BACKGROUND:Different methods and biomaterials have been applied in animal experiments and clinical practice to prevent the formation of epidural scars,Biodegradable and sticky semi-fluid gels are the most often used material.Salvia miltforrhiza radix (SMR) and carbomer have been clinically confirmed to be the safe and effective drugs and gel agents. OBJECTIVE:To observe the effect of SMR-gel on preventing epidural adhesion after laminectomy.DESIGN:A complete randomized grouping design, a controlled experiment. SETTING:Department of Orthopaedics,Shenzhen People's Hospital (Second Clinical College of Jinan University). MATERIALS:Thirty-six healthy pure New Zealand rabbits were used,either male of female,clean degree,2-3 years of age. They were randomly divided into four groups with 9 rabbits in each group:blank control group,gel contro group, HA group and SMR-gel group. Carbomer934 powder (Shanghai People's Pharmaceutical Factory, batch number: 20000510) , hyaluronic acid (HA) [Shandong Bausch & Lomb Freda Pharaceutical, Co., Ltd.,No.H10960136,2 mL (20 mg)].METHODS:The experiments were carried out in the animal laboratory of Shenzhen People's Hospital from April 2002 to August 2003.①Preparing SMR-gel:SMR was prepared into extract powder.Carbomer934 powder was added by water for dissolving and swelling and stayed overnight,then SMR-gel was prepared by dipping with triethanolamine,adding with SMR extract powder (2 g),then adding with purified water till 100.0 g and stirring uniformly.②The rabbits were anesthetized. and the lamina of vertebra was totally resected at L3 and L6 (reserving superior and Inferior articular processes).then defects of 10 mm×5 mm were made to expose the dura mater.The vertebral defects were added with 1 mL carbomer gel, 1 mL HA (20 g/L) and 1 mL SMR-gel in the gel control group,HA group and SMR-gel group respectively.whereas nothing was added in the blank control group.③Gross samples:Three rabbits were killed 4,6 and 8 weeks postoperatively in each group.vertebraI ventral fascia were stripped to remove the spinal segments (L3,L6) for operation completely,and totally 24 samples for each time.One sample was selected in each group 4 weeks postoperatively. and the samples were observed under H-600 transmission electron microscope (Hitachi). ④The adhesion compactness of scar tissue with dura mater was evaluated in the 24 samples of the 4 groups at 8 weeks postoperatively:There were 4 grades:No obvious adhesion between dural sac and scar tissue for grade O:Extensive and compact adhesion between dural sac and scar tissue. impossible blunt dissection between dural sac and scar tissue.incomplete dural sac after sharp dissection for grade Ⅲ.Each spinal segment was cut into 4 parts equally,and all were prepared into sections and stained,then the thickness of epidural scar was determined with Tiger2000 image analyzer. ⑤The rank sum test was used in the scar adhesion compactness grading evaluated with naked eyes,whereas analysis of variance.and two-two comparison were used in analyzing the thickness of epidural scar.P＜0.05 was considered as significant difference.MAIN OUTCOME MEASURES:①Results of gross scar adhesion compactness grading at 8 weeks and comparison of the thickness of epidural scar at 4.6 and 8 weeks;②Results of gross observation,histological examination and ultrastructure.RESULTS: All the 36 rabbits were involved in the analysis of results. ①Results of gross observation and pathohistological examination:There was compact adhesion at each time point in the blank control group,part adhesion in the gel control group and HA group, and no obvious adhesion in the SMR-gel group.②Results of quantitative analysis:The rabbits with lower scores of scar adhesion compactness grading In the blank control group,gel control group and HA group were obviously fewer than those in the SMR-gel group (W=45-52,P＜0.05-0.01).The scar thickness at 4 and 8 weeks in the SMR-gel group was obviously less than that in the other 3 groups(F=128.657,152.246,80.891,P＜0.01).③Results of observation under transmission electron microscope:The proliferation of fibroblasts at 4 week was active in the blank control group,gel control group and HA group,but inactive in the SMR-gel group.CONCLUSION:①SMR can inhibit the fibroblasts to proliferate,differentiate and synthetize into secretory collagens,and then inhibit the formation of epidural scar adhesion.②HA can be absorbed by organs very early,which reduces its role in preventing adhesion.Whereas carbomer gel can stay longer, and it plays a role in inhibiting and blocking adhesion in the whole process of wound repairing.

BACKGROUND: During recent years, mononucleotide polymorphism of some genes is possibly related to affectability of osteoarthritis (OA). However, previous researches mainly compare the gene expression of synoviocytes between OA and rheumatoid arthritis (RhA); therefore, the correlation of gene expression between synovioblast and fibroblast in other tissues should be further studied as compared with OA.OBJ ECTIVE: To observe the differences of gene expression between OA synovioblast and skin fibroblast.DESIGN: Observational contrast analysis.SETTING: People's Hospital of Shenzhen City.PARTICIPANTS: Synovium tissue was derived from OA patients who received replacement of knee joint in the Department of Orthopaedics, People's Hospital of Shenzhen City. All OA patients met the diagnostic criteria of osteoarthritis established by American College of Rheumatology in 1995. Three patients including 1 male and 2 females aged more than 65 years old and they did not have cardiac and pulmonary disease and diabetes mellitus. Three male normal volunteers who aged 25 to 35 years did not have rheumatic disease, osteoarthritis and dermatosis. All subjects provided a confirmed consent. The main reagents were detailed as follows: RPMI1640 culture medium, fetal bovine serum and TRIZOL agent (Invitrogen Life Technologies Company, USA); pGEM-T pUC (Progema Company, USA);Display PROFILE-BASIC and Display PROFILE Probe kits (Qbiogen Company, USA).METHODS: The experiment was carried out in People's Hospital of Shenzhen City from January to June 2005. Synovium of OA patients were treated with primary culture to obtain synovioblast; meanwhile, skin fibroblast treated with primary culture from normal subjects was regarded as the control group. Restricted enzyme section differential display was used to separate the different-expressed genes of synovioblast and skin fibroblast in OA patients. In addition, blast technique was used to compare the resulted ranks with Genbank ranks.MAIN OUTCOME MEASURES: Differences of gene expression between synovioblast and skin fibroblast in OA patients.RESULTS: Gene expressions of superoxide dismutase (SOD), TFPI2, CXCL2, CXCL6 and transforming growth factor (TGF) were high in synovioblast of OA patients as compared with those in skin fibroblast of normal subjects.CONCLUSION: Gene expressions of SOD, TFPI2, CXCL2, CXCL6 and TGF are high in synovioblast of OA patients as compared with those in skin fibroblast of normal subjects. This suggests that gene may play a certain role in onset of OA.

The multi-piece post-crown technique is more effective in restoring residual root than other restoration techniques. Various types of adhesives have different material properties that affect restoration. Therefore, the choice of adhesive is particularly important for patients. However, the effect of different kinds of adhesives was not too precise by experimental methods when concerning about individual differences of teeth. One tooth root can only be restored with one type of adhesive in experiment. After the mechanical test, this tooth root cannot be restored with other adhesives. With the help of medical imaging technology, reverse engineering and finite element analysis, a molar model can be reconstructed precisely and restored using different types of adhesives. The same occlusal and chewing loads were exerted on the same restored residual root models with different types of adhesives separately. Results of von Mises stress analysis showed that the adhesives with low Young's modulus can protect the root canal effectively. However, a root canal concentration is apparently produced around the root canal orifice when chewing. Adhesives with large Young's modulus can buffer the stress concentration of the root canal orifice. However, the root canal tissue may be destroyed because the adhesive is too hard to buffer the load.
Crowns ; Dental Bonding ; Dental Cements ; Dental Pulp Cavity ; Dental Stress Analysis ; methods ; Elastic Modulus ; Finite Element Analysis ; Humans ; Post and Core Technique ; Tooth, Nonvital

OBJECTIVESTo induce autologous bone marrow derived mesenchymal stem cell (aMSC) into chondrocyte, and to confirm the effects of 3 dimensional (3D) dynamic inducing in vitro and their long-term animal model repairing in vivo.

METHODSaMSC were separated from rabbits bone marrow aspirates, then respectively experienced 3D dynamic inducing in alginate drops in modified rotating wall bioreactor culture or in two dimensional (2D) inducing (culture flask) for 10 d. The induced cells were harvest and then mixed with fibrin sealant (FS) to repair rabbit knee femoral trochlea cartilage defects model. After 8, 12, 24, 48 weeks animals were euthanized. Gross appearance, histological appearances were examined.

RESULTSFlask culture groups showed a little chondrocyte differentiation, 3D inducing group showed obviously chondrocyte differentiation, improved collagen II and proteoglycan production. For 3D inducing ones in vivo, the cartilage defects were smoothly repaired by white translucent hard tissue with obvious hyaline-like cartilage histological appearance after 8, 12 weeks, and the defects boundary were hard to be identified with hyaline like cartilage with sustained histological appearance and score after 24, 48 weeks. For 2D ones in vivo, the cartilage defects were smoothly repaired after 8 weeks by hyaline like cartilage which showed accelerated degeneration after 24 weeks and lose cartilage performance completely after 48 weeks.

CONCLUSIONS3D dynamic inducing may assist aMSC on differentiating into chondrocyte, improve its long-term in vivo repairing effects, and enlighten its further applications in tissue engineering cartilage.

Animals ; Bone Marrow Cells ; cytology ; Cartilage, Articular ; injuries ; physiopathology ; surgery ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Chondrogenesis ; Disease Models, Animal ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Rabbits ; Tissue Engineering ; methods ; Transplantation, Autologous ; Wound Healing

BACKGROUNDRenal transplantation in sensitized candidates remains a highly significant challenge worldwide. The production of panel reactive antibody (PRA) against human leukocyte antigen (HLA) is a major risk factor in presensitized recipients. The aim of this study was to evaluate the impact of HLA matching and recipients' PRA on two-year outcome in presensitized renal allograft recipients.

METHODSWe determined the percentage of panel reactivity and specificity of anti-HLA immunoglobulin (Ig) G antibodies in 73 presensitized renal allograft recipients compared with 81 unsensitized recipients (control group). HLA genotyping of both recipients and corresponding donors was performed by PCR with sequence-specific primers (PCR-SSP). We analyzed the factors influencing the early graft outcome (two-year rejection rates and survival rates of the grafts), including HLA mismatching, class and degree of panel reactivity, and target antigen of donors.

RESULTSPresensitized recipients had a worse two-year outcome than unsensitized recipients (P = 0.019 for rejection rate, P = 0.01 for survival rate). The difference in number of HLA-mismatched alleles with either 6-antigen matching (Ag M) standard or amino acid residue matching (Res M) standard was not significant between the rejection and non-rejection groups of presensitized recipients or between the graft survival group and graft loss group. Compared with the control group, recipients with both PRA-I and PRA-II antibodies had a significantly worse two-year outcome (P = 0.001 for rejection rate, P = 0.002 for survival rate). The two-year outcomes of the peak PRA >/= 50% group and its subgroup, at-transplant PRA > or = 50% group, were significantly worse compared with the control group (P = 0.025 and P = 0.001 for rejection rate, P = 0.043 and P = 0.024 for survival rate). The rejection rates of the at-transplant target antigen positive group and its subgroup, HLA-I target antigen positive group, were significantly higher than the control group (P = 0.001 and P = 0.001), target antigen negative group (P = 0.003 and P = 0.001), and peak target antigen positive with negative at-transplant target antigen group (P = 0.024 and P = 0.002). Two-year graft survival rates of the target antigen positive group and HLA-I target antigen positive group were significantly lower than the control group (P = 0.012 and P = 0.001). The two-year outcome of target antigen unknown group was similar to that of the target antigen positive group. Presensitized recipients with pre-transplant plasmapheresis or immunoadsorption (PRA prepared group) had a better but non-significant two-year outcome than the control group. However, the PRA unprepared presensitized recipients were different to the control group (P = 0.004 for rejection rate and P = 0.005 for survival rate). Hyperacute rejection (HR) occurred in three recipients with positive HLA-I target antigen and without mismatch according to Res M and in one case with positive PRA-II (for an unknown target antigen). No HR occurred in eight cases with positive HLA-II target antigens.

CONCLUSIONSPre-transplant PRA preparations might improve the access of presensitized patients to renal donors. Avoiding antigen-positive donors remains a fundamental measure in preventing HR and early rejections.

Adult ; Enzyme-Linked Immunosorbent Assay ; Female ; Graft Rejection ; immunology ; Graft Survival ; immunology ; HLA Antigens ; immunology ; Histocompatibility Testing ; Humans ; Isoantibodies ; blood ; Kidney Transplantation ; adverse effects ; immunology ; mortality ; Male ; Middle Aged ; Transplantation, Homologous ; immunology ; Treatment Outcome

OBJECTIVETo investigate the effect of AMPK agonist 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) on proliferation, differentiation and apoptosis of U937 cells and explore its possible mechanism.

METHODSU937 cells were cultured with different concentrations of AICAR for 24 h and 48 h. Cell proliferation was evaluated. Cell growth curve was analyzed by CCK-8; cell apoptosis was analyzed by cell morphology, Annexin V/7-AAD double labeling. The differentiation of U937 cells was evaluated by expression of CD11b. The Bcl-xL, Bax, Bim, caspase-3 mRNA expressions of U937 cells were determined by real time PCR.

RESULTSAICAR significantly inhibited the growth of U937 cells in a time-and dose-dependent manner, with a 24 h IC50 value of 1.1 mmol/L and 48 h of 0.9 mmol/L. 1.0 mmol/L AICAR didn't induce differentiation of U937 cells with the increase of CD11b expression for 24 h (P > 0.05). The U937 cells apoptosis was confirmed by cell morphology and Annexin V/7-AAD labeling. AICAR induced apoptosis of U937 cells and the apoptosis rate was (6.81 ± 1.16)% at 1 mmol/L AICAR higher than control group (2.74 ± 0.32)% without AICAR for 24 h treatment (P < 0.05). The real time PCR assay revealed that as compared with control group, the expression of Bim and caspase-3 mRNA were increased, while Bcl-xL and Bax were unchanged on the AICAR treatment.

CONCLUSIONAICAR can effectively inhibit proliferation and induce apoptosis of U937 cells. However, it has no significant effect on differentiation of U937 cells. The mechanism may be related with up-regulating Bim and Caspase-3.

Aminoimidazole Carboxamide ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Ribonucleotides ; pharmacology ; U937 Cells

To identify the active constituents in vitro and blood-absorbed ingredients in vivo from Yin Chen Hao decoction provides scientific evidence for probing its prevention and treatment mechanism on acute liver injury. An ultrahigh performance liquid chromatography quadrupole-time of flight-mass spectrometry (UPLC-QTOF/MS) method was applied for analysis of Yin Chen Hao decoction and the serum samples of mice with con-A induced acute liver injury after preventive oral administration for 14 days (the use of all laboratory animals in this study was approved by the Ethics Committee of the Naval Medical University, 19YF1459400). A total of 90 chemical constituents were identified from Yin Chen Hao decoction, mainly were flavonoids, terpenoids, tannins, quinones. 5 prototype compounds were identified in the serum, including chrysophanol, deoxyrhapontin-8-O-gallate, mussaenosidic acid, herniarin, emodin. The established UPLC-QTOF/MS method could efficiently and sensitively identify the constituents in vitro and blood-absorbed ingredients of Yin Chen Hao decoction, primarily clarify the material basis of its hepatoprotective effect, and provided a scientific basis for the quality marker selection and the pharmacodynamic material basis research on the decoction.

OBJECTIVETo construct an eukaryotic expression vector for PRL-2 and evaluate its effect on the invasiveness and migration of a human hepatocellular carcinoma cell line.

METHODSRT-PCR was performed to amplify the complete PRL-2 open reading frame using the total mRNA of hepatocellular carcinoma HepG2 cells as the template. PRL-2 gene was inserted into the pGEM T easy vector and sequenced, and the correct PRL-2 sequence was subcloned into the mammalian expression vector pcDNA3.1+. The constructed PRL-2 vector was transfected into CL1 cells via lipofectamine, and the stable expression of PRL-2 mRNA was detected by RT-PCR, the expressed protein identified by immunohistochemistry and Western blotting, and the effect of PRL-2 on the adhesion ability of CL-1 cell evaluated with MTT assay 20 and 120 min after transfection. The effect of PRL-2 on the invasive migration of CL-2 cells was evaluated according to the number of cells penetrating the Matrigel layer of polycarbonate membrane of Boyden chamber.

RESULTSRT-PCR yielded a fragment of 504 bp and the inserted PRL-2 sequence was verified by sequence analysis. The subclones were identified by restriction endonuclease digestion, and a G418-resistant clone, PRL-2-CL1, was obtained after 8 weeks of selection. RT-PCR showed stable expression of PRL-2 mRNA, and Western blotting confirmed overexpression of PRL-2 protein in the transfected cells. PRL-2 increased the adhesion rate of CL-1 cells to fibronectin at 20 min and 120 min after transfection (P<0.05), and also the number of CL-1 cells penetrating the polycarbonate membrane from 10.0+/-3.7 to 44.8+/-2.6 (P<0.05).

CONCLUSIONAn eukaryotic expression vector of PRL-2 has been successfully constructed, which allows stable and efficient expression in CL-1 cell line. PRL-2 can promote cell adhesion and invasion activity of human hepatocellular carcinoma cells.

Animals ; Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Adhesion ; genetics ; Cell Line, Tumor ; Cell Movement ; genetics ; Eukaryotic Cells ; metabolism ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Humans ; Liver Neoplasms ; genetics ; pathology ; Neoplasm Invasiveness ; genetics ; Protein Tyrosine Phosphatases ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo investigate the clinical significance of vascular endothelial growth factor (VEGF) levels in serums of colorectal cancer patients at stage IV.

METHODSUsing enzyme linked immunosorbent assay (ELISA) to detect the VEGF levels in serums of 45 colorectal cancer patients at stage IV, and 20 healthy served as normal control.

RESULTSThe mean concentration of VEGF in 45 colorectal cancer patients at the 7 day after operation were significantly lower than that before operation (P<0.01). The mean concentration of VEGF in the patients who benefit from bevacizumab showed no statistical difference from the levels of who did not benefit (P=0.554).

CONCLUSIONThe VEGF levels in colorectal patients at stage IV are lowed as the load of tumor decrease. The circulating levels of VEGF seem not predict the response to bevacizumab in colorectal cancer patients at stage IV.

Adolescent ; Adult ; Aged ; Antibodies, Monoclonal ; administration & dosage ; Antibodies, Monoclonal, Humanized ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bevacizumab ; Colorectal Neoplasms ; blood ; drug therapy ; pathology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Vascular Endothelial Growth Factor A ; blood ; Young Adult

BACKGROUNDDiabetes has become one of the most common chronic diseases and the third leading cause of death in China. Many programs have been initiated at national and local levels to address the illness. However, the effect of these programs in daily outpatient clinics is still unclear. The objective of this study was to investigate the management status of type 2 diabetes mellitus (T2DM) and factors associated with it in diabetes clinics of tertiary hospitals in Beijing.

METHODSA cross-sectional survey was conducted in six tertiary hospitals in Beijing. Control criteria were defined based on 2007 China guideline for type 2 diabetes (CGT2D).

RESULTSA sample of 1151 patients, age (60.8 ± 9.2) years, and with a median disease duration of 7.3 years was included. The hemoglobin A1c (HbA1c) mean level was (7.15 ± 1.50)%, the percentage of patients achieving the targets for HbA1c was 37.8%, blood pressure 65.6%, triglyceride (TG) 48.8%, high-density lipoprotein (HDL) 59.2%, low-density lipoprotein (LDL) 34.0%, and total cholesterol (TC) 42.0%. The factors independently associated with glycemic control were diabetes duration (odds ratio (OR) = 0.95; 95% confidence interval (CI): 0.919 - 0.982, P < 0.01), body mass index (BMI) (OR = 0.914, 95%CI: 0.854 - 0.979, P = 0.01) and smoking (OR = 0.391, 95%CI: 0.197 - 0.778, P < 0.01). The factors independently associated with blood pressure control were BMI (OR = 0.915, 95%CI: 0.872 - 0.960, P < 0.01) and male gender (OR = 0.624, 95%CI: 0.457 - 0.852, P < 0.01). The factor independently associated with LDL control was education level (OR = 1.429, 95%CI: 1.078 - 1.896, P = 0.013).

CONCLUSIONSThe management status of T2DM patients in tertiary hospitals in Beijing has improved remarkably. However, there is still room for further improvement to reach the guideline target. Long diabetes duration, high BMI, smoking, male gender and low education level were independently associated with poor metabolic control.

Adolescent ; Adult ; Aged ; Blood Glucose ; metabolism ; Blood Pressure ; China ; Cholesterol ; blood ; Diabetes Mellitus, Type 2 ; blood ; metabolism ; Female ; Glycated Hemoglobin A ; metabolism ; Hospitals ; statistics & numerical data ; Humans ; Lipoproteins, HDL ; blood ; Lipoproteins, LDL ; blood ; Male ; Middle Aged ; Triglycerides ; blood ; Young Adult