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To explore the expressions of miR-132 in patients with chronic hepatitis B, posthepatitic cirrhosis and hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), and to investigate its possible mechanism affecting the function of the body.Among 125 patients with HBV, there were 44 cases of chronic hepatitis, 42 cases of liver cirrhosis and 39 cases of liver cancer. Their liver function and HBV-deoxyribonucleic acid (HBV-DNA) viral load as well as the expressions of micro ribonucleic acid-132 (miR-132), phosphoinositide 3-kinase (PI3K), phosphorylated-protein kinase B (p-Akt) and hepatitis B X protein (HBx), were detected.There were significant differences in some liver function indexes and the HBV-DNA level among the three groups of patients (p0.05). The HBV-DNA level was 6.91 Lg copies/mL in the liver cancer group and 5.34 Lg copies/mL in the chronic hepatitis B group. Differences in the expression level of miR-132 among the three groups were notable (p0.05), but this expression level had a negative correlation with the HBV-DNA level. The expressions of PI3K and p-Akt proteins and messenger ribonucleic acids (mRNAs) were markedly different among the three groups (p0.05). HBx was expressed in the three groups of patients, and liver cancer patients with the highest expression degree of HBx accounted for 46%.Differences in the expression of miR-132 among the three groups are evident, which may be associated with differences in liver function, the HBV-DNA level, HBx and the expressions of PI3K and p-Akt proteins to a certain degree.

Cholera toxin was investigated as an adjuvant for anti-virus antibody responses in the respiratory mucosa of mice. Two methods of applying cholera toxin were evaluated: oral administration and intranasal administration. Oral immunization with Sendai virus in the presence of cholera toxin effectively primed for respiratory anti-viral antibody responses, whereas oral immunization with Sendai virus alone was ineffective in this respect. In nasal washes, IgA was the predominant anti-viral antibody enhanced by oral cholera toxin; in bronchoalveolar washes, the enhanced anti-viral antibodies included IgG, IgA, and IgM. Effects of direct administration of cholera toxin to the respiratory mucosa on respiratory anti-viral antibody responses depended on the method of anesthesia used during immunization. With inhalation anesthesia (ether), cholera toxin had no adjuvant effect on respiratory antibody responses to coadministered Sendai virus. In contrast, under parenteral anesthesia (i.e., intraperitoneal ketamine), mice which received cholera toxin and Sendai virus via the respiratory tract showed significantly higher anti-viral IgA and IgG antibody titers in nasal washes and IgG antibody in bronchoalveolar washes than mice which received the virus only.

Based on the concept of a common mucosal immune system wherein mucosal associated lymphocytes traffic among the various mucous membranes, the murine gastrointestinal tract was immunized with Sendai virus antigens in order to elicit a virus-specific immune response in the respiratory tract. Multiple intragastric (oral) administration of live or killed Sendai virus induced IgA and IgG antiviral antibodies in both gastrointestinal secretions and serum. When cholera toxin as an adjuvant was included along with virus, gut IgA and IgG as well as serum IgA responses were enhanced. Antiviral antibodies induced in respiratory secretions by oral killed virus plus cholera toxin, however, were variable and protection from virus challenge was not demonstrated. Significantly higher levels of respiratory antiviral antibodies were induced if immunization with oral killed Sendai virus/cholera toxin was combined with intranasal administration of small amounts of killed virus. The combined immunization also resulted in protection of both the upper and lower respiratory tracts from virus infection. Protection of the upper respiratory tract was correlated with the presence of IgA antiviral antibodies in nasal washings. On the other hand, protection of the lower respiratory tract was correlated with IgG antiviral antibodies in bronchoalveolar lavage fluids. Immunization with intranasal killed virus alone conferred partial protection to the lower respiratory tract and no protection to the upper respiratory tract. Thus, oral immunization with killed virus antigen could prime for a protective immune response in the murine respiratory tract and this protective response included IgA antibodies.

Cholera toxin (CT), either mixed with or conjugated to unrelated protein Ag, is known to enhance the intestinal IgA response of rodents toward the unrelated Ag. Although relatively low doses of CT exert this gut mucosal adjuvant effect, the inherent toxicity of CT is a hindrance to its use in humans. Our report demonstrates that CT treated with 20 mM glutaraldehyde retains adjuvant properties but exhibits more than 1000-fold lower toxicity than untreated toxin. Glutaraldehyde was also used in a one-stage conjugation procedure to couple CT covalently to Sendai virus. Again, toxicity was reduced more than 1000-fold. This drop in toxicity is consistent with an observed 100-fold loss in binding capacity of the CT B subunit and a 20- to 50-fold reduction in adenylate cyclase activation by the CT A subunit. Oral administration of this virus-toxoid conjugate resulted in increased gut antiviral IgA titers compared with oral administration of either virus alone or of virus mixed with glutaraldehyde-treated toxin. This marked decrease in toxicity may afford a practical approach for the use of CT as a mucosal adjuvant.

Successful oral immunization to prevent infectious diseases in the gastrointestinal tract as well as distant mucosal tissues may depend on the effectiveness of an Ag to induce gut immune responses. We and others have previously reported that cholera toxin possesses strong adjuvant effects on the gut immune response to co-administered Ag. To explore further adjuvant effects of cholera toxin, the holotoxin or its B subunit was chemically cross-linked to Sendai virus. The resulting conjugates, which were not infectious, were evaluated for their capacity to induce gut immune responses against Sendai virus after oral administration to mice. Conjugating cholera toxin to virus significantly enhanced the adjuvant activity of cholera toxin compared to simple mixing. Cholera toxin B subunit, however, did not show an adjuvant effect either by itself or conjugated with the virus. Oral administration of the Sendai virus-cholera toxin conjugate was also able to prime for protective anti-viral responses in the respiratory tract. Mice that were orally immunized with the conjugate and intra-nasally boosted with inactivated virus alone showed virus-specific IgA titers in nasal secretions that correlated with protection against direct nasal challenge with live Sendai virus. For comparison, s.c. immunization was also studied. Systemic immunization with the virus-cholera toxin conjugate induced virus-specific antibody responses in serum as well as in the respiratory tract but failed to protect the upper respiratory tract against virus challenge. Systemic immunization plus an intra-nasal boost did, however, confer a variable degree of protection to the upper respiratory tract, which correlated primarily with bronchoalveolar lavage (lung) antibody titers.