Objective To investigate the neurotoxic effects of different concentrations of tetracaine and ropivacaine on the brachial plexus nerve in rats.Methods Forty-eight male Sprague-Dawley rats,weighing 410-430 g,were randomly divided into 8 groups (n =6 each):normal saline group (group NS),0.25％,0.50％ and 1.00％ tetracaine groups (groups T1-3 ),and 0.25％,0.50％,1.00％ and 2.00％ ropivacaine groups (groups R1-4 ).The rats received injection of normal saline 1.0 ml,0.25％,0.50％ and 1.00％ tetracaine 0.5 ml,0.25％,0.50％,and 1.00％ ropivacaine 1.0 ml and 2.00％ ropivacaine 0.5 ml in groups NS,T1-3 and R1-4 respectively through one side of the axillary sheath.The other side of the axillary sheath served as control side.Five days later,compound action potential and nerve conduction velocity (NCV) of the brachial plexus nerve were measured.Tne brachial plexus nerve was obtained as the specimen for microscopic examination with light and transmission electron microscope.Results Compared with the control side and group NS,the compound action potential and NCV of the brachial plexus nerve were significantly decreased in groups T2,3 and R3,4 ( P ＜ 0.05 ).The compound action potential and NCV of the brachial plexus nerve were gradually decreased with the increasing concentrations of tetracaine in groups T1 3 ( P ＜ 0.05 ).The compound action potential and NCV of the brachial plexus nerve were significantly decreased in group R4 as compared with groups R1-3 (P ＜ 0.05).The microscopic examination showed that the pathologic changes were more severe in groups T2,3 and R3,4 than those on the control side and than in group NS.Conclusion 0.50％ and 1.00％ tetracaine,and 1.00％ and 2.00％ ropivacaine can result in pathologic damage to the brachial plexus nerve in rats and the degree of damage is related to the concentration.

3a and 7a are nonstructural proteins of SARSCoV, which are encoded separately by ORF 3a and ORF 7a in SARSCoV genome. The expression of 3a has been founded in cells infected by virus in vivo or in vitro. Firstly, the pGL3Control vector was reconstructed , the pGL3Enhancer vector deletious of SV40 promoter gene was obtained . Then the IFN? promoter gene was cloned into the pGL3Enhancer vector and pGLIP21, the Luciferase reporter plasmid with IFN? promoter was established. The availability of pGLIP21 was verified by NDV ,the inductor of IFN?, the Luciferase activity was assayed in cells transfected with pGLIP21 by Luminometer. In order to see the function of 3a and 7a protein of SARSCoV,CHO cells expressing 3a or 7a protein were transfected with pGLIP21, the intensity of luciferase activity was analyzed . By analysis, in vitro, 3a and 7a protein of SARSCoV had the similar ability in triggering the expression of Luceferase gene, i.e 3a and 7a protein of SARSCoV could effectively activate the promoter fragment of IFN? gene. This result will help studying the function of 3a and 7a protein and provide a method to study the nosogenesis mechanism of SARSCoV.

OBJECTIVE: To study the application value of contrast vision in identifying the malingering decreased vision in the practice of clinical forensic medicine. METHODS: Thirty-one young and middle aged volunteers were selected and divided randomly into group 1 (16 persons with 32 eyes) and group 2 (15 persons with 30 eyes). The optotype contrast was 100%, 25%, 10% and 5%, respectively. The contrast vision of group 1 was tested. The contrast vision of group 2 was tested as follows: (1) the volunteers cooperated without inspector's interference; (2) the volunteers cooperated under inspector's interference; (3) the volunteers disguised decreased vision without inspector's interference; (4) the volunteers disguised decreased vision under inspector's interference. The data was then analyzed by statistics. RESULTS: There was a close correlation between contrast vision and contrast. As the contrast decreased, the vision acuity also decreased. The contrast vision curve of former two methods showed a good reproducibility while the contrast vision curve of latter two methods had a bad reproducibility. CONCLUSION The repetition of contrast curve with or without inspector's interference can be used to discriminate malingering vision. The acquired contrast curves can be provided to the court as direct evidence and can help enhance the verification conclusion.
Adult ; Contrast Sensitivity/physiology* ; Diagnosis, Differential ; Female ; Humans ; Male ; Malingering/psychology* ; Reproducibility of Results ; Sensitivity and Specificity ; Severity of Illness Index ; Vision Tests/statistics & numerical data* ; Vision, Binocular ; Vision, Low/psychology* ; Visual Acuity ; Young Adult

OBJECTIVE: To study the effect of pelvic rotation in three-dimensional direction on the actual placement angle of acetabular prosthesis in total hip arthroplasty. METHODS: Pelvic CT imaging data of normal adults were collected, and three-dimensional reconstruction of pelvic acetabulum was performed with computer software to simulate the rotation of the pelvis around X, Y and Z axes perpendicular to the sagittal, transverse and coronal planes of the human body. Radiographic inclination(RI) and radiographic anteversion (RA) of the acetabular cup were measured when the acetabular prosthesis was implanted at a standard angle. The correlation analysis was used to quantify the relationship between the rotation angle of each axis and the actual angle of acetabulum. RESULTS: The pelvic rotation along the X-axis and Y-axis had little effect on the RA of the acetabulum, but had a great influence on the RI and showed a linear correlation. The regression equations were RA=0.682 4X+10.256(²=0.308 4) and RA=-0.714 1Y+10.424(²=0.999 8). The pelvic rotation along the Z-axis had little effect on the RA, but had a linear correlation with the RI, and the regression equation was RI=1.0Z+46(²=1.0). CONCLUSIONS The anteroposterior rotation of the pelvis or the longitudinal rotation along the body significantly affected the acetabular anteversion, but had little effect on the abduction angle. On the contrary, the left and right deviation of the pelvis on the coronal plane could significantly affect the acetabular anteversion angle, but did not affect its anteversion angle.
Acetabulum ; Adult ; Arthroplasty, Replacement, Hip ; Hip Prosthesis ; Humans ; Pelvis ; Rotation

Objective Metabolites produced by metabolic imbalance such as free fatty acids and lipopolysaccharides can result in a state of chronic low-grade inflammation, or metabolic inflammation, which plays an important role in the pathogenesis of atherosclerosis, type 2 diabetes, non-alcoholic fatty liver disease, and obesity. The above metabolic disorders are closely related with the metabolic inflammation, which always coexist. Therefore, we proposed the concept ofmetabolic inflammatory syndrome ( MIS). According to our study, patients with two or more metabolic disorders above could be diagnosed as MIS. The current research is aimed to investigate the prevalence of MIS and its components, and to compare the clinical values of MIS and metabolic syndrome ( MS) . Methods 2 001 in patients with type 2 diabetes from 6 hospitals in Shanghai were recruited in the current multi-center cross-sectional study. The diagnostic rates of MIS and MS and their components of both syndromes were compared. Results In the patients with type 2 diabetes, the detective rate of MIS was 96. 2%, which was higher than that of MS (71. 3%). Among 4 components of MIS, atherosclerosis showed the highest detective rate (75.6%). MIS[OR=2.252(95%CI1.026-4.942),P=0.043],atherosclerosis[OR=2.726(95% CI1.953-3. 804),P<0. 001], and MS[OR=1. 915 (95%CI 1. 444-2. 540),P<0. 01] were the risk factors of coronary heart disease. Conclusion With atherosclerosis, type 2 diabetes mellitus, non-alcoholic fatty liver disease, and obesity as its 4 components, MIS has a high detective rate in patients with metabolic disorders, and seems to be more sensitive than MS to distinguish inflammation-related metabolic diseases. The concept of MIS will promote the screening and prevention of atherosclerosis in its early stage.

OBJECTIVETo study the tumor cell killing function of T lymphocytes stimulated by dendritic cells (DC) and to analyze the differences of protein contents of exosomes in each type of cell.

METHODSThe exosomes of hepatic cell lines with high (P group) or low (F group) metastatic potentials were isolated by a process of four-step centrifugation and the collected exosomes were observed under an electron microscope (EM). The tumor cell killing experiment was performed by adding T lymphocytes activated by DC loaded with exosomes from corresponding P and F group cells and was studied using 3H-TdR experiments. The proteomic analysis was performed by surface-enhanced laser desorption/ ionization time of flight mass spectrometry (SELDI-TOF-MS ) on the exosomes of P and F group cells.

RESULTSThe density distribution and content of exosomes in the P group were not equal to those in the F group observed by EM. The CD80, CD86, MHC-I and MHC-II in the P group were 64.27+5.00, 44.89+10.11, 84.35+19.89 and 59.03+19.37, and those in the F group were 71.53+4.85, 50.01+9.50, 80.68+29.87 and 58.86+21.11, respectively (P>0.05, compared with the control group). The counts per minute value in the P group was 528.40+179.06 and 78.80+24.44 in the F group after being loaded with exosomes (P<0.01, compared with the control group). There were significant differences between the proteins in the exosomes of hepatic cancer cell lines with high or low metastatic potentials.

CONCLUSIONExosomes have potential values of application in immunotherapy and in biotherapy for recurrences and metastases of hepatic carcinomas.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Dendritic Cells ; immunology ; metabolism ; Exosomes ; Liver Neoplasms ; metabolism ; pathology ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; immunology ; metabolism

OBJECTIVETo study the effects of gene transfer cytotoxic T lymphocyte associated antigen 4 immunoglobulin (CTLA4-Ig) and anti-cluster of differentiation 154 (CD154) mAb on the rejection of rat islet xenografts.

METHODSHuman islets were infected with the recombinant adenoviruses containing CTLA4-Ig gene. Transduced islets were transplanted under the left kidney capsule of diabetic rats. And then the animal model were treated with anti-CD154 monoclonal antibody. The changes of blood sugar were measured and the survival rates of grafts and transplantation rats were observed after transplantation. The morphological changes of grafts were observed. Expression of CTLA4-Ig and insulin were detected by immunohistochemical staining and cytokines were quantified by ELISA.

RESULTS(1) The blood glucose of transplantation rats decreased to normal level on 2nd day post-transplantation. The average level blood glucose of control group A, anti-CD154mAb treatment group B, transfected group C and associated treatment group D increased on day 8, 18, 25, 36, post-transplantation respectively. (2) The grafts of group A, B, C and D survived for (10.0 +/- 2.1) d, (22.0 +/- 8.2) d, (28.0 +/- 6.5) d and (37.0 +/- 9.3) d respectively. The survival of grafts in group D was significant longer than that in group A, B and C, respectively; The survival of group B and C were significantly prolonged compared with group A and the survival of group B was significantly different with group C (P < 0.05). The survival of transplantation rats were (21.0 +/- 5.7) d, (35.0 +/- 6.5) d, (48.0 +/- 8.5) d and (65.0 +/- 12.5) d in group A, B, C and D, respectively. The survival of transplantation rats compared each other among four groups were same as the survival of grafts (P < 0.05). (3) In control animals (group A), serum IL-2 and TNF-alpha concentration were elevated to a high level within seven days post-transplantation and significantly increased compared with that before transplantation (P < 0.01). (4) Hematoxylin-eosin staining of grafts showed a lot of islets under the kidney capsule of transplantation rats, no inflammatory cell infiltrate and immunohistochemical staining of grafts demonstrated expression of insulin protein at islets in group B, C and D. These grafts positively stained for CTLA4-Ig in group C and D.

CONCLUSIONSGene transfer CTLA4-Ig and anti-CD154mAb treatment can inhibit the rejection of rat islet xenografts and treatment Ad-CTLA4-Ig and anti-CD154 mAb could induce immune tolerance of islet xenografts.

Adenoviridae ; genetics ; Animals ; Antibodies, Monoclonal ; therapeutic use ; CD40 Ligand ; immunology ; Diabetes Mellitus, Experimental ; surgery ; Genetic Vectors ; Graft Rejection ; immunology ; prevention & control ; Humans ; Immunoconjugates ; genetics ; Islets of Langerhans Transplantation ; Rats ; Rats, Wistar ; Transfection ; Transplantation, Heterologous

OBJECTIVESTo study the relationship between lymphangiogenesis and lymphatic metastasis in mice bearing hepatic carcinoma and analyze the mechanism of the lymphatic metastasis.

METHODSHepatic carcinoma cell lines of high and low potentialities of lymphatic metastasis were injected into the footpads of Balb/c mice. Their metastases to lymph nodes were examined. The tumor tissues of each group were stained with 5'-nucleotidase-ALP to observe the lymphoangiogenesis. The total RNA of high and low metastatic potential cell lines were extracted for metastasis gene DNA array. The vascular endothelial cell growth factor C (VEGF-C) and VEGF-D of each cell line were detected using semi-quantitative RT-PCR and were further quantatively analyzed using real time PCR.

RESULTSThe para-common iliac a. and renal hilar lymph nodes metastases of the high metastatic potential cells were significantly higher than in the controls (P>0.05). The quantity of lymphatic vessels in the high metastasis group was significantly larger than that of the control group (P<0.05). The expressions of CD44, E-cadherin, HER2/neu, H-Ras and VEGF-C in the high metastasis group were higher than those in the low metastasis group shown by the cDNA micro array experiment but the expressions of nm23A, nm23-E4, p16ink4a, CD61 were lower. The VEGF-C expression was higher and the VEGF-D was lower in the high metastasis group compared to those of the low metastasis group shown by semi-quantitative RT-PCR. The secretion of VEGF-D was significantly lower and the ratio of VEGF-C/VEGF-D was significantly higher in the high metastasis group than the low metastasis group (P<0.05).

CONCLUSIONSThe lymphatic metastasis of hepatic carcinoma is related to lymphoangiogenesis. The changes of VEGF-C and VEGF-D expressions might be a cause influencing the lymphoangiogenesis. VEGF-C/VEGF-D might be an effective parameter in affecting lymphatic metastases.

Animals ; Liver Neoplasms, Experimental ; pathology ; Lymph Nodes ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

OBJECTIVETo investigate the effect of cytotoxic T lymphocyte-associated antigen 4 immunoglobulin (CTLA4-Ig) gene transfer on rejection of rat islet xenografts.

METHODSHuman islets were infected with the recombinant adenoviruses containing CTLA4-Ig gene, and the transduced islets were transplanted under the left kidney capsule of diabetic rats to establish rat models bearing human-rat islet xenografts. The blood sugar of the rats receiving the transplantation was measured and the xenografts and host survival were observed after transplantation. The morphological changes of grafts were examined, in which the expression of CTLA4-Ig and insulin were also detected by immunohistochemical staining and the cytokines were quantified using enzyme-linked immunosorbent assay (ELISA).

RESULTSThe blood glucose of the rats bearing the grafts decreased to the normal level on day 2 after transplantation. The average blood glucose level of the CTLA4-Ig gene transfected group and the control group increased on day 25 and 8 post-transplantation, respectively. The grafts of the transfected group survived for an average of 28-/+6 days, significantly longer than that in the control group (10-/+2 days, t=10.52, P<0.01), and the host survival were 48-/+8 and 21-/+6 days in the two groups, showing also significant difference between them (t=12.23, P<0.01). In the control rats, serum IL-2 and TNF-alpha concentration drastically increased within 7 days after transplantation to levels significantly higher than that before transplantation (P<0.01), but in the transfected group, the levels were decreased compared with the preoperative levels. In the transfected grafts, positive staining for CTLA4-Ig and insulin were detected.

CONCLUSIONCTLA4-Ig gene transfer may lower the rejection of rat islet xenografts and prolong the survival time of both the grafts and hosts.

Abatacept ; Adenoviridae ; genetics ; Animals ; Blood Glucose ; Graft Rejection ; prevention & control ; Graft Survival ; Humans ; Immunoconjugates ; genetics ; Interleukin-2 ; blood ; Islets of Langerhans Transplantation ; Rats ; Rats, Wistar ; Transduction, Genetic ; Transplantation, Heterologous ; Tumor Necrosis Factor-alpha ; blood

To increase the productivity and yield of recombinant protein in continual perfusion processing, Every amino acid consumption rate in continual perfusion culture of engineering CHO cell line which expressed recombinant TNFRp75: Fc fusion protein were analyzed. Then rational amino acids were accordingly added to improve its comprehensive utilizing. At the same time, glucose supply was controlled to make the concentration of glucose below 0.5 g/L for ameliorating the toxicity of lactate accumulation in order to decrease the perfusion rate. The result showed that the productivity of recombinant protein was 3.1 times (388mg/L) and the total yield was 4. 7 times (244. 4g) that of control cultures after nutrient compensation and metabolism control in 30 liter working volume, and the fermentation period was prolonged one week longer. The sialic acid content and bioactivity in vitro of recombinant TNFRp75: Fc were not changed after nutrient compensation and glucose control supply. Nutrient compensating and metabolic control in continual perfusion fermentation could significantly increase the productivity and yield of recombinant TNFRp75:Fc, and thus reduced relative industrialization costs.