1.Study on Cleavage in vitro of HPVll/E2 Gene by Specific Ribozyme
Hua, HOU ; De-zhong, LIU ; Guang-cai, YANG
Virologica Sinica 2001;16(2):146-150
In order to study the pathogenesis of HPVl1 and seek for a therapeutic approach of the disease caused by HPV1l. The HPVl1/E2 644bp was amplified by PCR with HPVll plasmid DNA. pGEM-T Easy was used as vector and a clone pTV-644 was obtained.The inserted DNA sequencing was carried out after selecting and identification. According to the hammerhead structure described by Symon's, the possible secondery cleavage sites on HPVll/E2 mRNA were analyzed and the secondery structure of substrate was predicted by computer and ribozyme, excluding analogous sequence of substrate combined with ribozyme were found in the mRNA.The hammerhead ribozyme of RZ2777 against HPV1l/E2 mRNA was seleted to carry out cleavage reaction in vitro. Results of the experiment showed that 644bp substrate derived from HPVll/E2 can be cleavaed site-specifically by ribozyme in vitro. The cleavage activity showed over 85% by choosing the optimun reaction condition, which was not affected by two cis-ribozymes on both 3′-and 5′-ends released by self cleavage,but two flan-king sequences of target RNA influenced the cleavage activity. Results demonstrated that the ribozyme will become a highly effective and specific therapy against HPVll infection.
2.CCR7 implications of spleen dendritic cells in multiple organ dysfunction syndrome in mice
Guocun HOU ; Jiangyang LU ; Hongwei WANG ; Qian LIU ; Guang TIAN ; Yi YANG ; Shaoran LI
Chinese Journal of Immunology 2009;25(12):1063-1066
Objective:To explore CCR7 expression in splenic dendritic cells and its role in migration and activity of splenic dendritic cells in multiple organ dysfunction syndrome (MODS) in mice.Methods:The MODS model of mice was reproduced by Zymosan injection into peritoneal cavity.The mice were randomly divided into groups of normal,3-6 hours,24-48 hours and 10-12 days post zymosan injection.CD11c and CD205 were analysed by immunohistochemistry;The expression of CD86 and CCR7 of DCs were studied by the flow cytometry analysis.Results:In normal mice,many DC were found at the margin between the red and white pulp.In the 3-6 h and 24-48 h groups,CD86 and CCR7 were strongly up-regulated in the DC,and they distributed in T cells areas.In the 10-12 d group,DC distributed at the margin by the immature form.Conclusion:The CCR7 expression level of DC has close correlations with the migration of DC,CCR7 can be used to evaluate the migration and functional activity of DC in MODS.
3.Decoration reconstruction for finger defect in emergency
Ji-Hui JU ; Rui-Xing HOU ; Yue-Fei LIU ; Qiang ZHAO ; Lei LI ; Guang-Zhe JIN ;
Chinese Journal of Microsurgery 2006;0(06):-
Objective To discuss methods of decoration reconstruction for finger defect in emergency and to observe the elinical effects.Methods Of the 41 cases of finger injuries of different degrees,15 were repaired with part of the skin flaps of the big toenails or skin flaps of the second toenalis,8 were repaired with part of the skin flaps of the big toenails,7 were reeonstructed with the second tiptoes,11 were repaired with the abdominal skin flaps of the big toes or lateral flaps of the second toes.Results All the 41 fingers sur- vived.One skin flap of the big toe was somewhat swelling and a decorating operation was performed.The 4~18 months of follow-up visitation of the rest cases revealed good function and shapes.No obvious functional ab- norality was found in the donating feet.Conclusion Various kinds of decoration reeonstruetion for finger defects are available to recover the hand shape and function as much as possible.
4.Study of biomechanical properties of mucosa scars after cleft palate surgery.
Min HOU ; Guang-Yu SHI ; Wei QIU ; Lan-Cheng ZHANG ; Tian-Ping YU ; Chun-Ming LIU
Chinese Journal of Plastic Surgery 2013;29(6):453-456
OBJECTIVETo explore biomechanical properties and stress-strain of mucosa scars after cleft palate surgery.
METHODSAfter the model of mucosa scars was made, the mucosa scars and normal mucosa were excised and examined immediately by tensionometry.
RESULTSThe mucosa scars after cleft palate surgery were compared with normal mucosa. The Poisson's ratio of mucosa scars and normal mucosa was 0.5 and 0.49, respectively, showing no significant difference between the two groups. The ultimate Young's modulus of mucosa scars was about 24.22 MPa, however, it declined to 3.32 Mpa in normal mucosa.
CONCLUSIONSThe mucosa scars after cleft palate surgery are biomechanically weaker than normal mucosa. It can be used for further research, such as maxillary orthognathic surgery, distraction osteogenesis, and orthodontic treatment.
Biomechanical Phenomena ; Cicatrix ; physiopathology ; Cleft Palate ; surgery ; Humans ; Mouth Mucosa ; physiopathology ; surgery ; Osteogenesis, Distraction ; Osteotomy, Le Fort
6.Effects of mycophenolate mofetil, valsartan and their combined therapy on preventing podocyte loss in early stage of diabetic nephropathy in rats.
Yan ZHANG ; Bing CHEN ; Xiang-hua HOU ; Guang-ju GUAN ; Gang LIU ; Hai-ying LIU ; Xue-gang LI
Chinese Medical Journal 2007;120(11):988-995
BACKGROUNDPodocyte has inflammatory role in the development of diabetic nephropathy (DN). Mycophenolate mofetil (MMF), an anti-inflammatory agent, can suppress macrophage infiltration and reduce renal injury in streptozotocin-induced diabetic rats. Angiotensin II receptor blocker (ARB), another renal protecting agent, can decrease podocyte loss in DN. In this study, we detected the expression levels of monocyte chemoattractant protein-1 (MCP-1) and nephrin to evaluate podocyte's role in inflammatory reaction in DN, observe and compare the effect of MMF alone and in combination with valsartan, on preventing podocyte loss in streptozotocin (STZ) induced diabetic rats.
METHODSDiabetic model was constructed in uninephrectomized male Wistar rats by single peritoneal injection of STZ (65 mg/kg). The successfully induced diabetic rats were randomly divided into four groups: diabetes without treatment group (DM), valsartan treated group (DMV), MMF treated group (DMM), and combined therapy group (DMVM). Normal rats of the same sibling were chosen as control (NC). At the end of the 8th week, serum biochemistry, 24-hour urinary protein (UP) and the ratio of kidney weight/body weight (RWK/B) were measured. The rats were sacrificed for the observation of renal histomorphology through light and electron microscope. Nephrin, desmin and MCP-1 levels were detected by semi-quantitative immunohistochemical assays. Real-time quantitative PCR was used to detect the mRNA levels of nephrin and MCP-1.
RESULTSCompared with group NC, serum glucose level, 24-hour UP and RWK/B in group DM were significantly higher (P < 0.01), and the nephrin mRNA level in DM group was significantly lower (P < 0.05). The nephrin mRNA expression levels in group DMV, DMM and DMVM were all higher than that of DM group (P < 0.05) and no significant differences were found among the three treatment groups (P > 0.05). Treatment with MMF, valsartan or their combination could significantly decrease the 24-hour UP and RWK/B, and suppress glomerulosclerosis and interstitial fibrotic lesions in diabetic rats. In diabetic rats, the high expressions of desmin and MCP-1 in kidney were suppressed by valsartan, MMF or their combination.
CONCLUSIONSPodocytes are involved in the inflammatory reaction of diabetic rats. MMF could suppress MCP-1 and desmin expression, enhance nephrin expression, and attenuate proteinuria in diabetic rats. The combined therapy of valsartan and MMF did not show any superiority over monotherapies on renal protection. MMF may have renoprotective effect in early stages of diabetic nephropathy through preventing podocytes loss and anti-inflammatory activity.
Animals ; Chemokine CCL2 ; analysis ; Desmin ; analysis ; Diabetic Nephropathies ; drug therapy ; pathology ; Drug Therapy, Combination ; Immunohistochemistry ; Male ; Membrane Proteins ; analysis ; Mycophenolic Acid ; administration & dosage ; analogs & derivatives ; therapeutic use ; Podocytes ; drug effects ; pathology ; Rats ; Rats, Wistar ; Tetrazoles ; administration & dosage ; therapeutic use ; Valine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Valsartan
7.Biomechanical characteristics of hip prosthesis in hip arthroplasty treating elderly patients with Evans I-III intertrochanteric fracture of femur.
Wen-guang LIU ; Sheng-hou LIU ; Qing-feng YIN ; Shi-peng XIAO ; Shao-jin WANG
Acta Academiae Medicinae Sinicae 2013;35(1):108-111
OBJECTIVETo investigate the feasibility of hip arthroplasty in the treatment of elderly patients with Evans I-III intertrochanteric fracture of femur by analyzing its biomechanics characters.
METHODSWe solved the CT digital image files with the graphics processing software Mimics at DICOM 3.0 standard, and reconstructed the three-dimensional entity of femur with CAD modeling software Unigraphics. Then the fracture line was defined in the model as the line between the tip of greater trochanter and inferior margin of small trochanter, above which the upper bone was removed. Afterwards the two prosthesises with different stem lengths (120 mm and 170 mm) were implanted into the fracture model respectively as hip arthroplasty with 3 mm bone cement layer between prosthesis and femur, and the bone defect was repatched with 5 mm bone cement layer. A three-dimensional finite element model was established with finite element analysis software ABAQUS 6.5. We formulated different material parameters under the stress condition standing with single leg to build the stress distribution map of the femur prosthesis, and took 5 loci of region of stress concentration to calculate the mean value of stress.
RESULTSThe stress distribution maps of the short and long stem length prothesises were similar. And there were two areas of stress concentration, including the upper portion and the lower portion close to the joint of the prosthesis stem, and the stress concentration in the junction part was obviously between the lower portion and the upper area of the small trachanter. The stress reached the first concentration area at the junction and then gradually reached the second concentration area at the interior terminal of the stem. While the stress gradually increased along the lateral prosthesis stem, and reached the stress concentration area at the end.
CONCLUSIONSThe stress distribution maps in the femur prosthesises are similar between hip arthroplasty in the treatment of intertrochanteric fracture of femur and the traditional hip arthroplasty surgery. The peak stress values are higher in the long stem prosthesis in the treatment of intertrochanteric fracture of femur than the short type, while they are under the rupture value of the metal.
Aged ; Arthroplasty, Replacement, Hip ; instrumentation ; methods ; Biomechanical Phenomena ; Bone Cements ; Computer Simulation ; Female ; Finite Element Analysis ; Hip Fractures ; surgery ; Hip Prosthesis ; Humans ; Image Processing, Computer-Assisted ; Software ; Stress, Mechanical
8.Biomechanical supporting effect of tantalum rods for the femoral head with various sized lesions: a finite-element analysis.
Wen-guang LIU ; Shao-jin WANG ; Qing-feng YIN ; Sheng-hou LIU ; Yan-jin GUAN
Chinese Medical Journal 2012;125(22):4061-4065
BACKGROUNDFeatures of necrotic lesions and various interventions could affect the biomechanics of the femoral head. A three-dimensional finite-element analysis was designed to demonstrate necrotic femoral head stress changes with various sizes of necrotic lesions, and evaluate the effect of tantalum rods on preventing femoral head cracking.
METHODSFemoral computed tomography scans were used to build a normal three-dimensional finite-element femoral head model in a computer. Based on the normal model, necrotic models of different lesion diameters (15 mm, 20 mm and 30 mm) were created, as were the repaired models with tantalum rods for each diameter. After a series of meshing and force loading, the von Mises stress distributions, simulating single-legged stance, and stresses on specific points under loaded conditions were determined for each model.
RESULTSDeep exploration into the burdened area of the femoral head indicated that higher stresses to the femoral head were observed with a larger necrotic lesion; the largest stress concentration, 91.3 MPa, was found on the femoral head with a lesion diameter of 30 mm. By contrast, topical stress on the surface of the necrotic regions was lowered following implantation of a tantalum rod, and the changes in stress were significant in models with lesions of 15 mm and 30 mm in diameter, with the best biomechanical benefit from the tantalum rod found with a lesion diameter of 15 mm.
CONCLUSIONSFemoral heads with larger necrotic lesions usually have a higher stress concentration and a higher risk of collapse. Various sized lesions on the femoral head can benefit from the mechanical support offered by the implantation of a tantalum rod; however, femoral heads with smaller sized lesions may benefit more. A thorough evaluation of the lesion size should be conducted prior to the use of tantalum rod implants in the treatment of femoral head necrosis.
Femur Head ; physiology ; Femur Head Necrosis ; physiopathology ; Finite Element Analysis ; Humans ; Stress, Mechanical
9.Impacts on vertebral arterial blood flow of cervical spondylosis of vertebral artery type treated by abdominal acupuncture.
Zhou AI ; Guang-De LIU ; Xiao-Cui XIONG ; Feng-Bin HOU
Chinese Acupuncture & Moxibustion 2013;33(7):601-604
OBJECTIVETo explore the therapeutic effect and mechanism of abdominal acupuncture for cervical spondylosis of vertebral artery type.
METHODSThirty cases of cervical vertigo, in which the color ultrasonography indicated vertebral arterial blood insufficiency, were treated with abdominal acupuncture therapy. The points were Zhongwan (CV 12), Qihai (CV 6), Guanyuan (CV 4), Xiawan (CV 10), Shangqu (KI 17) and Huaroumen (ST 24). The treatment was given once every day and five continuous treatments made one session. Separately, before treatment and in the 1st and 5th treatments, the cervical vertigo symptom and functional assessment scales were adopted for scoring. Simultaneously, the color ultrasonography was applied to observe the blood flow changes of the bilateral cervical arteries.
RESULTSExcept the score for headache in the 1st treatment, the scores in cervical vertigo and function assessment scale in the 1st and 5th treatments were all improved significantly in 30 patients as compared with those before treatment (P < 0.01, P < 0.05). In the 1st and 5th treatments, on the affected side, the vertebral artery diameter, mean velocity and blood flow per minute were all improved significantly as compared with those before treatment (all P < 0.01). In one session treatment, the total effective rate was 100.0% (30/30) and the curative rate was 60.0% (18/30).
CONCLUSIONAbdominal acupuncture therapy not only relieves the clinical symptoms, but also improves vertebral arterial blood supply for the patients of cervical spondylosis of vertebral artery type.
Abdomen ; Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Regional Blood Flow ; Spondylosis ; physiopathology ; therapy ; Treatment Outcome ; Vertebral Artery ; physiopathology ; Young Adult
10.Determination of platelet-associate autoantibodies against platelet-specific receptors by cytometric bead array and its clinical application in idiopathic thrombocytopenic purpura.
Xin-Guang LIU ; Yan-Ming WANG ; Ming HOU ; Ti ZHANG ; Yuan-Yuan ZHU ; Jun PENG
Chinese Journal of Hematology 2008;29(3):175-178
OBJECTIVETo establish a method for detecting platelet-associate autoantibodies against platelet-specific receptors using cytometric bead array, and compare the clinical usefulness of this method and modified indirect monoclonal antibody immobilization of platelet antigen technique (MAIPA) in the differential diagnosis of idiopathic thrombocytopenic purpura (ITP) from non-immune thrombocytopenic purpura (non-ITP).
METHODSThe microbeads were coated with monoclonal antibodies against glycoprotein IIb/IIIa (CD41a), platelets were isolated from blood samples, then platelet lysate was incubated with the coated microbeads, and R-Phycoerythrin-conjugated goat-antihuman IgG polyclonal antibodies, finally analyzed with flow cytometry. GP IIb/IIIa autoantibodies in sample plasma were measured by modified indirect MAIPA at the same time.
RESULTSThe individual fluorescence level was calculated as the ratio to the three controls. The mean ratios were 3.36 (range 0.84 - 22.94) in the ITP group, 1.16 (range 0.67 - 5.59) in the non-ITP patient and 1.08 (range 0.72 - 1.76) in the healthy controls. There was a highly significant difference (P <0.01) between the ITP patients and either the non-ITP patients or the normal controls. If the up limit of healthy controls was set as cutoff value, ratio of greater than 1.76 was considered positive. Cytometric bead array had a sensitivity of 71.43%, a specificity of 94.28% and a positive predictive value of 95.24% for the diagnosis of ITP, the sensitivity being higher than that of modified indirect MAIPA' s (5179%) (chi2 = 4.57, P <0.05). The ROC curve showed the discriminative validity of cytometric bead array was 0.916.
CONCLUSIONFlow cytometric bead method for detection of platelet-associate autoantibodies against platelet-specific receptors is a more rapid, better reproducibility and higher sensitivity than modified MAIPA, and has a potential value in promoting the diagnosis of ITP and guiding treatment.
Adult ; Autoantibodies ; blood ; Female ; Flow Cytometry ; methods ; Humans ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; immunology ; Purpura, Thrombocytopenic, Idiopathic ; blood ; diagnosis ; immunology