Objective: To study the cellular immune response and the anti-tumor effects induced by intramuscular DNA immu- nization with HER2/neu extracellular domain gene. Methods: HEH2/neu oncogene extracelltilar domain gene was isolated and inserted to pCDNA3 expression vector. The cellular immune response and its anti-tumor effects were analyzed after intramuscular DNA immunization. Results: A specific cytotoxic activity by spleen cells from HER2/neu ECD gene immunized mice was found in vitro. Tumor growth was inhibited after tumor challenge in vivo in immunized mice. Conclusion: Specific cellular immune re- sponse and anti-tumor effect were elicited by HEB2/neu extracellular domain gene immunization.

Objective: To induce an immune response by intramuscular immunization with HER2/neu oncogene extracellular do- main (ECD) DNA and observe the effect in vivo by an abortion model in mice.Methods: Human and rat HEH2/neu ECD gene were cloned and inserted into expressive vector pCDNA3. The DNA immunization, expression of HER2/neu ECD, antibody de- tection and abortion in immunized mice were studied. Results: HER2/neu ECD protein expressed on muscle cells in the injected site, specific antibody to HER2/neu was induced by DNA immunization. The average of the offspring per delivery from immu- nized mice was significantly lower than that from control group, and the abortive embryos were found in uterus from immunized mice.Conclusion: The results show that both human and rat HER2/neu ECD gene can induced effective immune response by DNA immunization.

Two reaction systems have been observed to ascertain the capability of ABC-ELISA on the detection of antibodies against particulate antigens.Mouse thymucytes as the antigens were coated on the enzyme immunoassay plate for one system,and rabbit anti- mouse brain serum was titrated.The titer of the serum in ABC-ELISA was 1:12,800, about 40-fold higher than that in complement dependent cytotoxicity assay.In the other system,the plate was coated with cultured human leukemia cells of T lymphocyte origin(Cem-T)and B cell origin(Daudi),where the ascites induced by an anti-human lymphocyte hybridoma was titrated.The results showed that ABC-ELISA was 8-16 times sensitive than the standard ELISA,while the specificity and reliability of ABC- ELISA seemed satisfectory.The experi mental condition that might reduce nonspeci- ficity of the reaction to a certain degree was discussed.

We have studied the effects of B7-1 on antitumor immunity induced in vivo by murine B7-1 gene transfected EL-4 lymphoma. At a lethal dose of wild - type(wt) or plasmid controlled (pc) B7-1 negative tumor cells. B7-1 gone transfected EL-4 cells inoculated in syngeneic mice were regressed. In contrast to the mice immunized with B7-1 tumor cells, immunization of mice with B7-l~EL-4 tumor cells showed significant protective effects against the sequence rechallenge of wt EL-4 tumor. Vaccination with X-irradiated B7-1 positive tumor cells at the early stage ( 7 days) after inoculation of wt EL-4 tumor cells showed some therapeutically effects but not at the late stage (14 days after inoculation). Vaccination with B7-l~+ EL-4 tumor cells delayed the occurrence of wt EL-4 tumors and prolonged the survival period of tumor-bearing mice. Our results indicate that the transfcction of B7-1 into tumor cells can increase the immunogenicity of EL-4 tumor and improve host response to wt EL-4 lymphoma. Immunization with B7-1 positive tumors can elicit effective antitumor immunity.

Objective To evaluate the clinical application of enzyme-linked immunospot(ELISPOT) assay for detection of M.tuberculosis antigen specific T cells in the rapid diagnosis of active tuberculosis.Methods Using the rapid enzyme-linked immunospot assay for detection of T cells with secretion of IFN-? specific for M.tuberculosis antigens in blood samples from 112 patients with tuberculosis, 24 patients with hepatitis C, 30 healthy persons, and 65 with other respiratory diseases.Results 107 of 112 tuberculosis patients had detectable M. tuberculosis antigen-specific T cells, whereas 2 of 30 healthy subjects and 65 patients with non-tuberculosis illnesses responded. This assay has a sensitivity of 95.5%, specificity of 93.3%.Conclusions M.tuberculosis specific T cells could serve as accurate markers of M.tuberculosis infection in an area of high tuberculosis prevalence.ELISPOT is a rapid, sensitive and specific method for detecting M.tuberculosis specific T cells.It also gives objective evidence to the diagnosis of active tuberculosis.

Objective To observe the inhibiting activities of fibronectin (FN) and citrullinated fibronectin (cFN) on disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4),and to explore the extracellular regulative mechanisms of ADAMTS-4.Methods FN was incubated with peptidylarginine deaminase type 4 (PADI4).Western blotting analysis was used to verify the citrullination of FN.The binding activity of FN and cFN to ADAMTS-4 were investigated by enzyme-linked immunosorbent assay (ELISA).The proteolytic ability of ADAMTS-4 after binding to FN and cFN were measured with the aggrecanase activity assay kit.One-way ANOVA,LSD-t test and t-test were used for statistical analysis.Results The immunosignal of citrulline was detected in FN after incubated with PADI4,but not in the absence of PADI4.A higher absorbance at 405 nm was detected when the full-length ADAMTS-4 protein was incubated with FN (2.182±0.042) than cFN (0.624±0.033; t=50.522,P＜0.01).Additionally,the recombinant ADAMTS-4 protein with a truncation at the C-terminus displayed low absorbance at 405 nm when the enzyme was incubated with both FN(0.971±0.024) and cFN(0.934±0.012; t=2.388,P＞0.05).Large amounts of ARGxx peptide were detected with full-length ADAMTS-4 in aggrecanase activity assay [(0.908±0.088) nmol/L],but significantly less when in the presence of FN and ADAMTS-4 [(0.573±0.000) nmol/L,P＜0.05].The production of this peptide was more when full-length ADAMTS-4 was incubated with cFN [(0.830±0.020) nmol/L,P＜0.05] than with FN.The reaction containing the truncated ADAMTS-4 without FN or cFN yielded the highest concentration of ARGxx peptide [(36.420±3.673) nmol/L],peptide production was not significantly altered when FN [(41.099±0.101) nmol/L] or eFN [(41.064±0.083) nmol/L] were added to the reaction.Conclusion FN could bind to ADAMTS-4 and inhibit its proteolytic activity.After citrullinated by PADI4,the binding activity of cFN is weakened and less inhibition to ADAMTS-4.

Objective To investigate the action mechanism of glutamate-mediated signaling pathway in malignant melanoma. Methods WM451LU melanoma cells in log phase were classified into 6 groups, negative control group treated with PBS (100 μl), MK801 group treated with the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (100 μmol/L), CPCCOEt group treated with non-competitive metabotropic glutamate receptor 1 (mGluR1) antagonist CPCCOEt, MAP2 group transfected with adenovirus vector containing microtubule associated protein 2a (Ad-MAP2a), MK801 + MAP2 group treated with MK801 of 100 μmol/L and transfected with Ad-MAP2a, CPCCOEt + MAP2 group treated with CPCCOEt of 10 μmol/L and transfected with Ad-MAP2a. Western blot was performed to detect the expression of an ionotropic glutamate receptor, i.e., N-methyl-D-aspartate receptor type 2A (NMDAR2A) in WM451LU cells transfected with Ad-MAP2a. Scratch motility assay and cell invasion assay were conducted in vitro to detect the changes in migration and invasion ability of WM451LU cells after treated with Ad-MAP2a, MK-801, CPCCOEt alone or in combination. In vivo study was carried out to compare the inhibitory effect of the above treatments on melanoma. Results Western blot revealed a decrease in the expression of NMDAR2A in WM451LU cells after transfected with Ad-MAP2a. The scratch motility assay showed that the number of migrating cells per high power field was 117.04 ± 2.76 in MAP2 group,107.64 ± 6.50 in MK801 group,97.36 ± 4.79 in CPCCOEt group, 43.28 ± 3.02 in MK801 + MAP2 group,30.76 ± 3.97 in CPCCOEt + MAP2 group,significantly different from that in the negative control group (152.3 ± 5.75,all P ＜ 0.01 ). Cell invasion assay demonstrated that the average number of invading cells per high power field in the negative control was significantly higher than that in MAP2 group, MK801 group, CPCCOEt group, MK801+MAP2 group and CPCCOEt + MAP2 group (170.43 ±8.72 vs. 98.26 ± 3.84, 97.22 ± 5.54, 112.23 ± 7.21, 42.89 ± 5.06, 58.25 ± 6.68, P ＜ 0.05, 0.05, 0.05, 0.01and 0.01, respectively).A significant decrease was observed in the average volume of experimental melanoma in mice of MAP2 group, MK801 group, MK801 + MAP2 group, CPCCOEt group and CPCCOEt + MAP2 group compared with the negative control group (224.02 ± 46.19 mm3, 160.33 ± 33.91 mm3, 91.49 ± 21.48 mm3,202.30 ± 52.37 mm3, 111.13 ± 69.81 mm3 vs. 342.70 ± 60.92 mm3, all P ＜ 0.01 ). Conclusions To block the glutamate signaling pathway in vitro can inhibit the invasion and migration of melanoma cells, and to block the pathway in vivo can inhibit the growth of malignant melanoma and alter the morphology of melanoma cells.

BACKGROUND: Facial nerve injury causes facial nerve paralysis (or facial palsy) and even results in psychosocial disturbances of the patients. Repair the injured facial nerve and reconstruction of the nerve function as early as possible have been the primary concern in clinical studies.OBJECTIVE: To investigate the timing and surgical approaches for repairing facial paralysis in order to provides evidences for its therapeutic and prognostic evaluation.DESIGN: Case analysis based on patients.SETTING: Hospital of Plastic Surgery of Chinese Academy of Medical Sciences.PARTICIPANTS: Nine patients with traumatic facial paralysis hospitalized in the Hospital of Plastic Surgery of Chinese Academy of Medical Sciences from December 1993 to November 2001.METHODS: Facial nerve anastomosis was performed microsurgically along with the implantation of the sural nerve graft into orbicular muscle of the eye 3 or 4 months after nerve injury in the 9 patients. The clinical data of the patients were retrospectively reviewed.MAIN OUTCOME MEASURES: The facial nerve function was evaluated with House-Brackmann scale and photographs of the patients' faces before and after surgical treatment were taken.RESULTS: In the follow-up of these patients lasting 6 months to 2 years,all the patients attained satisfactory outcome.CONCLUSION: Early operation is crucial for the treatment of traumatic facial paralysis,with facial nerve anastomosis as the primary choice. The implantation of the nerve graft into muscle is also indicated for repairing traumatic facial paralysis in some cases. The importance of individualized treatment choice is reiterated on the basis of cicatrectomy and the extent and specific features of the injury.

OBJECTIVE: To investigate the effect of operation on Benign thyroid nodules with hoarseness as primary symptom. METHOD: Twelve patients were underwent the operation of subtotal thyroidectomy and exposing of recurrent laryngeal nerve. We evaluating the effect by fibrolaryngoscope and voice acoustic analysis before and after operation. RESULT: All the 12 patients underwent surgery successfully. The hoarseness improved obviously and vocal cords were reactivate. Jitter, shimmer and dysphonia severity index showed significant difference pre and one month after surgery. CONCLUSION Benign thyroid nodules could also cause vocal cord paralysis and hoarseness, the effect can be satisfying by subtotal thyroidectomy and exposing of recurrent laryngeal nerve if it can be early diagnosed.
Adult ; Aged ; Female ; Follow-Up Studies ; Hoarseness ; etiology ; Humans ; Male ; Middle Aged ; Thyroid Nodule ; complications ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVE: To define the oncologic efficacy of transoral endoscopic CO2 laser surgery in early glottic carci noma. METHOD: A retrospective study of 112 patients with laryngocarcinoma treated. Surgical treatment included endoscopic CO2 laser cordectomies according to the classification of the European Laryngological Society. After the patients were given the general anesthesia and oral intubation, the tumors in the study group were resected along the margin of the tumor under larynscope, and the safety margin was reserved as 3-5 mm. All the patients were followed-up for 12-62 months. RESULT: Eight relapses were detected in 112 cases of glottic laryngeal carcinoma after CO2 laser surgery. The local recurernce rate was 7.14% (8/112),of the rate for T1a, T1b and T2 were 0.89%, 0.89% and 5.04% respectively, with significant differences among groups (chi2 = 5.306, P < 0.01) . The rate of local recurrence rate of anterior commissure involvement was 7.14% and that was 7.14% when this site was not compromised by the tumor, which has no statistically significant differences (chi2 = 0.000, P > 0.01). CONCLUSION According to our reaserch, endoscopic CO2 laser sur gery is an effective treatment for early laryngocarcinoma.
Adult ; Aged ; Aged, 80 and over ; Carbon Dioxide ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Laryngeal Neoplasms ; surgery ; Laser Therapy ; Lasers, Gas ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome