Identification of the cytochrome P450s responsible for the biosynthesis of two types of aporphine alkaloids and their de novo biosynthesis in yeast.

Aporphine alkaloids have diverse pharmacological activities; however, our understanding of their biosynthesis is relatively limited. Previous studies have classified aporphine alkaloids into two categories based on the configuration and number of substituents of the D-ring and have proposed preliminary biosynthetic pathways for each category. In this study, we identified two specific cytochrome P450 enzymes (CYP80G6 and CYP80Q5) with distinct activities toward (S)-configured and (R)-configured substrates from the herbaceous perennial vine Stephania tetrandra, shedding light on the biosynthetic mechanisms and stereochemical features of these two aporphine alkaloid categories. Additionally, we characterized two CYP719C enzymes (CYP719C3 and CYP719C4) that catalyzed the formation of the methylenedioxy bridge, an essential pharmacophoric group, on the A- and D-rings, respectively, of aporphine alkaloids. Leveraging the functional characterization of these crucial cytochrome P450 enzymes, we reconstructed the biosynthetic pathways for the two types of aporphine alkaloids in budding yeast (Saccharomyces cerevisiae) for the de novo production of compounds such as (R)-glaziovine, (S)-glaziovine, and magnoflorine. This study provides key insight into the biosynthesis of aporphine alkaloids and lays a foundation for producing these valuable compounds through synthetic biology.

Diagnosis and treatment of diabetic foot ulcer complicated with lower extremity vasculopathy: Consensus recommendation from the Chinese Medical Association (CMA), Chinese Medical Doctor Association (CMDA).

Diabetic foot ulcer complicated with lower extremity vasculopathy is highly prevalent, slow healing and have a poor prognosis. The final progression leads to amputation, or may even be life-threatening, seriously affecting patients' quality of life. The treatment of lower extremity vasculopathy is the focus of clinical practice and is vital to improving the healing process of diabetic foot ulcers. Recently, a number of clinical trials on diabetic foot ulcers with lower extremity vasculopathy have been reported. A joint group of Chinese Medical Association (CMA) and Chinese Medical Doctor Association (CMDA) expert representatives reviewed and reached a consensus on the guidelines for the clinical diagnosis and treatment of this kind of disease. These guidelines are based on evidence from the literature and cover the pathogenesis of diabetic foot ulcers complicated with lower extremity vasculopathy and the application of new treatment approaches. These guidelines have been put forward to guide practitioners on the best approaches for screening, diagnosing and treating diabetic foot ulcers with lower extremity vasculopathy, with the aim of providing optimal, evidence-based management for medical personnel working with diabetic foot wound repair and treatment.

Computational design of anisotropic stealthy hyperuniform composites with engineered directional scattering properties.

Disordered hyperuniform materials are an emerging class of exotic amorphous states of matter that endow them with singular physical properties, including large isotropic photonic band gaps, superior resistance to fracture, and nearly optimal electrical and thermal transport properties, to name but a few. Here we generalize the Fourier-space-based numerical construction procedure for designing and generating digital realizations of isotropic disordered hyperuniform two-phase heterogeneous materials (i.e., composites) developed by Chen and Torquato [Acta Mater. 142, 152 (2018)1359-645410.1016/j.actamat.2017.09.053] to anisotropic microstructures with targeted spectral densities. Our generalized construction procedure explicitly incorporates the vector-dependent spectral density function χ[over ̃]_{_{V}}(k) of arbitrary form that is realizable. We demonstrate the utility of the procedure by generating a wide spectrum of anisotropic stealthy hyperuniform microstructures with χ[over ̃]_{_{V}}(k)=0 for k∈Ω, i.e., complete suppression of scattering in an "exclusion" region Ω around the origin in Fourier space. We show how different exclusion-region shapes with various discrete symmetries, including circular-disk, elliptical-disk, square, rectangular, butterfly-shaped, and lemniscate-shaped regions of varying size, affect the resulting statistically anisotropic microstructures as a function of the phase volume fraction. The latter two cases of Ω lead to directionally hyperuniform composites, which are stealthy hyperuniform only along certain directions and are nonhyperuniform along others. We find that while the circular-disk exclusion regions give rise to isotropic hyperuniform composite microstructures, the directional hyperuniform behaviors imposed by the shape asymmetry (or anisotropy) of certain exclusion regions give rise to distinct anisotropic structures and degree of uniformity in the distribution of the phases on intermediate and large length scales along different directions. Moreover, while the anisotropic exclusion regions impose strong constraints on the global symmetry of the resulting media, they can still possess structures at a local level that are nearly isotropic. Both the isotropic and anisotropic hyperuniform microstructures associated with the elliptical-disk, square, and rectangular Ω possess phase-inversion symmetry over certain range of volume fractions and a percolation threshold ϕ_{c}≈0.5. On the other hand, the directionally hyperuniform microstructures associated with the butterfly-shaped and lemniscate-shaped Ω do not possess phase-inversion symmetry and percolate along certain directions at much lower volume fractions. We also apply our general procedure to construct stealthy nonhyperuniform systems. Our construction algorithm enables one to control the statistical anisotropy of composite microstructures via the shape, size, and symmetries of Ω, which is crucial to engineering directional optical, transport, and mechanical properties of two-phase composite media.

[Advances in research methods for biosynthetic pathway analysis of active ingredients in traditional Chinese medicine].

The active ingredients in traditional Chinese medicine(TCM)are the foundation for the efficiency of TCM and the key to the formation of Dao-di herbs. It is of great significance to study the biosynthesis and regulation mechanisms of these active ingredients for analyzing the formation mechanism of Daodi herbs and providing components for the production of active ingredients in TCM by synthetic biology. With the advancements in omics technology, molecular biology, synthetic biology, artificial intelligence, etc., the analysis of biosynthetic pathways for active ingredients in TCM is rapidly progressing. New methods and technologies have promoted the analysis of the synthetic pathways of active ingredients in TCM and have also made this area a hot topic in molecular pharmacognosy. Many researchers have made significant progress in analyzing the biosynthetic pathways of active ingredients in TCM such as Panax ginseng, Salvia miltiorrhiza, Glycyrrhiza uralensis, and Tripterygium wilfordii. This paper systematically reviewed current research me-thods for analyzing the biosynthetic functional genes of active ingredients in TCM, elaborated the mining of gene elements based on multiomics technology and the verification of gene functions in plants in vitro and in vivo with candidate genes as objects. Additionally, the paper summarized new technologies and methods that have emerged in recent years, such as high-throughput screening, molecular probes, genome-wide association studies, cell-free systems, and computer simulation screening to provide a comprehensive reference for the analysis of the biosynthetic pathways of active ingredients in TCM.

A modified mouse model for observational fear learning and the influence of social hierarchy.

Background: Indirectly experiencing traumatic events either by witnessing or learning of a loved one's suffering is associated with the highest prevalence rates of epidemiological features of PTSD. Social species can develop fear by observing conspecifics in distress. Observational fear learning (OFL) is one of the most widely used paradigms for studying fear contagion in mice. However, the impact of empathic fear behavior and social hierarchy on fear transfer in mice is not well understood. Methods: Fear emotions are best characterized in mice by using complementary tests, rather than only freezing behavior, and simultaneously avoiding behavioral variability in different tests across time. In this study, we modified the OFL model by implementing freezing (FZ), open field (OF), and social interaction (SI) tests in a newly designed experimental facility and applied Z-normalization to assess emotionality changes across different behaviors. Results: The integrated emotionality scores revealed a robustly increased emotionality of observer mice and, more importantly, contributed to distinguishing susceptible individuals. Interestingly, fos-positive neurons were mainly found in the interoceptive network, and mice of a lower social rank showed more empathy-like behaviors. Conclusion: Our findings highlight that combining this experimental model with the Z-scoring method yields robust emotionality measures of individual mice, thus making it easier to screen and differentiate between empathic fear-susceptible mice and resilient mice, and refining the translational applicability of these models.

Associations of BMI with all-cause mortality in normoglycemia, impaired fasting glucose and type 2 diabetes mellitus among an elderly Chinese population: a cohort study.

AIM: To explore the associations of body mass index (BMI) and mortality among people with normal fasting glucose (NFG), impaired fasting glucose (IFG), and type 2 diabetes mellitus (T2DM) in an elderly Chinese population.  METHODS: A retrospective cohort study was conducted that included 59,874 elderly people who were aged 60 and older at baseline. Data for the study came from a health check-up program in China between 2011 and 2019. Hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated using multivariable Cox proportional hazard models of BMI categories by glycemic status. RESULTS: During the median of 5.96 years of follow-up, 7928 participants died (6457/49057 with NFG, 712/5898 with IFG and 759/4919 with T2DM). In adjusted Cox models, risk of mortality showed a decreasing trend with BMI < 18.5 kg/m2, 24 ≤ BMI < 28 kg/m2, and BMI ≥ 28 kg/m2 compared to 18.5 ≤ BMI < 24 kg/m2: HR (95% CI): 1.33 (1.18 to 1.49), 0.88 (0.83 to 0.93), and 0.90 (0.82 to 0.98), respectively, for NFG; 0.89 (0.55 to 1.46), 0.84 (0.71 to 0.99), and 0.88 (0.70 to 1.11), respectively, for IFG; and 1.42 (0.88 to 2.29), 0.75 (0.64 to 0.89), and 0.76 (0.62 to 0.93), respectively, for T2DM. There were curvilinear-shaped associations between BMI and mortality in the NFG and T2DM groups (P overall < 0.001 and P overall < 0.001, respectively; P nonlinearity < 0.001 and P nonlinearity = 0.027, respectively) and no significantly association between BMI and all-cause mortality was observed in the IFG group (P overall = 0.170). CONCLUSION: High BMI compared to normal BMI was associated with decreased mortality, especially in the old populations with NFG and T2DM. Future studies are needed to explain the obesity paradox in elderly patients with T2DM.

Different Synaptic Plasticity After Physiological and Psychological Stress in the Anterior Insular Cortex in an Observational Fear Mouse Model.

Post-traumatic stress disorder (PTSD) can be triggered not only in people who have personally experienced traumatic events but also in those who witness them. Physiological and psychological stress can have different effects on neural activity, but little is known about the underlying mechanisms. There is ample evidence that the insular cortex, especially the anterior insular cortex (aIC), is critical to both the sensory and emotional experience of pain. It is therefore worthwhile to explore the effects of direct and indirect stress on the synaptic plasticity of the aIC. Here, we used a mouse model of observational fear to mimic direct suffering (Demonstrator, DM) and witnessing (Observer, OB) of traumatic events. After observational fear training, using a 64-channel recording system, we showed that both DM and OB mice exhibited a decreased ratio of paired-pulse with intervals of 50 ms in the superficial layers of the aIC but not in the deep layers. We found that theta-burst stimulation (TBS)-induced long-term potentiation (LTP) in OB mice was significantly higher than in DM mice, and the recruitment of synaptic responses occurred only in OB mice. Compared with naive mice, OB mice showed stronger recruitment and higher amplitude in the superficial layers of the aIC. We also used low-frequency stimulation (LFS) to induce long-term depression (LTD). OB mice showed greater LTD in both the superficial and deep layers of the aIC than naive mice, but no significant difference was found between OB and DM mice. These results provide insights into the changes in synaptic plasticity in the aIC after physiological and psychological stress, and suggest that different types of stress may have different mechanisms. Furthermore, identification of the possible causes of the differences in stress could help treat stress-related disorders.

Association of body mass index and waist circumference with type 2 diabetes mellitus in older adults: a cross-sectional study.

BACKGROUND: The prevalence of obesity and diabetes is rising. The aim of this study was to determine the association of body mass index (BMI) and waist circumference (WC) with type 2 diabetes mellitus (T2DM) in the elderly and to compare the discriminatory abilities of BMI, WC and other anthropometric indicators, including waist-to-height ratio (WHtR), body adiposity estimator (BAE) and body roundness index (BRI) for T2DM. METHODS: This cross-sectional study included 69,388 subjects aged ≥ 60 years living in Xinzheng, Henan Province, from January to December 2020. The data came from the residents' electronic health records of the Xinzheng Hospital Information System. Logistic regression was used to examine the relationships. Fully adjusted models adjusted for age, sex, place of residence, alcohol consumption, smoking, physical exercise, SBP and RHR. The area under the receiver operating characteristic curve (AUC) was used to compare the discriminatory ability of different anthropometric indicators for T2DM under the influence of potential risk factors. RESULTS: After adjusting for multiple covariates, compared with the first BMI quintile, the odds ratios (ORs) and 95% confidence intervals (CIs) from the second to fifth quintile for T2DM were 1.416 (1.335-1.502), 1.664 (1.570-1.764), 1.879 (1.774-1.990) and 2.156 (2.037-2.283), respectively. Compared with the first WC quintile, the ORs and 95% CIs from the second to fifth quintiles for T2DM were 1.322 (1.244-1.404), 1.549 (1.459-1.643), 1.705 (1.609-1.807) and 2.169 (2.048-2.297), respectively. Among men, compared with other anthropometric indicators (BMI, WHtR, BAE and BRI), WC showed the highest AUC (AUC: 0.629; 95% CI: 0.622-0.636). Among women, the AUCs of BMI (AUC: 0.600; 95% CI: 0.594-0.606), WC (AUC: 0.600; 95% CI: 0.593-0.606) and BAE (AUC: 0.600; 95% CI: 0.594-0.607) were similar, and the AUCs of BMI, WC and BAE were higher than WHtR, BRI. CONCLUSIONS: All anthropometric indicators were positively associated with T2DM. In men, WC with the strongest positive association with T2DM was the best predictor of T2DM. In women, BMI was most strongly associated with T2DM, and the predictive powers of BMI, WC and BAE were similar. After adjusting the potential confounding factors including age, sex, place of residence, alcohol consumption, smoking, physical exercise, SBP and RHR, the effect of these factors was eliminated, the findings were independent of the covariates considered.

Hypertension modifies the associations of body mass index and waist circumference with all-cause mortality among older Chinese: a retrospective cohort study.

BACKGROUND: The effect of baseline hypertension status on the BMI-mortality association is still unclear. We aimed to explore the relationships of body mass index (BMI) and waist circumference (WC) with all-cause mortality among older hypertensive and normotensive Chinese individuals. METHODS: This retrospective cohort study was conducted in Xinzheng, Henan Province, Central China. The data came from the residents' electronic health records of the Xinzheng Hospital Information System. A total of 77,295 participants (41,357 hypertensive participants and 35,938 normotensive participants) aged ≥ 60 years were included from January 2011 to November 2019. Cox proportional hazard regression model was used to examine the relationships. RESULTS: During a mean follow-up of 5.3 years, 10,755 deaths were identified (6,377 in hypertensive participants and 4,378 in normotensive participants). In adjusted models, compared with a BMI of 18.5-24 kg/m2, the hazard ratios (HRs) and corresponding 95% confidence intervals (CIs) of BMI < 18.5, 24-28 and ≥ 28 kg/m2 for mortality in hypertensive participants were 1.074 (0.927-1.244), 0.881 (0.834-0.931) and 0.856 (0.790-0.929), respectively, and 1.444 (1.267-1.646), 0.884 (0.822-0.949) and 0.912 (0.792-1.051), respectively, in normotensive participants. Compared with normal waist circumference, the adjusted HRs and 95% CIs of central obesity for mortality were 0.880 (0.832-0.931) in hypertensive participants and 0.918 (0.846-0.996) in normotensive participants. A sensitivity analysis showed similar associations for both hypertensive and normotensive participants. CONCLUSION: Low BMI and WC were associated with a higher risk of all-cause mortality regardless of hypertension status in older Chinese individuals. The lowest risk of death associated with BMI was in the overweight group in normotensive participants and in the obesity group in hypertensive participants.

Stronger Associations of Body Mass Index and Waist Circumference with Diabetes than Waist-Height Ratio and Triglyceride Glucose Index in the Middle-Aged and Elderly Population: A Retrospective Cohort Study.

Background: It remains controversial whether body mass index (BMI), waist circumference (WC), waist-to-height ratio (WHtR), or triglyceride glucose (TyG) index has a stronger association with diabetes. The aims of the study were to compare the magnitude of associations of four indicators with diabetes risk. Methods: Data collected from annual health examination dataset in the Xinzheng during 2011 and 2019. A total of 41,242 participants aged ≥ 45 years were included in this study. Cox proportional hazard regression models were used to examine associations between the four indicators and diabetes risk. Results: After 205,770 person-years of follow up, diabetes developed in 2,472 subjects. Multivariable-adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) of diabetes (highest vs reference group) were 1.92 (1.71-2.16) for BMI, 1.99 (1.78-2.23) for WC, 1.65 (1.47-1.86) for WHtR, and 1.66 (1.47-1.87) for TyG, respectively. In addition, the risk of diabetes increased with baseline BMI (HR: 1.30; 95% CI: 1.25, 1.35) and TyG (HR: 1.25; 95% CI: 1.20, 1.30), but the lowest HR was 0.78 (95% CI 0.65-0.92) when WC was approximately 72 cm, and 0.85 (95% CI 0.72-0.99) when WHtR was approximately 0.47 in women. In joint analyses, the highest risk was observed in participants with a high BMI combined with a high WC (HR: 2.26; 95% CI: 1.98, 2.58). Conclusions: In middle-aged and elderly Chinese population, BMI and WC were more strongly associated with diabetes than WHtR or TyG, especially the combined effect of BMI and WC.

Searching collaborative agents for multi-plane localization in 3D ultrasound.

3D ultrasound (US) has become prevalent due to its rich spatial and diagnostic information not contained in 2D US. Moreover, 3D US can contain multiple standard planes (SPs) in one shot. Thus, automatically localizing SPs in 3D US has the potential to improve user-independence and scanning-efficiency. However, manual SP localization in 3D US is challenging because of the low image quality, huge search space and large anatomical variability. In this work, we propose a novel multi-agent reinforcement learning (MARL) framework to simultaneously localize multiple SPs in 3D US. Our contribution is four-fold. First, our proposed method is general and it can accurately localize multiple SPs in different challenging US datasets. Second, we equip the MARL system with a recurrent neural network (RNN) based collaborative module, which can strengthen the communication among agents and learn the spatial relationship among planes effectively. Third, we explore to adopt the neural architecture search (NAS) to automatically design the network architecture of both the agents and the collaborative module. Last, we believe we are the first to realize automatic SP localization in pelvic US volumes, and note that our approach can handle both normal and abnormal uterus cases. Extensively validated on two challenging datasets of the uterus and fetal brain, our proposed method achieves the average localization accuracy of 7.03∘/1.59mm and 9.75∘/1.19mm. Experimental results show that our light-weight MARL model has higher accuracy than state-of-the-art methods.

Learn Fine-Grained Adaptive Loss for Multiple Anatomical Landmark Detection in Medical Images.

Automatic and accurate detection of anatomical landmarks is an essential operation in medical image analysis with a multitude of applications. Recent deep learning methods have improved results by directly encoding the appearance of the captured anatomy with the likelihood maps (i.e., heatmaps). However, most current solutions overlook another essence of heatmap regression, the objective metric for regressing target heatmaps and rely on hand-crafted heuristics to set the target precision, thus being usually cumbersome and task-specific. In this paper, we propose a novel learning-to-learn framework for landmark detection to optimize the neural network and the target precision simultaneously. The pivot of this work is to leverage the reinforcement learning (RL) framework to search objective metrics for regressing multiple heatmaps dynamically during the training process, thus avoiding setting problem-specific target precision. We also introduce an early-stop strategy for active termination of the RL agent's interaction that adapts the optimal precision for separate targets considering exploration-exploitation tradeoffs. This approach shows better stability in training and improved localization accuracy in inference. Extensive experimental results on two different applications of landmark localization: 1) our in-house prenatal ultrasound (US) dataset and 2) the publicly available dataset of cephalometric X-Ray landmark detection, demonstrate the effectiveness of our proposed method. Our proposed framework is general and shows the potential to improve the efficiency of anatomical landmark detection.

Deletion of HNF1A-AS1 Suppresses the Malignant Phenotypes of Breast Cancer Cells In Vitro and In Vivo Through Targeting miRNA-20a-5p/TRIM32 Axis.

Background: Hepatocyte nuclear factor 1 homeobox A-antisense RNA 1 (HNF1A-AS1) is a long noncoding RNA and controls human tumor development and progression. However, its expression and role in breast cancer, the most overwhelmingly occurring malignancy in women globally, remain poorly illuminated. Materials and Methods: Expression of HNF1A-AS1, miRNA (miR)-20a-5p, and tripartite motif containing 32 (TRIM32) was detected using quantitative real-time polymerase chain reaction and Western blotting. Cell proliferation, apoptosis, migration, and invasion were measured by cellTiter 96 AQueous one solution cell proliferation assay kit, flow cytometry, and transwell assays, respectively. Epithelial-mesenchymal transition (EMT) was evaluated by Western blotting, analyzing E-cadherin, N-cadherin, and vimentin expression. Mice xenograft model was generated to investigate tumor growth in vivo. The target binding among miR-20a-5p, HNF1A-AS1, and TRIM32 was confirmed by dual-luciferase reporter assay. Results: Expression of HNF1A-AS1 and TRIM32 was upregulated and miR-20a-5p was downregulated in breast cancer tumors and cell lines. Deletion of HNF1A-AS1 induced cell apoptosis rate, but suppressed cell proliferation, EMT, migration, and invasion in MDA-MB-231 and MCF-7 cells. Furthermore, HNF1A-AS1 downregulation impeded tumor growth in vivo. Interestingly, miR-20a-5p overexpression elicited the similar suppressive effects in MDA-MB-231 and MCF-7 cells, which was partially reversed by TRIM32 upregulation; besides, miR-20a-5p silencing could abolish the antitumor role of HNF1A-AS1 deletion. Notably, HNF1A-AS1 positively modulated TRIM32 expression through acting as a molecular "sponge" for miR-20a-5p. Conclusions: Knockdown of HNF1A-AS1 suppressed breast carcinogenesis presumably through targeting miR-20a-5p/TRIM32 axis, suggesting that HNF1A-AS1 might be a promising therapy target for breast cancer.

Pilot-scale expression, purification, and bioactivity of recombinant human TGF-ß3 from Escherichia coli.

TGF-ß3, a subtype of transforming growth factor-ß (TGF-ß), is essential to various biological processes, including endoderm development, organogenesis, epithelial hyperplasia, synthesis of extracellular matrix, and immune response. Essentially, TGF-ß3 engages the TGF-ß1/Smad signaling pathway to stimulate mesenchymal lineage cells, inhibit epithelial or neuroectodermal lineage cells, and regulate repair, remodeling, and potential scarring after cutaneous wounding. We have now expressed recombinant human TGF-ß3 in Escherichia coli Origami B (DE3), with yield 300 ±â€¯17 mg/L monomeric protein at pilot scale. Identity was confirmed by western blot and HPLC-based peptide mapping. After purification and refolding, dimeric proteins were found to induce chondro-related genes in adipose-derived stem cells, and to suppress scarring in injured rabbit ears. Thus, the recombinant protein has excellent potential for medical applications.

The novel cannabinoid receptor GPR55 mediates anxiolytic-like effects in the medial orbital cortex of mice with acute stress.

The G protein-coupled receptor 55 (GPR55) is a novel cannabinoid receptor, whose exact role in anxiety remains unknown. The present study was conducted to explore the possible mechanisms by which GPR55 regulates anxiety and to evaluate the effectiveness of O-1602 in the treatment of anxiety-like symptoms. Mice were exposed to two types of acute stressors: restraint and forced swimming. Anxiety behavior was evaluated using the elevated plus maze and the open field test. We found that O-1602 alleviated anxiety-like behavior in acutely stressed mice. We used lentiviral shRNA to selective ly knockdown GPR55 in the medial orbital cortex and found that knockdown of GPR55 abolished the anxiolytic effect of O-1602. We also used Y-27632, a specific inhibitor of ROCK, and U73122, an inhibitor of PLC, and found that both inhibitors attenuated the effectiveness of O-1602. Western blot analysis revealed that O-1602 downregulated the expression of GluA1 and GluN2A in mice. Taken together, these results suggest that GPR55 plays an important role in anxiety and O-1602 may have therapeutic potential in treating anxiety-like symptoms.

Gastrodin relieved complete Freund's adjuvant-induced spontaneous pain by inhibiting inflammatory response.

The analgesic effects of gastrodin (GAS), an active component derived from the Chinese herb Tian ma (Gastrodia elata Blume), on chronic inflammatory pain of mice and the involved molecular mechanisms were investigated. GAS significantly attenuated mice chronic inflammatory pain induced by hindpaw injection of complete Freund's adjuvant (CFA) and the accompanying anxiety-like behaviors. GAS administration reduced CFA-induced up-regulation of GluR1-containing α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, GluN2A- and GluN2B-containing N-methyl-d-aspartate (NMDA) receptors, and Ca2+/calmodulin-dependent protein kinase II-alpha (CaMKII-α) in the anterior cingulate cortex (ACC). The GluN2A and GluN2B subunits of NMDA receptors, the GluR1 type of AMPA receptor, and CaMKII-α are key molecules responsible for neuroplasticity involved in chronic pain and the accompanying anxiety. Moreover, GAS administration reduced the activation of astrocyte and microglia and the induction of TNF-α and IL-6 in the ACC of the CFA-injected mice. Therefore, GAS administration relieved chronic pain, exerted anxiolytic effects by regulating neuroplasticity molecules, and attenuated the inflammatory response by reducing the induction of TNF-α and IL-6 in the ACC of the CFA-injected mice.

A moderate static magnetic field enhances TRAIL-induced apoptosis by the inhibition of Cdc2 and subsequent downregulation of survivin in human breast carcinoma cells.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) exhibits its potent antitumor activity via membrane receptors on cancer cells without deleterious side effects for normal tissue. However, as many other cancer types, breast cancer cells develop a resistance to TRAIL. In the present study, we reported that exposure to 3.0 mT static magnetic field (SMF) mediated the sensitization of breast cancer cells to TRAIL-induced apoptosis. This effect was significantly reduced by the forced expression of survivin, suggesting the sensitization was mediated at least in part through the inhibition of survivin expression. In addition, SMF alone or in combination with TRAIL induced a cell cycle arrest within the G2 /M phase, and the reduction in the survivin protein level was associated with the downregulated expression of Cdc2, a cyclin B-dependent kinase that is necessary for the entry into the M phase. Taken together, our results demonstrated that SMF promoted TRAIL-induced apoptosis by inhibiting the expression of Cdc2 and, subsequently, survivin. Of note, SMF did not sensitize untransformed human mammary epithelial cells to TRAIL-mediated apoptosis. Therefore, the combined treatment of SMF and TRAIL may offer an attractive strategy for safely treating resistant breast cancers.

Ethanolic cofermentation with glucose and xylose by the recombinant industrial strain Saccharomyces cerevisiae NAN-127 and the effect of furfural on xylitol production.

Saccharomyces cerevisiae strain NAN-127 (2n, prototroph), which contains the xylose reductase-xylitol dehydrogenase (XR-XDH) xylose metabolic pathway was used for the cofermentation of glucose and xylose. Oxygen supply was the most important factor for xylose fermentation and pH 4.5 and a ventilation rate of 0.04 vvm were optimal. The xylose utilization ratio reached 0.655 at an initial xylose concentration of 50 gL(-1) and was 0.9 at an initial concentration of 20 gL(-1). Addition of furfural at late logarithmic phase as electron acceptor to a final concentration of 3.0 gL(-1) decreased the xylitol yield by 17% under micro-aeration conditions without inhibiting cell growth, but also without an increase in ethanol yield. The results are important to the application of strain NAN-127 in the lignocellulosic ethanol process.

Establishment of a xylose metabolic pathway in an industrial strain of Saccharomyces cerevisiae.

To produce an industrial strain of Saccharomyces cerevisiae that metabolizes xylose, we constructed a rDNA integration vector and YIp integration vector, containing the xylose-utilizing genes, XYL1 and XYL2, which encode xylose reductase (XR) and xylitol dehydrogenase (XDH) from Pichia stipitis, and XKS1, which encodes xylulokinase (XK) from S. cerevisiae, with the G418 resistance gene KanMX as a dominant selectable marker. The rDNA results in integration of multiple copies of the target genes. The industrial stain of S. cerevisiae NAN-27 was transformed with the two integration vectors to produce two recombinant strains, S. cerevisiae NAN-127 and NAN-123. Upon transformation, multiple copies of the xylose-utilizing genes were integrated into the genome rDNA locus of S. cerevisiae. Strain NAN-127 consumed twice as much xylose and produced 39% more ethanol than the parent strain, while NAN-123 consumed 10% more xylose and produced 10% more ethanol than the parent strain over 94 h.