BACKGROUND:In anoxic environment, invasive ability of tumor cels significantly increased. The increase in above invasive ability is possibly associated with the changes in urokinase or matrix metaloproteinases. Studies on hypoxia and tumor therapy especialy the effect on chemotherapy have made great progress, and found that hypoxia-inducible factor 1 is a key nuclear factor mediating celular hypoxia response, and also an important factor for regulating cel apoptosis during hypoxia. OBJECTIVE:To observe the effects of cyclooxygenase-2 inhibitor NS-398 on invasive ability of MG-63 cels under hypoxic condition in cel models of hypoxia osteosarcoma. METHODS: Chemical hypoxia inducer cobalt chloride was used to establish chemical hypoxia environment of concentration gradient of 0, 100, 200, 400 μmol/L. Under 200 μmol/L cobalt chloride, concentration gradient of 30, 60, 90 μmol/L of NS-398 were set. Osteosarcoma cels were determined after 48 hours of culture. Quantitative PCR was used to detect the expression of matrix metaloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA. Western blot assay was applied to measure hypoxia-inducible factor 1α expression. Flow cytometry was utilized to examine cel apoptosis in each group. RESULTS AND CONCLUSION:PCR and western blot assay results demonstrated that with increased concentration of cobalt chloride, matrix metaloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA and protein expression increased (P < 0.05), showing a concentration-dependent manner. Under cobalt chloride concentration of 200 μmol/L and hypoxia, with increased concentration of NS-398, matrix metaloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA and protein expression gradualy reduced, showing a concentration-dependent manner (P < 0.05). Flow cytometry results showed that under 400 μmol/L cobalt chloride concentration, cel apoptosis noticeably reduced. Under cobalt chloride concentration of 200 μmol/L and hypoxia, with increased concentration of NS-398, cel apoptosis increased (P < 0.05). Results confirm that anaerobic environment is an important factor for increased invasive ability of osteosarcoma. Under the anaerobic environment, NS-398 could effectively suppress the invasive ability of osteosarcoma and promote cel apoptosis.

To evaluate the inhibitive effect of plant protein MAP30 (momordica anti-HIV protein of 30kDa), AZT (3′-azido-2′,3′-dideoxythymidine) , ACV (acyclovir) and IFN-?2a (interferon-?2a) against HIV-1 in vitro, MT4 was used as the target cell and the inhibitive effects of these drugs on HIV-1 P24 expression were investigated by ELISA. The inhibitory effect of these drugs on HIV-1-induced cytopathy was also evaluated. The 50% inhibition concentration (IC 50) of MAP30 was 0.9?mol/L. In comparison, the IC 50 for AZT , a commonly used anti-HIV drug, was 0.7?mol/L. The cytopathic effect induced by HIV-1 was also inhibited by MAP30 and AZT. ACV and IFN-?2a showed little effect on HIV-1. All these results strongly indicated that plant protein MAP30 could obviously inhibit HIV-1 replication in vitro.

Objective To construct the nucleic acid vaccine(DNA vaccine) plasmid of the gag gene of the prevalent HIV-1 strains in China so as to develop a therapeutic AIDS vaccine against the HIV-1 strains, and to investigate the immune responses induced by HIV-1 DNA vaccine in mice. Methods The recombinant expression vector pCI-neo GAG was constructed by inserting HIV gag gene into the eukaryotic expression vector pCI-neo, which was identified by restrictive enzymes (Xba Ⅰ/Sal Ⅰ) digestion analysis and DNA sequencing. Balb/c mice were immunized with pCI-neo GAG or pCI-neo. Their sera were collected for the assay of the titer of anti-HIV antibody and IFN-? level by ELISA, and their splenic cells were isolated for detecting antigen-specific lymphoproliferative responses and specific cytotoxic T lymphocyte(CTL) response by MTT assay and LDH assay, respectively. Results Restrictive enzymes digestion analysis and DNA sequencing results revealed that the recombinant expression vector pCI-neo GAG had been constructed successfully. Both the anti-HIV antibody titer and the IFN-? level of mice immunized with the pCI-neo GAG were higher than those of mice immunized with pCI-neo (P

Objective To discuss the apparatus of inspired oxygen in hyperbaric oxygen(HBO)chamber for patients after tracheotomy.Methods Many kinds of apparatus of inspired oxygen were analyzed,including composition,usage and characteristics.Results The apparatus of inspired oxygen play an active role in treating patients.But in the above apparatus,there are great differences in serving range,efficiency of inspired oxygen,curative effect,reliability and so on.Conclusions When serious patients after tracheotomy are treated in HBO chamber earlier,it is very important to select reasonable apparatus.

MicroRNAs ( miRNAs ) , a highly conserved and non coding RNA, modulates gene expression via inhibiting the translation of protein and thereby influences cell fate and function.A large body of literature demonstrates that miRNAs plays a critical role in the maintenance of immune homeostasis through modulating immune cell differentiation, function as well as apoptosis.As important antigen presentation cells, dendritic cells ( DCs) play a central role in the initiation and control of the adaptive immune response in the peripheral lymphatic system.Uncontrolled inflammatory response and immunosuppressive response followed by tissue and organ dysfunction were associated with DC loss and apoptosis after septic challenge.This review will focus on the effect of miRNAs on DC function and apoptosis, and its potential role in sepsis.

This paper introduces the current situation of hyperbaric oxygen (HBO) therapy, the environment characteristics in HBO chamber and the qualification of sputum aspirators. What's more, this paper also describes the construction, principles and characteristics of all kinds of sputum aspirators in HBO chamber. When critical patients are treated in HBO chamber, it is very important to select reasonable sputum aspirators.

Objective:To construct the eukaryotic expression vector of interleukin-18 gene and carry out analysis of pVAX1-IL-18 sequence . Methods:The recombinant eukaryotic expression vector pVAX1-IL-18 was constructed by inserting interleukin-18 gene into the eukaryotic expression vector pVAX1 with molecular cloning technique . It was confirmed by restrictive enzymes(BamHⅠ/ EcoRⅠ) digestion and analysis of DNA sequencing . Similar nucleotide sequences encoding IL-18 of pVAX1-IL-18 were looked up by Blast means in NCBI GenBank. Results:Restrictive enzymes digestion analysis and DNA sequencing results revealed that the interleukin-18 gene was cloned into the eukaryotic expression vector pVAX1 successfully .The sequence of encoding IL-18 of pVAX1-IL-18 was exactly the same as the reported sequence of encoding human IL-18 in GenBank . Conclusion: The recombinant eukaryotic expression vector pVAX1-IL-18 has been constructed successfully, which lay the foundation for pVAX1-IL-18 as a genetic adjuvant of DNA vaccine.

Objective To detect the expressions of Wnt2 and dishevelled (Dvl) protein in esophageal squamous carci-noma, and analyze their relationship with the occurrence and development of esophageal squamous cell carcinoma. Meth-ods The expression levels of Wnt2 and Dvl protein were detected by Western blot assay in 60 samples of esophageal carci-noma and adjacent non-carcinomatous esophageal tissues, and their relationship with clinical pathological features were ana-lyzed. Results The relative expression levels of Wnt2 and Dvl protein were higher in esophageal squamous carcinoma tis-sue (0.512 ± 0.406, 1.218±1.082) than those of esophageal tissue adjacent to carcinoma (0.153 ± 0.189, 0.505±0.358). There were significant differences in the expression levels of Wnt2 and Dvl protein between different infiltration depth, different TNM stages, and lymph node metastasis (P<0.01). There was a positive correlation between Wnt2 and Dvl protein in esopha-geal squamous carcinoma (r=0.718, P<0.01). Conclusion The high expression levels of Wnt2 and Dvl protein promote the development and metastasis of esophageal squamous cell carcinomas collaboratively via Wnt2 signal transduction path-ways.

Objective To detect the expression of CD44v5 in esophageal squamous carcinoma (ESCC) and analyze its relationship with clinical pathological features,so as to explore the value of detecting CD44v5 in serum for screening ESCC. Methods The CD44v5mRNA and protein in ESCC tissues and its corresponding adjacent non-carcinomatous tissues were detected by RT-PCR and Western-Blot. Enzyme-linked immunosorbent assay (ELISA) was applied to detect the protein in serum of 100 ESCC patients and 60 healthy subjects. Results (1)The expression of CD44v5 in esophageal squamous carcinoma tissues is higher than the tissue adjacent to carcinoma, and CD44v5 expression in different TNM stages, invasion depth, presence of lymph node metastasis and differentiation is with statistical differences(P < 0.05). (2)CD44v5 proteins in patients with ESCC (31.308 ± 10.123) μg/L is higher than healthy subjects (19.364 ± 1.680) μg/L, the difference is statistically significant (P < 0.01). (3)In the trial of using ELISA method to detect CD44v5 content in serum in the diagnosis of ESCC, the area under the ROC curve is 0.865. If the healthy subjects' serum content of the upper limit of 95%confidence interval was taken as a positive judgment standard, the diagnosis effect: sensitivity is 91.0%and speciality is 60.0%. Conclusion The expression of CD44v5 was related to the occurrence and lymph node metastasis of ESCC. Using ELISA method to detect the contents of CD44v5 proteins in serum, with a high sensitivity, can be used to screen ESCC.