1.c-Cbl Mediated Ubiquitination and Degradation of hSef
Yongming REN ; Zhili RONG ; Zhiyong LI ; Long CHENG ; Yinghua LI ; Yinyin WANG ; Fangli REN ; M.irwin DAVID ; Zhijie CHANG
Progress in Biochemistry and Biophysics 2008;35(1):43-49
Sef (similar expression to fgf genes) was identified as a feedback antagonist of FGF signaling in zerbrafish, mouse and human. Sefhas been reported to function in different ways, however the regulation of Sef stability remains unknown. The possible role of c-Cbl in the regulation of Sef protein degradation was investigated. Results from coimmunoprecipitation and immunostaining assays reveal that hSef colocalizes and interacts with c-Cbl. Data suggest that the interaction between hSef and c-Cbl results in the ubiquitination and subsequent degradation of the hSef protein. It was proposed that c-Cbl may serve as a modulator to regulate Sef protein stability during FGF signal transduction.
2.Total arch replacement combined with stented elephant trunk implantation for Stanford type A aorticdissection
Zhiping LIU ; Xianming ZHU ; Shuzhen LI ; Yulong ZHANG ; Jian WNAG ; Junxiao GUO ; Rong GAO ; Jie REN ; Long ZHAO ; Min WANG
Journal of Chinese Physician 2011;13(8):1060-1062,1066
ObjectiveTo improve the long term outcomes of the surgery for Stafford type A aorticdissection, we performed ascending aorta and total aortic arch replacement combined with transaorticstented graft implantation into the descending aorta for acute type A aortic dissection.MethodsFrom May 2005 to February 2011,36 consecutive patients with acute Stanford type A aorticdissection underwent this procedure.Right axillary artery cannulation was routinely used forcardiopulmonary bypass and selected cerebral perfusion.The stented elephant trunk was implanted through the aortic arch under hypothermic circulatory arrest.The stented elephant trunk was a 10 cm long selfexpandable graft.34 patients were followed up for 2 ~36 months.ResultsCardiopulmonary bypass time was (160 ± 31)min, average cross clamp time was (101 ±26)min, and average selective cerebral perfusion and lower body arrest time was (31 ± 16)min.The in-hospital mortality was 5.5% (2/36).One patient died of multi-organ failure postoperatively and another died of cerebral infarction 9 day after surgery.No one suffered from spinal cord injury perioperatively.There was no late death during follow up.ConclusionsAscendingaorta and total aortic arch replacement combined with transaortic stented graft implantation into the descending aorta is an effective way in closing the residual false lumen of the descending aorta and might contribute to better long term outcomes of type A aortic dissection.
3.Expression of ki-67 and intimal hyperplasia of the irradiated human umbilical artery incorporated with nofloxacin and silver grafts
Zhiping LIU ; Jian WANG ; Xianming ZHU ; Yulong ZHANG ; Shuzhen LI ; Long ZHOU ; Junxiao GUO ; Jie REN ; Rong GAO ; Nengyong QIU
Journal of Chinese Physician 2009;11(5):599-602
Objective To investigate the expression of ki-67 and the development of the intimal hyperplasia(IH) of the irradiated human umbilical artery incorperated with nofloxacin and silver(IHUAINS) grafts into the carotid arteries of the rabbit. Methods Twenty IHUAINSs were sterilely produced. Thirty rabbit were performed bilateral carotid bypass grafting. The IHUAINS(experimental group)and the left carotid arteries (control group) were implanted in the left and right carotid arteries respectively. Graft patency was checked at the 2nd and 6th week after implantation, and the grafts were studied with standard histological techniques and immunohistochemieal method for meas-urement of intimal thickness and the expression of ki-67. Results The total patency rate of the grafts was 89.6%. Light microscopic exami-nation of the grafts revealed intimal and media proliferation, cellular in-filtration. The endothelial cells covered the vascular lumen. There was no significant difference of the intimal thickness between two groups at the 2nd week after grafting (P>0.05). The intimal thickness of the experimental group was larger than that in control group at the 6th week after implantation without statistical significance (P>0.05). At the same time, immunocytochemical analysis showed that the expression of ki-67 in the experimental group was stronger than that in control group without statistical significance(P>0.05). Conclusion The IH of the IHUAINS was larger than that of the autologous artery, but there is no difference between these two groups. Thus, IHUAINS may be an ideal graft in the field of coronary surgery.
4.Transcription factor Sp1 in regulation of transcription of VEGF in hypoxia-induced hepatocellular cells
rong Yu WANG ; yan Yan LI ; long Ren LU
Military Medical Sciences 2017;41(7):572-575
Objective To investigate the role of Sp1 in the regulation of transcription of vascular endothelial growth factor ( VEGF ) in hypoxia-induced HepG2 cells.Methods Quantitative real-time polymerase chain reaction and immunoblot assay were used to examine the effect of hypoxia on Sp1 protein and VEGF mRNA expression .Mithramycin A, the selective inhibitor of Sp1 and knock-out Sp1 gene with siRNA were used to examine the effect on VEGF mRNA expression induced by hypoxia in HepG2 cells.Results Hypoxia induced Sp1 protein and VEGF mRNA expression in HepG2 cells.Mithramycin A produced a concentration-dependent decrease of hypoxia-induced VEGF mRNA expression . After inhibition of Sp1 RNAi, VEGF mRNA expression was significantly repressed in HepG 2 cells.Conclusion Hypoxia can increase the expression of Sp1 protein and VEGF mRNA in HepG2 cells induced by hypoxia.The transcription of VEGF is regulated by Sp1 in hypoxia-induced HepG2 cells.
5.Posterior vertebral column resection osteotomy combined with step correction in treatment of stiff angular kyphosis:a biomechanical analysis
Huasong MA ; Xiaoping WANG ; Rong TAN ; Zhiming CHEN ; Ming LU ; Wei YUAN ; Qiming XU ; Dongyun REN ; Wei MA ; Long LI ; Jing ZHANG ; Rui ZHENG ; Xin XIN
Chinese Journal of Tissue Engineering Research 2014;(35):5647-5653
BACKGROUND:Severe spinal angular kyphosis aggravated spinal cord injury and early degeneration, even caused incomplete paralysis or complete paralysis. Surgical treatment is the only solving approaches and method, but it is difficult, exhibits high risk, and easily affects postoperative complications. OBJECTIVE:To analyze the science and effectiveness of posterior vertebral column resection osteotomy combined with step correction in treatment of stiff angular kyphosis based on biomechanical principle. METHODS:A total of 90 cases underwent posterior vertebral column resection osteotomy combined with bilateral pedicle screw spinal cord gradual y shortening echelon tight closure and orthopedic fixation were selected, including 37 males and 52 females, at the average age of 47 years. Kyphotic angle, spinal sagittal imbalance, trunk side offset rate, operation time, intraoperative blood loss were compared and analyzed before and after treatment. RESULTS AND CONCLUSION:The kyphotic angles were 31°-138° (averagely 90.1°) preoperatively and 10°-90° (averagely 41.6°) postoperatively, with an improvement rate of 65%. The distance from C 7 plumb line to the S 1 upper edge was averagely 5.2 mm, with a correction rate of 73%. Intraoperative blood loss was 1 200-6 000 mL, averagely 2 089 mL. Operation time was 212-470 minutes, averagely 326 minutes. The patients were fol owed up for 20 to 35 months after the surgery. Osteotomy segments had achieved bone fusion in al patients, and no complications of spinal cord injury or orthopedic angle loss appeared. These data verified that in the accordance with cellbiomechanics and spinal biomechanical principles, bilateral pedicle screw spinal cord gradual y shortening echelon tight closure and orthopedic fixation protected utmost spinal cord cells against injury in the correction of thoracolumbar angular kyphosis. There is sufficient basis for cellphysiology and it accorded biomechanical and physiological characteristics. During the surgery, we should pay attention to protection and release of nerve root and avoid postoperative corresponding nerve root irritation. Ful fusion ensures kyphosis correction and avoids spine lateral offset, is an effective safeguard for the recovery of spinal function and postoperative orthopedic effect.
6.Bilateral pedicle screw and echelon tight closure spinal cord technique combined with implant fixations for correcting stiff spinal angular kyphosis
Huasong MA ; Xiaoping WANG ; Rong TAN ; Zhiming CHEN ; Ming LU ; Wei YUAN ; Qiming XU ; Dongyun REN ; Wei MA ; Long LI ; Jiajian WU ; Jing ZHANG ; Rui ZHENG ; Shen XIN
Chinese Journal of Tissue Engineering Research 2014;(31):4992-4997
BACKGROUND:The treatment difficulties of thoracolumbar angular kyphosis surgery are:low correction rate, hard to rebuild sagittal plane, easily induce neurological complications, postoperative loss of balance, high incidence of pseudarthrosis and postoperative loss of correction degree.
OBJECTIVE:To explore the safety and efficacy of modified posterior vertebral column resection osteotomy and bilateral pedicle screw combined with echelon tight closure spinal cord technique and implant fixation for severe spinal angular kyphosis.
METHODS:A total of 87 severe spinal angular kyphosis patients, 36 males and 51 females, who were treated in the Department of Orthopedics, the 306 Hospital of Chinese PLA from January 2006 to December 2013, were enrol ed in this study. They underwent posterior vertebral column resection, bilateral pedicle screw combined with echelon tight closure spinal cord, and implant fixation. Kyphosis, spinal sagittal imbalance, offset rate towards trunk side, operation time and intraoperative blood loss were observed before and after treatment.
RESULTS AND CONCLUSION:The preoperative average kyphosis was 90.1° (31°-138°). The postoperative average kyphosis was 27.9° (15°-57°). The improvement rate was 76%. The improvement rate of trunk sagittal offset was 76%. Intraoperative blood loss was 800-3 000 mL, and average blood loss was 2 300 mL. The operation time was 5-7 hours, averagely 5.9 hours. Before treatment, two patients affected neurologic symptoms in double lower extremity, and their Frankel classification was grade C and became grade E after treatment. Al patients were fol owed up for 9-57 months. Bony fusion was achieved in al patients. No complications of spinal cord injury appeared, and no orthopedic angle missing occurred. These results indicate that during posterior vertebral column resection for treating severe angular stiffness of the thoracic kyphosis, blood vessels could be maintained greatly. Blood vessel injury-induced ischemic changes in spinal cord and ischemic reperfusion injury could be avoided. To reduce hemorrhage and to keep effective blood volume in patients with low body mass are effective for early recovery after treatment. Bilateral pedicle screw combined with echelon tight closure spinal cord technique greatly protected spinal cord cells against injury. We should pay attention to the protection and loose of nerve root to avoid postoperative nerve root irritation. Sufficient bone fusion ensures kyphosis correction, avoids spine lateral offset, and plays a key role in spinal function and postoperative orthopedic effect.
7.Preparation and in vitro evaluations of topically applied capsaicin transfersomes.
Xiao-ying LONG ; Jia-bo LUO ; Li-ren LI ; Dan LIN ; Hui-si RONG ; Wei-min HUANG
China Journal of Chinese Materia Medica 2006;31(12):981-984
OBJECTIVECapsaicin transfersomes were prepared and its quality specifications were evaluated.
METHODCapsaicin transfersomes were prepared by high shear dispersing machine and evaluated on the entrapment efficiency, drugs release rate and in vitro skin permeation.
RESULTCapsaicin transfersomes is composed of single unilamellar vesicles, with average size of 150.6 nm. Capsaicin entrapment efficiency achieved 96.7% while concentration of lecithin used was 8%. cumulative release amount of capsaicin was in direct proportion to the ethanol concentration in the medium. The in vitro rate cumulative penetration rate of capsaicin was higher in transfersomes than in cream and suspension in rats. Adomen skin cumulative penetration rate in vitro of capsaicin transfersomes in mouse was significantly higher than that from rat and men. In the same way,cumulative penetration rate in vitro of capsaicin transfersomes through abdomen skin epidermal membrance was significantly higher than that with derma and full skin in men.
CONCLUSIONEntrapment efficiency of capsaicin transfersomes reached 96.7%, meeting the criterion of China pharmacopia( > 80%), skin penetration of capsaicin was enhanced by a capsaicin transfersomes preparation and was affected by diverse characters and levels of skin.
Administration, Cutaneous ; Analgesics, Non-Narcotic ; administration & dosage ; pharmacokinetics ; Animals ; Capsaicin ; administration & dosage ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; methods ; Humans ; In Vitro Techniques ; Male ; Mice ; Particle Size ; Phosphatidylcholines ; administration & dosage ; chemistry ; pharmacology ; Rats ; Skin ; drug effects ; metabolism ; Skin Absorption ; drug effects
8.An optimized method for extracting corosolic acid from loquat leaves.
Jian-rong HE ; Qian LIU ; Zhi-long BAI ; Ren-quan HUANG ; Zeng-lu WANG
Journal of Southern Medical University 2010;30(11):2533-2535
OBJECTIVETo extract corosolic acid from loquat leaves for medical use.
METHODSLoquat leaves were boiled in water to remove the water-soluble substances followed by 3 cycles of extraction with 25% aqueous methanol for 30 min and then by 95% aqueous methanol for 1 h at 80 degrees celsius;. After cooling at room temperature and filtration, the extract was treated with activated carbon to remove chlorophyll, and the liquid was filtered and concentrated to allow precipitation. The sediment was washed to obtain the total crude triterpene acid, which was further dissolved with methanol and purified by high-performance liquid chromatography (HPLC). The fractions including corosolic acid were collected, concentrated with vacuum distillation, and dried to obtain corosolic acid product, which was analyzed with HPLC.
RESULTSHPLC analysis of the extracts showed that the percentages of corosolic acid were 4.66%, 2.42%, and 24.18% in crude corosolic acid extracted with methanol, boiling water, and 95% aqueous methanol, respectively. After purification with HPLC, the purity of corosolic acid in the product exceeded over 80%.
CONCLUSIONThe optimal extraction method, which is convenient and cost-effective, is established for extracting corosolic acid from loquat leaves for medical use.
Chromatography, High Pressure Liquid ; methods ; Eriobotrya ; chemistry ; Plant Leaves ; chemistry ; Technology, Pharmaceutical ; Triterpenes ; isolation & purification
9.Repair of rabbit tendon by autologous bone marrow mesenchymal stem cells.
Jian-hong LONG ; Min QI ; Xiao-yuan HUANG ; Shao-rong LEI ; Li-cheng REN
Chinese Journal of Burns 2005;21(3):210-212
OBJECTIVETo investigate the effect of rabbit bone marrow mesenchymal stem cells (MSCs) as seed cells for the repair of tendon defect.
METHODSThe MSCs were isolated, amplified and identified by detection of surface protein CD44 mRNA. A 3 cm long defect was made in the Achilles tendon of the rabbit. The rabbits were divided into experimental (E) and control (C) groups. The autologous MSCs were implanted into a collagen-polyglycolic acid (PGA) scaffold to form a tissue-engineered tendon, which was then transplanted to bridge the defect in the E group, while only collagen-PGA was transplanted to bridge the defect in the C group. The transplanted tendon was observed grossly and microscopically at 4, 8, 12 weeks after operation.
RESULTSThe cultured MSCs exhibited positive staining of CD44 on 11 days after in vitro culture. A tendon-like tissue could be discerned at the operation site in the E group 4 weeks after operation. Tendon-like cells similar to normal tendon tissue, being axially arranged in collagen matching the mechanical direction, with uniform morphology could be seen in E group 12 weeks after operation. The newly regenerated tissue in C group adhered to the adjacent tissue and was smaller than that in E group. The collagen fibers in the regenerated tissue were loose with reticular and filiform structure, and the cells were arranged disorderly 12 weeks after the transplantation.
CONCLUSIONIt is feasible to repair the tendon defect with autologous MSCs as seed cells.
Achilles Tendon ; injuries ; Animals ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Mesenchymal Stem Cell Transplantation ; Rabbits ; Tendon Injuries ; surgery ; Tissue Engineering ; methods
10.Analysis of impurity in caderofloxacin.
Ren-ling MA ; Hong-hua ZHOU ; Rong LIU ; Er-hua WANG ; Long-sheng SHENG
Acta Pharmaceutica Sinica 2003;38(12):950-952
AIMTo analyse the main impurity of caderofloxacin.
METHODSThe impurity of caderofloxacin was analysed and determinated by RP-HPLC/ESI/MS with a Zorbax SB-C18 (150 mm x 4.6 mm ID, 5 microns) column. The mobile phase was acetonitrile-0.5% acetic acid solution (17:83). A compound was synthesized: 1-cyclopropyl-8-(difluoromethoxy)-6-fluoro-1, 4-dihydro-7-(1-piperazinyl)-4-oxo-3-quinoline carboxylic acid (DMCA). Its HPLC chromatogram, UV and MS spectrum were compared with those of the impurity in caderofloxacin.
RESULTSThe molecular weight of the impurity was 14 less than that of caderofloxacin. It means the impurity was a CH2-group less than caderoflixacin. The tR, UV and MS of DMCA were the same as those of the impurity in caderofloxacin.
CONCLUSIONBased on the tR (HPLC), UV and MS, the impurity of caderofloxacin is confirmed as DMCA.
Anti-Infective Agents ; chemistry ; Carboxylic Acids ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Contamination ; Fluoroquinolones ; chemistry ; Molecular Structure ; Piperazines ; chemistry ; Quinolines ; analysis ; chemistry ; Spectrometry, Mass, Electrospray Ionization