Quantifying depressive symptoms on incidence of common chronic diseases and multimorbidity patterns in middle-aged and elderly Chinese adults.

BACKGROUND: Depressive symptoms are highly prevalent and increase risks of various morbidities. However, the extent to which depressive symptoms could account for incidence of these chronic conditions, in particular multimorbidity patterns, remains to be examined and quantified. METHODS: For this cohort analysis, we included 9024-14,093 participants aged 45 years and older from the China Health and Retirement Longitudinal Study (CHARLS). Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for the longitudinal associations between depressive symptoms and 13 common chronic diseases and 4 multimorbidity patterns. Population attributable fractions (PAFs) combining the information on both exposure prevalence and risk association were estimated to quantify the magnitude of the burden of these conditions attributable to depressive symptoms. RESULTS: Depressive symptoms were associated with increased risks of liver disease, stroke, heart problem, asthma, diabetes, arthritis, kidney disease, chronic lung disease, digestive disease, dyslipidemia, and memory-related disease, and the adjusted HRs (95% CIs) and PAFs (95% CIs) ranged from 1.15 (1.05-1.26) to 1.64 (1.38-1.96) and 5% (0-10%) to 17% (6-28%), respectively. In addition, individuals with depressive symptoms had elevated risks of the cardiometabolic-cancer pattern, the cerebrovascular-memory pattern, the articular-visceral organ pattern, and the respiratory pattern, with respective HRs (95% CIs) of 1.26 (1.11-1.42), 1.34 (1.07-1.69), 1.45 (1.29-1.63), and 2.01 (1.36-2.96), and respective PAFs (95% CIs) of 5% (0-10%), 8% (-4-21%), 12% (7-17%), and 20% (5-35%). CONCLUSION: Depressive symptoms contribute substantially to the burden across a broad range of chronic diseases as well as different multimorbidity patterns in middle-aged and older Chinese.

Transiently impaired endothelial function during thyroid hormone withdrawal in differentiated thyroid cancer patients.

Purpose: Endothelial dysfunction, which was associated with chronic hypothyroidism, was an early event in atherosclerosis. Whether short-term hypothyroidism following thyroxine withdrawal during radioiodine (RAI) therapy was associated with endothelial dysfunction in patients with differentiated thyroid cancer (DTC) was unclear. Aim of the study was to assess whether short-term hypothyroidism could impair endothelial function and the accompanied metabolic changes in the whole process of RAI therapy. Methods: We recruited fifty-one patients who underwent total thyroidectomy surgery and would accept RAI therapy for DTC. We analyzed thyroid function, endothelial function and serum lipids levels of the patients at three time points: the day before thyroxine withdrawal(P1), the day before 131I administration(P2) and 4-6 weeks after RAI therapy(P3). A high-resolution ultrasound named flow-mediated dilation (FMD) was used to measure endothelial function of the patients. Results: We analyzed the changes of FMD, thyroid function and lipids at three time points. FMD(P2) decreased significantly compared to FMD(P1) (P1vsP2, 8.05 ± 1.55vs 7.26 ± 1.50, p<0.001). There was no significant difference between FMD(P3) and FMD(P1) after restoring TSH (thyroid stimulating hormone) suppression therapy (P1 vs P3, 8.05 ± 1.55 vs 7.79 ± 1.38, p=0.146). Among all parameters, the change of low-density lipoprotein (ΔLDL) was the only factor correlated negatively with the change of FMD (ΔFMD) throughout the RAI therapy process (P1-2, r=-0.326, p=0.020; P2-3, r=-0.306, p=0.029). Conclusion: Endothelial function was transiently impaired in DTC patients at short-term hypothyroidism state during the RAI therapy, and immediately returned to the initial state after restoring TSH suppression therapy.

Predicting 18F-FDG SUVs of metastatic pulmonary nodes from CT images in patients with differentiated thyroid cancer by using a convolutional neural network.

Purpose: The aim of this study was to predict standard uptake values (SUVs) from computed tomography (CT) images of patients with lung metastases from differentiated thyroid cancer (DTC-LM). Methods: We proposed a novel SUVs prediction model using 18-layer Residual Network for generating SUVmax, SUVmean, SUVmin of metastatic pulmonary nodes from CT images of patients with DTC-LM. Nuclear medicine specialists outlined the metastatic pulmonary as primary set. The best model parameters were obtained after five-fold cross-validation on the training and validation set, further evaluated in independent test set. Mean absolute error (MAE), mean squared error (MSE), and mean relative error (MRE) were used to assess the performance of regression task. Specificity, sensitivity, F1 score, positive predictive value, negative predictive value and accuracy were used for classification task. The correlation between predicted and actual SUVs was analyzed. Results: A total of 3407 nodes from 74 patients with DTC-LM were collected in this study. On the independent test set, the average MAE, MSE and MRE was 0.3843, 1.0133, 0.3491 respectively, and the accuracy was 88.26%. Our proposed model achieved high metric scores (MAE=0.3843, MSE=1.0113, MRE=34.91%) compared with other backbones. The predicted SUVmax (R2 = 0.8987), SUVmean (R2 = 0.8346), SUVmin (R2 = 0.7373) were all significantly correlated with actual SUVs. Conclusion: The novel approach proposed in this study provides new ideas for the application of predicting SUVs for metastatic pulmonary nodes in DTC patients.

TSH ≥30 mU/L may not be necessary for successful 131I remnant ablation in patients with differentiated thyroid cancer.

Purpose: To determine whether thyroid-stimulating hormone level ≥ 30 mU/L is necessary for radioiodine (131I) remnant ablation (RRA) in patients with differentiated thyroid cancer (DTC), as well as its influencing factors and predictors. Methods: A total of 487 DTC patients were retrospectively enrolled in this study. They were divided into two groups (TSH < 30 and ≥ 30 mU/L) and further divided into eight subgroups (0-<30, 30-<40, 40-<50, 50-<60, 60-<70, 70-<80, 80-<90, and 90-<100 mU/L). The simultaneous serum lipid level, successful rate of RRA and its influencing factors in different groups were analyzed. The receiver operating characteristic curves derived from pre-ablative thyroglobulin (pre-Tg) and pre-Tg/TSH ratio were compared for RRA success prediction performance. Results: There was no statistical difference in success rates of RRA between the two groups (P = 0.247) and eight subgroups (P = 0.685). Levels of total cholesterol (P < 0.001), triglyceride (P = 0.006), high-density lipoprotein cholesterol (P = 0.024), low-density lipoprotein cholesterol (P = 0.001), apolipoprotein B (P < 0.001), and apolipoprotein E (P = 0.002) were significantly higher while apoA/apoB ratio (P = 0.024) was significantly lower at TSH ≥ 30 mU/L group. Pre-Tg level, gender, and N stage were influencing factors for RRA. The area under the curve of pre-Tg level and pre-Tg/TSH ratio was 0.7611 (P < 0.0001) and 0.7340 (P < 0.0001) for all enrolled patients and 0.7310 (P = 0.0145) and 0.6524 (P = 0.1068) for TSH < 30 mU/L, respectively. Conclusion: TSH ≥ 30 mU/L may not be necessary for the success of RRA. Patients with higher serum TSH levels prior to RRA will suffer from severer hyperlipidemia. Pre-Tg level could be used as a predictor for the success of RRA, especially when TSH < 30 mU/L.

[Role of IRE1 in autophagy induced by vitamin E succinate in human gastric cancer cell].

OBJECTIVE: To investigate the role of inositol-requiring enzyme 1(IRE1) in autophagy of human gastric cancer cells induced by vitamin E succinate(VES). METHODS: Human gastric cancer SGC-7901 cells were cultured in vitro and divided into solvent control group(0.1% ethanol absolute), different doses(5, 10, 15 and 20 µg/mL) VES group, 4µ8C group, and VES + 4µ8C group. The endoplasmic reticulum stress-related molecules glucose regulated protein 78(GRP78) and C/EBP homologous protein(CHOP), autophagy marker microtubule associated Protein1 light chain 3(LC3), Beclin-1, unfolded protein response branching pathway Inositol-requiring enzyme 1(IRE1), X box-binding protein 1(XBP1), c-Jun n-terminal kinase(JNK) and p-JNK were detected by Western blot in the solvent control group and different doses of VES group. IRE1 was inhibited by 4µ8C. The expressions of IRE1, XBP1, JNK, p-JNK, GRP78 and CHOP were detected by Western blot, and the expressions of LC3 and Beclin-1 were detected. RESULTS: The expression of GRP78(1.16±0.06) and CHOP(1.36±0.11) in 20 µg/mL VES group were significantly higher than those in solvent control group GRP78(0.36±0.10) and CHOP(0.48±0.05)(P<0.001). The expression of Beclin-1(1.09±0.20) and LC3-Ⅱ/LC3-Ⅰ(1.29±0.03) in 20 µg/mL VES group were significantly higher than those in solvent control group(0.27±0.07) and LC3-Ⅱ/LC3-Ⅰ(0.43±0.06)(P<0.001). The expression levels of IRE1(1.07±0.20), XBP1(1.33±0.07) and p-JNK/JNK(1.19±0.31) in 20 µg/mL VES group were significantly higher than those in the solvent control group(P<0.01). After IRE1 is inhibited: The expression level of IRE1(0.63±0.27), XBP1(0.74±0.09), p-JNK/JNK(0.35±0.04), GRP78(0.66±0.02), CHOP(0.51±0.02), LC3-Ⅱ/LC3-Ⅰ(0.72±0.01), Beclin-1(0.70±0.15) was significantly lower than that of VES group(P<0.05). CONCLUSION: VES may participate in the regulation of autophagy in gastric cancer cells by upregulating IRE1 pathway.

Change in Antithyroglobulin Antibody Levels is a Good Predictor of Responses to Therapy in Antithyroglobulin Antibody-Positive Pediatric Papillary Thyroid Carcinoma Patients.

Objective: Antithyroglobulin antibodies (TgAbs) could be used as a surrogate tumor marker of TgAb-positive-differentiated thyroid carcinoma. This study aims to determine whether the change in TgAb levels over time could be used as a predictor of responses to therapy in pediatric papillary thyroid carcinoma (PTC) patients. Methods: We retrospectively analyzed the records of 48 pediatric PTC patients with TgAb levels ≥50 IU/ml 6 months after initial 131I treatment. Suppressed thyroglobulin (Tg) levels 6 months after initial 131I treatment were used to divide the patients into positive Tg (P-Tg, Tg ≥ 0.2 ng/ml) and negative Tg (N-Tg, Tg < 0.2 ng/ml) groups. Responses to therapy were classified as the acceptable response (AR) group and the not acceptable response (NAR) group. Results: Of 48 enrolled patients with 58 months (range, 24-143 months) of follow-up, 28 patients had NAR and 20 patients had AR. TgAb levels were decreasing ≥50% in 28 patients, decreasing <50% in 8 patients, and increasing in 12 patients. Multivariate analysis showed that high initial risk stratification and TgAb levels decreasing <50% or increasing were significantly associated with NAR (p < 0.05). Changes in Tg levels were also associated with NAR in the P-Tg group (p < 0.05). Conclusion: Changes in TgAb levels over time could be used as a predictor of responses to therapy in TgAb-positive pediatric PTC patients. Changes in Tg levels over time are also associated with NAR to therapy in both TgAb-positive and Tg-positive pediatric PTC patients.

Circular noncoding RNA circ_0007865, serves as a competing endogenous RNA, targeting the miR-214-3p/FKBP5 axis to regulate oxygen-glucose deprivation-induced injury in brain microvascular endothelial cells.

BACKGROUND: Ischemic stroke (IS) is a major cause of permanent morbidity and lifelong disability worldwide. Circular RNA (circRNA) circ_0007865 has been reported to be upregulated in acute ischemic stroke (AIS) patients. Also, AIS patients exhibited increased death of human brain microvascular endothelial cells (HBMECs). This study is designed to explore the role and mechanism of circ_0007865 in the oxygen-glucose deprivation (OGD)-induced cell damage in AIS. METHODS: Circ_0007865, microRNA-214-3p (miR-214-3p), and FK506-binding protein 5 (FKBP5) levels were detected by real-time quantitative PCR. Cell proliferative angiogenesis, migration, and apoptosis were assessed by Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine, colony formation, tube formation, wound healing, transwell, and flow cytometry assays. B-cell lymphoma-2 (Bcl-2), Bcl-2-related X protein (Bax), cleaved caspase-3, and FKBP5 protein levels were determined by western blot assay. The binding relationship between miR-214-3p and circ_0007865 or FKBP5 was predicted by StarBase, and verified by a dual-luciferase reporter, RNA pull-down assay. RESULTS: Circ_0007865 and FKBP5 were increased, and miR-214-3p was decreased in OGD-treated HBMECs. Furthermore, the silencing of circ_0007865 could promote cell proliferative angiogenesis, migration, and inhibit apoptosis in OGD-triggered HBMECs in vitro. Mechanically, circ_0007865 acted as a sponge of miR-214-3p to regulate FKBP5. CONCLUSION: According to these results, circ_0007865 deficiency could attenuate OGD-induced HBMEC damage by modulating the miR-214-3p/FKBP5 axis, hinting at a promising therapeutic target for future acute IS therapy.

Association between fetal famine exposure and risk of type 2 diabetes: a prospective cohort study.

The objective of this study was to explore the effects of fetal experience of famine on the onset of type 2 diabetes mellitus (T2DM) in adults. The analysis included 16 594 participants from the Kailuan Study who were free of diabetes at baseline (2006). According to the date of birth, the individuals born on October 1, 1962 - September 30, 1964, were divided into the non-exposed group (used as the reference group), individuals born on October 1, 1959 - September 30, 1961, were divided into the fetal exposure group, and the early childhood exposure group included those born on October 1, 1956 - September 30, 1958. The cumulative incidence of T2DM for each group was calculated and compared among the 3 groups, and the Cox regression model was used to analyze the effects of fetal famine experience on the risk of diabetes. During a median 10.27 years (170 358 person-years) (2006-2017), 3509 incident T2DM cases were identified, with a cumulative incidence rate of 19.46%. The cumulative incidences of T2DM in the non-exposed, fetal exposure, and early childhood exposure groups were 17.38%, 20.85%, and 20.65%, respectively (P < 0.01). After adjusting for confounding factors, the hazard ratio (HR) of T2DM in the fetal exposure group was 1.222 (95% confidence interval: 1.087-1.374, P < 0.01), compared with the reference group. The association was modified by sex and hypertension (both P interaction less than 0.05). Fetal famine exposure may increase the risk of developing T2DM in adults. This association was more pronounced among women and those with hypertension. Novelty: The association was modified by sex and hypertension. Long follow-up time.

Circ_0001955 Acts as a miR-646 Sponge to Promote the Proliferation, Metastasis and Angiogenesis of Hepatocellular Carcinoma.

BACKGROUND: Circular RNA (circRNA) exerts a crucial role in the progression of many cancers, including hepatocellular carcinoma (HCC). However, the function of circ_0001955 in HCC progression has been poorly studied. AIMS: Elucidating the role and molecular mechanism of circ_0001955 in HCC progression. METHODS: Quantitative real-time PCR was employed to detect the expression of circ_0001955 and miR-646. Cell counting kit 8 assay, Ethynyl-2-deoxyuridine assay, flow cytometry, transwell assay, and tube formation assay were conducted to measure cell proliferation, metastasis, angiogenesis and apoptosis. Dual-luciferase reporter assay and biotin-labeled RNA pull-down assay were performed to analyze the interactions among circ_0001955, miR-646 and frizzled class receptor 4 (FZD4). Moreover, animal experiments were performed to examine the influence of circ_0001955 on HCC tumor growth in vivo. RESULTS: Circ_0001955 was a highly expressed circRNA in HCC tissues and cells. Circ_0001955 knockdown inhibited the proliferation, metastasis, angiogenesis, and enhanced the apoptosis of HCC cells. Meanwhile, miR-646 could be sponged by circ_0001955, and its inhibitor could reverse the negative regulation of circ_0001955 knockdown on HCC progression. Further, FZD4 was a target of miR-646, and its overexpression could invert the inhibition effect of miR-646 mimic on HCC progression. Besides, our results also indicated that circ_0001955 promoted FZD4 expression by sponging miR-646. Animal experiment results showed that circ_0001955 silencing restrained HCC tumor growth in vivo. CONCLUSION: Our findings suggested that circ_0001955 might play a positive role in HCC progression via regulating the miR-646/FZD4 axis, indicating that circ_0001955 might be a potential therapeutic target for HCC.

Long-term outcomes and prognostic factors in papillary thyroid microcarcinoma patients with distant metastases.

PURPOSES: Distant metastasis from papillary thyroid microcarcinoma (PTMC) is extremely rare and the long-term outcomes and independent prognostic factors remain unclear. The present study aimed to investigate clinicopathological characteristics and evaluate the long-term outcomes and prognostic factors of PTMC patients with distant metastases (DM) who underwent surgery and radioactive iodine (131I) treatment. METHODS: We retrospectively reviewed the medical records of 13,441 patients with thyroid cancer (including 1697 cases with PTMC) who underwent 131I treatment at our institution between January 2008 and December 2019. PTMC patients with distant metastases with sufficient clinical follow-up data were enrolled in this cohort study. The overall survival (OS) and progression-free survival (PFS) were analyzed by the Kaplan-Meier method and the prognostic factors were assessed by Cox proportional hazards. RESULTS: Thirty-three PTMC patients with DM were enrolled in this study. The median follow-up was 75 months (range: 5-151 months). The 5-year and 10-year OS rates were 96.97 and 81.41%, respectively, and the 5-year and 10-year PFS rates were 90.46 and 69.68%, respectively. Multivariate analysis showed that male sex (P = 0.005), radioactive iodine refractory PTMC (P = 0.033), and symptomatic DM (P = 0.022) were significantly associated with worse 10-year PFS in PTMC patients with DM. No independent predictor related to poor 10-year OS was found in the present study. CONCLUSIONS: The prognosis of PTMC patients becomes worse after the development of DM. Male sex, radioactive iodine refractory PTMC, and symptomatic DM were identified as independent factors associated with PFS.

Unexpected Retrosternal Goiter Interpreted as Hyperfunctioning Metastasis on the Posttherapeutic 131I SPECT/CT.

ABSTRACT: We present a 38-year-old man who underwent total thyroidectomy with radical right neck dissection due to papillary thyroid cancer was referred for 131I treatment. The patient was in subclinical hypothyroidism with remarkable stimulated Tg level after 4 weeks of l-thyroxine withdrawal before 131I treatment, which indicated hyperfunctioning metastasis. Posttherapeutic 131I whole-body scan combined with 131I SPECT/CT performed on day 3 after 131I administration revealed an elevated 131I uptake mass in cervicothoracic region. To our surprise, the mass was histologically confirmed to be a retrosternal goiter.

Influencing factors of supernormal vascular aging in Chinese population.

OBJECTIVES: Previous studies have analysed the epidemic characteristics of supernormal vascular aging (SUPERNOVA), and found that SUPERNOVA were significantly associated with lower risk of cardiovascular disease. However, the influencing factors of SUPERNOVA are still unclear. The aim of this study was to investigate the characteristics and influencing factors of SUPERNOVA. METHODS: A total of 42 196 participants of the Kailuan Study were enrolled in the study. SUPERNOVA was defined as the lowest 2.5% of the age-quintile brachial-ankle pulse wave velocity (baPWV), early vascular aging was defined as the highest 2.5% of the age-quintile baPWV. Multivariable logistic regression analysis was applied to investigate the influencing factors of SUPERNOVA. RESULTS: The population with SUPERNOVA was mostly women, nonsmokers, nondrinkers, and those with higher education. They had lower levels of cardiovascular disease risk factors and healthier lifestyles. The results of logistics regression showed that the influencing factors of SUPERNOVA include age, sex, hypertension, diabetes, resting heart rate, hypersensitive C-reactive protein, and uric acid. However, the effects of these factors were different across age groups. We also observed that in addition to the unalterable factors (age and sex), only resting heart rate above 80 bpm (OR = 0.396, 95% CI: 0.231-0.681) and SBP (OR = 0.945, 95% CI: 0.932-0.958) were significantly associated with odds of SUPERNOVA in participants without cardiovascular risk factors. CONCLUSION: This study investigated the characteristics of the population with SUPERNOVA and the factors influencing it, which provided a basis for different populations to take preventive measures to slow down the process of vascular aging.

MicroRNA-497-5p attenuates IL-1ß-induced cartilage matrix degradation in chondrocytes via Wnt/ß-catenin signal pathway.

Osteoarthritis (OA) is a degenerative joint disease. Degradation of extracellular matrix (ECM) in chondrocytes is closely related to joint destruction in OA progression. MicroRNAs (miRNAs) have been reported to play important roles in progression of OA. However, the roles of miR-497-5p in OA process and its underlying mechanism remain not been well established. Chondrocytes were obtained from articular cartilage and stimulated with IL-1ß. The expression of miR-497-5p and Wnt3a was detected by qRT-PCR. Western blot analysis was performed to measure the proteins of Wnt3a, collagen II, aggrecan matrix metalloproteinase (MMP) 13 and ADAMTS4. Cell apoptosis was detected by flow cytometry. The putative binding sites of miR-497-5p and Wnt3a were predicted by Targetscan and verified through luciferase report assay. We found that miR-497-5p expression was reduced and Wnt3a expression was enhanced in OA cartilage and IL-1ß-stimulated chondrocytes. Moreover, Wnt3a was a direct target of miR-497-5p, and expression of miR-497-5p was negatively correlated with Wnt3a level in OA cartilage. Furthermore, overexpression of miR-497-5p prominently increased the expression of cartilage matrix molecules collagen II and aggrecan, and reduced the expression of matrix-degrading enzymes MMP13 and ADAMTS4 while overexpression of Wnt3a reversed these effects, whereas addition of DKK-1attenuated the Wnt3a-mediated functions in IL-1ß-stimulated chondrocytes. In conclusion, miR-497-5p attenuated IL-1ß-induced cartilage matrix degradation in chondrocytes via Wnt/ß-catenin signal pathway, providing a potential therapeutic target for treatment of OA.

Akt/AMPK/mTOR pathway was involved in the autophagy induced by vitamin E succinate in human gastric cancer SGC-7901 cells.

Vitamin E succinate (VES), a derivative of vitamin E, is a promising cancer chemopreventive agent that inhibits tumor promotion by inducing apoptotic cell death. The effects of VES on autophagy, an intricate programmed process which helps cells survive in some stressed situations by degrading some cytoplasmic material, are unclear. When human gastric cancer cells SCG-7901 were exposed to VES, both the level of microtubule-associated protein 1 light chain 3 and the yeast ATG6 homolog Beclin-1 increased, and related autophagy genes were activated, thereby suggesting that autophagy was induced by VES. We also observed that VES-induced autophagy was accompanied by the activation of AMP-activated protein kinases (AMPK). VES-induced autophagy decreased when AMPK was inhibited by using small interfering RNA (siRNA), thereby suggesting that VES-induced autophagy is mediated by AMPK. Moreover, further studies revealed that the decreased activity of mammalian target of rapamycin (mTOR) and its downstream targets P70S6K and 4EBP-1 were involved in VES-activated autophagy associated with AMPK activation. The experiments also showed that the activity of protein kinases B (Akt)-mTOR axis was inhibited by VES. VES-induced AMPK activation could be attenuated by Akt activation. Overall, our studies demonstrated that AMPK was involved in the VES-induced autophagy. Crosstalk exists between AMPK and the Akt/mTOR axis. The results elucidated the mechanism of VES-induced autophagy in human gastric cancer cells.

Vitamin E succinate induces apoptosis via the PI3K/AKT signaling pathways in EC109 esophageal cancer cells.

Esophageal cancer is the fourth most common gastrointestinal cancer, it generally has a poor prognosis and novel strategies are required for prevention and treatment. Vitamin E succinate (VES) is a potential chemical agent for cancer prevention and therapy as it exerts anti­tumor effects in a variety of cancers. However, the role of VES in tumorigenesis and progression of cancer remains to be elucidated. The present study aimed to determine the effects of VES in regulating the survival and apoptosis of human esophageal cancer cells. EC109 human esophageal cancer cells were used to investigate the anti­proliferative effects of VES. The MTT and Annexin V­fluorescein isothiocyanate/propidium iodide assays demonstrated that VES inhibited cell proliferation and induced apoptosis in esophageal cancer cells. Furthermore, VES downregulated constitutively active basal levels of phosphorylated (p)­serine­threonine kinase AKT (AKT) and p­mammalian target of rapamycin (mTOR), and decreased the phosphorylation of AKT substrates Bcl­2­associated death receptor and caspase­9, in addition to mTOR effectors, ribosomal protein S6 kinase ß1 and eIF4E­binding protein 1. Phosphoinositide­3­kinase (PI3K) inhibitor, LY294002 suppressed p­AKT and p­mTOR, indicating PI3K is a common upstream mediator. The apoptosis induced by VES was increased by inhibition of AKT or mTOR with their respective inhibitor in esophageal cancer cells. The results of the present study suggested that VES targeted the PI3K/AKT signaling pathways and induced apoptosis in esophageal cancer cells. Furthermore, the current study suggests that VES may be useful in a combinational therapeutic strategy employing an mTOR inhibitor.

Effect of vitamin E succinate on the expression of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor in gastric cancer cells and CD4(+) T cells.

Gastric malignancy, which shows poor prognosis, is one of the most frequent causes of cancer-associated deaths. Vitamin E succinate (VES) inhibits cell proliferation and induces apoptosis in a concentration- and time-dependent manner. We explored the effect of VES on the expression of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor in gastric cancer cells and CD4(+) T cells. On one hand, VES dose-dependently regulated the expression of the TRAIL receptor in gastric cancer cells. Moreover, the activation of the TRAIL receptor, death receptor 4 (DR4), and death receptor 5 (DR5) in gastric cancer cells increased for up to 12 h. On the other hand, the expression of TRAIL protein in human CD4(+) T cells was obviously upregulated in the presence of VES. On the basis of these findings, we combined VES and human CD4(+) T cells to induce apoptosis of MKN28 human gastric cancer cells. The results showed that VES induced higher gastric cancer cell apoptosis when combined with human CD4(+) T cells than when applied alone. We conclude that VES can induce the expression of TRAIL receptor in gastric cancer cells, as well as the expression of TRAIL in CD4(+) T cells. Overall, our results provide a theoretical basis for future immunotherapy studies.

Protective Macroautophagy Is Involved in Vitamin E Succinate Effects on Human Gastric Carcinoma Cell Line SGC-7901 by Inhibiting mTOR Axis Phosphorylation.

Vitamin E succinate (VES), a potential cancer therapeutic agent, potently induces apoptosis and inhibits the growth of various cancer cells. Autophagy has been supposed to promote cancer cell survival or trigger cell death, depending on particular cancer types and tumor microenvironments. The role of autophagy in the growth suppressive effect of VES on gastric cancer cell is basically unknown. We aimed to determine whether and how autophagy affected the VES-induced inhibition of SGC-7901 human gastric carcinoma cell growth. SGC-7901 cells were treated with VES or pre-treated with autophagy inhibitor, chloroquine (CQ) and 3-methyladenine (3-MA). Electron microscopy, fluorescence microscopy and Western blot were used to study whether VES induced autophagy reaction in SGC-7901 cells. Western blot evaluated the activities of the mammalian target of rapamycin (mTOR) axis. Then we used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry to detect the level of cell viability and apoptosis. Collectively, our data indeed strongly support our hypothesis that VES treatment produced cytological variations that depict autophagy, increased the amount of intracellular green fluorescent protein-microtubule associated protein 1 light chain 3 (GFP-LC3) punctate fluorescence and the number of autophagic vacuoles. It altered the expression of endogenous autophagy marker LC3. VES activated the suppression of mTOR through inhibiting upstream regulators p38 MAPK and Akt. mTOR suppression consequently inhibited the activation of mTOR downstream targets p70S6K and 4E-BP-1. The activation of the upstream mTOR inhibitor AMPK had been up-regulated by VES. The results showed that pre-treatment SGC-7901 with autophagy inhibitors before VES treatment could increase the capacity of VES to reduce cell viability and to provoke apoptosis. In conclusion, VES-induced autophagy participates in SGC-7901 cell protection by inhibiting mTOR axis phosphorylation. Our findings not only strengthen our understanding of the roles of autophagy in cancer biology, but may also be useful for developing new treatments for gastric cancer patients.

RRR-α-tocopheryl succinate induces apoptosis in human gastric cancer cells via the NF-κB signaling pathway.

To investigate the effects of the nuclear factor (NF)-κB signaling pathway on the induction of apoptosis by vitamin E succinate (RRR-α-tocopheryl succinate; VES) in human gastric carcinoma cells. Human gastric carcinoma SGC-7901 cells were treated with temperate concentrations of VES and pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB. Cell viability and apoptosis were respectively estimated by methylthiazol tetrazolium (MTT) assay and the Annexin V­FITC method. Western blot analysis was used to evaluate the protein expressions of NF-κBp65 and Bcl-2 family members Bcl-2, Bax and cleavage of caspase-3, caspase-9, and poly (ADP-ribose) polymerase (PARP). The DNA-binding activity of NF-κBp65 was measured by electrophoretic mobility shift assay (EMSA). Reverse transcription and polymerase chain reaction (RT-PCR) was implemented to evaluate the transcription of inhibitor of apoptosis (IAP) genes. Apoptosis assessment showed that VES induces apoptotic cell death in human gastric carcinoma cells. In the following experiments, PDTC (100 µM) was used in cell treatment 2 h before VES. The decreased ratio of the nuclear and cytosolic NF-κBp65 protein level was induced by VES and PDTC reinforced this trend. PDTC treatment significantly enhanced the decrease of NF-κB-DNA binding activity induced by VES in human gastric SGC-7901. The decrease in protein expression of Bcl-2 as well as the increase in the protein expression of Bax were induced by VES treatment. The cleavage of caspase-9, caspase-3 and PARP was induced. There was no effect on the gene transcription of c-IAP-1, c-IAP-2, and x-linked IAP (XIAP) compared with the control group, whereas mRNA levels of survivin and the neuronal apoptosis inhibitory protein (NAIP) markedly decreased. Notably, pretreatment with PDTC reinforced all the above VES-induced effects. In conclusion, VES-induced apoptosis in SGC-7901 cells is accompanied by the inhibition of the NF-κB signaling pathway, including changes in Bcl-2 family members, cleavage of caspases and gene transcription of survivin and NAIP.

Crosstalk between endoplasmic reticulum stress and oxidative stress in apoptosis induced by α-tocopheryl succinate in human gastric carcinoma cells.

α-Tocopheryl succinate (α-TOS) has been shown to be a potent apoptosis inducer and growth inhibitor in a variety of cancer cells. Our previous studies showed the important role of endoplasmic reticulum (ER) stress and reactive oxygen species (ROS) generation in the apoptosis induced by α-TOS. However, the relationship of oxidative stress with ER stress is still controversial. The objective of the present study was to investigate the interplay between the two stress responses induced by α-TOS in SGC-7901 human gastric cancer cells. In response to α-TOS, cytological changes typical of apoptosis, induction of glucose-regulated protein 78 (GRP78) and CCAAT/enhancer-binding protein (C/EBP) homologous protein transcription factor (CHOP), and activation of caspase-4 were observed. And the antioxidant N-acetyl-l-cysteine inhibited induction of both GRP78 and CHOP by α-TOS transcriptionally and translationally. Furthermore, knocking down CHOP by RNA interference decreased ROS generation, increased glutathione level and induced glutathione peroxidase mRNA expression in α-TOS-treated cells, whereas catalase and superoxide dismutases mRNA expression were not altered. The results imply that α-TOS induces ER stress response through ROS production, while CHOP perturbs the redox state of SGC-7901 cells treated with α-TOS.

[Effect of Baoganning on serum and hepatic leptin and its receptor levels in rats with liver fibrosis].

OBJECTIVE: To investigate the mechanism underlying the protective effects of a traditional Chinese medicinal formula, Baoganning, against liver fibrosis. METHODS: Male Wistar rats were subjected to injection of carbon tetrachloride- peanut oil mixture and given daily 5% alcoholic beverage, and 2 days after the injection, Baoganning was administered intragastrically at two different doses for 6 weeks. Radioimmunoassay was used to detect serum leptin level, and immunohistochemistry employed to examine the effect of Baoganning on expressions of leptin and its receptor in the liver tissue of the rats. RESULTS: Compared with the normal control group, the rats in the liver fibrosis model group and Baoganning-treated groups showed significantly increased serum leptin levels (P<0.01), and the serum leptin level was significantly lower in Baoganning group than in the liver fibrosis model group (P<0.01). Baoganning significantly reduced the hepatic expression of leptin and OB-Rb in rats with liver fibrosis in comparison with their expression in the model group (P<0.01). CONCLUSION: Baoganning can effectively ameliorate liver fibrosis in rats possibly through reducing serum leptin level and inhibiting hepatic leptin and its receptor expressions.