Citrus maxima and tea regulate AMPK signaling pathway to retard the progress of nonalcoholic fatty liver disease.

Background: Nonalcoholic fatty liver disease (NAFLD) is a chronic metabolic disease that easily induces hepatitis, cirrhosis, and even liver cancer. The long-term use of NAFLD therapeutic drugs produces toxicity and drug resistance. Therefore, it is necessary to develop high efficiency and low-toxicity active ingredients to alleviate NAFLD. Objective: This study aimed to reveal the role and mechanism of a new functional food CMT in alleviating NAFLD. Results: In the ob/ob fatty liver mice models, the CMT extracts significantly inhibited the weight gain of the mice and reduced the accumulation of white fat. The anatomical and pathological results showed that CMT relieved fatty liver in mice and reduced excessive lipid deposition and inflammatory infiltration. Serological and liver biochemical indicators suggest that CMT reduced dyslipidemia and liver damage caused by fatty liver. CMT obviously activated the adenosine 5'-monophosphate-activated protein kinase (AMPK)/acetyl-coA carboxylase (ACC) and AMPK/fatty acid synthase (FAS) signaling pathways, promoted fat oxidation, and inhibited synthesis. Moreover, CMT regulated the expression of inflammatory factors to relieve hepatitis caused by NAFLD. Conclusion: The study explained the role and mechanism of CMT in alleviating NAFLD and suggested that the active ingredients of CMT might be beneficial in NAFLD therapy.

Discovery of a Colon-Targeted Azo Prodrug of Tofacitinib through the Establishment of Colon-Specific Delivery Systems Constructed by 5-ASA-PABA-MAC and 5-ASA-PABA-Diamine for the Treatment of Ulcerative Colitis.

To mitigate the systemic adverse effects of tofacitinib, 5-ASA-PABA-MAC and 5-ASA-PABA-diamine colon-specific delivery systems were constructed, and tofacitinib azo prodrugs 9 and 20a-20g were synthesized accordingly. The release studies suggested that these systems could effectively release tofacitinib in vitro, and the 5-ASA-PABA-diamine system could successfully realize the colon targeting of tofacitinib in vivo. Specifically, compound 20g displayed a 3.67-fold decrease of plasma AUC(tofacitinib, 0-∞) and a 9.61-fold increase of colonic AUC(tofacitinib, 0-12h), compared with tofacitinib at a molar equivalent oral dose. Moreover, mouse models suggested that compound 20g (1.5 mg/kg) could achieve roughly the same efficacy against ulcerative colitis compared with tofacitinib (10 mg/kg) and did not impair natural killer cells. These results demonstrated the feasibility of compound 20g as an effective alternative to mitigate the systemic adverse effects of tofacitinib, and 5-ASA-PABA-MAC and 5-ASA-PABA-diamine systems were proven to be effective for colon-specific drug delivery.

Small breast epithelial mucin promotes the invasion and metastasis of breast cancer cells via promoting epithelial­to­mesenchymal transition.

The aim of the present study was to observe the influence of the small breast epithelial mucin (MUCL1) (also known as SBEM) gene on migration and invasion ability of breast cancer cells and to explore the potentially involved mechanism. SBEM­interference plasmid and SBEM­overexpressing plasmid were constructed. SBEM­knockdown or SBEM­â€‹overexpressing MCF­7 and MDA­MB­231 breast cancer cells were established by lentivirus­mediated stable transfection method. The scratch wound­healing assay and Transwell chamber experiment were used to detect the influence of the SBEM gene on the migration and invasion abilities of MCF­7 and MDA­MB­231 cells. Real­time PCR (polymerase chain reaction) and western blotting were used to detect the expression of epithelial­to­mesenchymal transition (EMT)­related markers and regulators. The cell morphology was observed after transfection. The SBEM­knockdown or SBEM­overexpressing MCF­7 and MDA­MB­231 cells were established successfully. The migration and invasion abilities were decreased after SBEM was downregulated, and were increased after SBEM was overexpressed both in MCF­7 and MDA­MB­231 cell lines. The mRNA and protein expressions of N­cadherin, Twist and vimentin were elevated following SBEM overexpression, while the expression of E­cadherin and claudin­1 were found to be decreased following SBEM overexpression. In conclusion, SBEM has the potential to promote migration and invasion ability of breast cancer cells via promoting epithelial­to­mesenchymal transition.

Mixtures of Tea and Citrus maxima (pomelo) Alleviate Lipid Deposition in HepG2 Cells Through the AMPK/ACC Signaling Pathway.

Tea and citrus maxima are natural, medicinal homologous plants, typically used for making beverages, which have anticancer, antiobesity, and antioxidation properties. Green tea, yellow tea, and black tea were combined with citrus maxima to obtain green tea and Citrus maxima (GTCM), yellow tea and Citrus maxima (YTCM), and black tea and Citrus maxima (BTCM). The biochemical components of these mixtures were analyzed, and their possible effects and mechanisms on relieving liver lipid deposition were explored. The tea polyphenols, free amino acids, phenolamine ratio, and caffeine were comparable in YTCM and GTCM, being significantly higher than those in BTCM. In addition, the content of esterified catechins, nonesterified catechins, and total catechins in YTCM was significantly higher than those in GTCM and BTCM. All three mixtures of Citrus maxima tea significantly reduced lipid deposition in HepG2 cells, with GTCM and YTCM being slightly more effective than BTCM. Regarding the possible mechanism, Western blot analysis revealed that the three Citrus maxima tea mixtures could activate the AMPK/ACC signaling pathway, upregulate the expression of p-AMPK, p-ACC, and CPT-1 proteins, and downregulate the expression of SREBP1c and fatty acid synthase proteins to inhibit fat synthesis, thereby relieving lipid deposition in liver cells. In conclusion, as a novel and healthy beverage, Citrus maxima tea has the potential to alleviate liver lipid deposition, and further could be responsible for obesity treatment.

[Comparison of Functional Groups of Phytoplankton in FG, MFG, and MBFG: Taking Three Reservoirs as an Example in Guizhou Plateau].

Phytoplankton classification is the foundation for revealing the succession rules of phytoplankton communities and for conducting water ecology investigation. At present, the classification methods of functional group (FG), morpho-functional group (MFG), and morphology-based functional group (MBFG) are widely used, although great differences occur among the classification methods of different FGs. In order to compare the differences among FG, MFG and MBFG classification methods, FG classification and redundant analysis (RDA) were used to sample and analyze the phytoplankton community structures and the physical and chemical indicators of water environment in April 2017 (normal seasons) and August 2017 (wet seasons). The following results were noted. ①The MFG and FG had overlapping parts, whereas the MBFG was differed significantly from FG and MFG. ②The FG considers the physiological and ecological characteristics of phytoplankton, which can better explain the correlation with the environment and can accurately reflect the relationship between environmental factors and the phytoplankton niche. Moreover, the environmental factors and eutrophication status of the reservoir can be predicted according to the dominant FGs of phytoplankton; however, the FG needs to have systematic knowledge of phytoplankton identification and a large workload. The MFG classification method is simple and easily understood, which can better explain the relationship between phytoplankton and environmental factors; however, it is weakly connected with environmental factors. The MBFG classification method is the simplest and most efficient, although its interpretation degree to the environment is not sufficient. ③Each of the three classification methods has advantages and disadvantages; therefore, appropriate classification methods are selected according to different research purposes and their actual needs.

[Spatial and Temporal Distribution of Chlorophyll a and Its Relationship to Algae and Environmental Factors in Aha Reservoir].

As one of the drinking water sources for Guiyang City in southwest China, the Aha Reservoir has an area of 190 km2 and a volume of 5.42×108 m3. The water depth is less than 30 m, with an average depth of 13 m. Regulated by subtropical humid monsoon climate, it has cool summers and warm winters, with an annual mean air temperature of about 15.3℃ and an annual normal rainfall of approximately 1,129 mm. Impacted heavily by human activity (e.g., untreated industrial and domestic sewage and agricultural non-point pollution sources), the eutrophication problem in the Aha Reservoir has become more serious each year. In order to explore the spatial and temporal distribution characteristics of chlorophyll a (Chl-a) and its relationship to algae and the driving factors in the Aha Reservoir, phytoplankton and water samples were collected in the dry period, normal period, and flood period. The results showed a significant seasonal variation in Chl-a, same as biomass, but not the same as the algal abundance. Highest Chl-a concentration (91 µg·L-1) occurred in the mean season with the dinoflagellate bloom but during dry and wet seasons, they were only 8 µg·L-1 and 16 µg·L-1, respectively. During the dry and flood periods, the Chl-a concentrations in surface waters were slightly higher than the other layers caused by sufficient light and dissolved oxygen. But in the normal period, the Chl-a concentrations in surface waters were far higher than the other layers because of the dinoflagellate bloom assembling in surface waters. Located at the reservoir entrance of Jinzhong River, Dam sampling point owned higher Chl-a concentration than Kuzhong as a result of higher nutrients. Correlation analysis indicated that Dinoflagellate was positively correlated with Chl-a (R=0.798, P<0.01). Chl-a was positively associated with total phosphorus, dissolved oxygen, pH value, and total nitrogen (R=0.762, P<0.01; R=0.792, P<0.01; R=0.658, P<0.01; R=0.388, P<0.05) and it had a negative correlation with the N/P ratio and nitrate nitrogen (R=-0.37, P<0.05; R=-0.435, P<0.05). Stepwise regression analysis showed that TP, N/P ratio, and DO were the most important factors influencing the temporal and spatial distribution of Chl-a. Thermal stratification and water temperature were also the significant factors that could not be ignored.

[Seasonal stratification and eutrophication characteristics of a deep reservoir, Longtan Reservoir in subtropical area of China].

In order to reveal the seasonal stratification and eutrophication characteristics of the subtropical large deep reservoir--Longtan Reservoir, the spatial and temporal distribution of environmental factors and eutrophic index were investigated during November (dry period) 2012, April (level period) and July (wet period) 2013. The results suggested that: (1) The stratification structure of Longtan Reservoir was meromictic lake, it had a single thermocline structure in the dry season, the surface layer to the 60 m was a mixomolimnion, 60-80 m was a thermocline, deeper more than 80 m was a monimolimnion. It had a double thermocline structure in flow period and wet period, the surface to 10 m was a mixed layer, 10-20 m was a thermocline, 20-40 m was a mixed layer, 40-60 m was a thermocline, deeper more than 60 m was a mixed layer. (2) The thermal stratification dominated the structure of other environmental factors, the stratification structure limited the water convection, especially the monimolimnion reduced the harm of the endogenous pollution. (3) The trophic level index (TLI) was 23.4-32.8 in the dry period, 27.1-38.6 in the flow period and 26.0-45.1 in the wet period, which were all Mesotropher. The trophic state index of total nitrogen was 60.3-72.5, which was eutropher to hyper eutropher, N: P was 107:1, which was phosphorus limited.

[Response of Phytoplankton Functional Groups to Eutrophication in Summer at Xiaoguan Reservoir].

Hydrology and Water Resources Bureau of Guizhou Province, Guiyang 550002, China) Abstract: In order to explore the distribution characteristics of phytoplankton functional groups, eutrophication characteristics and response of phytoplankton functional groups to eutrophication in Xiaoguan Reservoir, phytoplankton and water samples were taken once a week from 25th July 2014 to 27th September 2014. The results showed that there were 22 phytoplankton functional groups, groups S1, D, J, B, G, MP, L0, SN, X1, Y, Xph, F, T and W1 were comparatively common functional groups, Wherein, S1, D and J were the dominant functional groups. Weekly dynamics of phytoplankton functional groups were: S1-->S1-->S1-->S1-->S1--S1-->S1-->J/D/S1-->Sl1- >/1D. group Sl1dominated over other groups, the cell abundance of S1 appeared two peaks at week 5 and week 7 respectively, but there was a slump at week 8, and rose again at last, compared to two peaks before, the cell abundance had dropped from 108cells · L⁻¹ to 107cells · L⁻¹ Water flush caused by discharge gate opening artificially was the main reason. Based on the three methods of eutrophication evaluation, the water was in moderately eutrophic and eutrophic states in Xiaoguan Reservoir in the summer of 2014. Multivariate analysis (RDA) indicated transparency was the main factor affecting the distribution of phytoplankton functional groups, and nutrients were no longer the limiting factor. The study suggested that phytoplankton functional groups could make a good response to eutrophication: groups S1 and J adapted to the turbid eutrophic water bodies, D adapted to shallow turbid waters and was sensitive to nutrient depletion. Also, common functional groups like G, X1, WW1 F etc. mostly adapted to eutrophic water bodies.

[Eutrophication control in local area by physic-ecological engineering].

An integrated physical and ecological engineering experiment for ecological remediation was performed at the Maixi River bay in Baihua Reservoir Guizhou Province, China. The results show that eutrophic parameters, such as total nitrogen, total phosphorus, chlorophyll a and chemical oxygen demand from the experimental site (enclosed water) were significantly lower than those of the reference site. The largest differences between the sites were 0.61 mg x L(-1), 0.041 mg x L(-1), 23.06 microg x L(-1), 8.4 mg x L(-1) respectively; experimental site transparency was > 1.50 m which was significantly higher than that of the reference site. The eutrophic index of the experimental site was oligo-trophic and mid-trophic, while the control site was mid-trophic state and eutrophic state. Phytoplankton abundance was 2 125.5 x 10(4) cells x L(-1) in June, 2011 at the control site,but phytoplankton abundance was lower at the experimental site with 33 x 10(4) cells x L(-1). Cyanobacteria dominated phytoplankton biomass at both sites, however the experimental site consisted of a higher proportion of diatoms and dinoflagellates. After more than one year of operation, the ecological engineering technology effectively controlled the occurrence of algae blooms, changed phytoplankton community structure, and controlled the negative impacts of eutrophication. Integrating physical and ecological engineering technology could improve water quality for reservoirs on the Guizhou plateau.

[Endometrial microvessel density for assessing endometrial receptivity during the peri-implantation period].

OBJECTIVE: To evaluate the value of endometrial microvessel density (MVD) in assessing the endometrial receptivity during the peri-implantation period. METHODS: A total of 104 patients undergoing in vitro fertilization and embryo transfer (IVF-ET) treatment were analyzed retrospectively. The subjects were divided into clinical pregnancy group (50 cases) and nonpregnant group (54 cases) according to the IVF-ET outcome. Endometrial tissues were collected 7 days after the natural ovulation prior to IVF-ET for measurement of the endometrial MVD using electron microscopy, which was analyzed in relation to the clinical outcome of the treatment. RESULTS: The endometrial MVD was significantly higher in the clinical pregnancy group than in the nonpregnant group [(4.12∓1.84)% vs (3.46∓1.26)%, t=-2.127, P=0.036). ROC curve analysis showed that the MVD had an area under the curve slightly over 0.5 (0.598) for predicting clinical pregnancy, suggesting a poor specificity in predicting the clinical outcome of the treatment. CONCLUSION: In IVF-ET cycles, the endometrial MVD during the peri-implantation period is helpful for assessing the endometrial receptivity, but the specificity remains low.

[Association between cell apoptosis and the quality of early mouse embryos].

OBJECTIVE: To investigate the relationship between cell apoptosis and the quality of early mouse embryos, understand the significance of apoptosis-regulatory genes in early embryonic development, and explore a new approach to improving the embryo quality. METHODS: The levels of cell apoptosis and proliferation in early mouse embryos in different developmental status (morphologically normal embryos, arrested embryos and fragmented embryos) were analyzed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), caspase in situ fluorescence and Bcl-2 immunofluorescence, and immunofluorescent detection of proliferating cell nuclear antigen (PCNA). RESULTS: The cells in arrested embryos and embryonic fragments showed positive results in TUNEL assay with enhanced caspase activity and lowered expressions of Bcl-2 and PCNA. CONCLUSION: Cell apoptosis in early mouse embryos may be closely related to embryonic arrest and fragmentation.

[Analysis of the clinical outcomes of 160 patients undergoing repeated IVF/ICSI-ET treatment].

OBJECTIVE: To study the clinical factors affecting the outcomes of repeated assisted reproductive technology (ART) cycles. METHODS: A retrospective analysis of the clinical data and outcomes was conducted among 160 patients undergoing repeated IVF/ICSI-ET treatment between January 2006 and April 2009. RESULTS: The patients with successful clinical pregnancy after two ART cycles (group A) had a younger age and shorter duration of infertility, and had more antral follicles (AFC), more eggs and good-quality embryos with more transferred embryos available and higher good-quality embryo rate (P<0.05) than those who failed to have pregnancy after the cycles (group B). In the second cycle, the patients in group A had higher doses of short-acting GnRHa, r-HCG and HMG and at the same time more good eggs and embryos than in the first cycle. CONCLUSIONS: Female age is one of the most important factors affecting the pregnancy rate after repeated ART cycles. The clinical pregnancy rate can be enhanced by administering short-acting GnRHa, HMG, oral contraceptives and adjusting the dose of Gn as well as changing the culture medium of embryos.