Preliminary observation of thyroid function changes in subclinical thyroid diseases.

BACKGROUND: It is estimated that 1.95% and 5.55% of adults in China suffer from subclinical thyroid diseases, which is difficult to diagnose and treat. OBJECTIVE: This study aimed to explore the development and prognosis of subclinical thyroid diseases to provide a reference from our single center experience. METHODS: A total of 240 cases from April 2019 to August 2021 in the laboratory information system database of Huanghua Development Boai Hospital were retrospectively analyzed. Binary logistic regression was conducted to analyze odds ratio (OR) of subclinical thyroid disease types returning to a normal state. RESULTS: Among the patients hypothyroidism Ia and hyperthyroidism Ia were the most common type with conversion to the normal state (P< 0.001). TSH level of patients with conversion to a normal state was significantly lower than that of those who developed to abnormal disease (P= 0.015). The OR values of hyperthyroidism Ia and hypothyroidism Ia that returned to a normal state compared with hyperthyroidism Ib were 2.659 (1.159 ∼ 6.096, P= 0.021) and 3.138 (1.1.278 ∼ 7.709, P= 0.013), respectively. The OR value of hypothyroidism Ib that returned to normal compared with hyperthyroidism Ib was 0.629 (0.131 ∼ 3.010, P= 0.561). Thyroid hormone levels, age, and gender at first diagnosis were not impact factor for prognosis of subclincal thyroid disease (P> 0.05). CONCLUSION: Cases with grade hypothyroidism Ia and hyperthyroidism Ia are more likely to revert to normal state than other subclinical thyroid diseases. TSH reference range should be explored for diagnosis and treatment.

Hepatitis B virus X protein promotes liver cell pyroptosis under oxidative stress through NLRP3 inflammasome activation.

OBJECTIVE: Hepatitis B virus X protein (HBx) is a pivotal factor for HBV-induced hepatitis. Herein, we sought to investigate HBx-mediated NLR pyrin domain containing 3 (NLRP3) inflammasome activation and pyroptosis under oxidative stress. METHODS: The effect of HBx on the NLRP3 inflammasome was analyzed by enzyme-linked immunosorbent assays, quantitative reverse transcription-polymerase chain reaction, western blotting, and immunofluorescence in hepatic HL7702 cells. Pyroptosis was evaluated by western blotting, lactate dehydrogenase release, propidium iodide staining, and transmission electron microscopy. NLRP3 expression in the inflammasome from liver tissues was assessed by immunohistochemistry. RESULTS: In hydrogen peroxide (H2O2)-stimulated HL7702 cells, HBx triggered the release of pro-inflammatory mediators apoptosis-associated speck-like protein containing a CARD (ASC), interleukin (IL)-1ß, IL-18, and high-mobility group box 1 (HMGB1); activated NLRP3; and initiated pro-inflammatory cell death (pyroptosis). HBx localized to the mitochondria, where it induced mitochondrial damage and production of mitochondrial reactive oxygen species (mitoROS). Treatment of HL7702 cells with a mitoROS scavenger attenuated HBx-induced NLRP3 activation and pyroptosis. Expression levels of NLRP3, ASC, and IL-1ß in liver tissues from patients were positively correlated with HBV DNA concentration. CONCLUSIONS: The NLRP3 inflammasome was activated by elevated mitoROS levels and mediated HBx-induced liver inflammation and hepatocellular pyroptosis under H2O2-stress conditions.

HBx promotes the proliferative ability of HL­7702 cells via the COX­2/Wnt/ß­catenin pathway.

Hepatitis B virus X protein (HBx) has been termed a viral oncoprotein, and is involved in the initiation and progression of hepatocellular carcinoma (HCC). Cyclooxygenase­2 (COX­2) and ß­catenin have been attributed to the oncogenic activity of HBx in HBV­associated HCC. The present study aimed to determine whether there is crosstalk between COX­2 and the Wnt/ß­catenin signaling pathway during HL­7702­HBx cell proliferation, and to investigate the associated underlying molecular mechanism.  In the present study, cell proliferation assay, colony formation assay and flow cytometric analysis were used to detect the proliferative ability of cells. Reverse transcription­quantitative polymerase chain reaction and western blotting were performed to examine the mRNA and protein expression of COX­2, ß­catenin, cyclin­D1 and c­myc. The results demonstrated that HL­7702­HBx exhibited increased cell proliferation, higher colony formation efficiency and a shortened G1 period of the cell cycle. In addition, the mRNA and protein expression levels of COX­2 were increased, and this was associated with HL­7702­HBx cell growth. Furthermore, the expression of ß­catenin and its target genes, cyclin­D1 and c­myc proto­oncogene protein, was upregulated by HBx via COX­2. Finally, HBx promoted HL­7702 cell proliferation through the Wnt/ß­catenin signaling pathway. In conclusion, the primary finding of the present study was that HBx may promote HL­7702 cell proliferation via the COX­2/Wnt/ß­catenin pathway. Thus, it may be helpful to further investigate the molecular mechanism of HBV­associated hepatocellular carcinoma.

Hepatitis B Virus X protein elevates Parkin-mediated mitophagy through Lon Peptidase in starvation.

Hepatocellular Carcinoma (HCC) is the fifth most prevalent cancer worldwide. Specially, Hepatitis B viurs X protein (HBx) is a leading factor in the progression of Hepatitis B viurs-related HCC. Nutrient-deprived tumor microenvironment also contributes to tumor development. However, the role of HBx in nutrient-deprived HCC has received little investigation. Here, we show that HBx elevates PINK1-Parkin mediating mitophagy in starvation. HBx not only increases the PINK1/Parkin gene expression but also accelerates Parkin recruitment to partial mitochondria. Further analysis indicates that, HBx either promotes mitochondrial unfolded protein response, with remarkable mitochondrial LONP1 increases, or reduces LONP1 expression in cytosol inducing LONP1-Parkin pathway, both consequently enhancing mitophagy. Moreover, the enhanced mitophagy lowers mitochondrial apoptosis in starved hepatoma cells, and Bax is implied in the machinery. In addition, we define differential centrifuge, 3000 g or 12,000 g to pellet mitochondria, as an effective method to obtain distinct mitochondria. In collect, HBx regulates diverse aspects of LONP1 and Parkin, enhancing mitophagy in starvation. This study may shed new insights into the machinery development of hepatocellular carcinoma.

Hepatitis B virus X protein sensitizes HL-7702 cells to oxidative stress-induced apoptosis through modulation of the mitochondrial permeability transition pore.

Chronic hepatitis B virus (HBV) infection is a leading cause of liver cirrhosis and cancer. Among the pathogenic factors of HBV, HBV X protein (HBx) is attracting increased attention. Although it is documented that HBx is a multifunctional regulator that modulates cell inflammation and apoptosis, the exact mechanism remains controversial. In the present study, we explored the effect of HBx on oxidative stress-induced apoptosis in normal liver cell line, HL-7702. Our results showed that the existence of HBx affected mitochon-drial biogenesis by modulating the opening of the mitochondrial permeability transition pore (MPTP). Notably, this phenomenon was associated with a pronounced translocation of Bax from the cytosol to the mitochon-dria during the period of exposure to oxidative stress with a release of cytochrome c and activation of cleaved caspase-3 and PARP. Moreover, MPTP blockage with cyclosporin A prevented the translocation of Bax, and inhibited oxidative stress-induced apoptotic killing in the HBx-expressing HL-7702 cells. Our findings suggest that HBx exhibits pro-apoptotic effects upon normal liver cells following exposure to oxidative stress by modulating the MPTP gateway.