[Clinical characteristics and prognostic features of 63 HIV-associated diffuse large B-cell lymphoma: a single-center real-world study in China].

Objective: This study aimed to look into the clinical characteristics and prognosis of patients with human immunodeficiency virus (HIV) -associated diffuse large B-cell lymphoma (DLBCL) . Methods: Retrospective review of the clinical data of 63 HIV-infected patients with DLBCL diagnosed at Chongqing University Cancer Hospital between July 2008 and August 2021. The Kaplan-Meier method was used to calculate survival curves, and the log-rank test method was used to compare survival between groups. The Cox proportional hazards model was used for multivariate analysis. Results: In 63 patients with HIV-associated DLBCL, 57 (90.5% ) were men, and the median age was 49 (23-87) years. The most common pathological subtype was the germinal center B-cell-like lymphoma (74.6% ) ; 46.0% (29/63) were combined with extranodal lesions. Seventeen of 63 (27.0% ) patients had large masses (≥7.5 cm) . Twenty of 63 (31.7% ) patients had B symptoms. The median CD4(+) T cell count was 203 (4-1022) ×10(6)/L. A total of 49% (25/51) patients had CD4(+) cell count <200×10(6)/L, 56.9% (33/58) had high (3-5) International Prognostic Index (IPI) scores, and 43.1% (25/58) had low (0-2) IPI scores. Further, 78% (46/59) were diagnosed with Ann Arbor Stage Ⅲ/Ⅳ, and 25.4% (16/63) didn't receive chemotherapy. A total of 22.2% (14/63) of patients received less than four cycles of chemotherapy, and 52.4% (33/63) received four or more cycles of chemotherapy. Among patients undergoing chemotherapy, 61.7% (29/47) received R-CHOP-like regimens, and 38.3% (18/47) used CHOP-like regimens. The 1-, 2-, 3-, and 5-year overall survival (OS) rates were 65.0% , 53.8% , 47.1% , and 43.5% , respectively. Univariate analysis revealed that age ≥ 60 years (P=0.012) , Eastern Cooperative Oncology Gruop Performance Status (ECOG-PS) score 2-4 points (P=0.043) , IPI score 3-5 points (P=0.001) , ß(2)-MG elevation (≥5.5 mg/L) (P=0.007) , and systemic chemotherapy cycles less than four times (P<0.001) were the negative prognostic factors affecting the OS of patients. The Cox multivariate analysis depicted that age ≥60 years (HR=2.272, 95% CI 1.110-4.651, P=0.025) , IPI score 3-5 points (HR=3.562, 95% CI 1.794-7.074, P<0.001) , ECOG-PS score 2-4 points (HR=2.675, 95% CI 1.162-6.153, P=0.021) , and number of cycles of chemotherapy<4 (HR=0.290, 95% CI 0.176-0.479, P<0.001) were independent risk factors for adverse prognosis of OS. Conclusion: HIV-associated DLBCL is the most common HIV-related tumor, is most commonly seen in men, and has a high 1-year mortality rate. Chemotherapy combined with antiretroviral therapy can improve patient prognosis.

Expressions of MiR-132 in patients with chronic hepatitis B, posthepatitic cirrhosis and hepatitis B virus-related hepatocellular carcinoma.

OBJECTIVE: 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. PATIENTS AND METHODS: 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. RESULTS: There were significant differences in some liver function indexes and the HBV-DNA level among the three groups of patients (p < 0.05). 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 (p < 0.05), 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 (p < 0.05). HBx was expressed in the three groups of patients, and liver cancer patients with the highest expression degree of HBx accounted for 46%. CONCLUSIONS: 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.

[Peripheral nerve injury in LAMA2-related congenital muscular dystrophy patients].

Objective: To explore the injury pattern and features of peripheral nerve in congenital muscular dystrophy patients caused by LAMA2 gene mutation. Method: Seventeen patients genetically or molecular pathologically diagnosed as LAMA2-related congenital muscular dystrophy were recruited in Peking University First Hospital between 2002 and 2015. All the patients received nerve conduction velocity (NCV) and needle electromyography tests. Clinical and laboratory examination data of the patients was retrospectively analyzed. The correlation between the NCV and disease course was determined by Pearson correlation analysis. Additionally, one patient underwent a sural nerve biopsy. Result: Among these 17 identified patients (13 male and 4 female), all of them were diagnosed as congenital muscular dystrophy, and all of them underwent electrophysiological examination at ages between 1 month to 6 years. Electromyogram indicated seventeen patients of myogenic damage, of whom 10 cases were complicated with reduced NCV. Twenty-six of 95 analyzed nerves showed NCV slower than the normal average of contemporary in 17%-47%. Correlation analysis between NCV and the disease course indicated that NCV of median nerves, ulnar nerves, tibial nerves and common peroneal nerves were negatively associated with the disease course (r=-0.737, -0.771, -0.540 and -0.682, respectively; all P<0.05). Sural nerve biopsy revealed peripheral neuropathy changes of myelin. Conclusion: There is peripheral nerve injury in LAMA2-related muscular dystrophy patients. It mainly manifests as demyelinating lesions. Moreover, the NCV of peripheral nerve will decrease with the increase of the course of the disease.

Cooled, ultrahigh Q, sapphire dielectric resonators for low-noise, microwave signal generation.

Ultra-high Q, X-band resonators, used in a frequency discriminator for stabilization of a low-noise signal generator, can provide a means of obtaining significant reduction in phase noise levels. Resonator unloaded Qs on the order of 500 K can be obtained in sapphire dielectric resonator (DR) operating on a low-order (i.e. TE(01)) mode at 77 K and employing high-temperature superconducting (HTS) films installed in the DR enclosure covers. Rigorous analysis for the determination of resonator frequency, modes, and unloaded Q have been carried out using mode matching techniques. Trade-off studies have been performed to select resonator dimensions for the optimum mode yielding highest unloaded Q and widest spurious mode separation. Field distributions within the resonator have been computed to enable practical excitation of the required mode. The results of both analysis and prototype device evaluation experiments are compared for resonators fabricated using enclosures consisting of conventional, metal sidewalls and covers employing HTS films as a function of cover conductivity.

Pseudorabies virus gIII and bovine herpesvirus 1 gIII share complementary functions.

The gIII glycoproteins of bovine herpesvirus 1 (BHV-1) and of pseudorabies virus (PRV) are structurally homologous. Both proteins also play preeminent roles in mediating virus attachment to permissive cells. To directly compare the functional relation between these glycoproteins, we constructed a recombinant BHV-1 in which the BHV-1 gIII coding sequence was replaced by the PRV gene homolog. The resultant recombinant virus efficiently expressed PRV gIII and then incorporated it into its envelope. The levels of PRV gIII expression and incorporation were equivalent to those achieved by the wild-type virus for BHV-1 gIII. The recombinant virus was fully susceptible to neutralization by anti-PRV gIII neutralizing antibody. In addition, the virus attachment and penetration functions, as well as the virus replication efficiency, which were lost by deleting the BHV-1 gIII gene, were restored by expressing the PRV gIII homolog in its place. These results demonstrated that PRV gIII and BHV-1 gIII share complementary functions.

Bovine herpesvirus 1 attachment to permissive cells is mediated by its major glycoproteins gI, gIII, and gIV.

A bovine herpesvirus 1 (BHV-1) gIII deletion mutant (gIII-) was produced by means of recombinant DNA that retained the ability to replicate in cell culture. However, the gIII- mutant was functionally defective, showing impaired attachment to permissive cells, a delay in virus replication, and reduced extracellular virus production. The attachment defect exhibited by the gIII- mutant is an indication of the role played by gIII in the normal infection process. This was shown by dramatically decreased binding of radiolabelled gIII- virus to permissive cells and a slower adsorption rate, as measured by plaque formation, than the wild-type (wt) virus. Furthermore, treatment of the gIII- virus with neomycin increased virus adsorption and plaque formation by severalfold, whereas neomycin treatment had no effect on the wt virus. This observation showed that the gIII- mutant was strictly defective in adsorption but fully competent to produce productive infections once induced to attach. The gIII- mutant showed greater sensitivities than did the wt virus to anti-gI and anti-gIV antibody-mediated neutralization. Analyses with panels of monoclonal antibodies to gI and gIV revealed that the epitopes gI-IV and gIV-III were the main targets for enhanced neutralization. This provided evidence that gI and gIV may also participate in virus attachment. Finally, when affinity-purified gI, gIII, and gIV were tested for their ability to inhibit virus adsorption, gIII had the most pronounced inhibitory effect, followed by gI and then gIV. gIII was able to completely inhibit wt virus adsorption, and at a high concentration, it also partially inhibited the gIII- mutant. gI and gIV inhibited wt and gIII- mutant adsorption to a comparable extent. Our results collectively indicate that gIII plays a predominant role in virus attachment, but gI and gIV also contribute to this process. In addition, a potential cooperative mechanism for virus attachment with these three proteins is presented.

Cholera toxin as a mucosal adjuvant. Glutaraldehyde treatment dissociates adjuvanticity from toxicity.

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.

Cholera toxin as a mucosal adjuvant for respiratory antibody responses in mice.

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.

Transport of serum IgA into murine respiratory secretions and its implications for immunization strategies.

Both nonlabeled and radiolabeled IgA mAb with specificity toward Sendai virus, a respiratory pathogen, were used to investigate the transport of serum polymeric and monomeric IgA into murine upper and lower respiratory secretions as well as into the gut. After purification by affinity chromatography, IgA mAb were fractionated into monomers and polymers by gel filtration and radiolabeled with 125I. Mice were injected i.v. with either 125I-monomer and 131I-albumin or 125I-polymer and 161I-albumin. At various times after injection, serum and gut, nasal and bronchoalveolar lavage samples were collected. The TCA precipitable radioactivities were determined and the selective transport indices calculated. The results indicated selective transport of polymeric IgA but not monomeric IgA from serum into upper respiratory and intestinal secretions. The degree of TCA precipitability in nasal lavage and to a lesser extent gut secretions suggested significant degradation of the antibody during or after transport. To investigate further the integrity of the IgA in mucosal secretions, ELISA viral binding activity of nonradiolabeled IgA was determined for both IgA incubated with nasal secretions in vitro and polymeric IgA recovered by nasal lavage 4 h after i.v. injection. Although reconstitution experiments indicated no significant loss of antibody binding activity after incubation of antibody with lavage fluid in vitro, only negligible ELISA binding activity was detected in nasal washes after i.v. injection of antibody. The data overall suggest that although there is a quantitatively small, but selective transport of polymeric IgA into the upper respiratory tract, this transport results in minimal functional antibody activity. Implications of these and other findings for strategies of oral immunization in prophylaxis against respiratory infections are discussed.

Oral immunization with Sendai virus in mice.

We have shown that in mice cholera toxin can be an effective adjuvant for gastrointestinal immune responses against a virus. The adjuvant properties can be increased and even dissociated from the toxic properties if virus and toxoid are covalently linked. Finally, oral immunization with these preparations of cholera toxin/toxoid and Sendai virus can be used to prime for respiratory immune responses to Sendai virus in which protection from infection correlates with IgA in the upper and with IgG in the lower respiratory tract.

Oral administration of cholera toxin-Sendai virus conjugate potentiates gut and respiratory immunity against Sendai virus.

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.

Combined oral/nasal immunization protects mice from Sendai virus infection.

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.