SPOP negatively regulates mTORC1 activity by ubiquitinating Sec13.

The mammalian target of rapamycin complex1 (mTORC1) can response to amino acid to regulate metabolism and cell growth. GATOR2 act as important role in amino acid mediated mTORC1 signaling pathway by repressing GTPase activity (GAP) of GATOR1. However, it is still unclear how GATOR2 regulates mTORC1 signaling pathway. Here, we found that K63-ubiquitination of Sce13, one component of GATOR2, suppresses the mTORC1 activity by lessening the inter-interaction of GATOR2. Mechanistically, the ubiquitination of Sec13 was mediated by SPOP. Subsequently, the ubiquitination of Sec13 attenuated its interaction with the other component of GATOR2, thus suppressing the activity of mTORC1. Importantly, the deficiency of SPOP promoted the faster proliferation and migration of breast cancer cells, which was attenuated by knocking down of Sec13. Therefore, SPOP can act as a tumor suppressor gene by negatively regulating mTORC1 signaling pathway.

WFDC21P promotes triple-negative breast cancer proliferation and migration through WFDC21P/miR-628/SMAD3 axis.

Long non-coding RNAs (lncRNAs) modulate cell proliferation, cycle, and apoptosis. However, the role of lncRNA-WFDC21P in the tumorigenesis of triple-negative breast cancer (TNBC) remains unclear. Results of this study demonstrated that WFDC21P levels significantly increased in TNBC, which was associated with the poor survival of patients. WFDC21P overexpression significantly promoted TNBC cell proliferation and metastasis. WFDC21P interacted with miR-628-5p, which further suppressed cell proliferation and metastasis by negatively regulating Smad3-related gene expression. Recovery of miR-628-5p weakened the roles of WFDC21P in promoting the growth and metastasis of TNBC cells. Moreover,N6-methyladenosine (m6A) modification upregulated WFDC21P expression in the TNBC cells. WFDC21P and its m6A levels were increased after methyltransferase like 3 (METTL3) overexpression but reduced after METTL3 silencing. The proliferation and metastasis of TNBC cells were promoted by METTL3 overexpression but suppressed by METTL3 silencing. This study demonstrated the vital roles of WFDC21P and its m6A in regulating the proliferation and metastasis of TNBC cells via the WFDC21P/miR-628/SMAD3 axis.

Structural, Physicochemical, and Functional Properties of Wheat Bran Insoluble Dietary Fiber Modified With Probiotic Fermentation.

Insoluble dietary fiber (IDF) were isolated from wheat bran (WB) after microbial fermentation with single or mixed strain [Lactobacillus plantarum, Lactobacillus acidophilus, Bacillus subtilis or mixed lactic acid bacteria (L. plantarum and L. acidophilus with ration of 1:1)]. Structure, physicochemical, functional properties, and antioxidant activity of the wheat bran insoluble dietary fiber (W-IDF) modified by fermentation were studied. Fourier transformed infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM) analysis suggested the successful modification of W-IDF. After fermentation with L. plantarum and mixed lactic acid bacteria, the water retention capacity (WRC), oil retention capacity (ORC), and water swelling capacity (WSC) of W-IDF were improved. The sodium cholate adsorption capacity (SCAC), and cation exchange capacity (CEC) of W-IDF modified with L. acidophilus fermentation were significantly increased. Although the cholesterol adsorption capacity (CAC) of W-IDF decreased after modification with probiotic fermentation, nitrite ion adsorption capacity (NIAC), and total phenolic content (TPC) were enhanced. Additionally, W-IDF modified by fermentation with B. subtilis or mixed lactic acid bacteria exhibited superior antioxidant capacity verified by DPPH, ABTS and total reducing power assays. Results manifested that microbial fermentation is a promising methods to modify the W-IDF to provide high-quality functional IDF for food processing and human health management.

Activation of C3 and C5 May Be Involved in the Inflammatory Progression of PCM and GM.

Plasma cell mastitis (PCM) and granulomatous mastitis (GM) are the most common inflammatory diseases constituting nonbacterial mastitis (NBM). However, the pathogenesis of NBM remains unclear. In this study, risk factors for NBM were assessed, as well as the pathological features of PCM and GM. The levels of C3/C3a-C3aR and C5/C5a-C5aR1 of tissues were detected by IHC and WB. Exosomes were isolated from serum and identified by transmission electron microscopy. Then, C3 and C5 levels were detected in peripheral blood, and exosomes were assessed by flow cytometry and immunoelectron microscopy. Obesity and prolonged lactation were risk factors for NBM. The infiltration of plasma cells and lymphocytes around the dilated catheter in PCM and the formation of granulomatous structures in GM were the respective pathological features. C3/C3a-C3aR and C5/C5a-C5aR1 levels were elevated in PCM and GM tissue samples. There were no differences in peripheral blood levels of C3 and C5, while C3a and C5a were highly expressed in exosomes. These results suggest that the complement family is activated in PCM and GM, exosomes enrich C3a and C5a, and mediate the spread of inflammation. These findings provide new insights into the molecular mechanisms of PCM and GM and identify therapeutic targets.

A novel and effective approach to generate germline-like monoclonal antibodies by integration of phage and mammalian cell display platforms.

Phage display technology allows for rapid selection of antibodies from the large repertoire of human antibody fragments displayed on phages. However, antibody fragments should be converted to IgG for biological characterizations and affinity of antibodies obtained from phage display library is frequently not sufficient for efficient use in clinical settings. Here, we describe a new approach that combines phage and mammalian cell display, enabling simultaneous affinity screening of full-length IgG antibodies. Using this strategy, we successfully obtained a novel germline-like anti-TIM-3 monoclonal antibody named m101, which was revealed to be a potent anti-TIM-3 therapeutic monoclonal antibody via in vitro and in vivo experiments, indicating its effectiveness and power. Thus, this platform can help develop new monoclonal antibody therapeutics with high affinity and low immunogenicity.

XRCC5/6 polymorphisms and their interactions with smoking, alcohol consumption, and sleep satisfaction in breast cancer risk: A Chinese multi-center study.

BACKGROUND: X-ray repair cross-complementary 5 (XRCC5) and 6 (XRCC6) are critical for DNA repair. Few studies have assessed their association with breast cancer risk, and related gene-environment interactions remain poorly understood. This study aimed to determine the influence of XRCC5/6 polymorphisms on breast cancer risk, and their interactions with cigarette smoking, alcohol consumption, and sleep satisfaction. METHODS: The study included 1039 patients with breast cancer and 1040 controls. Four single-nucleotide polymorphisms of XRCC5 and two of XRCC6 were genotyped. Information about smoking, alcohol consumption, and sleep satisfaction was collected through questionnaires. Odds ratios (OR) and related 95% confidence intervals (95% CI) were assessed using unconditional logistic regression models. Gene-environment interactions were analyzed using logistic regression with multiplicative interaction models. RESULTS: XRCC5 rs16855458 was associated with increased breast cancer risk in the co-dominant (ptrend  = 0.003) and dominant (CA + AA vs. CC, OR = 1.29, 95% CI = 1.07-1.56, p = 0.008) genetic models after Bonferroni correction. The CG + GG genotype of XRCC6 rs2267437 was associated with an increased risk of estrogen receptor-negative/progesterone receptor-negative (ER-/PR-) breast cancer (CG + GG vs. CC: OR = 1.54, 95% CI = 1.12-2.13, p = 0.008) after Bonferroni correction. Moreover, an antagonistic interaction between XRCC5 rs16855458 and alcohol consumption (pinteraction  = 0.017), and a synergistic interaction between XRCC6 rs2267437 and sleep satisfaction were associated with breast cancer risk (pinteraction  = 0.0497). However, these interactions became insignificant after Bonferroni correction. CONCLUSION: XRCC5 rs16855458 was associated with breast cancer risk, and XRCC6 rs2267437 was associated with the risk of ER-/PR- breast cancer. Breast cancer risk associated with XRCC5 and XRCC6 polymorphisms might vary according to alcohol consumption and sleep satisfaction, respectively, and merit further investigation.

Relationship Between Lifestyle Habits and Health-Related Quality of Life of Recently Diagnosed Breast Cancer Patients: A Comparison Between Younger and Older Women in China.

Objective: The aim of this study was to evaluate the relationship between lifestyle habits and health-related quality of life (HRQoL) among different ages who were initially diagnosed with breast cancer (within the first 2 weeks) and to determine the contribution of lifestyle habits factors on HRQoL. Methods: Patients with breast cancer were recruited from 22 hospitals in 11 provinces or municipalities in northern and eastern China. The Functional Assessment of Cancer Therapy-Breast Cancer (FACT-B) was used to measure HRQoL. Chi-square test, ANOVA, and multivariable generalized linear models were conducted to identify the differences in HRQoL between two age groups (age <50 years and ≥50 years) and to evaluate the contribution of lifestyle habits factors on HRQoL of patients with breast cancer. Results: About 1,199 eligible patients with breast cancer were used for analysis. Younger women (aged <50 years) appeared to show lower scores than older women (aged ≥50 years) in HRQoL subscales, including emotional well-being (p = 0.003), functional well-being (p = 0.006), breast cancer subscale (p = 0.038), and FACT-B Total scores (p = 0.028). Tea and alcohol consumption and being very satisfied with sleep and current life were the strongest predictors of higher HRQoL in younger group. Meanwhile, no coffee consumption, frequent participation in physical activities, high sleep satisfaction, and current life satisfaction were the key predictors of higher HRQoL in older women with breast cancer. Conclusion: The relationship of the nine lifestyle habit items with HRQoL differed among younger and older women. The associated variable of low HRQoL can help clinicians take intervention early in order to improve the prognosis of patients with breast cancer.

Using KISS Flaps in a Chest Wall Reconstruction After Mastectomy for Locally Advanced Breast Cancer: A New Technique.

This study described a technique for the reconstruction of large lateral thoracic defects after local advanced breast cancer resection that allows for complete cover of the defect and primary closure of the donor site. The authors performed reconstruction using the newly designed KISS flap in 2 women for coverage of their large skin defect (15 × 13 cm each) following mastectomies with extensive tissue resection. The KISS flap consisting of 2 skin islands (marked Flap A and Flap B; 15 × 6 cm each) was designed and transferred to the thoracic defect through the subcutaneous tunnel, and based on the same vessel. The flap covered properly without causing excessive tension and allowed primary closure of chest wound and donor defect. The security it brings is comparable with that of classical radical mastectomy, and its success rate is similar to that of single skin flap transplantation. Compared with the conventional pedicled latissimus-dorsi-musculocutaneous flap, we believe that the donor zone tension decreases, wherein the KISS flaps can reduce the incidence of incision dehiscence and nonhealing complications to some extent. The study reported good results from this technique and discussed the techniques that referenced previous reports.

Molecular binding mode of PF-232798, a clinical anti-HIV candidate, at chemokine receptor CCR5.

The chemokine receptor CCR5 is an important anti-HIV (human immunodeficiency virus) drug target owning to its pivotal role in HIV-1 viral entry as a co-receptor. Here, we present a 2.9 Å resolution crystal structure of CCR5 bound to PF-232798, a second-generation oral CCR5 antagonist currently in phase II clinical trials. PF-232798 and the marketed HIV drug maraviroc share a similar tropane scaffold with different amino (N)- and carboxyl (C)- substituents. Comparison of the CCR5-PF-232798 structure with the previously determined structure of CCR5 in complex with maraviroc reveals different binding modes of the two allosteric antagonists and subsequent conformational changes of the receptor. Our results not only offer insights into the phenomenon that PF-232798 has higher affinity and alternative resistance profile to maraviroc, but also will facilitate the design of new anti-HIV drugs.

[Enhanced Nitrogen and Carbon Removal Performance of Simultaneous ANAMMOX and Denitrification (SAD) with Trehalose Addition Treating Saline Wastewater].

Enhanced nitrogen and carbon removal performance of simultaneous ANAMMOX and denitrification (SAD) process with trehalose addition treating saline wastewater was investigated in a sequencing batch reactor (SBR). The optimal nitrogen removal was achieved at 0.25 mmol·L-1 trehalose, during which NH4+-N, NO2--N, NO3--N, and COD could be completely removed. Compared to no addition of trehalose, ammonium removal efficiency (ARE), nitrite removal efficiency (NRE) and total nitrogen removal efficiency (TNRE) increased by 50%, 43% and 46%. Ammonium removal rate (ARR) and nitrite removal rate (NRR) increased by 81.25% and 75%, respectively. With increasing concentration of trehalose to 0.5 mmol·L-1, ARE was only 58.82% and the effluent concentration of NH4+-N was 33.25 mg·L-1. Compared to the Haldane model and the Aiba model, the Luong model was the most suitable to simulate the nitrogen removal performance of SAD with trehalose addition treating saline wastewater. The NRRmax, KS, Sm, and n fitted from Luong model were 0.954 kg·(m3·d)-1, 0 mg·L-1, 184.785 mg·L-1, and 0.718, respectively. Compared to the modified Logistic model and the modified Boltzman model, the modified Gompertz model was the most suitable to describe the degradation of a substrate in a single cycle.

Single-incision surgery for gynecomastia using TriPort: A case report.

Gynecomastia is a common benign condition of the male breast, frequently observed in newborns, adolescents and the elderly. Surgical excision remains one of the most effective methods for the management of patients with gynecomastia, particularly those with fibrosis or a course of >1 year. The present study describes two cases of patients with gynecomastia and presents a novel method of treatment, combining endoscopic surgery and liposuction using a TriPort, which may effectively remove breast tissue and shorten hospitalization time compared with a direct excision method.

Crystal structure-based comparison of two NAMPT inhibitors.

Inhibition of nicotinamide phosphoribosyltransferase (NAMPT) is a novel strategy for cancer therapy, but only two inhibitors of NAMPT (FK866 and CHS828) have progressed into clinical trials. This study seeks to compare a novel potent NAMPT inhibitor, MS0, with a classical inhibitor FK866 in their biological activity and molecular binding mode, thereby contributing to future chemical optimization and a further understanding of the action mode of NAMPT inhibitors. The IC50 values of MS0 and FK866 in inhibition of recombinant human NAMPT activity were 9.08±0.90 and 1.60±0.32 nmol/L, respectively. Consistently, FK866 exerted better antiproliferation in 6 human cancer cell lines (HepG2, A2780, 95-D, A549, U2OS and U266) than MS0 with IC50 values nearly 12-fold to 225-fold lower than those of MS0. Co-crystal structures of wild-type human NAMPT complexed with MS0 or FK866 were elucidated, which revealed that MS0 did not interact with Ser241. The hydrogen bond mediated by crystallographic water between MS0 and His191 or Val350 of NAMPT did not exist in FK866. Instead, FK866 exhibited hydrophobic interactions with Arg349. Based on the activity assays and crystal structure analyses, we elaborate the reason why the antiproliferation activity of MS0 was not as good as that of FK866, which would contributes to the current understanding of the mode of action of NAMPT inhibitors and will also contribute to further development of anticancer drugs in the future.

A case-control study on risk factors of breast cancer in Han Chinese women.

This study aimed to investigate risk factors associated with breast cancer among Han Chinese women in northern and eastern China. A matched case-control study involving 1489 patients with breast cancer and 1489 controls was conducted across 21 hospitals in 11 provinces in China, from April 2012 to April 2013. We developed a structured questionnaire to record information from face-to-face interviews with participants. Student's t-tests, Pearson's chi-square tests, and univariate and multivariate conditional logistic regression analyses were used to identify variables with significant differences between the case and control groups. Ten variables were identified (P<0.05): location, economic status, waist-to-hip ratio, menopause, family history of breast cancer, present life satisfaction, sleep satisfaction, milk products, behavior prevention scores, and awareness of breast cancer. We identified a comprehensive range of factors related to breast cancer, among which several manageable factors may contribute to breast cancer prevention. Further prospective studies concerning psychological interventions, sleep regulation, health guidance, and physical exercise are required. A screening model for high-risk populations should be put on the agenda.

[Enrichment and Nitrogen Removal Characteristics of Marine Anaerobic Ammonium Oxidizing Bacteria].

In this work, enrichment and nitrogen removal characteristics of marine anaerobic ammonium oxidizing bacteria were investigated by seeding sediment sludge from the Jiaozhou bay. Experimental results showed that the whole process could be divided into four phases:bacterial lysis phase (1-15 d), lag phase (16-152 d), exponential growth phase (153-183 d) and stationary phase (184-192 d). Compared to freshwater anaerobic ammonium oxidizing bacteria, the lag phase (137 d) was longer. However, the exponential growth phase (30 d) was shorter. Besides, marine anaerobic ammonium oxidizing bacteria were more sensitive to variation in substrate concentration and HRT. The activity delay time caused by inflow and effluent was 5 h, which was far longer than fresh anaerobic ammonium oxidizing bacteria. As a result, the adaptive ability of marine anaerobic ammonium oxidizing bacteria was weaker and it was harder to enrich. After successful enrichment of 192 d, the removal rates of ammonia nitrogen and nitrite nitrogen were 96.98% and 95.66%, respectively. n(NH4+-N):n(NO2--N):n(NO3--N) was 1:(1.2±0.2):(0.22±0.06), which was close to the theoretical ratio. NRRNH4+-Nwas 0.080 kg·(m3·d)-1, which indicated that the activity of marine anaerobic ammonium oxidizing bacteria increased significantly. The enrichment of marine anaerobic ammonium oxidizing bacteria was achieved. The sludge characteristics changed from black to red. SEM observation confirmed that the red granule was cenobium, which consisted of closely spaced micrococcus with smooth surface and crateriform shape.

Circulating High-Molecular-Weight (HMW) Adiponectin Level Is Related with Breast Cancer Risk Better than Total Adiponectin: A Case-Control Study.

The level of total adiponectin, a mixture of different adiponectin forms, has been reported associated with breast cancer risk with inconsistent results. Whether the different forms play different roles in breast cancer risk prediction is unclear. To examine this, we measured total and high molecular weight (HMW) adiponectin in a case-control study (1167 sets). Higher circulating HMW adiponectin was negatively associated with breast cancer risk after adjusting for menopausal status and family history of breast cancer (P=0.024). We analyzed the relationship between adiponectin and breast cancer risk in 6 subgroups. Higher circulating HMW adiponectin was also negatively associated with breast cancer risk (P=0.020, 0.014, 0.035) in the subgroups of postmenopausal women, negative family history of breast cancer, BMI>=24.0. Total adiponectin was positively associated with breast cancer (P=0.028) in the subgroup of BMI<=24.0. Higher HMW/total adiponectin ratio was negatively associated with breast cancer (P=0.019) in the subgroup of postmenopausal women. Interestingly, in the subgroup of women with family history of breast cancer, higher circulating total and HMW adiponectin were positively associated with breast cancer risk (P=0.034, 0.0116). This study showed different forms of circulating adiponectin levels might play different roles in breast cancer risk. A higher circulating HMW adiponectin is associated with a decreased breast cancer risk, especially in postmenopausal, without family history of breast cancer or BMI>=24.0 subgroups, whereas higher circulating HMW adiponectin levels is a risk factor in women with a family history of breast cancer. Further investigation of different forms of adiponectin on breast cancer risk is needed.

Effect of bFGF on the MCF-7 Cell Cycle with CD44(+)/CD24(-): Promoting the G0/G1→G2/S Transition.

PURPOSE: Few cells with stem cell characteristics possess capabilities of self-renewal and differentiation, which leads to high tumorigenesis and resistance to standard chemotherapeutic agents. These cells are mostly quiescent, and arrest occurs at the mitotic G0/G1 phase in mitosis. We explored the effects of basic fibroblast growth factor (bFGF) on the MCF-7 cell cycle with CD44(+)/CD24(-). METHODS: Cancer-initiating cells were propagated as mammospheres. The CD44(+)/CD24(-) subpopulation was sorted by a fluorescence activating cell sorter-Vantage flow cytometer. A cell cycle analysis was performed with different bFGF concentrations. RESULTS: Differences in the CD44(+)/CD24(-) cell proliferation under different bFGF concentrations were observed (p=0.001). When the bFGF concentration was increased, the proportion of CD44(+)/CD24(-) at G0/G1 decreased (p=0.023). CONCLUSION: We conclude that bFGF may sustain CD44(+)/CD24(-) cell proliferation and could promote cell progression through the G0/G1→G2/S phase transition.

Lymphotactin enhances the in-vitro immune efficacy of dendritoma formed by dendritic cells and mouse hepatocellular carcinoma cells.

OBJECTIVE: To investigate the in-vitro antitumor immune responses of dendritoma formed by mouse hepatocellular carcinoma (HCC) cells and lymphotactin (Lptn) gene modified dendritic cells (DCs). METHOD: DCs prepared from mouse bone marrow were genetically modified by lymphotactin adenovirus, and fused with H22 cells by polyethylene glycol (PEG). RT-PCR and ELISA were employed to identify lymphotactin expression at mRNA and protein level. Cell phenotypes and fusion efficiency was detected by FACS. The stimulatory effect of DC on T cells was detected by mixed lymphocyte reaction. The cytotoxicity activity against H22 cells was assayed by LDH method. RESULTS: Lymphotactin could be efficiently expressed by DCLptn/H22 hybridoma. DCLptn/H22 cells could induce potent T cell proliferation effect and generate strong cytotoxic T lymphocyte (CTL) reaction against allogenic H22 cells. CONCLUSION: Lymphotactin genetic modification could enhance the in vitro immune activity of the dendritoma.

Antitumor effect of immunization with fusion of dendritic cells and hepatocellular carcinoma cells in mice.

BACKGROUND: Important advances have made within the past few years in the treatment of hepatocellular carcinoma (HCC), however, the long-term prognosis after resection of HCC remains unsatisfactory as a result of a high incidence of recurrence. This study was to investigate immunization with fusions of DCs and HCC cells against HCC tumors transplanted to mice. METHODS: Fusion cells of dendritic cells (DCs) and H22 cells were prepared with polyethylene glycol. Expression of MHC and costimulatory molecules by dendritomas were determined by FACs. To study the antitumor immune preventitive and therapeutic effects, fusions were subcutaneously injected into naive or tumor-bearing mice; the CTL activity was assumed by the LDH method, and the expression of tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma(IFN-gamma) in tumors were assayed by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The hybridomas of DCs and H22 cells acquired both DCs and H22 cells phenotypes. Immunization of BALB/C mice with DC/H22 fusions induced protective immunity against a high dose of H22 tumor challenge. After treatment with hybridomas, the survival time of tumor-bearing mice was extended. The expression level of TNF-alpha and IFN-gamma mRNA was markedly increased. CONCLUSION: The hybridomas of DCs and H22 cells could induce effective antitumor immune responses and may be potentially used in prevention and management of recurrence and metastasis of HCC.

What has luck got to do with economic development? An interpretation of resurgent Asia's growth experience.

This paper critically reexamines the belief, currently gathering strength in the literature, that economic development depends on good luck rather than on good policy, and that Prometheus is "unchained by chance". While it is impossible to disprove the role of luck in growth, we argue that luck is endogenous, and good luck is a function of good policy. Luck favours those who strive. Again contrary to common belief, we show that resurgent Asian economies have endured more, not less, than their fair share of economic volatility. They learned their lessons by success and failures, and luck is endogenous through learning-by-investing.