OBJECTIVE To study the expressions of heat shock protein 70 and human papillomavirus16E7 protein in human laryngeal squamous cell carcinoma, and their relationship in the genesis of human laryngeal squamous cell carcinoma. METHODS The expressions of HSP70 and HPV16E7 protein were detected by the immunohistochemical method in 78 specimens with laryngeal squamous cell carcinoma, 24 specimens with vocal cord polyps and 10 specimens of normal laryngeal tissues. RESULTS In human laryngeal squamous cell carcinoma, vocal cord polyps and normal laryngeal tissues, the positive expression rates of HSP70 were 69.2 % , 8.3 % and 0 % respectively, with those of HPV16E7 protein being 43.6 % 4.2% and 0 % respectively. There was a significant difference of the expression rate of HSP70 or HPV16E7 protein between the laryngeal squamous cell carcinoma and the vocal cord polyps(P

OBJECTIVE To construct a prokaryotic expression plasmid encoding HPV16E7-HSP70 fusion gene for further study on the immunity of HPV16E7- HSP70 fusion protein against laryngeal carcinoma. METHODS HPV16E7 was PCR-amplified,digested by NheI and SacI,and ligated into pET28a. HSP70 was cloned into pGEMTeasy,then recut from the vector by SalI and NotI and ligated into pET28a-HPV16E7. PCR amplification, restrict enzyme digestion, DNA sequencing, IPTG induction and Western Blot were used to identify the recombinant plasmid. RESULTS Double digestion and PCR amplification of the recombinant plas- mid have shown that the size of the inserted fragment is as expected. Sequence analysis has demonstrated that the inserted fragment encodes for the HPV16E7- HSP70 fusion gene. IPTG induction and Western Blot have shown that the fusion protein is expressed suc- cessfully in the prokaryotic expression plasmid. CONCLUSION The recombinant prokaryotic expression plasmid pET28a-HPV16E7-HSP70 has been con- structed successfully.

Objective:To evaluate the toxic effect of 3 different gingival retraction cords.Methods:DMEMextraction of DL-adren-aline HCl,aluminium sulphate and non-drug retraction cords with the extraction time of 5,10,15 and 30 min were respectively pre-pared and were used to culture human gingival fibroblasts(HGFs)in vitro respectively.Cell proliferation was tested by MTT assay. Cell apoptosis was examined by Annexin/PI method.Results:The 3 gingival retraction cord extractions inhibited the roliferation,pro-moted the apoptosis of HGFs(P <0.05),the effects were related to the extraction time.Conclusion:The 3 retraction cords have time-dependant cytotoxity.

Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Child ; Cytarabine ; administration & dosage ; Daunorubicin ; administration & dosage ; adverse effects ; Female ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Male ; Middle Aged ; Mitoxantrone ; administration & dosage ; adverse effects ; Young Adult

Background The neovascular form of the disease usually causes severe vision loss in a number of eye diseases.Special-domain optical coherence tomography (SD-OCT) provides high-quality in retinal imaging and the possibility of the measurement in vivo.Objective This study aimed to investigate the feasibility of SD-OCT dynamically measuring choroidal neovascularization (CNV).Methods CNV was induced in 30 left eyes of 30 clean Brown Norway(BN)rats by retinal photocoagulation with the laser parameter as follows: wavelength 532 nm,exciting power 200 mW,spot diameter 100 μm and irradiating time 50 ms.Bubble or less retinal bleeding was thought as Brunch membrane breakage and CNV model establishment.Fundus fluorescein angiography(FFA) was performed to determine the establishment of CNV model and scored based on the fluorescein leakage on 3,7,14,21 days after photocoagulation.Meanwhile,CNV memberane thickness (CMT) was dynamically measured in vivo as the maxiume value from retinal inner limiting membrane through choroidal vessel layer in various time points.Histopathologic examination was used in the 14th day to evaluate and verify the result of SD-OCT.The right eyes were as controls.Results FFA examination showed that disc-like leakage of fluorescein appeared in 7 days and extended in 14 days after photocoagulation with the scores of 1.6±0.4,2.5±0.6 and 2.4±0.5 in 7,14 and 21 days,showing a significant difference among them(F=13.11,P＜0.01).The fluorescein leakage score was significantly higher in 14 and 21 days than that of 7 days(both P＜0.05).CMT measured by SD-OCT was(76.33±10.09),(102.03±14.21)and(98.03±13.76) μm in 7,14 and 21 days after photocoagulation respectively,with a significant difference among 3 time points (F=23.25,P＜0.01),and that in 14 and 21 days was significantly declined in comparison with 7 days(both P＜0.05).The results of SD-OCT showed a consistent tendency with that of FFA.Histopathological examination showed CNV formation in 14 days after photocoagulation.Conclusions Experimental CNV model was successfully induced by laser photography.SD-OCT technology allows excellent visualization of CNV in vivo.

PurposeTo study the effect of gasoline on the extracellular matrix of dermis in rat. Methods45 male Wistar rats were divided into control group, 3 different exposure level groups, which were exposed to gasoline on 2 cm× 3 cm skin area with the dose of 250 mg/cm2 for 1 (group A) ,4(group B)and 8 days (group C) respectively,and an intervention group exposed to 250 mg/cm2 × 8 days gasoline after the application of protective agent on skin. After the treatment, collagen, elastin and glycosaminoglycan (GAG) were assayed in skin departed from intoxicated area.ResultsCompared with control group, collagen was decreased in group C(P<0.05); elastin was decreased in group B and C (P<0.05); glycosaminoglycan was decreased in all exposure groups and intervention group ( P < 0.05 or P < 0.01 )。 Collagen, elastin and glycosaminoglycan(GAG) were significantly lower in group C than in intervention group ( P < 0.05 ). ConclusionsThe extracellular matrix of dermis,including collagen,elastin and GAG were decreased in rat dermal exposed to gasoline.

To cultivate applied capability of bioengineering students, five aspects should be done: optimization of experiment teaching system, reformation of experiment teaching mode, intensifying management of experiment teaching, increase of new experiments related tightly to scientific research, and training students’ self-study capability.

OBJECTIVE: To observe the effects of intermedin preconditioning on hypoxic injury in rat's cardiac myocytes and to provide the hypothetical mechanism of sudden cardiac death in the field of forensic pathology. METHODS: The H9c2 cultured rat cardiac myocytes were randomly divided into control group, hypoxia group and IMD group. The myocardial cell viability, cellular ultrastructure, intracellular calcium concentration and apoptosis rate were determined by MTT assay, transmission electron microscopy, laser scanning confocal microscope and flow cytometry, respectively. RESULTS: Compared with the control group, cell viability obviously decreased with inner ultrastructure injury in the hypoxia group (P<0.05), while cell viability significantly increased in the IMD group by reducing the hypoxia injury of cardiac myocytes (P<0.05). Compared with the control group, [Ca2+]i (fluorescence intensity) and apoptosis rate significantly increased in the hypoxia group, but decreased in the IMD group (P<0.05). CONCLUSION IMD increases the cell survival rate and decreases the cell apoptosis inhibited by intracellular calcium overload from hypoxia. This finding may reveal the mechanism of protective effects of myocardial hypoxia, and provide a scientific basis for the identification sudden cardiac death.
Animals ; Apoptosis ; Calcium ; Cell Hypoxia ; Cell Survival ; Hypoxia ; Myocardial Ischemia ; Myocardium/cytology* ; Myocytes, Cardiac/physiology* ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To investigate the relation between injury time and the expression of cytochrome c oxidase subunit VIc (COX6C) mRNA in skeletal muscle of rat after contusion. METHODS: A total of fifty-four SD rats were divided into the control group and the contusion groups (0.5, 1, 6, 12, 18, 24, 30, and 36 h after contusion), randomly. The contusion model was established by free fall drop of gravity hammer. At corresponding time point after contusion, the regular histology was examined and expression level of COX6C mRNA was tested by real-time PCR after extraction of total RNA from the tissues. RESULTS: The main pathological features of 6 h after injury included edema and hemorrhage in myocytes with no inflammatory cells found. After 6 hours, the findings included myocyte degeneration and necrosis, inflammatory cells infiltration, and fibrous connective tissue proliferation in the contused zone. The expression level of COX6C mRNA was higher than that of the control group within 6 h after contusion. The expression level was lower than that of the control group from 6-36 h after contusion. CONCLUSION The level of COX6C mRNA expresses in a regular way after contusion. It may be useful for estimating wound age in combination with the results of pathological features.
Animals ; Contusions/metabolism* ; Electron Transport Complex IV/metabolism* ; Muscle, Skeletal/metabolism* ; RNA ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Time Factors

Background Angiogenesis is an essential event for the growth and mobility of malignant tumors,but there is little literature about the effects of anoxia and angiogenesis-related factors on angiogenesis in choroidal melanoma.Objective The aim of this study was to investigate the expression of angiogeneic factors such as hypoxia inducible factor-1α(HIF-1α),inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),and vascular endothelial growth factor(VEGF)in choroidal melanoma and their clinical significance.Methods The specimens from 44 cases of choroidal melanoma and 16 cases of eyelid nevi identified by pathology were collected.Immunohistochemistry SP was used to examine the expression of HIF-1α,iNOS,COX-2 and VEGF in the samples.The expression of HIF-1 α,iNOS,COX-2 and VEGF in tumors with different sizes,different pathological types and different growth patterns was evaluated.The correlations among these four proteins in tumor growth and development were assessed.Results The expression of HIF-1α,iNOS,COX-2 and VEGF in choroidal melanoma were significantly higher than those in the eyelid nevi group(x2 =6.58,4.105,8.350,13.240,P＜0.05).The expression of VEGF was associated with tumor size(x2 =9.389,P＜0.05),but not associated with pathological types(x2 =1.186,P＞0.05)and infiltration depth(x2 =5.398,P＞ 0.05).The expression of HIF-1α was associated with tumor size(x2=8.664,P＜0.05)and pathological type(x2=6.622,P ＜ 0.05),but not associated with infiltration depth(x2=4.914,P＞0.05).The expression of iNOS was associated with tumor size(x2 =8.609,P＜0.05),but was not associated withpathological type(x2 =4.789,P＞0.05)and infiltration depth(x2 =4.309,P＞0.05).The expression of COX-2 was associated with tumor size(x2 =7.320,P＜0.05),but was not associated with pathological type(x2 =2.772,P＞0.05)and infiltration depth(x2 =2.103,P＞0.05).The expression of HIF-1 α,iNOS,and COX-2 were significantly correlated with the expression of VEGF(r =0.943,1.000,0.986,P ＜ 0.05).The expression of COX-2 was significantly correlated with the expression of iNOS(r =0.986,P＜0.05).The expression of HIF-1 α was significantly associated with the expression of COX-2(r=0.986,P＜O.05).The expression of HIF-1α was significantly associated with the expression of iNOS(r=0.943,P＜0.05).Conclusions The expressions of the HIF-1 α,iNOS,COX-2 and VEGF protein are up-regulated in choroidal melanoma.They may play important roles in the angiogenesis of the choroidal melanoma.Assessment of the expression of HIF-1α,iNOS and COX-2 may be useful for the diagnosis and therapy of choroidal melanoma.