHENG ; Hongfang, TAO ; Hui, ZHAO ; Jianzhuang, REN ; Gansheng, FENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(1):113-8
In order to investigate the inhibitory effects of all trans-retinoitc acid (ATRA) on differentiation and apoptosis of Walker-256 hepatocellular carcinoma cells and the therapeutic effects of ATRA combined with transarterial chemoembolization (TACE) on rat Walker-256 transplanted hepatocarcinoma, Walker-256 hepatocarcinoma cell lines were treated with ATRA at different concentrations. After culture for 48 h, the inhibitory rate of cell proliferation was determined by MTT assay; the changes of Fas and Bcl-2 mRNA expression were determined by RT-PCR, and the expression levels of Caspase3 and Caspase8 proteins were detected by Western blot. Twenty-seven Wistar rat models of hepatocarcinoma were set up successfully by implanting Walker-256 cell lines. The tumor volume at the 11th day after implantation (V(preoperation)) was measured by magnetic resonance imaging (MRI). The 27 rats were randomly and equally divided into three groups, and the therapy scheme was performed as follows: group A (ATRA 0.1 mg+mitomycin 0.05 mL+lipiodol 0.05 mL+gelfoam powder 0.025 mg); group B (mitomycin 0.05 mg+lipiodol 0.05 ml+gelfoam 0.025 mg; group C (0.9% NaCl 0.2 mL). After another 11 days, MRI was performed once again to measure the tumor volume (V(postoperation)). The expression of factor and Ki VIII -67 in the tumor tissues was detected by immunohistochemistry. The results showed that ATRA could suppress proliferation of Walker-256 cell lines. After treatment of Walker-256 cell lines with ATRA, the expression of Fas mRNA was significantly up-regulated and the Bcl-2 mRNA was significantly down-regulated by ATRA at the concentration of 10 mumol/L as compared with the control group (P<0.05). After treatment with 10 mumol/L ATRA for 48 h, the Caspase3 and Caspase8 were significantly activated as compared with the control group (P<0.05). Significant difference existed in growth rate among the three groups (P<0.01) and between either two groups (P<0.05). The expression rate of factor VIII and Ki-67 was gradually increased from group A, group B to group C. The study suggests that ATRA could inhibit the proliferation of Walker-256 cells and the effectiveness of the combined therapy (ATRA+TACE) for treating transplanted hepatoma of rats is superior to that of TACE alone.
9.En bloc resection of semi-facet and lamina for thoracic ossification of ligamentum flavum with epidural adhesion
Zhicheng ZHANG ; Tiansheng SUN ; Fang LI ; Dajiang REN ; Kai GUAN ; Guangmin ZHAO ; Jianlin SHAN ; Tianlin WEN
Chinese Journal of Orthopaedics 2010;30(11):1039-1043
Objective To explore the strategy and outcomes of surgical treatment of thoracic ossification of ligamentum flavum(OLF),especially combined with ossification of posterior longitudinal ligament,thoracic kyphosis and epidural adhesion.Methods Fifty-three cases of thoracic OLF from January 2003 to December 2009 were reviewed retrospectively.All patients were treated by the methods of en bloc resection of semi-facet and lamina.All patients were followed up for more than half an year,including 32 males and 21 females,aged from 43 to 73 years(average 54.7 years).The lesions located in upper thoracic for 18 patients,and in thoracolumbar for 35 patients.For multi-level or jumping OLF patients,the responsible levels were determined by combination of images and clinical symptoms.For multi-level OLF with ossification of posterior longitudinal ligament(OPLL)or thoracic kyphosis(>50°),multi-level pedicle screw fixation and correction of kyphosis were performed.For dural adhesion patients,part of cerebrospinal fluid was released with a caudal incision of dural sac resulting in collapse and epidural arachnoid separation.Ossific and adhesion dura mater were removed with integrity of arachnoid.The surgical outcomes were evaluated with preoperative and postoperative thoracic Japanese Orthopaedic Association(JOA)score,Nurick grade and neurologic functional recovery ratio.Results Fifty-three cases were followed up for 6 months to 6 years,with an average of 18 months.The average preoperative JOA score was 4.3±2.3,which significantly increased to 8.3±1.8 after operation.Postoperative neurologic functional recovery rates were 11% to 80%(average 65.8%),including excellent in 18 cases,good in 20,fair in 10,and poor in 5.The excellent or good rate was 71.7%.The mean preoperative Nurick grade was 3.7(2-5 grade)and decreased to 2.3 grade after operation.Conclusion En bloc resection of semi-facet and lamina is a safe and effective method for treatment of thoracic OLF.For the patients with OPLL or kyphosis,pedicle screws fixation and kyphosis correction was beneficial for recovery of neurologic function of thoracic OLF patients.
10.Anatomic basis and application experience of the puncture technique through anterolateral cervical approach
Jianlin SHAN ; Heng JIANG ; Chongwei WANG ; Yang ZHANG ; Fei WANG ; Fang LI
Chinese Journal of Orthopaedics 2013;(1):76-81
Objective To explore the anatomical basis,safty and manipuility of the puncture technique through anterolateral cervical approach.Methods Twenty two embalmed cadavers and 50 patients who underwent operation through anterolateral cervical approach were used to observe the cervical anatomical characteristics.During the cadaver dissection and operation,the following information was observed:the movement of the carotid sheath while the visceral sheath was pushed left,the features of interspace between the two sheaths at each cervical level,the movement of the esophagus and the interfascial space in which the esophagus moved while the trachea was pushed medially,and the movement of the carotid sheath while it was pushed medially and laterally.From January 2007 to December 2011,206 patients diagnosed as cervical discogenic pain or cervical vertigo were treated by radiofrequency using puncture technique through anterolateral cervical approach,including 93 males and 113 females,aged from 22 to 71 years (average,48years).A total of 434 discs were involved.Results Both in cadaver dissection and in operation,the following results were observed:there was no natural interspace between the carotid sheath and visceral sheath below the C4 level,as a result,manual separation was needed to create a interspace between the two sheaths for the puncture; the two layers of the prevertebral fascia could be separated easily; keeping the visceral sheath intact,the esophagus moved following the trachea when the latter was pushed medially,and the movement of the visceral sheath relative to the cervical vertebra occurred between the two layers of prevertebral fascia; the carotid sheath was primarily constituted by continuation of the prevertebral fascia,as a result,the movement of the carotid sheath was limited by its own tension and the prevertebral fascia.A total of 434 cases of puncture to cervical disc were performed smoothly with no complications related to the carotid artery,esophagus and cervical disc.Conclusion The puncture technique through anterolateral cervical approach requires a correct,thorough understanding of the anterior cervical anatomy,based on which this technique is safe and feasible.