Previous work showed that there was high ratio of Mycoplasma hyorhinis infection in human gastric carcinoma. P37 protein is a membrane lipoprotein of Mycoplasma hyorhinis. It was reported that P37 inhibited cell adhesion and induced tumor invasiveness. To investigate the function of P37 in cancer development, the p37 gene was subcloned into shuttle vector of pAdTrack-CMV. Linearized pAdTrack-CMV-p37 was co-transformed into BJ5183 cells with adenoviral genomic plasmid pAdEasy-1. The identified recombinant DNA was transfected into 293 cells to package adenovirus. From the supernatant and cell lysis, the presence of recombinant adenovirus P37 was proved by PCR. And then BICR cells with this recombinant adenovirus of P37 were successfully infected. RT-PCR and Western blot demonstrated the transcription and expression of P37 in infected BICR cells. And the soluble P37 protein can be secreted into the culture supernatant of infected BICR cells. In the migration assay in vitro, the number of migration BICR cells infected with Ad-p37 was 52.03% more than that of control. The construction of recombinant adenovirus provided a good tool for studying P37 function and its molecular mechanism, which will be further used for in vivo migration and invasion experiments.

The efficacy of injecting sclerosing agent next to transverse process of cervical vertebra to induce vertebral artery type of cervical syndrome (CSA) was observed. Twenty rabbits were randomly divided into two groups: the model group and the control group. The rabbits in the model group were injected with sclerosing agent next to transverse process of cervical vertebray, on the contrary, the rabbits in the control group were injected with nothing. Transcranial Doppler (TCD) was used to detect the average speed of blood (Vm), pulsatility index (Pi) and the resistant index (Ri) of the vertebral artery, hematoxylin and eosin (HE) staining was used to observe the morphological changes, and immunohistochemistry was used to detect the expression of alpha-smooth muscle actin (alpha-SMA) and matrix metalloproteinase-2(MMP-2). TCD showed increased Pi, Ri and decreased Vm in the model group (P<0.05) compared with the control group. HE staining revealed hyperplasia and hypertrophied smooth muscle cells in the model group (P<0.05). Immunohistochemistry displayed up-regulation of alpha-SMA and MMP-2 in the model group (P<0.05). It was concluded that injecting sclerosing agent next to transverse process of cervical vertebra induces remodeling of vertebral artery in rabbits, suggesting it is a practical method to establish CSA animal model.

Aim The kinetics of nicotinic acetylcholine receptor (N-AChR) was stadied. MethodsThe saturation experiments of Torpedo electric organ N-AChR were taken with125I-?-BuTX. The results were calculated by Scatfit program. Experiments were thenperformed to study the kinetics of dissociation of ?-BuTX and nicotine from N-AChR,respectively. When their binding reached saturation, an excess of 1000 times ?-BuTX ornicotine was added. The saturation binding properties of the N-AChR extracted fromleg muscles of the 13th day chick embroys and optic lobes of the 20th day chick em-broys were studied. The results were calculated by Scatfit program. The othe experi-ments were taken to observe the competition between mcotine and 125I-?-BuTX forbinding with Torpedo, optic lobe and skeletal muscle of the chick N-AChRs. ResultsThe saturation experiments of Torpedo electric organ N-AChR resulted in a Scatchardplot of hyperbola which responded to the model of two kinds of receptor with on eli-gand.The difference of and B max between high affinity binding site and low affnitybineing site was significant. The dissaciation experiment showed that the fast dissocia-tion rate of tow ligands was 500 times more than that of the slow dissociation rate. Thisresult suggested that there may be two subtype N-AChRs.The saturation bindingproperties of N-AChR of leg muscles and optic lobe of the chick embroys revealed aScatchard plot of two kinds of N-AChR indicating a single type of site. The bindingaffinity of receptor of optic lobe was 100 times more than that of muscles. In competi-tion for Torpedo receptor by nicotine and 125I-?-BuTX, the values of IC50 were different:which suggested that two kinds of receptor sites were existent. In competition ofNicotine and 125I-?-BuTX for optic lobe and skeletal muscles of the chick N-AChR, thevalue IC50 of skeletal muscles N-AChR was 7. 7 times higher than that of optic lobe. Itindicated that two kinds of N-AChR subtype existed in the optic lobe and in theskeletal muscles of chick respectively. Conclusion N-AChR of Torpedo electric organcontains two kinds of subtype receptors. N-AChR of optic lobe of chick embroys is onesubtype receptor, and N-AChR of skeletal muscles of chick embroy is another subtypereceptor.

Career Mobility ; China ; Humans ; Occupational Diseases ; epidemiology ; prevention & control ; Occupational Exposure ; prevention & control ; statistics & numerical data ; Occupational Health

Objective To investigate the related high-risk factors of the occurrence of progressive hemorrhagic injury (PHI) after acute traumatic brain injury ,and to provide the basis for early clinical diagnosis and treatment .Methods Retrospective analysis the clinical data of 398 cases of traumatic brain injury patients .According to whether PHI occurred ,the patients were divided into the progress group and non-progress group .Relevant factors with progressive hemorrhagic injury were assessed .Results The univari-ate analysis showed that ,the age ,gender ratio ,injury to first CT time ,GCS score when admitted in hospital ,mean arterial pressure , combined with skull fracture ,combined with epidural hematoma ,combined with cerebral contusion ,bilateral injury ,subarachnoid hemorrhage ,disturbance of consciousness ,mydriasis ,volume of intracranial hematoma more than 10 mL and volume of hematoma at the first CT scanning ,Platelets ,plasma fibrin concentration and D-dimer influenced the development of progressive hemorrhagic in-jury(P<0 .05) .Logistic regression showed that ,injury to first CT time ,GSC score less than 12 ,disturbance of consciousness ,my-driasis ,volume of hematoma more than 10 mL at the first CT scanning ,combined with cerebral contusion ,combined with subarach-noid hemorrhage ,platelet and D-dimer were the independent risk factors for PHI (P<0 .05) .Conclusion Patients with acute brain injury should be promptly head CT .Patients with GCS score less than 12 ,disturbance of consciousness ,mydriasis ,volume of in-tracranial hematoma more than 10 mL at the first CT scanning ,combined with cerebral contusion ,subarachnoid hemorrhage ,platelet and D-dimer were the independent risk factors of the progressive hemorrhagic injury after traumatic brain injury ,Should closely ob-serve the illness progress ,regularly review the head CT as soon as possible .

Adult ; Ear Neoplasms ; Eustachian Tube ; pathology ; Humans ; Male ; Teratoma

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Female ; Herbicides ; poisoning ; toxicity ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Paraquat ; poisoning ; toxicity ; Risk Factors ; Time Factors ; Young Adult

Objective: Overexpression of the HER2/neu oncogene is a frequent molecular event in multiple human cancers. Being a cancer antigen, p185 erbB2 is an ideal target for immunotherapy. In order to decrease the immunogenicity of mouse anti-p185 erbB2 monoclonal antibody in human cancer therapy, we constructed the eukaryotic expression vector of anti-p185 erbB2 chimeric monoclonal antibody and verified expression of the chimeric antibody in CHO-dhfr - cell. Methods: The variable regions of light chain and heavy chain were amplified with RT-PCR and inserted into the chimeric antibody vector pWSD2. After CHO-dhfr - cells were transfected with recombination plasmid by lipofectAMINE, the chimeric antibody expressing level was identified with RT-PCR, indirect-ELISA, and Western blot. The specificity of the anti-p185 erbB2 chimeric antibody was testified with ELISA assay and immunoprecipitation. Moreover, the effects of chimeric antibody on the proliferation of breast cancer cell line SKBR3, which is overexpressing p185 erbB2 , were measured with MTT assay in vitro. Results: The anti-p185 erbB2 chimeric antibody eukaryotic expression vector was constructed successfully and the expression of the chimeric antibody in CHO-dhfr - was verified by RT-PCR, indirect-ELISA, and Western blot. ELISA assay showed that chimeric antibody reacted with cells overexpressing p185 erbB2 specifically, but did not react with that non-overexpressing p185 erbB2 . Immunoprecipitation test confirmed that the chimeric antibody could bind to p185 erbB2 specifically. The MTT assay demonstrated that the chimeric antibody could inhibit the growth of SKBR3 cells overexpressing p185 erbB2 . Conclusion: The anti-p185 erbB2 mouse/human chimeric antibody that was expressed in CHO-dhfr - cells can bind to p185 erbB2 specifically and inhibit proliferation of SKBR3 cells overexpressing p185 erbB2 . It has a potential application in biotherapy of cancer.

BACKGROUD: It is not consistent on whether melatonin correlates with the adolescent idiopathic scoliosis(AIS) demonstrated by the domestic and foreign scholars.OBJECTIVE: To study the pathogenesis of AIS and its correlation with melatonin.DESIGN: A nonrandomized age-matched controlled study screened with two tests.SETTING: The Second Department of Orthopaedics, Haikou People' s Hospital.PARTICIPANTS: The experiment was completed in the Second Department of Orthopaedics, Haikou People' s Hospital. Totally 8 198 in-school students.from over 10 schools of towns in Haifu District, 4 423 males and 3 775 females, aged 7 to 16 years were surveyed.METHODS: Forty-two adolescents who have been diagnosed with AIS were selected and 50 healthy age-matched adolescents were selected as controls. Melatonin was assayed with the radioimmunology and the data were processed statistically.MAIN OUTCOME MEASURES: The levels of serum melatonin in the two groups.RESULTS: The serum melatonin of the preadolescents below 10 years old in AIS group, especially of females, was less than that of adolescents in the control group, with significant differencein statistics.CONCLUSION: The serum melatonin correlates with pathogenesis of AIS.

Objective To validate the analytical performance of three Cys C reagents with particle-enhanced turbidimetric immunoassay(PETIA) method used on the automatic biochemistry analyzer for preliminary clinical application.Methods The performance of three Cys C reagents (labeled as A, B, C) with PETIA method from Shanghai Jing Yuan Co., Beijing Leadman Co. and Beijing Jiuqiang Co. on OlympusAU2700 automatic biochemistry analyzer were assessed.According to the standard of CLSI EP6-A, EP15-A and EP7-P, the precision, linearity range, disturbance (bilirubin, hemoglobin, chyle) were assessed, and compared with those of Cys C reagent based on particle-enhanced nephelometric immunoassay(PENIA) from Dade Behring Co.. The reference ranges for Cys C in serum of 120 healthy individual were evaluated.Results The within-run CVs of the three reagents (A, B and C) were 3.08%-3.2%, 2.3%-4.15% and 1.38%-1.53% respectively.The total CV in A, B and C were 3.29%-3.44%, 2.65%-5.18% and 1.67%-1.69% respectively, lower than the stated.Limits of quantitative determination (LOQ) of the three reagents were 0.41, 0.23 and 0.07 mg/L, basically meeting the testing requirement.The linearity range was 0.22-7.26 mg/L(r=0.996), 0.20-7.72 mg/L(r=0.999)and 0.20-7.62 mg/L(r=0.997)in the three reagents, which demonstrated a sound linear correlation. For interference tests, no remarkable interference (<±10%) of reagent C was detected when bilirubin≤684 μmol/L, hemoglobin≤9.7 g/L and Chyle turbidity≤6 200 FTU; and no significant interference of reagent B was found when bilirubin≤684 μmol/L, hemoglobin≤6.79 g/L and Chyle turbidity≤6 200 FTU; when bilirubin≤684 μmol/L, hemoglobin≤4.85 g/L and Chyle turbidity≤1 240 FTU reagent A was not interfered significantly. The comparison afte and before the high-speed centrifugation reveals that the average percentage of bias for reagents A, B, C measured Cys C in chylous serum samples of patients was -8.31%, 1.52%, 1.32%, respectively.In method comparison tests, the regression equations of the three reagents compared with Dade Behring PENIA Cys C reagent were as follows:Y=0.787X+0.492 (R2=0.976), Y=1.098X+0.137 (R2=0.982) and Y=1.037X+0.249 (R2=0.996), respectively. Agreement rates of the high Cys C in reagent A, B, C and Dade Behring Cys C reagent were 80% (Kappa=0.615,P=0.000), 100% (Kappa=1.000,P=0.000), 91.2% (Kappa=0.824,P=0.000); While for reference range of preliminary clinical assessment, diagnosis coincidence rate of reagent A increased to 98.8% (Kappa=0.974,P=0.000). Conclusions When used in automatic biochemical analyzer, the three Cys C reagent with PETIA showed high precision,sensitivity, and sound correlation with Dade Behring PENIA reagents.The three reagents are all able to meet clinical test requirements, nevertheless, anti-interference capability were diffierent and the reference range should be further validated.