Exploring the Effect of Milk Fat on Fermented Milk Flavor Based on Gas Chromatography-Ion Mobility Spectrometry (GC-IMS) and Multivariate Statistical Analysis.

Milk fat is a premium nutritional health product, yet there is a lack of high-fat dairy products for daily consumption in the current market. This study investigated the influence of different milk fat contents on the physicochemical and textural properties of fermented milk. The research revealed that an increase in milkfat content significantly improved the water-holding capacity, syneresis, color, hardness, springiness, gumminess, and chewiness of fermented milk, while showing minimal changes in pH and total titratable acidity. Response surface analysis indicated that fermented milk with 25% milk fat, 2.5% inoculum, a fermentation time of 16 h, and a fermentation temperature of 30 °C exhibited the highest overall acceptability. Using GC-IMS technology, 36 volatile compounds were identified, with an increase in milk fat content leading to elevated levels of ketone compounds, and 14 compounds were defined as key aroma compounds (ROAV > 1). Electronic nose distinguished samples with different milk fat contents. The results demonstrate that an increase in milk fat content enhances the physicochemical and flavor attributes of fermented milk. This work provides theoretical references for the production and development of high-fat fermented milk.

Improved Cycling Stability of Ni-Rich Cathode Material by In Situ Introduced TM-B-O Amorphous Surface Structure.

Current research has found the amorphous/crystal interface has some unexpected electrochemical behaviors. This work designed a surface modification strategy using NaBH4 to induce in situ conversion of the surface structure of Ni-rich LiNi0.8Co0.1Mn0.1O2 (NCM811) into TM-B-O amorphous interface layer. Oxidizing the surface from transition metals (TM) with high valence and reductive BH4- in a weak polar medium of ethanol results in an easy redox reacton. A TM-B-O amorphous structure is formed on NCM811 surface. The action of reactive wetting ensures a complete and uniform structure evolution of the surface crystals. The complete coverage protects the outer crystal and the heterogeneous interface impedance between the modified layer and bulk is reduced. More importantly, this amorphous interface layer through in situ conversion enhances the heterogeneous link at interface and its own structural stability. The modified NCM811 (TB2@NCM) treated with 1 wt % NaBH4 shows excellent electrochemical performance, especially cyclic stability. At a high cutoff voltage of 4.5 V, the capacity retention was 72.5% at 1 C after 500 cycles. The electrode achieves 173.7 mAh·g-1 at 10 C. This work creates a modifying strategy with potential application prospect due to simple technology with low-cost raw material under mild operating conditions.

The Age-Male-Albumin-Bilirubin-Platelets (aMAP) Risk Score Predicts Liver Metastasis Following Surgery for Breast Cancer in Chinese Population: A Retrospective Study.

Objective: The current study is conducted to investigate the potential prognostic value of the age-male-albumin-bilirubin-platelets (aMAP) score in breast cancer patients with liver metastasis after surgery. Methods: This is a retrospective study of 178 breast cancer patients who developed liver metastasis after surgery. These patients were treated and followed up from 2000 to 2018 at our hospital. The aMAP risk score was estimated in accordance with the following formula: . The optimal cutoff value of the aMAP was evaluated via X-tile. Kaplan-Meier, Log-rank and Cox proportional hazards regression models were applied to determine the clinical influence of the aMAP score on the survival outcomes. The nomogram models were established by multivariate analyses. The calibration curves and decision curve analysis were applied to evaluate the estimated performance of the nomogram models. Results: A total of 178 breast cancer patients were divided into low aMAP score group (<47.6) and high aMAP score group (≥47.6) via X-tile plots. The aMAP score was a potential prognostic factor in multivariate analysis. The median disease free survival (p=0.0013) and overall survival (p=0.0003) in low aMAP score group were longer than in high aMAP score group. The nomograms were constructed to predict the DFS with a C-index of 0.722 (95% CI, 0.673-0.771), and the OS with a C-index of 0.708 (95% CI, 0.661-0.755). The aMAP-based nomograms had good predictive performance. Conclusion: The aMAP score is a potential prognostic factor in breast cancer with liver metastasis after surgery. The aMAP score-based nomograms were conducive to discriminate patients at high risks of liver metastasis and develop adjuvant treatment and prevention strategies.

Physical and Chemical Properties, Flavor and Organoleptic Characteristics of a Walnut and Purple Rice Fermented Plant Drink.

In recent years, green and healthy foods have attracted much attention. Plant-based foods have become an alternative to animal-derived foods. In this study, we used walnut and purple rice as the primary raw materials to produce a fermented plant drink. The process included boiling, mixing, grinding, inoculation, fermentation, and sterilization. We then analyzed the similarities and differences between the resulting walnut and purple rice fermented plant drink and an unfermented walnut and purple rice plant drink, as well as dairy-based yoghurt, in terms of physical chemistry, flavor, and sensory characteristics. We also examined the similarities and differences between the walnut and purple rice fermented plant drink and room-temperature yoghurt. The study results revealed that the walnut and purple rice fermented plant drink exhibited greater viscosity than the walnut and purple rice unfermented plant drink and room-temperature yoghurt. Additionally, the former displayed enhanced stability and recovery ability. Notably, distinguishable differences were observed between the three samples in terms of the presence of unknown volatiles and the umami signal, as indicated by electronic nose/tongue and GC-IMS analyses. The umami flavor of the walnut and purple rice fermented plant drink surpasses that of room-temperature yoghurt, while its taste is less salty than that of the walnut and purple rice plant drink. Despite possessing a weaker aroma than dairy-based yogurt, it is more potent than the walnut and purple rice plant drink. Additionally, its relative abundance of olefins, ketones, and alcohols enhances its unique flavor profile, surpassing both other options. Based on sensory analysis, it can be deduced that walnut and purple rice fermented plant drink has the highest overall acceptance rate.

Assessment of the albumin-bilirubin score in breast cancer patients with liver metastasis after surgery.

Objective: This study aims to investigate the potential prognostic value of albumin-bilirubin (ALBI) score in breast cancer patients with liver metastasis after surgery. Methods: This was a retrospective study of 178 breast cancer patients with liver metastasis after surgery. ALBI score was calculated by the following formula: (log10 bilirubin × 0.66) - (albumin × 0.085). The optimal cutoff value of ALBI score was assessed by X-tile. The clinical influence of ALBI score on survival outcomes using Kaplan-Meier method, Log-rank test, Cox proportional hazards regression model. The calibration curves, decision curve analysis and time-dependent ROC curve were used to assess the predictive performance of the nomogram's models. Results: The classifications of 178 breast cancer patients with liver metastasis after surgery were as follows: low ALBI score group (＜-3.36) vs. high ALBI score group (≥-3.36). The Cox proportional hazards regression model indicated that ALBI score was a potential predictor. Kaplan-Meier survival curve performed that the median disease free survival (p = 0.0029) and overall survival (p＜0.0001) in low ALBI score group were longer than in high ALBI score group. The ALBI-based nomograms had good predictive performance. Conclusions: The ALBI score has high prognostic ability for survival time in breast cancer with liver metastasis after surgery. These models will be valuable in discriminating patients at high risks of liver metastasis.

The association between intermediate-term sulfur dioxide exposure and outpatient visits for Parkinson's disease: a time-series study in southwestern China.

Parkinson's disease (PD) is the second most common human neurodegenerative disorder, and the pathogenesis of it remains poorly understood. Limited studies have shown that both long- and short-term exposure to air pollutants may be associated with increased risk of PD while lacking evidence on the effects of intermediate-term exposure. In this study, over-dispersed Poisson generalized additive models (GAMs) were applied to explore the association between intermediate-term sulfur dioxide (SO2) exposure and outpatient visits for PD in Chongqing, China, and further stratified analyses were performed by age and gender. A total of 39,984 PD cases from January 1, 2014, to December 31, 2019 (2191 days) were included. The association of intermediate-term SO2 exposure with outpatient visits for PD was statistically significant: per 1 µg/m3 increase of SO2 corresponded to 2.34% (95% CI: 0.88%, 3.80%) elevation in monthly PD outpatient visits at lag 0 (the concurrent month). Stratified analyses showed that the associations between SO2 and PD outpatient visits were stronger in younger (≤ 60 years) and female patients. In conclusion, intermediate-term SO2 exposure can be associated with an increased risk of PD outpatient visits. Our results highlight the importance of recognizing the role of intermediate-term SO2 exposure in the development of PD. In addition to focusing on the effects of long-term or short-term air pollutants, it is necessary to pay more attention to the health effects of intermediate-term exposure time windows of air pollutants, which will facilitate policy formulation and public health interventions for health risks.

The Effects of Ultrasonic and Gamma Irradiation on the Flavor of Potato Wines Investigated by Sensory Omics.

Aroma is one of the most fascinating and least-known mysteries of Baijiu research. The volatile compounds (VOCs) of potato wine were evaluated by sensory omics techniques in order to comprehend their overall flavor characteristics and investigate the effects of ultrasonic treatment and gamma irradiation therapy on the aroma of the wine. The findings revealed that a total of 14 flavor compounds were identified by GC-MS. Isoamyl alcohol, ethyl octanoate, and 1,1-diethoxyethane were the key aroma components, according to GC-O analysis. A total of 50 volatile substances were identified by GC-IMS. After being subjected to irradiation and ultrasonic treatment, the alcohol level of the potato wine reduced while the esters content increased. By calculating the relative odor activity value, a total of 29 aroma components were classified as key aroma compounds (ROAV > 1). According to the results of the sensory evaluation-fruity, Fen-flavor, and sweet-and the acceptability of the irradiated and ultrasonicated potato wine were improved. Therefore, the use of ultrasonic and irradiation therapy in potato wine, as well as the overall aroma building of potato wine, can be supported theoretically by this study.

Association between short-term ambient nitrogen dioxide and type 2 diabetes outpatient visits: A large hospital-based study.

Type 2 diabetes (T2DM) as a non-communicable disease imposes heavy disease burdens on society. Limited studies have been conducted to assess the effects of short-term air pollution exposure on T2DM, especially in Asian regions. Our research aimed to determine the association between short-term exposure to ambient nitrogen dioxide (NO2) and outpatient visits for T2DM in Chongqing, the largest city in western China, based on the data collected from November 28, 2013 to December 31, 2019. A generalized additive model (GAM) was applied, and stratified analyses were performed to investigate the potential modifying effects by age, gender, and season. Meanwhile, the disease burden was revealed from attributable risk. Positive associations between short-term NO2 and daily T2DM outpatient visits were observed. The strongest association was observed at lag 04, with per 10 µg/m3 increase of NO2 corresponded to increased T2DM outpatient visits at 1.57% [95% confidence interval (CI): 0.48%, 2.65%]. Stronger associations were presented in middle-aged group (35-64 years old), male group, and cool seasons (October to March). Moreover, there were 1.553% (8664.535 cases) of T2DM outpatient visits attributable to NO2. Middle-aged adults, males, and patients who visited in cool seasons suffered heavier burdens. Conclusively, short-term exposure to NO2 was associated with increased outpatient visits for T2DM. Attention should be paid to the impact of NO2 on the burden of T2DM, especially for those vulnerable groups.

Short-term ambient air pollution exposure and adult primary insomnia outpatient visits in Chongqing, China: A time-series analysis.

Growing evidence suggest that air pollutants can be associated with sleep disorders. However, no study has explored the association of short-term air pollution exposure with primary insomnia, a specific type of sleep disorders. To evaluate the correlation of short-term air pollution exposure with adult primary insomnia outpatient visits in Chongqing, China, we collected data of adult primary insomnia outpatient visits and air pollutants' concentrations between 2013 and 2019 and the associations were estimated with single-day lags as well as moving average lags using a generalized additive model. Totally, 23,919 outpatient visits for adult primary insomnia were identified. The daily data of adult insomnia outpatient visits, air pollutants (NO2, CO, SO2, O3, PM10 and PM2.5) and meteorological conditions (daily mean temperature and relative humidity) were gathered. Short-term exposure to multiple air pollutants, especially NO2 and SO2, was associated with adult primary insomnia visits. A 10 µg/m3 increase in NO2 and SO2 at lag 05 corresponded to increased primary insomnia outpatient visits 3.87% (95% CI: 1.50%-6.24%) and 7.22% (95% CI: 2.10%-12.35%), respectively. Moreover, stronger links were presented in females and cool seasons for NO2 while in the elderly for SO2. Collectively, this time-series study suggested that short-term exposure to air pollutants, especially to NO2 and SO2, was associated with higher risk of adult primary insomnia outpatient visits, and such association could to be sex-, age-, and season-modified.

Stabilized Cathode Interphase for Enhancing Electrochemical Performance of LiNi0.5Mn1.5O4-Based Lithium-Ion Battery via cis-1,2,3,6-Tetrahydrophthalic Anhydride.

The continuous degradation of carbonate electrolytes and the dissolution of transition metal cations due to parasitic reactions on the cathode-electrolyte interphase (CEI) block the practical application of LiNi0.5Mn1.5O4-based lithium-ion batteries (LNMO-based LIBs) at a high voltage. cis-1,2,3,6-Tetrahydrophthalic anhydride (CTA) has been used as a functional additive in a carbonate baseline electrolyte (BE) for constructing the CEI film to enhance the cyclic stability of LNMO-based LIBs. The LNMO/Li cell with CTA exhibits a preponderant capacity retention of 83.3% compared with those of propionic anhydride (PA) (46.5%) and BE (13.6%) after 500 cycles at the current density of 1 C from 3.5 to 4.9 V. Additionally, the LNMO/graphite full cell with CTA still has a higher capacity retention of 95.46% even after 300 cycles at 1 C. By characterizations, it is reasonably demonstrated that CTA was oxidated to participate in the construction of a CEI film. An unsaturated aromatic group was introduced into the composition of the CEI film along with CTA in the formation process of the CEI film, which further improved the antioxidative activity of the CEI film under the influence of field-effect. Specifically, the CEI film obtains appreciable stability because of its higher antioxidative activity under the influence of field-effect. The stabilized CEI can significantly suppress the parasitic reactions of electrolytes, decrease the consumption of active-Li+, and protect the LNMO cathode structure, thereby enhancing the cyclic compatibility of LNMO-based LIBs with the carbonate electrolytes from 3.5 to 4.9 V.

MicroRNA-221 promotes tumor progression by targeting HHIP in human glioblastoma.

BACKGROUND: MicroRNAs are found to be aberrantly expressed in multiple cancers, including glioblastoma (GBM), and microRNA-221 (miR-221) has been verified as an oncogene in various human cancers. Nevertheless, the role of miR-221 in GBM is unclear. This study aimed to investigate the miR-221 expression level in GBM and to evaluate its function and underlying mechanisms. METHODS: Western blotting and qPCR were used to determine the expression of human hedgehog-interacting protein (HHIP) and miR-221 levels. MiR-221-inhibited cell models were constructed, and siRNA was used for HHIP silencing. Cell proliferation was analyzed by MTT and colony formation assays and a subcutaneous xenograft model. Cell migration and invasion was analyzed by wound healing and Transwell invasion assays. A dual luciferase reporter assay system was used to clarify the relationship between miR-221 and HHIP. RESULTS: The results of this study revealed that miR-221 expression was upregulated in GBM tissues and A172, U251, as well as T98G cells, as detected by real-time PCR analysis. MTT, Transwell, and colony formation assays revealed that miR-221 knockdown could suppress GBM cells from proliferating, migrating, and invading in vitro. Moreover, animal experiments showed that tumor growth in vivo was inhibited when miR-221 expression decreased. Furthermore, HHIP was predicted and verified to be a target of miR-221 by bioinformatics analysis, and luciferase and western blot assays. In addition, HHIP silencing rescued the suppressive effect of a miR-221 inhibitor on the proliferation, migration, and invasion of GBM cells. CONCLUSIONS: Our results indicated that miR-221 is upregulated in GBM and enhances tumor progression by targeting HHIP, which suggests this may be a potential therapeutic target for GBM.

A green, efficient, closed-loop direct regeneration technology for reconstructing of the LiNi0.5Co0.2Mn0.3O2 cathode material from spent lithium-ion batteries.

Lithium nickel manganese cobalt oxide in the spent lithium ion batteries (LIBs) contains a lot of lithium, nickel, cobalt and manganese. However, how to effectively recover these valuable metals under the premise of reducing environmental pollution is still a challenge. In this work, a green, efficient, closed-loop direct regeneration technology is proposed to reconstruct LiNi0.5Co0.2Mn0.3O2 (NCM523) cathode materials from spent LIBs. Firstly, the failure mechanism of NCM523 cathode materials in the spent LIBs is analyzed deeply. It is found that the spent NCM523 material has problems such as the dissolution of lithium and transition metals, surface interface failure and structural transformation, resulting in serious deterioration of electrochemical performance. Then NCM523 material was directly regenerated by supplementing metal ions, granulation, ion doping and heat treatment. Meanwhile, PO43- polyanions were doped into the regenerated NCM material in the recovery process, showing excellent electrochemical performance with discharge capacity of 189.8 mAh g-1 at 0.1 C. The recovery process proposed in this study puts forward a new strategy for the recovery various lithium nickel cobalt manganese oxide (e.g., LiNi1/3Co1/3Mn1/3O2, LiNi0.5Co0.2Mn0.3O2, LiNi0.6Co0.2Mn0.2O2 and LiNi0.8Co0.1Mn0.1O2) and accelerates the industrialization of spent lithium ion battery recycling.

FeSe2@C Microrods as a Superior Long-Life and High-Rate Anode for Sodium Ion Batteries.

Transition-metal selenides have emerged as promising anode materials for sodium ion batteries (SIBs). Nevertheless, they suffer from volume expansion, polyselenide dissolution, and sluggish kinetics, which lead to inadequate conversion reaction toward sodium and poor reversibility during the desodiation process. Therefore, the transition-metal selenides are far from long cycling stability, outstanding rate performance, and high initial Coulombic efficiency, which are the major challenges for practical application in SIBs. Here, an efficient anode material including an FeSe2 core and N-doped carbon shell with inner void space as well as high conductivity is developed for outstanding rate performance and long cycle life SIBs. In the ingeniously designed FeSe2@NC microrods, the N-doped carbon shell can facilitate mass transport/electron transfer, protect the FeSe2 core from the electrolyte, and accommodate volume variation of FeSe2 with the help of the inner void of the core. Thus, the FeSe2@NC microrods can maintain strong structural integrity upon long cycling and ensure a good reversible conversion reaction of FeSe2 during the discharge/charge process. As a result, the as-prepared FeSe2@NC microrods exhibit excellent sodium storage performance and ultrahigh stability, achieving an excellent rate capability (411 mAh g-1 at 10.0 A g-1) and a long-term cycle performance (401.3 mAh g-1 after 2000 cycles at 5.0 A g-1).

Enhancing the Electrochemical Performance of a High-Voltage LiNi0.5 Mn1.5 O4 Cathode in a Carbonate-Based Electrolyte with a Novel and Low-Cost Functional Additive.

Butyric anhydride (BA) is used as an effective functional additive to improve the electrochemical performance of a high-voltage LiNi0.5 Mn1.5 O4 (LNMO) cathode. In the presence of 0.5 wt % BA, the capacity retention of a LNMO/Li cell is significantly improved from 15.3 to 88.4 % after 200 cycles at 1 C. Furthermore, the rate performance of the LNMO/Li cell is also effectively enhanced, and the capacity goes up to 112 mAh g-1 even at 5 C, which is considerably higher than that of a LNMO/Li cell in electrolyte without BA additive (95.4 mAh g-1 at 5 C). Linear sweep voltammetry and cyclic voltammetry results reveal that the BA additive can be preferentially oxidized to construct a stable cathode electrolyte interphase (CEI) film on the LNMO cathode. Subsequently, the BA-derived CEI film can alleviate the decomposition of the electrolyte and the dissolution of Mn and Ni ions from the LNMO cathode as well as maintain the structural stability of LNMO during the cycling process; this leads to outstanding electrochemical performance. Thus, this work provides an effective and low-cost functional electrolyte for high-voltage LNMO-based LIBs.

FOXP1-induced lncRNA CLRN1-AS1 acts as a tumor suppressor in pituitary prolactinoma by repressing the autophagy via inactivating Wnt/ß-catenin signaling pathway.

As the commonest type of functional pituitary tumor, prolactinoma takes up around 40-60% of functional pituitary tumors. Despite dedications attributed to the treatment of prolactinoma, complete cure remains difficult. Hence, it is of significance to bring to light the underlying mechanism of prolactinoma. Long noncoding RNAs (lncRNAs) are a group of transcripts which can regulate various biological processes. In the present study, we explored an lncRNA that was differentially downregulated in prolactinoma samples. LncRNA clarin 1 antisense RNA 1 (CLRN1-AS1) was downregulated in 42 patient samples and inactivated the Wnt/ß-catenin signaling pathway. Functionally, CLRN1-AS1 suppressed cell proliferation, promoted apoptosis, and inhibited autophagy. Subcellular fractionation assay revealed that CLRN1-AS1 was located in the cytoplasm of prolactinoma cells. Based on bioinformatics analysis and mechanism experiments, we determined that CLRN1-AS1 acted as a competing endogenous RNA (ceRNA) by sponging miR-217 to upregulate the dickkopf WNT signaling pathway inhibitor 1 (DKK1). Furthermore, Forkhead box P1 (FOXP1) was verified to be a transcription suppressor of CLRN1-AS1. In summary, this study revealed that FOXP1-induced CLRN1-AS1 regulated cellular functions in pituitary prolactinoma by sponging miR-217 to release the DKK1/Wnt/ß-catenin signaling pathway.

Estrogen receptor antagonist fulvestrant inhibits proliferation and promotes apoptosis of prolactinoma cells by regulating the IRE1/XBP1 signaling pathway.

The aim of the present study was to evaluate the effects of an estrogen receptor antagonist, fulvestrant, on proliferation and apoptosis of prolactinoma cells, and to reveal potential regulatory mechanisms. Prolactinoma GH3 cells were treated with 10­6 mol/l fulvestrant for 2, 4, 8, 12 and 24 h. GH3 cell growth was observed under a microscope and cell viability was detected by MTT assay. Morphological changes of the nuclei in GH3 cells were observed by Hoechst 33258 staining and apoptotic rates were detected by flow cytometry. Preprolactin (PPL) and prolactin (PRL) secretion levels from GH3 cells were measured using ELISA. In addition, the protein expression levels of inositol­requiring enzyme 1 (IRE1), X­box binding protein (XBP)­1 and glucose­regulated protein, 78 kDa (GRP78) in GH3 cells were detected by western blot analysis. Cell density and cell viability of GH3 cells were significantly reduced in a time­dependent manner following treatment with fulvestrant (P<0.05). GH3 cells treated with fulvestrant also acquired an apoptotic morphology and the apoptotic rate of GH3 cells was significantly increased by fulvestrant in a time­dependent manner (P<0.05). PPL and PRL secretion levels were significantly reduced by fulvestrant treatment in a time­dependent manner (P<0.05). The protein expression levels of IRE1, XBP1 and GRP78 were also significantly reduced in a time­dependent manner following treatment with fulvestrant (P<0.05). Therefore, fulvestrant may inhibit proliferation and promote apoptosis of GH3 cells by downregulating the IRE1/XBP1 signaling pathway.

Enhancement of second-harmonic generation from silicon nitride with gold gratings.

We report strong enhancement of second-harmonic generation in a hybrid nanostructure with gold gratings embedded in a silicon nitride film. Compared to a flat silicon nitride film, the enhancement factor can be as large as 102 to 103 for transverse magnetic and electric polarizations, respectively in good agreement with numerical results calculated using finite element method. For both polarizations, the enhancement arises from a resonance between the waveguide modes and grating.

MicroRNA-133b inhibits cell migration and invasion by targeting matrix metalloproteinase 14 in glioblastoma.

Increasing evidence has suggested that microRNA-133b (miR-133b) is important in regulating the genesis of different types of cancer. However, the effects and the underlying mechanisms of miR-133b in the development of glioblastoma (GBM) remain largely unknown. The aim of the present study was to investigate the role of miR-133b in GBM and to determine the molecular mechanisms underlying its action. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression levels of miR-133b in 21 human GBM samples and 9 normal brain tissue samples. A wound healing assay, and Transwell migration and invasion assays were used to evaluate the effects of miR-133b on cell migration and invasion. Western blotting and a luciferase reporter assay were used to identify the target genes of miR-133b. It was found that miR-133b suppressed GBM cell migration and invasion, and matrix metalloproteinase 14 (MMP14) was identified as a direct target gene. In conclusion, miR-133b may suppress GBM migration and invasion through directly targeting MMP14, highlighting its potential as a novel agent for the treatment of GBM invasion.

Effects of resveratrol on cell growth and prolactin synthesis in GH3 cells.

Resveratrol (RE), a phytoestrogen, has antiestrogenic properties. Estrogen plays a key role in the development and progression of pituitary prolactinoma. Moreover, RE is a potent cancer chemopreventive agent that inhibits the initiation, promotion and progression of carcinogenesis. The present study investigated the antitumor effects of RE on GH3 pituitary tumor cells. A concentration- and treatment duration-dependent biphasic effect of RE on the proliferation of the GH3 cells was demonstrated. After three days of treatment, RE stimulated proliferation at low concentrations and inhibited proliferation at high concentrations. However, when the treatment duration was reduced to 6 h, RE inhibited proliferation in a concentration-dependent manner. In addition, RE induced apoptosis with the activation of caspase-3 and -8, and decreased the percentage of prolactin (PRL)-immunopositive GH3 cells. Furthermore, RE suppressed expression of the PRL gene and inhibited the cell proliferation and PRL synthesis induced by 17ß-estradiol (E2). In GH3 cells, the proliferation response exhibited higher sensitivity to E2 compared with the PRL response; by contrast, the PRL response was more sensitive to RE than the proliferation response was. These results indicate that RE, an antiestrogenic compound, exerts its antitumor effect on GH3 cells through the suppression of GH3 cell growth and through the inhibition of PRL synthesis. The RE-induced cell apoptosis was shown to be caspase-dependent. Therefore, the present study provides support for the use of RE in the chemoprevention and chemotherapy of pituitary prolactinoma.

The prosurvival role of autophagy in resveratrol-induced cytotoxicity in GH3 cells.

In a previous study, we reported that resveratrol exerts antitumor effects through the estrogen receptor in prolactinoma. The autophagy/lysosomal degradation pathway plays an important role in damage control and energy efficiency. In this study, we investigated the involvement of autophagy and the related signaling pathways in resveratrol-induced apoptosis of GH3 cells. We demonstrate that resveratrol inhibits cell proliferation and induces apoptosis in a dose-dependent manner in GH3 cells. The cleavage of PARP was also observed, and the activation of caspase-3 and caspase-8 was detected. Consistent with this finding, the inhibition of caspase activation effectively attenuated resveratrol-induced cell apoptosis. In addition, the decreased level of Bcl-2 was also observed. The induction of autophagy was confirmed by the detection of the formation of autophagic vacuoles, and the increase in microtubule-associated protein 1 light chain 3 (LC3)-II and beclin-1 levels, two hallmarks of autophagy. Pre-treatment with bafilomycin A1 or 3-methyladenine, inhibitors of autophagy, enhanced the resveratrol-mediated caspase activation and cell death. Moreover, resveratrol induced the activation of ERK1/2, as well as the downregulation of Akt and mTOR phosphorylation. Taken together, these findings indicate that resveratrol induces caspase-dependent apoptosis and decreases Bcl-2 levels. In addition, resveratrol-induced autophagy is regulated by the PI3K/Akt/mTOR and ERK1/2 pathways. Furthermore, the inhibition of autophagy increases the cytotoxicity of resveratrol to GH3 cells.