Myricetin exhibits anti-glioma potential by inducing mitochondrial-mediated apoptosis, cell cycle arrest, inhibition of cell migration and ROS generation.

The Editors of JBUON issue an Expression of Concern to 'Myricetin exhibits anti-glioma potential by inducing mitochondrial-mediated apoptosis, cell cycle arrest, inhibition of cell migration and ROS generation', by Hu-Guang Li, Jun-Xia Chen, Jun-Hui Xiong, Jin-Wei Zhu, JBUON 2016;21(1):182-190; PMID:27061547. Following the publication of the above article, readers drew to our attention that part of the data was possibly unreliable. We sent emails to the authors with a request to provide the raw data to prove the originality, but received no reply. Therefore, as we continue to work through the issues raised, we advise readers to interpret the information presented in the article with due caution. We thank the readers for bringing this matter to our attention. We apologize for any inconvenience it may cause.

Specific determination of hepatitis B e antigen by antibodies targeting precore unique epitope facilitates clinical diagnosis and drug evaluation against hepatitis B virus infection.

Hepatitis B e antigen (HBeAg) is a widely used marker both for chronic hepatitis B (CHB) clinical management and HBV-related basic research. However, due to its high amino acid sequence homology to hepatitis B core antigen (HBcAg), most of available anti-HBe antibodies are cross-reactive with HBcAg resulting in high interference against accurate measurement of the status and level of HBeAg. In the study, we generated several monoclonal antibodies (mAbs) targeting various epitopes on HBeAg and HBcAg. Among these mAbs, a novel mAb 16D9, which recognizes the SKLCLG (aa -10 to -5) motif on the N-terminal residues of HBeAg that is absent on HBcAg, exhibited excellent detection sensitivity and specificity in pairing with another 14A7 mAb targeting the HBeAg C-terminus (STLPETTVVRRRGR, aa141 to 154). Based on these two mAbs, we developed a novel chemiluminescent HBeAg immunoassay (NTR-HBeAg) which could detect HBeAg derived from various HBV genotypes. In contrast to widely used commercial assays, the NTR-HBeAg completely eliminated the cross-reactivity with secreted HBcAg from precore mutant (G1896A) virus in either cell culture or patient sera. The improved specificity of the NTR-HBeAg assay enables its applicability in cccDNA-targeting drug screening in cell culture systems and also provides an accurate tool for clinical HBeAg detection.

Clinical significance and prognostic value of tumor necrosis factor-α and dickkopf related protein-1 in ankylosing spondylitis.

BACKGROUND: Ankylosing spondylitis (AS) frequently occurs in people aged 30-45 years, and its prevalence is generally believed to be between 0.1% and 1.4% globally. At present, the "gold standard" for diagnosis of AS requires the provision of pelvic X-rays, which makes it more difficult to perform in population-based epidemiological studies. Therefore, the identification of serological indicators related to the diagnosis, treatment, and prognosis of AS patients is of great significance. AIM: To analyze the therapeutic, diagnostic significance and prognostic value of dickkopf-related protein-1 (DKK-1) and tumor necrosis factor-α (TNF-α) in AS. METHODS: A total of 113 patients with active AS were selected as the research group, and 100 healthy subjects who underwent physical examination were selected as the control group. The levels of DKK-1 and TNF-α in peripheral blood in the two groups were compared. The diagnostic and predictive values of DKK-1 and TNF-α for AS were analyzed with ROC curves, and the factors influencing AS recurrence were analyzed with COX regression. RESULTS: Before treatment, the research group showed lower DKK-1 levels but higher TNF-α levels than the control group (both a P < 0.05). In the research group, DKK-1 was up-regulated and TNF-α was down-regulated after 12 wk of treatment (a P < 0.05). The area under the curve, sensitivity and specificity of DKK-1 combined with TNF-α for diagnosing AS were 0.934, 82.30% and 97.00%, respectively. Before treatment, the area under the curve, cutoff value, sensitivity and specificity of DKK-1 for predicting the curative effect were 0.825, 68.42 pg/mL, 73.68% and 80.00%, respectively, and those of TNF-α were 0.863, 32.79 ng/L, 92.11% and 77.33%, respectively. DKK-1 and TNF-α levels after treatment were closely related to the curative effect (a P < 0.05). C-reactive protein, the Bath Ankylosing Spondylitis Disease Activity Index, DKK-1, and TNF-α were risk factors for AS recurrence (a P < 0.05). CONCLUSION: DKK-1 and TNF-α are effective in the diagnosis and treatment of AS and are risk factors for its recurrence. In addition, DKK-1 may be a potential target for the diagnosis of AS.

Myricetin exhibits anti-glioma potential by inducing mitochondrial-mediated apoptosis, cell cycle arrest, inhibition of cell migration and ROS generation.

PURPOSE: To study the antiproliferative effects of myricetin in human glioma U251 cells together with assessing its effects on cell cycle, apoptosis, apoptosis-related proteins, reactive oxygen species (ROS) generation and cell migration. METHODS: Cell viability of human glioma cells after myricetin treatment was assessed by MTT assay. Phase-contrast and confocal fluorescence microscopies were used to assess the morphological changes that occured in these cells following myricetin treatment. Flow cytometry using propidium iodide (PI) and Annexin-V FITC as probes was employed to evaluate the effects on cell cycle arrest and apoptosis induction, respectively. The effect of myricetin on intracellular ROS production was measured by flow cytometry with a fluorescent probe CM-DCFH2-DA. RESULTS: Myricetin induced a dose-dependent as well as time-dependent growth inhibitory effect in U251 human glioma cells. Myricetin treatment resulted in U251 cells detachment from adjacent cells making clusters of cells floating in the medium. Detached cells had irregular shape and incapable to maintain their membranes intact. Apoptotic cell death was induced by myricetin treatment as witnessed by fluorescence microscopy. The percentage of early and late apoptotic cells increased from 0.41% and 8.2% to 23.1% and 10.2%, 25.2% and 19.4%, to finally 36.2% and 28.4% after treatment with 15 µM, 60 µM and 120 µM of myricetin, respectively. We also observed a dose-dependent increase in Bax and Bad levels and a dose-dependent decrease in Bcl-2 and Bcl-xl expression levels following myricetin treatment. Cell cycle arrest in G2/M phase of the cell cycle was also induced by the drug treatment. A concentration-dependent ROS generation was also witnessed and a 3-fold increase of ROS production was seen after 60 µM myricetin treatment. CONCLUSION: Myricetin exerts anticancer effects in U251 human glioma cells by inducing mitochondrial-mediated apoptosis, G2/M phase cell cycle arrest, ROS generation and inhibition of cell migration.

Expression of nuclear factor -κBp65 in mononuclear cells in Kawasaki disease and its relation to coronary artery lesions.

OBJECTIVE: To assess the association of nuclear factor-kappa B (NF-κB) and complications of Kawasaki disease (KD) in Chinese children. METHODS: Based on color Doppler examination results, 86 affected children in the KD group were divided into two groups: 39 cases in coronary artery lesion group (CALs subgroup) and 47 cases in non-coronary artery lesion group (Non-CALs subgroup). Infection control group consisted of 65 cases of hospitalized infected children with fever, having same age as the affected children. Healthy control group consisted of 102 cases of healthy children of the same age, visiting the hospital for physical examination. Western blot was used to detect the expression of NF-кBp65 and IкBα proteins in periphery blood mononuclear cells (PBMC); reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of TNF-α and MCP-1 mRNA. RESULTS: The value of NF-kBp65 (optical density) in the PBMC cell nuclei in the KD group was significantly higher than that in the two control groups (p < 0.01). The value of NF-κBp65 in the CALs subgroup was significantly higher than that in the Non-CALs subgroup (p < 0.05). The value of NF-κBp65 inhibitor IκBα in the KD group was significantly lower than that in the infection control group and the healthy control group (p < 0.01). There was a positive correlation between the ratio nucleus NF-κBP65/ IκBα and the severity degree of CALs(r = 0.536, p < 0.05). The value of TNF-α mRNA (O.D ratio) in the KD group was significantly higher than that in the two control groups (P < 0.01), and the value of TNF-α mRNA in the CALs subgroup was significantly higher than that in the Non-CALs subgroup (P < 0.05). The value of MCP-1 mRNA in the KD group was significantly higher than that in the two control groups (P < 0.01), and the value of MCP-1 mRNA in the CALs subgroup was significantly higher than that in the Non-CALs subgroup (P < 0.05). CONCLUSIONS: NF-κBp65 participates in the pathogenesis of vasculitis of KD in acute stage, and may aggravate the vasculitis in KD and plays a part in the formation of CALs.

Prevalence of hepatitis E virus in Chinese blood donors.

A point prevalence study of hepatitis E virus (HEV) in Chinese blood donors was conducted, and the prevalences of antibodies against HEV immunoglobulin G (IgG) and IgM among Chinese blood donors were 32.60% and 0.94%, respectively. HEV viremia was 0.07%.

Randomized-controlled phase II clinical trial of a bacterially expressed recombinant hepatitis E vaccine.

The candidate recombinant hepatitis E vaccine, HEV 239, protect monkeys against infection by hepatitis E virus (HEV). The safety and immunogenicity of the vaccine for humans was assessed in a randomized controlled phase II clinical trial. The study was conducted in an endemic area of southern China and consisted of a dose scheduling, involving 457 adults and a dose escalation component involving 155 high school students. The results showed that the vaccine is safe and immunogenic for humans and suggest that it could prevent new HEV infection.

[The preliminary analysis of the recognition epitopes of anti-HEV monoclonal antibodies on HEV ORF2].

Western blot, capture-PCR, blocking ELISA and synthetic polypeptides were used to systematically study the recognition epitopes on HEV ORF2 of 23 anti-HEV monoclonal antibodies(McAbs) which were previously generated in our laboratory directed against HEV ORF2. Results showed that seven McAbs recognized linear epitopes that located at aa408-458 of HEV ORF2 and 16 conformation-dependent McAbs, most of which recognized the surface epitopes of native HEV, located at aa459-606 of HEV ORF2. The systematical study of the recognition epitopes of anti-HEV McAbs on HEV ORF2 provides important information for the investigation of virus receptor and HEV infection mechanism, as well as its vaccine and diagnostics development.

[The molecular differences between genotype 1 and genotype 4 of HEV in their neutralization region].

HEV is classified into H (human) group and Z (zoonosis) group according to its compatible host. H group contains genotype 1 and genotype 2 HEV isolates which infect human only; Z group contains genotype 3 and genotype 4 HEV isolates which infect both human and animals. After analysis of amino acid sequences between ORF2 aa368 and aa606, four group-conserved sites that were all located in the neutralization region of ORF2 were identified. They are aa483, aa492, aa497 and aa599. Mutation analysis and capture PCR were then performed on these sites with a group of monoclonal antibodies. Results showed that the difference of the aa497 between H and Z groups was responsible for the maintenance of their group-specific immunodominant epitopes, probably through confirmation-dependent epitope changes. Thus, aa497 and its related change on the surface structure of HEV may play important roles in host selection by H and Z groups of HEV.

[Establishment of a new combined enzyme immunoassay for detection of HBV preS1 and core antigens and the consistency with HBV DNA test].

In this study, a new combined enzyme immunoassay(NRAg ELISA) for detection of HBV PreS1 and core antigens which was highly consistent with serum HBV DNA test was established. The serial serum dilution test indicated that the average sensitivity of the assay was 10(3.2) genome copies/mL (95% CI: 10(2.2-4.2) genome copies/mL), which was notably higher than the test performed on Pre S1 or core antigen alone. The test with sera from 994 blood donors whose HBsAg were negative demonstrated that the specificity of this assay was 99.7% (95% CI: 99.1%-99.9%). 271 serum samples from chronic hepatitis patients were also examined and the result showed that the total consistent rate between NRAg ELISA and HBV DNA was 96.3% (95% CI: 93.3%-98.2%). The NRAg ELISA S/CO(signal/cutoff) was closely correlated with HBV genome copies (R = 0.9158, n=231). Furthermore,by using this assay,we found a patient whose HBsAg was negative but HBV DNA was positive. Sequencing result showed that HBV genome from this patient had a point mutation in the "a"epitope of S gene. Our results indicate that HBV NRAg ELISA has a high relativity with HBV DNA test, and can effectively detect the mutation of HBsAg,it is expected to be a potent tool for screening HBsAg mutant and is a convenient method for substituting HBV DNA test.