Noninvasive Strategies for the Treatment of Tiny Liver Cancer: Integrating Photothermal Therapy and Multimodality Imaging EpCAM-Guided Nanoparticles.

Surgical resection and ablation therapy have been shown to achieve the purpose of a radical cure for liver cancer with a size of less than 3 cm; however, tiny liver cancer lesions of diameters smaller than 2 cm remain challenging to diagnose and cure due to the failure of the generation of new blood vessels within tumors. Emerging evidence has revealed that optical molecular imaging combined with nanoprobes can detect tiny cancer from the perspective of molecular and cellular levels and kill cancer cells by the photothermal effect of nanoparticles in real time, thereby achieving radical goals. In the present study, we designed and synthesized multicomponent and multifunctional ICG-CuS-Gd@BSA-EpCAM nanoparticles (NPs) with a potent antineoplastic effect on tiny liver cancer. Using subcutaneous and orthotopic liver cancer xenograft mouse models, we found that the components of the NPs, including ICG and CuS-Gd@BSA, showed synergistic photothermal effects on the eradication of tiny liver cancer. We also found that the ICG-CuS-Gd@BSA-EpCAM NPs exhibited triple-modal functions of fluorescence imaging, magnetic resonance imaging, and photoacoustic imaging, with targeted detection and photothermal treatment of tiny liver cancer under near-infrared light irradiation. Together, our study demonstrates that the ICG-CuS-Gd@BSA-EpCAM NPs in combination with optical imaging technique might be a potential approach for detecting and noninvasively and radically curing tiny liver cancer by the photothermal effect.

Effect of DAP and glutamine supplementation on sulfur-containing volatiles and sensory properties of Chardonnay wine fermented with Saccharomyces cerevisiae yeast.

Volatile compounds in wine have a critical impact on the consumers' senses. In this study, the effect of diammonium phosphate (DAP) and glutamine on sulfur-containing volatiles and sensory properties of Chardonnay wine fermented with Saccharomyces cerevisiae yeast were evaluated. Fermentation kinetics was determined by monitoring reducing sugar consumption rates during fermentation. The volatile profile of wines was analyzed by headspace solid phase microextraction (HS-SPME) coupled with gas-chromatography-mass spectrometry (GC-MS). The volatile sulfur compounds (VSCs) were analyzed by HS-SPME-GC-MS/MS. Flavor attributes of wines were assessed by a sensory panel with quantitative descriptive analysis. A total of 53 volatiles, including 6 VSCs, were identified and quantified in the Chardonnay wine. The results suggested that glutamine supplementation at the beginning of fermentation could help to initiate fermentation earlier and promote the formation of isoamyl acetate, phenethyl acetate, ethyl nonanoate, methyl decanoate, diethyl succinate and phenethyl alcohol, isobutanol, while DAP supplementation had no obvious effect on the volatile composition of the resulting wine and fermentation kinetics. PRACTICAL APPLICATION: Suitable nitrogen source is helpful to a healthy fermentation, and can also prevent the off-flavor and regulate aroma profile of wine. This study provides insights on the volatile and sensory characteristics of Chardonnay wines affected by different nitrogen source addition.

The impact of Traditional Chinese Medicine on mouse gut microbiota abundances and interactions based on Granger causality and pathway analysis.

Objectives: The intestinal microbiota is essential in absorbing nutrients and defending against pathogens and is associated with various diseases, including obesity, type 2 diabetes, and hypertension. As an alternative medicine, Traditional Chinese Medicine (TCM) has long been used in disease treatment and healthcare, partly because it may mediate gut microbiota. However, the specific effects of TCM on the abundance and interactions of microbiota remain unknown. Moreover, using TCM ingredients and data detailing changes in the abundance of gut microorganisms, we developed bioinformatic methods that decipher the impact of TCM on microorganism interactions. Methods: The dynamics of gut microorganisms affected by TCM treatments is explored using a mouse model, which provided the abundance of 70 microorganisms over time. The Granger causality analysis was used to measure microorganism interactions. Novel "serial connection" and "diverging connection" models were used to identify molecular mechanisms underlying the impact of TCM on gut microorganism interactions, based on microorganism proteins, TCM chemical ingredients, and KEGG reaction equations. Results: Codonopsis pilosula (Dangshen), Cassia twig (Gui Zhi), Radices saussureae (Mu Xiang), and Sijunzi Decoction did not cause an increase in the abundance of harmful microorganisms. Most TCMs decreased the abundance of Bifidobacterium pseudolongum, suggesting a Bifidobacterium pseudolongum supplement should be used during TCM treatment. The Granger causality analysis indicated that TCM treatment changes more than half the interactions between the 70 microorganisms, and "serial connection" and "diverging connection" models suggested that changes in interactions may be related to the reaction number connecting species proteins and TCM ingredients. From a species diversity perspective, a TCM decoction is better than a single herb for healthcare. The Sijunzi Decoction only significantly increased the abundance of Bifidobacterium pseudolongum and did not cause a decrease in the abundance of other species but was found to improve the alpha diversity with the lowest replacement rate. Conclusions: Because most of the nine TCMs are medicinal and edible plants, we expect the methods and results presented can be used to optimize and integrate microbiota and TCMs into healthcare processes. Moreover, as a control study, these results can be combined with future disease mouse models to link variations in species abundance with particular diseases.

Study on the Potential Mechanism of Tonifying Kidney and Removing Dampness Formula in the Treatment of Postmenopausal Dyslipidemia Based on Network Pharmacology, Molecular Docking and Experimental Evidence.

Background: Management of menopausal dyslipidemia is the main measure to reduce the incidence of cardiovascular disease in postmenopausal women. Tonifying Kidney and Removing Dampness Formula (TKRDF) is a traditional Chinese medicine (TCM) formula that ameliorates dyslipidemia in postmenopausal women. This study applied network pharmacology, molecular docking, and in vitro and in vitro experiments to investigate the underlying mechanism of TKRDF against postmenopausal dyslipidemia. Methods: Network pharmacology research was first conducted, and the active compounds and targets of TKRDF, as well as the targets of postmenopausal dyslipidemia, were extracted from public databases. Protein-protein interaction (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to identify the potential targets and signaling pathways of TKRDF in postmenopausal dyslipidemia. Molecular docking was then performed to evaluate the combination of active compounds with principal targets. Finally, an ovariectomized rat model was used for the in vivo experiment and alpha mouse liver 12 (AML12) cells treated with palmitic acid were used for the in vitro experiments to provide further evidence for the research. Results: Based on network pharmacology analysis, we obtained 78 active compounds from TKRDF that acted on 222 targets of postmenopausal dyslipidemia. The analysis results indicated that IL6, TNF, VEGFA, AKT1, MAPK3, MAPK1, PPARG and PIK3CA, etc., were the potentially key targets, and the PI3K/AKT signaling pathway was the possibly crucial pathway for TKRDF to treat postmenopausal dyslipidemia. Molecular docking suggested that the active compounds have good binding activity with the core targets. The in vivo and in vitro experiments demonstrated that TKRDF ameliorates postmenopausal dyslipidemia by regulating hormone levels, inhibiting inflammation, promoting angiogenesis and inhibiting lipid synthesis, which appear to be related to TKRDF's regulation of the ERK1/2 and PI3K/AKT signaling pathways. Conclusion: This study clarified the active ingredients, potential targets, and molecular mechanisms of TKRDF for treating postmenopausal dyslipidemia. It also provided a feasible method to uncover the scientific basis and therapeutic mechanism for prescribing TCM in the treatment of diseases.

Effects of Angelica Fritillaria Kushen Pill on Renal Function and Immune Function after Laparoscopic Radical Nephrectomy for Patients with Renal Carcinoma.

Objective: To investigate the effect of Danggui Beimu Kushen Pills on renal cell carcinoma patients undergoing laparoscopic radical resection and their effects on renal function and immune function. Methods: 106 patients with renal cell carcinoma who underwent retroperitoneal laparoscopic radical surgery in our hospital from March 2015 to February 2017 were selected, and they were divided into control group and treatment group. The control group was treated with interferon and the treatment group was treated with Danggui Beimu Kushen pills on the basis of the control group. The clinical efficacy, incidence of adverse reactions, survival rate, and the relationship with clinical stages were detected in the two groups. The renal function index levels, immune function index levels, and quality of life levels were measured in the two groups after treatment. Results: Compared with the control group, the total clinical effective rate was increased, the abnormal renal function, abnormal liver function, and platelet decline were decreased, and the survival rate was significantly elevated in the treatment group. The patients with clinical stage I∼II' survival rate was increased, relative to patients with clinical stage III. After treatment, the levels of ß2-MG, SCr, BUN, and CD8+ in the two groups were decreased, while the levels of CD3+, CD4+, and CD4+/CD8+ were increased. Moreover, the scores of physical function, social function, emotional function, role function, cognitive function, and total score of the two groups were improved. Compared with the control group, the treatment group changes were more obvious. Conclusion: The Danggui Beimu Kushen Pill has significant clinical effects on the treatment of renal cancer, which can improve the clinical symptoms, enhance the immune function, restore the health renal function, improve the quality of life, prolong the survival time of patients, reduce the incidence of adverse reactions, and provide high safety.

Protective effect of Huyang Yangkun Formula on ovarian function in premature ovarian insufficiency rats based on apoptotic mechanism.

ETHNOPHARMACOLOGICAL RELEVANCE: Huyang Yangkun Formula(HYF) is a traditional Chinese medicine formula based on the traditional theory of Yin and Yang. It could consolidate the Qi of Yin and Yang, adjust the balance of Qi and blood. It has shown clinical efficacy for patients with Premature Ovarian Insufficiency(POI). AIM OF THE STUDY: Aim to access the effect of Huyang Yangkun formula (HYF) on premature ovarian insufficiency rat model and explores the mechanism related to aquaporins(AQPs) and apoptosis. MATERIALS AND METHODS: Female SD rats were injected with 4-vinylcyclonhexenediepoxide(VCD, 160 mg/kg/day) for 15 days. Then, HYF (0.297 g/kg)/estradiol valerate (0.1 mg/kg) was administered for 105 days in the HYF/estradiol valerate treatment(EVT) group. Serum AMH, FSH and E2 were detected by ELISA, and the developing follicles were counted in each group.The TUNEL assay was used to detect positive apoptotic signals. IHC and western blots were used to verify differentially expressed AQPs and apoptosis-related regulators potentially associated with HYF. RESULTS: Total follicles were increased significantly in the HYF group. Serum AMH was increased in the HYF group compared with MOD group. Serum FSH and E2 showed no obvious difference between HYF group and MOD group. Apoptosis occurred in POI model was proved by TUNEL and Caspase3/9 and HYF could rescue this apoptosis, besides the anti-apoptotic effect may be better than EVT. Distribution of AQPs in rat ovaries based on developmental stages of follicle was observed, and AQP8 was obviously expressed in the developing follicles and corpus luteum, particularly in granulosa cells. Upregulation of AQP8 in the MOD group and downregulation by HYF were observed. BCL-XL was significantly upregulated in the HYF group and EVT group; BCL-2 was upregulated in the HYF group with no statistical difference; MCL-1 was downregulated in the HYF group. BAX and BIM were significantly upregulated in the MOD group. The ratio of BCL-2/BAX and BCL-XL/BAX were decreased in the MOD group, and BCL-XL/BAX was increased in the HYF group and EVT group. CONCLUSION: This study evaluated the treatment effect of HYF on POI in rats. It showed that HYF could promote the follicles development by regulating AQP8/Bcl-2 family-related mitochondrial apoptosis, which provides basic evidence for TCM as an alternative therapy for POI.

Licorice (Glycyrrhiza spp.) and jujube (Ziziphus jujuba Mill.) formula for menopausal symptoms: Classical records, clinical evidence and experimental data.

OBJECTIVES: This study sought to determine the most common oral herbal formula for menopausal symptoms in classical Chinese medicine textbooks and investigate its clinical effectiveness and potential mechanisms of action. METHODS: The most common formula used for menopause-like symptoms in past eras was identified from the Encyclopedia of Traditional Chinese Medicine. A systematic review of randomized controlled trials (RCTs) was undertaken and findings from relevant experimental studies were summarized. RESULTS: Licorice (Glycyrrhiza spp.) and jujube (Ziziphus jujuba Mill.) formula (LJF) was used in 63 of the 175 citations (36%) in the classical literature. Evidence from four RCTs showed that while LJF may improve sleep symptoms, there is insufficient evidence to provide recommendations for clinical practice. Experimental studies showed sedative, antidepressant-like, estrogenic and antiprogestogenic actions. CONCLUSIONS: LJF has a long history of use for menopause-like symptoms, but further research is needed to confirm its clinical effects and guide clinical decision-making.

Danggui Buxue Tang Rescues Folliculogenesis and Ovarian Cell Apoptosis in Rats with Premature Ovarian Insufficiency.

Premature ovarian insufficiency (POI) is a common female endocrine disease that is closely linked to ovarian function. Danggui Buxue Tang (DBT) is a classic prescription of traditional Chinese medicine that is helpful for rescuing ovarian function. However, the mechanism by which DBT rescues ovarian function remains unclear. This study explored the molecular mechanism of DBT with respect to apoptosis and related signals in ovarian cells. The quality control of DBT was performed by HPLC. After DBT intervention in the POI rat model, serum AMH/FSH/LH/E2 levels were detected by ELISA, follicles at various developmental stages were observed by HE staining, apoptosis was detected by TUNEL, and the expression profiles of Bcl-2 family proteins and key proteins in the Jak2/Foxo3a signaling pathway were evaluated by western blot. The results demonstrated that DBT could encourage the development of primary/secondary/antral follicles and increase the secretion of AMH. Moreover, DBT might inhibit Foxo3a by upregulating Jak2, thereby mediating Bcl-2 family activities and inhibiting apoptosis in ovarian cells. In conclusion, DBT promotes follicular development and rescues ovarian function by regulating Bcl-2 family proteins to inhibit cell apoptosis, which could be related to the Jak2/Foxo3a signaling pathway.

Manganese dioxide driven the carbon and nitrogen transformation by activating the complementary effects of core bacteria in composting.

This study revealed core bacterial metabolic mechanisms involved in carbon (C) and nitrogen (N) in composting with adding MnO2. Two tests (control group (CK), adding MnO2 (M)) were performed. The results indicated that the MnO2 accelerated the transformation of carbon and nitrogen in composting. Core bacteria involved in the C and N conversion were identified, the complementarity effects of core bacteria were stimulated in M composting. Additionally, the influence of core bacteria on the C and N conversion could be divided into two pathways in M composting. One was that core bacteria promoted C and N conversion by accelerating the flow of amino acids into the tricarboxylic acid cycle. Another was that the complementarity effects of core bacteria increased the overall bacterial diversity, which contributed to C and N conversion. These findings showed that the addition of MnO2 to composting was a promising application to treat agricultural organic waste.

New Insights into the Mechanisms of Polyphenol from Plum Fruit Inducing Apoptosis in Human Lung Cancer A549 Cells Via PI3K/AKT/FOXO1 Pathway.

Recent studies have been found that polyphenols from plums fruits can inhibit the proliferation of multiple cancer cells, while the molecular mechanism was unclear. This study aimed to investigate the molecular mechanism underlying the pro-apoptotic effect of purified plum polyphenols (PPP) on human lung cancer A549 cells. Quercitrin (quercetin-3-O-glucoside, 814.19 ± 40.71 mg/g) was identified as the primary polyphenol in PPP via ultra high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS). PPP showed a strong capacity for inhibiting the proliferation of the A549 cells by inducing apoptosis, which was reflected by an increase in the Bax/Bcl-2 ratio. Additionally, the inhibitory rate of PPP on the A549 cells were higher than that of vitamin C when the treatment dose exceeded 160 µg/mL. Transcriptome analysis suggested that PPP-induced apoptosis was closely associated with regulating the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/forkhead box protein O 1 (FOXO1) pathway in the A549 cells. Subsequently, as an activator of AKT, SC79 was applied to confirm that the inhibition of AKT phosphorylation play an important role in the PPP-induced apoptosis of the A549 cells. These results illustrated the potential of PPP as a dietary compound for the prevention of cancer or for use during chemotherapy.

Synergistic effect of Huyang Yangkun Formula and embryonic stem cells on 4-vinylcyclohexene diepoxide induced premature ovarian insufficiency in mice.

BACKGROUND: Huyang Yangkun Formula (HYYKF) was developed based on theory of traditional Chinese medicine as well as clinical experience and used to improve ovarian function of premature ovarian insufficiency (POI) patients. Transplantation of embryonic stem cells (ESCs) has great potential in improving POI, and studies have confirmed that traditional Chinese medicine promoted the treatment effect of ESCs. In the present study, we compared the effect of combining HYYKF and ESCs, single HYYKF treatment and single ESCs intervention on POI mice to explore the effect of combination of HYYKF and ESCs in improving ovarian function. METHODS: C57BL/6 mice were used to create a POI model by 15-day intraperitoneal injection of 160 mg/kg of 4-vinylcyclonhexene diepoxide (VCD) and then treated with HYYKF, ESCs transplantation and combination of ESCs and HYYKF. When the treatments were finished, estrus cycle, ovarian follicle counting, serum sex hormone level, and expression of key nodes in the transforming growth factor beta/transforming growth factor beta-activated kinase 1 (TGF-ß/TAK1) signaling pathway were determined. RESULTS: Combination therapy brought down the abnormal estrus cycle rate to 5.26%, significantly lower than that of HYYKF or ESCs alone (30%, 25%, respectively). The numbers of follicles at all levels were increased significantly in the combination ESCs with HYYKF group (P < 0.05), especially that of antral follicles (P < 0.01), which was not increased significantly when HYYKF or ESCs was single used. The level of anti-Mullerian hormone (AMH) was more significantly increased in the combination ESCs with HYYKF group (P < 0.01) than that of HYYKF or ESCs alone (both P < 0.05). The expression of the key nodes TGF-ß1, TAK1, JNK, Smad4 and FSHR in the TGF-ß/TAK1 pathway were obviously affected in the SCHY group. CONCLUSION: Both HYYKF and ESCs improve the ovarian function of POI induced by VCD, and a combination of HYYKF and ESCs has the advantage that they work together to promote follicles developing probably by inhibiting expression of the TGF-ß1/TAK1 pathway.

Nonhormonal Hot Flash Management for Breast Cancer Survivors: A Systematic Review and Network Meta-Analysis.

MATERIALS AND METHODS: We conducted a systematic literature search in PubMed, Cochrane Central Register of Controlled Trials, Embase, Chinese Biomedicine Database (CBM), China National Knowledge Infrastructure (CNKI), Wan Fang, and VIP up to May 2018. Randomized controlled trials (RCTs) reporting nonhormonal hot flash treatments for breast cancer survivors were included. Primary outcome measurements were hot flash frequency and hot flash score of posttreatment. The methodological quality of each study was assessed with Cochrane's risk of bias tool. RESULTS: 16 RCTs involving 2,349 participants were included. The nonhormonal therapies used in the included studies were classified as follows: lifestyle changes, mind-body techniques, dietary/supplements, SSRIs/SNRIs, other medications, and other therapies. Pairwise meta-analysis showed that the general effect of nonhormonal management was statistically more effective than no treatment/placebo/sham in reducing hot flash frequency (SMD = -0.60, 95% CI [-1.13, -0.06]; P=0.03)) and hot flash score (SMD = -0.38, 95% CI [-0.68, -0.08]). For hot flash frequency, results from the NMA showed that there was no statistically significant difference between any two of the nonhormonal treatments. Another NMA result indicated that acupuncture (other therapies) was 16.05 points more effective in reducing hot flash scores than no treatment/waitlist (SMD = -16.05, 95% CI [-30.2, -1.99]). These results were statistically significant. Acupuncture was also ranked the optimal nonhormonal therapy for both hot flash frequency and hot flash score. The safety analysis showed that there were few related adverse events during acupuncture and that drug related adverse reactions could have also occurred in studies using drug interventions. CONCLUSIONS: This network meta-analysis comparing nonhormonal treatments suggested that acupuncture might be more effective in improving hot flashes for breast cancer survivors. A pronounced placebo response was found during hot flash treatments. The evidence of safety for nonhormonal therapies was also insufficient. Therefore, at present, we cannot make confirmative recommendations of nonhormonal hot flash management for breast cancer survivors. This study is registered with PROSPERO (CRD42018082008).

Effect of Chinese herbal medicine on serum lipids in postmenopausal women with mild dyslipidemia: a randomized, placebo-controlled clinical trial.

OBJECTIVE: Previous studies have shown the association between menopause and dyslipidemia. This study was aimed to evaluate the effect of Chinese herbal medicine, tonifying kidney and descending turbidity (TKDT) granule, on serum lipid profiles in postmenopausal women with dyslipidemia. METHODS: A double-blind randomized, placebo-controlled trial was conducted among 104 postmenopausal Chinese women with mild dyslipidemia. Participants were randomized into treatment group (n = 53) and control group (n = 51). The participants in the treatment group received TKDT granule once per day for 24 weeks, whereas the control group received placebo in the same manner. All participants were subjected to healthy lifestyle during the study. Serum lipid profiles, body weight, waist circumference, and safety indicators were measured both at baseline and 24 weeks after admission. RESULTS: Compared with placebo, significant improvements in total cholesterol, low-density lipoprotein cholesterol, apolipoprotein-B (Apo-B), weight, and waist circumference in the TKDT group (P < 0.05) were observed after 24 weeks of treatment. Total cholesterol, low-density lipoprotein cholesterol, and Apo-B were decreased by 0.84 (0.77) mmol/L, 0.72 (0.77) mmol/L, and 0.12 (0.27) mmol/L, mean ±â€ŠSD respectively. The weight, waist circumference, and body mass index were decreased by 4.07 (3.01) kg, 3.10 (2.95) cm, 1.60 (1.13), respectively. There were no significant differences in triglycerides, high-density lipoprotein cholesterol, and Apo-A between the two groups. Seven participants in the treatment group and six participants in the placebo group had mild or moderate adverse reactions. CONCLUSION: TKDT granule improved the lipid profile and reduced the related metabolic abnormalities in postmenopausal women with mild dyslipidemia based on lifestyle changes.

α-Linolenic acid but not linolenic acid protects against hypertension: critical role of SIRT3 and autophagic flux.

Although dietary α-linolenic acid (ALA) or linolenic acid (LA) intake was reported to be epidemiologically associated with a lower prevalence of hypertension, recent clinical trials have yielded conflicting results. Comparable experimental evidence for the roles of these two different fatty acids is still lacking and the underlying mechanisms need to be further elucidated. Our data showed that ALA but not LA supplementation alleviated systolic blood pressure elevation and improved ACh-induced, endothelium-dependent vasodilation in both spontaneously hypertensive rats (SHRs) and AngII-induced hypertensive mice. In addition, SHRs displayed reduced vascular Sirtuin 3 (SIRT3) expression, subsequent superoxide dismutase 2 (SOD2) hyperacetylation and mitochondrial ROS overproduction, all of which were ameliorated by ALA but not LA supplementation. In primary cultured endothelial cells, ALA treatment directly inhibited SIRT3 reduction, SOD2 hyperacetylation, mitochondrial ROS overproduction and alleviated autophagic flux impairment induced by AngII plus TNFα treatment. However, these beneficial effects of ALA were completely blocked by silencing SIRT3. Restoration of autophagic flux by rapamycin also inhibited mitochondrial ROS overproduction in endothelial cells exposed to AngII plus TNFα. More interestingly, SIRT3 KO mice developed severe hypertension in response to a low dose of AngII infusion, while ALA supplementation lost its anti-hypertensive and endothelium-protective effects on these mice. Our findings suggest that ALA but not LA supplementation improves endothelial dysfunction and diminishes experimental hypertension by rescuing SIRT3 impairment to restore autophagic flux and mitochondrial redox balance in endothelial cells.

Identifying a wild allele conferring small seed size, high protein content and low oil content using chromosome segment substitution lines in soybean.

KEY MESSAGE: A wild soybean allele conferring 100-seed weight, protein content and oil content simultaneously was fine-mapped to a 329-kb region on Chromosome 15, in which Glyma.15g049200 was predicted a candidate gene. Annual wild soybean characterized with small 100-seed weight (100SW), high protein content (PRC), low oil content (OIC) may contain favourable alleles for broadening the genetic base of cultivated soybeans. To evaluate these alleles, a population composed of 195 chromosome segment substitution lines (SojaCSSLP4), with wild N24852 as donor and cultivated NN1138-2 as recurrent parent, was tested. In SojaCSSLP4, 10, 9 and 8 wild segments/QTL were detected for 100SW, PRC and OIC, respectively. Using a backcross-derived secondary population, one segment for the three traits (q100SW15, qPro15 and qOil15) and one for 100SW (q100SW18.2) were fine-mapped into a 329-kb region on chromosome 15 and a 286-kb region on chromosome 18, respectively. Integrated with the transcription data in SoyBase, 42 genes were predicted in the 329-kb region where Glyma.15g049200 showed significant expression differences at all seed development stages. Furthermore, the Glyma.15g049200 segments of the two parents were sequenced and compared, which showed two base insertions in CDS (coding sequence) in the wild N24852 comparing to the NN1138-2. Since only Glyma.15g049200 performed differential CDS between the two parents but related to the three traits, Glyma.15g049200 was predicted a pleiotropic candidate gene for 100SW, PRC and OIC. The functional annotation of Glyma.15g049200 indicated a bidirectional sucrose transporter belonging to MtN3/saliva family which might be the reason that this gene provides a same biochemical basis for 100SW, PRC and OIC, therefore, is responsible for the three traits. This result may facilitate isolation of the specific gene and provide prerequisite for understanding the other two pleiotropic QTL.

Huyang yangkun formula protects against 4-Vinylcyclohexene diepoxide-induced premature ovarian insufficiency in rats via the Hippo-JAK2/STAT3 signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Huyang Yangkun Formula (HYF) has been prescribed for premature ovarian insufficiency (POI) for decades in the clinical setting. Little is known regarding its underlying molecular mechanism. This study was conducted to elucidate the possible mechanism of the protective potential of HYF against POI induced by the industrial chemical 4-vinylcyclohexene diepoxide (VCD) in rats. AIM OF THE STUDY: Quality control of HYF was conducted via HPLC and UPLC-MS. Female rats were injected with VCD (160 mg/kg) daily for 15 days. Then, 1.35 g/kg (low dose) or 0.235 g/kg (high dose) HYF was administered once/day for 25 days. Serum AMH, FSH, E2, ALT, AST, BUN and Cr levels were detected through ELISA and HE-stained follicles were counted in ovarian sections. Additionally, RNA-seq profiling analysis and functional assays were used to screen for differentially expressed genes and key regulators with potentially important roles associated with HYF. RESULTS: The ovaries of POI rats contained fewer antral and maturing follicles (p < 0.05) than those of control rats, whereas atretic follicles were increased significantly (p < 0.05), and AMH levels were significantly lower in the VCD group than in the control group (p < 0.05). These conditions showed some improvement after low- and high-dose HYF treatment. Low- and high-dose HYF increased AMH levels by 42.4% and 25.9% and decreased FSH levels by 17.5% and 24.1%, respectively, in comparison to the VCD group. The two HYF dosage groups showed significantly increased numbers of antral and maturing follicles but a reduced number of atretic follicles (p < 0.05). HYF down-regulation of JAK, Lats2 and YAP mRNA expression gene expression (p < 0.05) compared with the VCD group. HYF resulted in a strongly attenuated VCD-induced phosphorylation of JAK2 and STAT3 (p < 0.01) and YAP (p < 0.001), but induced an increase in protein levels of LATS2 (p < 0.05). CONCLUSION: Our findings demonstrated the treatment efficacy of HYF in POI rats and showed that HYF repairs the dysfunction and enhances the ovarian function of POI rats through the Hippo-JAK2/STAT3 signaling pathway.

Polyphenol-Rich Loquat Fruit Extract Prevents Fructose-Induced Nonalcoholic Fatty Liver Disease by Modulating Glycometabolism, Lipometabolism, Oxidative Stress, Inflammation, Intestinal Barrier, and Gut Microbiota in Mice.

Fructose as a daily sweetener is widely recognized as a risk catalyst for nonalcoholic fatty liver disease (NAFLD). The aim of current study is to evaluate the effects and molecular mechanism by which polyphenol-rich loquat fruit extract (LFP) prevents NAFLD in mice fed 30% fructose water (HF) for 8 weeks. Administration of LFP to HF-fed mice mitigated abnormal body weight, disordered lipid metabolism, oxidative stress, and inflammation through a mechanism regulated by the AKT, ChREBP/SREBP-1c, Nrf2, and TLR4/MyD88/TRIF pathways. LFP caused a significant decrease in the endotoxin content (16.67-12.7 EU/mL) in the liver of HF-fed mice. LFP not only improved HF-induced breakage of the intestinal barrier via interacting with tight junction proteins (ZO-1, occludin), mucin, and immunoreaction in the colon but also maintained normal colonic Firmicutes/Bacteroidetes ratios and the relative abundance of Veillonella in HF-fed mice. Our results suggest that LFP may serve as a nutritional agent for protecting liver in HF-fed mice.

Pectin Alleviates High Fat (Lard) Diet-Induced Nonalcoholic Fatty Liver Disease in Mice: Possible Role of Short-Chain Fatty Acids and Gut Microbiota Regulated by Pectin.

Consumption of pectin contributes to changes in the gut microbiota and the metabolism of short-chain fatty acids (SCFAs). We aimed to investigate the effects of and mechanism by which pectin prevented nonalcoholic fatty liver disease (NAFLD) in mice that were fed a high-fat diet containing 30% lard (HF). HF-fed mice that orally ingested pectin for 8 weeks exhibited improvements in lipid metabolism and decreased oxidative stress and inflammation through a mechanism regulated by the mitogen-activated protein kinase pathway. Pectin dose-dependently generated an increase in acetic acid (from 566.4 ± 26.6 to 694.6 ± 35.9 µmol/mL, p < 0.05) and propionic acid (from 474.1 ± 84.3 to 887.0 ± 184.7 µmol/mL, p < 0.05) contents and significantly increased the relative abundance of Bacteroides (from 0.27% to 11.6%), Parabacteroides (from 3.9‰ to 5.3%), Olsenella (from 2.9‰ to 1.3%), and Bifidobacterium (from 0.03% to 1.9%) in the gut of HF-fed mice. Intestinal microbiota and SCFAs may thus contribute to the well-established link between pectin consumption and NAFLD.

Diversity of endophytic fungi from roots of Panax ginseng and their saponin yield capacities.

Endophytes of medicinal plants have the capacity to synthesis same or similar active substances with their hosts. To investigate the diversity and capacity to produce saponins of endophytic fungi of Panax ginseng, thirty-eight strains of were isolated. Polymerase chain reaction (PCR) and sequencing were used to identify the isolates, and saponins concentrations in the cultures were measured. Agar diffusion method was used to test antimicrobial activity. High-performance liquid chromatography (HPLC) was used to analyze ginsenosides produced by representative strains. Nectria, Aspergillus, Fusarium, Verticillium, Engyodontium, Plectosphaerella, Penicillium, Cladosporium, and Ascomycete species were isolated. Overall, 18.4% of the isolates belonged to Nectria (Nectria haematococca), 13.2% belonged to Aspergillus, and 10.5% belonged to Penicillium. The highest concentration of triterpenoid saponin was 0.181 mg/ml (Pg27), followed by 0.144 mg/ml (Pg30 and Pg42-1). According to the results of the phylogenetic results, these isolates were species of Fusarium, Aspergillus and Verticillium, respectively. The culture filtrate of Pg30 exhibited its antibacterial activity Staphylococcus aureus. Pg 27 and Pg30 could excrete the substances to inhibit the growth of Rhizoctonia solani. Pg42-1 showed strong inhibition against Klebsiella pneumoniae. From HPLC results, the ginsenoside Rb2 was detected in both Pg27 and Pg30 cultures. The ginsenoside Rc was found in Pg42-1 cultures. In conclusion, thirty-eight endophytic fungal strains were isolated and Pg27 (Fusarium sp.) has a potential application value in saponins production.

L-3-n-butylphthalide reduces tau phosphorylation and improves cognitive deficits in AßPP/PS1-Alzheimer's transgenic mice.

L-3-n-butylphthalide (L-NBP), an extract from seeds of Apium graveolens Linn (Chinese celery), has been shown to have neuroprotective effects on cerebral ischemic, vascular dementia and amyloid-ß (Aß)-induced animal models by inhibiting oxidative injury, neuronal apoptosis and glial activation, regulating amyloid-ß protein precursor (AßPP) processing and reducing Aß generation. The aim of the present study was to examine the effect of L-NBP on learning and memory in AßPP and presenilin 1 (PS1) double-transgenic AD mouse model (AßPP/PS1) and the mechanisms of L-NBP in reducing Aß accumulation and tau phosphorylation. Twelve-month old AßPP/PS1 mice were given 15 mg/kg L-NBP by oral gavage for 3 months. L-NBP treatment significantly improved the spatial learning and memory deficits compared to the vehicle-treated AßPP/PS1 mice, whereas L-NBP treatment had no effect on cerebral Aß plaque deposition and Aß levels in brain homogenates. However, we found an L-NBP-induced reduction of tau hyperphosphorylation at Ser199, Thr205, Ser396, and Ser404 sites in AßPP/PS1 mice. Additionally, the expressions of cyclin-dependent kinase and glycogen synthase kinase 3ß, the most important kinases involved in tau phosphorylation, were markedly decreased by L-NBP treatment. The effects of L-NBP on decreasing tau phosphorylation and kinases activations were further confirmed in neuroblastoma SK-N-SH cells overexpressing wild-type human AßPP695 (SK-N-SH AßPPwt). L-NBP shows promising candidate of multi-target neuronal protective agent for the treatment of Alzheimer's disease.