"Toxicity Testing in the 21st Century—A Vision and Strategy"proposed by the National Research Council of US has brought innovative directives and objectives for toxicity evaluation and risk assessment,pushing forward the next generation of toxicity testing and risk assessment.In this initiative,the concept of adverse outcome pathways(AOPs)has emerged as a prominent methodology,capturing the attention of toxicologists and researchers due to its promising applications in recent years.The quantitative AOP(qAOP)is an extension of the adverse outcome pathway,which is built upon the foundational qualitative adverse outcome pathway model and leverages mathematical frame-works to depict dose-response and/or response-response relationships.This article reviews the princi-ples and advancement surrounding qAOP,introduceds two prevalent methodologies for constructing qAOP,Bayesian network models and regression models,and demonstrates diverse applications of qAOP.Actual cases are used to underscore the transformative role of qAOP in contemporary toxicology and risk assessment practices.

Objective:To investigate the effects of 30% ethanol elution fraction of Artemisia absinthium extract with macroporous resin (AAEM-30%) on the dendritic cell (DC) and immunity of mice. Methods:AAEM-30% was obtained from the alcoholic extracts of A. absinthium by AB-8 macroporous resin, and its polysaccharide, flavonoid, and terpenoid contents were determined. The expressions of AAEM-30% on DC surface molecular cluster of differentiation (CD) 40, CD80 and CD86 were detected in vitro by flow cytometry, and the expressions of DC cytokines IL-6 and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). The effect of AAEM-30% on the immune function of ICR mice was measured in vivo with different doses (50 and 100 mg/kg) and different administration methods (subcutaneous injection, intraperitoneal injection, and gavage). Results:The contents of polysaccharides, flavonoids, and terpenoids in AAEM-30% were 24.30%, 22.50% and 28.19%, respectively. AAEM-30% significantly enhanced the expression of CD40, and CD86 and the secretion of IL-6 and TNF-α (all P<0.001). Compared with the control group, no statistically significant differences were found in the body mass of mice compared with the three administration methods (all P>0.05). The thymus index in the 50 and 100 mg/kg AAEM-30% intraperitoneal injection groups and the spleen index in the 50 mg/kg AAEM-30% gavage group were increased (all P<0.05). CD19 + cells increased in the 100 mg/kg AAEM-30% intraperitoneal injection group ( P<0.01) and in the 50 mg/kg AAEM-30% gavage group ( P<0.05). The CD11b + and CD11c + counts increased in the 100 mg/kg AAEM-30% gavage group ( P<0.05). The number of CD4 + and CD8 + T lymphocytes was increased by both gavage and intraperitoneal administration (all P<0.05). Conclusions:AAEM-30% can promote the maturation of DC and enhanced the immunity of mice without obvious side effects.

Objective To explore the clinical significance of monitoring drainage fluid parathyroid hormone (dPTH) for estimating the in situ reserves and function of the parathyroid by analyzing the change of serum calcium,serum parathyroid hormone(sPTH) and dPTH after thyroid surgery.Methods According to the operative method,the total of 144 patients with thyroid disease were divided into five groups:unilateral lobectomy,unilateral lobectomy plus isthmectomy with unilateral lymph node dissection,total thyroidectomy,total thyroidectomy with unilateral lymph node dissection,and total thyroidectomy with bilateral lymph node dissection group.The blood calcium,sPTH and dPTH level of patients were tested before operation and on the 1st,2nd,3rd and 4th day after operation.The depression of serum calcium,hypocalcemia and hypoparathyroidism were observed after operation.The serum calcium,serum PTH and dPTH level were summarized and analyzed statistically in order to evaluate the in situ reserves and postoperative function of the parathyroid.Results Among the 114 cases,the decline of serum calcium level mostly happened on the 2nd day after operation(70 cases,61.4％).There were 36 patients with hypocalcemia (31.58％) and 34 patients with hypoparathyroidism (29.82％).Serum calcium level increased gradually in all of the patients.Although sPTH level swung,it had a rising trend on the whole.The level of serum calcium and sPTH was positively correlated.The level of dPTH was discrete and decreased along with time.The decline level of dPTH among different groups had statistical difference.Conclusions It is a promising method to evaluate the in situ reserves and function of the parathyroid by monitoring the level and changes of dPTH after thyroid surgery,and it is of value for preventive calcium supplementation after thyroid surgery.

PURPOSE: The importance of long noncoding RNAs (lncRNAs) in tumorigenesis has recently been demonstrated. However, the role of lncRNAs in development of thyroid cancer remains largely unknown. MATERIALS AND METHODS: Using quantitative reverse transcription polymerase chain reaction, expression of three lncRNAs, including BRAF-activated long noncoding RNA (BANCR), papillary thyroid cancer susceptibility candidate 3 (PTCSC3), and noncoding RNA associated with mitogen-activated protein kinase pathway and growth arrest (NAMA), was investigated in the current study. RESULTS: Of the three lncRNAs (BANCR, PTCSC3, and NAMA), expression of BANCR was significantly up-regulated while PTCSC3 and NAMA were significantly down-regulated in papillary thyroid carcinoma (PTC) compared to that in normal tissue. BANCR-knockdown in a PTC-derived cell line (IHH-4) resulted in significant suppression of thyroid stimulating hormone receptor (TSHR). BANCR-knockdown also led to inhibition of cell growth and cell cycle arrest at G0/G1 phase through down-regulation of cyclin D1. In addition, BANCR was enriched by polycomb enhancer of zeste homolog 2 (EZH2), and silencing BANCR led to decreased chromatin recruitment of EZH2, which resulted significantly reduced expression of TSHR. CONCLUSION: These findings indicate that BANCR may contribute to the tumorigenesis of PTC through regulation of cyclin D1 and TSHR.
Carcinogenesis ; Cell Cycle Checkpoints ; Cell Line ; Cell Proliferation* ; Chromatin ; Cyclin D1 ; Down-Regulation ; Polymerase Chain Reaction ; Protein Kinases ; Receptors, Thyrotropin* ; Reverse Transcription ; RNA, Long Noncoding* ; RNA, Untranslated ; Thyroid Gland* ; Thyroid Neoplasms* ; Thyrotropin*

Objective To evaluate the effects of geraniol(GOH) on neointima hyperplasia in rat carotid artery balloon injury model, and explore the potential molecular mechanisms associated with this effect. Methods Totally 20 male Sprague-Dawley (SD) rats were randomly divided into sham operation group (without balloon injury), control group (with balloon injury), low concentration group (with 50 mg/kg GOH intervention after balloon injury) and high concentration group (with 200 mg/kg GOH intervention after balloon injury). The intima to media (I/M) area ratio of neointima was measured by hematoxylin- eosin (HE) staining. The expression of proliferating cell nuclear antigen (PCNA) was measured by immunohistochemical staining at 14th day after operation. As the marker of oxidative stress, the levels of 8-hydroxy-2’-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) were measured by enzyme linked inmmnosorbent assay (ELISA). Results The I/M ratio, IOD, 8-OHdG and MDA values were increased in control group compared with those of sham group. The I/M ratio, IOD and 8-OHdG values were reduced in low concentration group compared with those of control group. But there was no significant difference in MDA level between low concentration group and control group. The I/M ratio, IOD, 8-OHdG and MDA values were significantly reduced in high concentration group compared with those of control group, which showed a more significant inhibitory effect than that of low concentration group (P<0.05). Conclusion GOH could attenuate balloon iniury induced neointima hyperplasia, which might be related to its effect on inhibiting expression of PCNA and decreasing oxidative stress.

Objective To explore the effects of fasudil on inhibiting vascular smooth muscle cells (VSMCs) proliferation in vitro,increasing cell apoptosis,and inhibiting the Ras-MEK 1/2-ERK 1/2 pathway.Methods Healthy male SD rats (80～100 g) were selected.VSMCs were separated by the thoracoabdominal aortic vascular membrane dissection.Cultured VSMCs were randomly divided into 5 groups:serum-free group; serum group; serum + 1 μmol/L fasudil intervention group; serum + 10 μmol/L fasudil intervention group; serum + 100 μmol/L fasudil intervention group.The proliferation and migration of VSMCs were detected by MTT method and wound healing assay.Cell cycle and apoptosis of VSMCs were examined by flow cytometric analysis.The mRNA expressions of pro-apoptotic protein (Bax) and anti-apoptotic protein(Bcl-2) were determined by RT-PCR method,and the ratio of Bax/Bcl 2 was calculated.Western blot were performed to detect the protein expressions of Ras,MEK1/2,ERK1/2 and Akt in VSMCs.Results Fasudil inhibited rat VSMCs proliferation and migration,and blocked FBS-induced progression from the G0/G1 phase to S phase in a dose-dependent manner.Fasudil inhibited the early and late apoptosis in VSMCs,increased Bax mRNA expression and inhibited Bcl 2 mRNA expression.Fasudil significantly inhibited the protein expressions of FBS-stimulated intracellular Ras,phosphorylated MEK1/2,ERK1/2 in a dosedependent manner,but did not affect the protein expression of phosphorylated Akt.Conclusions Fasudil can attenuate VMSCs proliferation by blocking Ras-MEK1/2-ERK1/2 pathway and increasing cell apoptosis.