Theabrownin inhibits obesity and non-alcoholic fatty liver disease in mice via serotonin-related signaling pathways and gut-liver axis.

INTRODUCTION: Non-alcoholic fatty liver disease (NAFLD) with obesity seriously threats public health. Our previous studies showed that dark tea had more potential on regulating lipid metabolism than other teas, and theabrownin (TB) was considered to be a main contributor to the bioactivity of dark tea. OBJECTIVES: This in vivo study aims to reveal the effects and molecular mechanisms of TB on NAFLD and obesity, and the role of the gut-liver axis is explored. METHODS: The histopathological examinations, biochemical tests, and nuclear magnetic resonance were applied to evaluate the effects of TB on NAFLD and obesity. The untargeted metabolomics was used to find the key molecule for further exploration of molecular mechanisms. The 16S rRNA gene sequencing was used to assess the changes in gut microbiota. The antibiotic cocktail and fecal microbiota transplant were used to clarify the role of gut microbiota. RESULTS: TB markedly reduced body weight gain (67.01%), body fat rate (62.81%), and hepatic TG level (51.35%) in the preventive experiment. Especially, TB decreased body weight (32.16%), body fat rate (42.56%), and hepatic TG level (42.86%) in the therapeutic experiment. The mechanisms of action could be the improvement of fatty acid oxidation, lipolysis, and oxidative stress via the regulation of serotonin-related signaling pathways. Also, TB increased the abundance of serotonin-related gut microbiota, such as Akkermansia, Bacteroides and Parabacteroides. Antibiotics-induced gut bacterial dysbiosis disrupted the regulation of TB on serotonin-related signaling pathways in liver, whereas the beneficial regulation of TB on target proteins was regained with the restoration of gut microbiota. CONCLUSION: We find that TB has markedly preventive and therapeutic effects on NAFLD and obesity by regulating serotonin level and related signaling pathways through gut microbiota. Furthermore, gut microbiota and TB co-contribute to alleviating NAFLD and obesity. TB could be a promising medicine for NAFLD and obesity.

Brain-specific loss of Abcg1 disturbs cholesterol metabolism and aggravates pyroptosis and neurological deficits after traumatic brain injury.

Based on accumulating evidence, cholesterol metabolism dysfunction has been suggested to contribute to the pathophysiological process of traumatic brain injury (TBI) and lead to neurological deficits. As a key transporter of cholesterol that efflux from cells, the ATP-binding cassette (ABC) transporter family exerts many beneficial effects on central nervous system (CNS) diseases. However, there is no study regarding the effects and mechanisms of ABCG1 on TBI. As expected, TBI resulted in the different time-course changes of cholesterol metabolism-related molecules in the injured cortex. Considering ABCG1 is expressed in neuron and glia post-TBI, we generated nestin-specific Abcg1 knockout (Abcg1-KO) mice using the Cre/loxP recombination system. These Abcg1-KO mice showed reduced plasma high-density lipoprotein cholesterol levels and increased plasma lower-density lipoprotein cholesterol levels under the base condition. After TBI, these Abcg1-KO mice were susceptible to cholesterol metabolism turbulence. Moreover, Abcg1-KO exacerbated TBI-induced pyroptosis, apoptosis, neuronal cell insult, brain edema, neurological deficits, and brain lesion volume. Importantly, we found that treating with retinoid X receptor (RXR, the upstream molecule of ABCG1) agonist, bexarotene, in Abcg1-KO mice partly rescued TBI-induced neuronal damages mentioned above and improved functional deficits versus vehicle-treated group. These data show that, in addition to regulating brain cholesterol metabolism, Abcg1 improves neurological deficits through inhibiting pyroptosis, apoptosis, neuronal cell insult, and brain edema. Moreover, our findings demonstrate that the cerebroprotection of Abcg1 on TBI partly relies on the activation of the RXRalpha/PPARgamma pathway, which provides a potential therapeutic target for treating TBI.

The cytokine profiles in follicular fluid and reproductive outcomes in women with endometriosis.

PROBLEM: Endometriosis patients undergoing in vitro fertilization-embryo transfer (IVF-ET) treatment suffer from poor oocyte quality, a reduced likelihood of the fertilization rate, and low embryo quality. The dysregulation of immune cells and cytokine profiles in the follicular fluid (FF) may play an important role in the competence of the oocyte and the development of the embryo, but the mechanism remains largely unknown. METHOD OF STUDY: A total of 40 proved advanced staged endometriosis patients were enrolled in this study. The pregnancy results were followed until all the embryos collected by the first oocyte retrieval cycle were used up. The immune cells subtypes in FF and serum collected on the day of oocyte retrieval were detected by flow cytometry and 27 cytokines were determined using the Bio-Plex Pro Human Cytokine 27-Plex Immunoassay. The specific effect of cytokine on the gene expression of human granulosa cells was determined by RT-qPCR. RESULTS: The fertilization rate and the cumulative live birth rate were significantly lower in the endometriosis group. The ratio of CD4+ /CD8+ T cells in FF was significantly lower, while the level of IP-10, RANTES and G-CSF were statistically higher in the endometriosis group. The level of IP-10 correlated with the IVF outcome. Moreover, treated by IP-10, the mRNA level of FSHR and CYP19A1 the human granulosa cells were downregulated in vitro. CONCLUSION: These results suggest that alterations of the lymphocyte subsets and cytokines in women with advanced endometriosis may have an impact on the oocyte development and resulting in poorer IVF outcomes.

Krüppel-like factor 4 promotes the proliferation and osteogenic differentiation of BMSCs through SOX2/IGF2 pathway.

OBJECTIVES: Human bone marrow mesenchymal stem cells (BMSCs) have multi-lineage differentiation potential and have been widely researched in regenerative medicine. The purpose of this research was to explore whether Krüppel-like factor 4 (KLF4) can regulate the osteogenic differentiation of BMSCs. METHODS: We transfected human BMSCs with KLF4 overexpression plasmid and si-KLF4 to study the effects of KLF4. We performed cell proliferation assay, flow cytometry and Alizarin Red staining on BMSCs. Quantitative real-time PCR and western blot was performed to determined mRNA and protein expression of osteogenic differentiation markers, KLF4, SOX2 and IGF2. Bone defect animal model was created and the adenovirus containing KLF4 overexpression or knockdown plasmid was injected. Finally, HE staining was performed on tibia to assess the new bone formation. RESULTS: Our results showed that KLF4 promotes not only the growth of BMSCs, but also their osteogenic differentiation. Also, it mediated these effects through SOX2/IGF2 signaling pathway. In addition, KLF4 overexpression could increase the bone regeneration in in-vivo model, whereas KLF4 knockdown decreased the bone regeneration. CONCLUSIONS: KLF4 regulates BMSC's osteogenic differentiation via SOX2/IGF2 pathway.

Decidual-derived RANKL facilitates macrophages accumulation and residence at the maternal-fetal interface in human early pregnancy.

PROBLEM: During the first trimester, the accumulation of macrophages, which is the second largest decidual leukocyte population (~20%) at the maternal-fetal interface, is quite vital for a successful pregnancy, including embryo implantation, trophoblast invasion, and vascular remodeling. The mechanism of the enrichment and redistribution of macrophages in the uterine decidua of early pregnancy is largely unclear. METHOD OF STUDY: A total of 37 women with normal early pregnancies were included. Primary decidual macrophages (dMφs) (n = 37) and primary decidual stromal cells (DSCs) (n = 37) were isolated, and the adhesion molecules were analyzed by flow cytometry (FCM). Adhesive experiment was carried out to evaluate the adhesion capacity by counting cell numbers of dMφs adhered to DSCs in a co-culture system. RESULTS: We found that RANK+ dMφs was the dominating subtype at the maternal-fetal interface. The expression of adhesion molecules (eg, CD29, CD31, CD54, and CD62L) on the surface of RANK+ dMφs was higher than that of RANK- dMφs. After co-culture with DSCs, the expression of adhesion molecules on dMφs was up-regulated in a RANKL-dependent manner. Meanwhile, dMφs promoted the releasing of RANKL on DSCs after co-culture. Consistently, dMφs exhibited the lessoned capacity of adhesion to DSCs when blocking the crosstalk of RANKL-RANK between the DSCs and dMφs in vitro. CONCLUSION: These results suggest that the interaction of RANKL-RANK up-regulates the expression of adhesion molecules on the surface of dMφs, contributing to the accumulation and residence of dMφs in human early pregnancy.

Zero-knowledge identity authentication for internet of vehicles: Improvement and application.

The popularity of Internet of Vehicles (IoV) has made people's driving environment more comfortable and convenient. However, with the integration of external networks and the vehicle networks, the vulnerabilities of the Controller Area Network (CAN) are exposed, allowing attackers to remotely invade vehicle networks through external devices. Based on the remote attack model for vulnerabilities of the in-vehicle CAN, we designed an efficient and safe identity authentication scheme based on Feige-Fiat-Shamir (FFS) zero-knowledge identification scheme with extremely high soundness. We used the method of zero-one reversal and two-to-one verification to solve the problem that FFS cannot effectively resist guessing attacks. Then, we carried out a theoretical analysis of the scheme's security and evaluated it on the software and hardware platform. Finally, regarding time overhead, under the same parameters, compared with the existing scheme, the scheme can complete the authentication within 6.1ms without having to go through multiple rounds of interaction, which reduces the additional authentication delay and enables all private keys to participate in one round of authentication, thereby eliminating the possibility that a private key may not be involved in the original protocol. Regarding security and soundness, as long as private keys are not cracked, the scheme can resist guessing attacks, which is more secure than the existing scheme.

Anonymous-authentication scheme based on fog computing for VANET.

Privacy protection in vehicular ad hoc networks (VANETs) has always been a research hotspot, especially the issue of vehicle authentication, which is critical to ensure the safe communication of vehicles. However, using the real identity in the process of authentication can easily result in a leak of the privacy information of the vehicles. Therefore, most existing privacy-protection schemes use anonymous authentication and require one-to-one communication between vehicles and the trusted authority (TA). However, when the number of vehicles is too large, network congestion can take place. In addition, the process of updating the anonymous by the TA or the vehicle itself, can result in both poor real-time performance and leakage of the system master key. To solve these problems, this study proposes a fog-computing-based anonymous-authentication scheme for VANETs; the scheme reduces the communication burden of the TA by enabling self-authentication between vehicles and road-side units (RSUs), thus improving the vehicle-authentication efficiency. For updating the anonymous, we design a fog-computing-based pseudonym-updating and -tracking strategy, which guarantees real-time communication and reduces the instances of re-authentication interactions for legitimate vehicles. The experimental results show that the scheme not only meets the privacy-protection requirements of VANETs but also offers better performance than that of the existing anonymous-authentication schemes.

Arabidopsis SINAT Proteins Control Autophagy by Mediating Ubiquitylation and Degradation of ATG13.

In eukaryotes, autophagy maintains cellular homeostasis by recycling cytoplasmic components. The autophagy-related proteins (ATGs) ATG1 and ATG13 form a protein kinase complex that regulates autophagosome formation; however, mechanisms regulating ATG1 and ATG13 remain poorly understood. Here, we show that, under different nutrient conditions, the RING-type E3 ligases SEVEN IN ABSENTIA OF ARABIDOPSIS THALIANA1 (SINAT1), SINAT2, and SINAT6 control ATG1 and ATG13 stability and autophagy dynamics by modulating ATG13 ubiquitylation in Arabidopsis (Arabidopsis thaliana). During prolonged starvation and recovery, ATG1 and ATG13 were degraded through the 26S proteasome pathway. TUMOR NECROSIS FACTOR RECEPTOR ASSOCIATED FACTOR1a (TRAF1a) and TRAF1b interacted in planta with ATG13a and ATG13b and required SINAT1 and SINAT2 to ubiquitylate and degrade ATG13s in vivo. Moreover, lysines K607 and K609 of ATG13a protein contributed to K48-linked ubiquitylation and destabilization, and suppression of autophagy. Under starvation conditions, SINAT6 competitively interacted with ATG13 and induced autophagosome biogenesis. Furthermore, under starvation conditions, ATG1 promoted TRAF1a protein stability in vivo, suggesting feedback regulation of autophagy. Consistent with ATGs functioning in autophagy, the atg1a atg1b atg1c triple knockout mutants exhibited premature leaf senescence, hypersensitivity to nutrient starvation, and reduction in TRAF1a stability. Therefore, these findings demonstrate that SINAT family proteins facilitate ATG13 ubiquitylation and stability and thus regulate autophagy.

[Association of Ureaplasma urealyticum with the types of antisperm antibody in infertile men].

OBJECTIVE: To investigate the prevalence of Ureaplasma urealyticum (UU) infection in infertile men, its influence on routine semen parameters and the distribution of antisperm antibody (AsAb) and its types in infertile patients with UU infection. METHODS: We detected the positive rate of UU infection, semen parameters, and the distribution of AsAb and its types in 662 infertile men and 25 normal fertile male controls followed by comparison of the obtained data between the two groups of subjects. RESULTS: The positive rate of UU infection was significantly higher in the infertile men than in the normal controls (52.87% ï¼»350/662ï¼½ vs 16.00% ï¼»4/25ï¼½, χ2 = 11.68, P <0.05). The semen volume, sperm count, sperm concentration and percentage of progressively motile sperm were remarkably lower in the UU-positive infertile males than in the control group (P <0.05). No statistically significant difference was observed between the UU-positive and UU-negative groups in the positive rates of total AsAb (43.4% vs 36.5%, χ2 = 3.25, P >0.05) and AsAb IgA, IgM and IgG in the seminal plasma, or in the percentages of serum AsAb IgM (16.9% vs 20.5%, χ2 = 1.22, P >0.05) and IgG (32.7% vs 28.9%, χ2 = 0.99, P >0.05) except in that of serum AsAb IgA (23.6% vs 17.0%, χ2 = 4.03, P <0.05). CONCLUSIONS: The UU infection rate is high in infertile males, which decreases the semen volume, total sperm count, motile sperm concentration and percentage of progressively motile sperm and increases the positive rate of serum AsAb IgA.

Multi-Attributed Graph Matching With Multi-Layer Graph Structure and Multi-Layer Random Walks.

This paper addresses the multi-attributed graph matching problem, which considers multiple attributes jointly while preserving the characteristics of each attribute for graph matching. Since most of conventional graph matching algorithms integrate multiple attributes to construct a single unified attribute in an oversimplified manner, the information from multiple attributes is often not completely utilized. In order to solve this problem, we propose a novel multi-layer graph structure that can preserve the characteristics of each attribute in separated layers, and also propose a multi-attributed graph matching algorithm based on random walk centrality with the proposed multi-layer graph structure. We compare the proposed algorithm with other state-of-the-art graph matching algorithms based on a single-layer structure using synthetic and real data sets and demonstrate the superior performance of the proposed multi-layer graph structure and the multi-attributed graph matching algorithm.

Protective effect of cornel iridoid glycoside on hepatocytes injured by D-galactosamine/tumor necrosis factor-α / 中国药理学通报

Aim To determine the effect of cornel iri-doid glycoside ( CIG ) on human hepatocyte cell line (L-02) injured by D-galactosamine (GalN) and tumor necrosis factor-α( TNF-α) .Methods Firstly, CIG was extracted , separated and purified . Cell lesion model injured by D-GalN/TNF-αwas tested by MTT method.T-AOC, SOD, MDA and calcium ion concen-tration were taken as indicators to study the effects of CIG on L-02 cell injured by D-GalN/TNF-α.The ex-pression of p-PERK, p-eIF-2α, caspase-3 protein were detected by Western blot .Results 44 mg · L-1 D-GalN and 100 μg · L-1 TNF-αwere suitable for L-02 cell lesion model.CIG high, middle, low concentra-tion group could significantly increase the L-02 cell ac-tivity by 21％, 13％, 8％, respectively and SOD activity and T-AOC ability as well compared with model group.At the same time, they markedly reduced the MDA activity except the low concentration .Three con-centrations of CIG could reduce the expression of endo-plasmic reticulum stress related protein PERK , eIF-2αand apoptosis-associated protein caspase-3. Conclu-sions CIG could protect L-02 cells injured by D-GalN/TNF-α.Increasing the cellular antioxidant abili-ty, reducing the damage of endoplasmic reticulum stress and the expression of apoptosis-associated protein may be the possible mechanism .

A wellness platform for stereoscopic 3D video systems using EEG-based visual discomfort evaluation technology.

Recent advances in three-dimensional (3D) video technology have extended the range of our experience while providing various 3D applications to our everyday life. Nevertheless, the so-called visual discomfort (VD) problem inevitably degrades the quality of experience in stereoscopic 3D (S3D) displays. Meanwhile, electroencephalography (EEG) has been regarded as one of the most promising brain imaging modalities in the field of cognitive neuroscience. In an effort to facilitate comfort with S3D displays, we propose a new wellness platform using EEG. We first reveal features in EEG signals that are applicable to practical S3D video systems as an index for VD perception. We then develop a framework that can automatically determine severe perception of VD based on the EEG features during S3D video viewing by capitalizing on machine-learning-based braincomputer interface technology. The proposed platform can cooperate with advanced S3D video systems whose stereo baseline is adjustable. Thus, the optimal S3D content can be reconstructed according to a viewer's sensation of VD. Applications of the proposed platform to various S3D industries are suggested, and further technical challenges are discussed for follow-up research.

[Sperm chromatin structure assay versus sperm chromatin dispersion test in detecting sperm DNA integrity and correlation of sperm DNA fragmentation with semen parameters].

OBJECTIVE: Sperm DNA fragmentation (SDF) is widely used to predict male infertility and the methods of detecting SDF are varied. This study aimed to compare two methods of SDF detection and investigate the correlation between SDF and sperm quality. METHODS: Using sperm chromatin structure assay (SCSA) and sperm chromatin dispersion test (SCD), we detected SDF in 108 semen samples collected in the Center of Reproduction and Genetics of Suzhou Municipal Hospital. We compared the results of the two methods and analyzed the correlations of SDF routine semen parameters, sperm morphology and the age of the patients. RESULTS: A significant consistency was found in the SDF index (DFI) between the two methods (P<0.01). The DFI was correlated negatively with sperm motility, the percentage of progressively motile sperm, and that of morphologically normal sperm (P <0.01), but positively with the teratozoospermia index (P <0.01 in SCSA and P <0.05 in SCD). The DFI measured by SCSA showed a significantly positive correlation with the patients' age (P <0.01), but not that obtained by SCD. CONCLUSIONS: The results of both SCSA and SCD play an important role in predicting sperm quality. As a clinical index, the DFI has a predictive value for male infertility. However, the results of different detecting methods vary widely, which calls for further studies on their standardization.

Fish Oil-Rich Diet Promotes Hematopoiesis and Alters Hematopoietic Niche.

The self-renewal and differentiation of hematopoietic stem cells (HSCs) in bone marrow are essential to replenish all blood cell types, but how this process is influenced by diet remains largely unclear. Here we show that a diet rich in fish oils promotes self-renewal of HSCs and extramedullary hematopoiesis. Chronic intake of a fish oil-rich diet increases the abundance of HSCs, alters the hematopoietic microenvironment, and, intriguingly, induces the expression of matrix metalloproteinase 12 (MMP12) in the bone marrow. Pointing to a direct effect of fish oil on MMP12 expression, omega-3 polyunsaturated fatty acids induce the expression of MMP12 in a dose-dependent manner in bone marrow cells. Importantly, down-regulation of MMP12 activity using an MMP12-specific inhibitor attenuates diet-induced myelopoiesis in both bone marrow and spleen. Thus, a fish oil-rich diet promotes hematopoiesis in the bone marrow and spleen, in part via the activity of MMP12. Taken together, these data provide new insights into diet-mediated regulation of hematopoiesis.

Fed-batch cultures of Escherichia coli cells with oxygen-dependent nar promoter systems.

The recombinant proteins produced from Escherichia coli as a host cell need to be made at as low a cost as possible because of the end of the monopoly following expiry of the patent on early pharmaceutical proteins, and thus expanding applications to non-pharmaceutical large-scale products. We review in this article how the various promoters used in recombinant E. coli could affect its protein products, especially with emphasis on relatively new oxygen-dependent nar promoters for beta-galactosidase production. Several studies carried out in the authors' laboratory show that the nar promoter does not require any chemicals except 1% nitrate and oxygen for protein production. And according to recent work with the modified strains it is possible to produce the enzyme (beta-galactosidase) even without the nitrate ions at 45% of its total protein content when its cell density reached OD = 176.

Comparison of the Epidemiology and Clinical Characteristics of Primary Hepatocellular Carcinoma Between HBsAg and Anti-HCV Positive Group

BACKGROUND: Primary hepatocellular carcinoma(HCC) is the second cause of cancer death in our country. Hepatitis B virus(HBV) and hepatitis C virus(HCV) are important risk factors for hepatocellular carcinoma. The mechanism of HCC development and the epidemiology in HCV infected individuals are still unclear. In this study, we investigated the epidemiolgical and clinical features of HCC in relation to viral infection. METHODS: We reviewed the medical records of 160 HCC patients retrspectively who had been admitted to one University Hospital located in Seoul between January 1991 and December 1995. Among these patients, 113 patients were positive for HBsAg(B group), 24 for anti-HCV(C group). We compared epidemiological and clinical data between B group and C guoup. RESULTS: Anti-HCV positivity was significantly higher in HBsAg negative patients than in HBsAg positive patients(53.3:1.7%, p<0.01). The mean age of patients in B group was significantly lower than that in C group(52:62yr, p<0.01). In C group, the proportion of Child-Pugh class B and C was significantly larger than that of B group(35.4: 75.0%, p<0.01). In C group, the proportion of transfusion history was significantly larger than that in B group(4.4: 16.7%, p<0.05), and the proportion of drug abuse hestory was significantly larger than that in B group (31.0:62.5%, p<0.01). In C group, the albumin, cholesterol, Gamma-glutamyl transferase leves were significantly lower than those in B group. In B group, the proportion of metastasis was significantly larger than that in C group(31.9:4.2%, p<0.01). Alpha fetoprotein levels greater than 400ng/ml are much more prevalent in group B significantly(67:39.1%, p<0.05). No significant differences in cumulative survival rate(1yr, 2yr) and median survival time were observed between the two groups. CONCLUSIONS: We ascertain that the HBV and HCV are inportant factors in HCC. In epidemiology and clinical features of HCC, there were some difference between the HBsAg and anti HCV positive group. Therefore, on primary health care settings, it is necessary to test for hepatitis C as well as hepatitis B in order to prevent and manage HCC and chronic liver desease.