OBJECTIVETo observe the effect of high-mobility group box-1 protein (HMGB1) on the proliferation of human nasopharyngeal carcinoma cell line C666-1 and explore the possible underlying mechanisms.

METHODSCultured C666-1 cells were treated with a siRNA targeting HMGB1 gene. The changes in the cell proliferation were detected by CCK8 analysis, the cell cycle distribution was assayed with flow cytometry, and the expressions of cyclin D1, CDK6 and related pathway proteins were detected with Western blotting. The effect of a HMGB1 plasmid carrying the reporter gene GFP on the proliferation of C666-1 cells was tested with CCK8 and EDU analysis.

RESULTSCompared with the control cells, the cells transfected with the siRNA targeting HMGB1 showed obviously suppressed cell proliferation (P<0.001), cell cycle arrest in G1 phase (P<0.001), and down-regulated expressions of cyclin D1, CDK6, STAT3 and P-STAT3. Overexpression of HMGB1 in cells transfected with the HMGB1 plasmid showed a significantly increased ratio of S phase cells (P<0.05) and obviously enhanced cell proliferation (P<0.001).

CONCLUSIONHMGB1 can promote the proliferation of human nasopharyngeal carcinoma cell line C666-1 by up- regulating cyclin D1 and CDK6 via the STAT3 signaling pathway.

Carcinoma ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Division ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 6 ; metabolism ; Down-Regulation ; HMGB1 Protein ; genetics ; Humans ; Nasopharyngeal Neoplasms ; pathology ; RNA, Small Interfering ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Transfection ; Up-Regulation

Objective To determine the impact of hypertension on the outcomes of patients with COVID-19. Methods This matched cohort study was conducted among a total 442 patients with COVID-19 admitted in Honghu People's Hospital and First Affiliated Hospital of Nanchang University between January 1 to March 18, 2020, including 61 patients with hypertension and 381 normotensive patients. To minimize the effects of the confounding factors including age, gender and other comorbidities, we excluded patients with comorbidities other than hypertension, and matched the patients with and without hypertension for age and gender at a 1:1 ratio. We analyzed the clinical characteristics, laboratory findings and clinical outcomes of in 32 matched pairs of patients with and without hypertension. Results Compared with the normotensive patients, COVID-19 patients with hypertension were more likely to develop bacterial infections (P=0.002) and had higher neutrophil counts (P=0.007), neutrophil/lymphocyte ratio (P=0.045), and lactate dehydrogenase levels (P=0.035). A greater proportion of patients had bilateral patchy opacities on chest CT (P=0.012) in the hypertension group than in the normotensive group. COVID-19 patients with hypertension group were more likely to receive antibiotics (P=0.035) and corticosteroid therapies (P=0.035). Conclusion Hypertension increases the risk of bacterial infection in patients with COVID-19. Hypertensive patients with COVID-19 have higher neutrophil counts and neutrophil/lymphocyte ratios and are more likely to require treatment with antibiotics. Hypertensive patients with COVID-19 should therefore take cautions to avoid bacterial infections.

Objective To determine the impact of hypertension on the outcomes of patients with COVID-19. Methods This matched cohort study was conducted among a total 442 patients with COVID-19 admitted in Honghu People's Hospital and First Affiliated Hospital of Nanchang University between January 1 to March 18, 2020, including 61 patients with hypertension and 381 normotensive patients. To minimize the effects of the confounding factors including age, gender and other comorbidities, we excluded patients with comorbidities other than hypertension, and matched the patients with and without hypertension for age and gender at a 1:1 ratio. We analyzed the clinical characteristics, laboratory findings and clinical outcomes of in 32 matched pairs of patients with and without hypertension. Results Compared with the normotensive patients, COVID-19 patients with hypertension were more likely to develop bacterial infections (P=0.002) and had higher neutrophil counts (P=0.007), neutrophil/lymphocyte ratio (P=0.045), and lactate dehydrogenase levels (P=0.035). A greater proportion of patients had bilateral patchy opacities on chest CT (P=0.012) in the hypertension group than in the normotensive group. COVID-19 patients with hypertension group were more likely to receive antibiotics (P=0.035) and corticosteroid therapies (P=0.035). Conclusion Hypertension increases the risk of bacterial infection in patients with COVID-19. Hypertensive patients with COVID-19 have higher neutrophil counts and neutrophil/lymphocyte ratios and are more likely to require treatment with antibiotics. Hypertensive patients with COVID-19 should therefore take cautions to avoid bacterial infections.

RNA interference (RNAi) is useful for selective gene silencing. Cytochrome P450 3A4 (CYP3A4), which metabolizes approximately 50% of drugs in clinical use, plays an important role in drug metabolism. In this study, we aimed to develop a short hairpin RNA (shRNA) to modulate CYP3A4 expression. Three new shRNAs (S1, S2 and S3) were designed to target the coding sequence (CDS) of CYP3A4, cloned into a shRNA expression vector, and tested in different cells. The mixture of three shRNAs produced optimal reduction (55%) in CYP3A4 CDS-luciferase activity in both CHL and HEK293 cells. Endogenous CYP3A4 expression in HepG2 cells was decreased about 50% at both mRNA and protein level after transfection of the mixture of three shRNAs. In contrast, CYP3A5 gene expression was not altered by the shRNAs, supporting the selectivity of CYP3A4 shRNAs. In addition, HepG2 cells transfected with CYP3A4 shRNAs were less sensitive to Ginkgolic acids, whose toxic metabolites are produced by CYP3A4. These results demonstrate that vector-based shRNAs could modulate CYP3A4 expression in cells through their actions on CYP3A4 CDS, and CYP3A4 shRNAs may be utilized to define the role of CYP3A4 in drug metabolism and toxicity.

Atropa belladonna seedlings were used as experimental materials and cultivated by soil culture method. Different concentrations(0,0.05,0.1,0.2,0.5 mmol·L~(-1))of NO donor sodium nitroprusside(SNP) were sprayed on the leaves. The effects of different concentrations of SNP and different treatment time(4,8,12,16 d) on nitrogen metabolism, secondary metabolite content, precursor content of tropane alkaloid synthesis pathway and expression of key enzyme genes under 100 mmol·L~(-1) NaCl stress were studied. The results showed that with the prolongation of salt stress, the nitrogen metabolism and the accumulation of secondary metabolites of A. belladonna were inhibited to some extent. After treatment with different concentrations of exogenous SNP, the ammonium nitrogen content decreased dramatically, and the contents of nitrate nitrogen, free amino acid, soluble protein and the activities of key enzymes of nitrogen metabolism(NR, GS, GDH) were all greatly improved; the contents of precursor amino acids(ornithine, arginine) and polyamines(Put, Spd, Spm) in the secondary metabolic pathway have increased to varying degrees. The qRT-PCR analysis showed that exogenous SNP treatment can effectively promote the high expression of key enzyme genes PMT, TRⅠ and H6H in the secondary metabolic pathway of A. belladonna, and the production of hyoscyamine and scopolamine were increased notably. In summary, the application of appropriate concentration of SNP can effectively alleviate the inhibition of salt stress on the nitrogen metabolism and secondary metabolism of Atropa belladonna, and enhance its salt tolerance. Overall, 0.1 mmol·L~(-1) and 0.2 mmol·L~(-1) SNP treatment achieved the most remarkable effect.
Atropa belladonna/metabolism* ; Hyoscyamine/analysis* ; Nitrogen/metabolism* ; Nitroprusside ; Scopolamine/analysis* ; Secondary Metabolism ; Sodium Chloride ; Stress, Physiological