Possible molecular exploration of herbal pair Haizao-Kunbu in the treatment of Graves' disease by network pharmacology, molecular docking, and molecular dynamic analysis.

Objective: To promote the development and therapeutic application of new medications, it is crucial to conduct a thorough investigation into the mechanism by which the traditional Chinese herb pair of Haizao-Kunbu (HK) treats Graves' disease (GD). Materials and methods: Chemical ingredients of HK, putative target genes, and GD-associated genes were retrieved from online public databases. Using Cytoscape 3.9.1, a compound-gene target network was established to explore the association between prosperous ingredients and targets. STRING, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes pathway analyses visualized core targets and disease pathways. Additionally, we conducted a refined analysis of the binding interactions between active ingredients and their respective targets. To visualize these findings, we employed precise molecular docking techniques. Furthermore, we carried out molecular dynamics simulations to gain insights into the formation of more tightly bound complexes. Results: We found that there were nine key active ingredients in HK, which mainly acted on 21 targets. These targets primarily regulated several biological processes such as cell population proliferation, protein phosphorylation, and regulation of kinase activity, and acted on PI3K-AKT and MAPK pathways to treat GD. Analysis of the molecular interaction simulation under computer technology revealed that the key targets exhibited strong binding activity to active ingredients, and Fucosterol-AKT1 and Isofucosterol-AKT1 complexes were highly stable in humans. Conclusion: This study demonstrates that HK exerts therapeutic effects on GD in a multi-component, multi-target, and multi-pathway manner by regulating cell proliferation, differentiation, inflammation, and immunomodulatory-related targets. This study provides a theoretical foundation for further investigation into GD.

Effect of ultrasonic degradation on the physicochemical property and bioactivity of polysaccharide produced by Chaetomium globosum CGMCC 6882.

Similar to the enzymatic process, there might also be an active fragment in polysaccharides, how to obtain is important for investigating the bioactivity and pharmacological mechanism of polysaccharides. Presently, a Gynostemma pentaphyllum endophytic fungus Chaetomium globosum CGMCC 6882 polysaccharide [Genistein Combined Polysaccharide (GCP)] was degraded by ultrasonic treatment, two polysaccharide fragments of GCP-F1 and GCP-F2 were obtained. Physicochemical results showed that GCP-F1 and GCP-F2 had the same monosaccharide composition of arabinose, galactose, glucose, xylose, mannose, and glucuronic acid as compared to GCP with slightly different molar ratios. However, weight-average molecular weights of GCP-F1 and GCP-F2 decreased from 8.093 × 104 Da (GCP) to 3.158 × 104 Da and 1.027 × 104 Da, respectively. In vitro scavenging assays illustrated that GCP-F1 and GCP-F2 had higher antioxidant activity against 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, superoxide anions, and hydroxyl radical than GCP, the order was GCP < GCP-F1 < GCP-F2. Meanwhile, antibacterial tests showed that ultrasonic degradation increased the antibacterial activity of GCP-F1 as compared to GCP, but GCP-F2 almost lost its antibacterial activity with further ultrasound treatment. Changes in the antioxidant and antibacterial activities of GCP-F1 and GCP-F2 might be related to the variation of their molecular weights.

Effect of Polysaccharide Extracted From Gynostemma Pentaphyllum on the Body Weight and Gut Microbiota of Mice.

Researchers have investigated the role of polysaccharides in disease treatment via gut microbiota regulation but ignore their function in disease prevention and physique enhancement. In this work, a Gynostemma pentaphyllum polysaccharide (GPP) was tested by methyl thiazolyl tetrazolium (MTT) assay and proved to be safe to Caco-2 cells. Animal experiments showed that the administration of GPP for 3 weeks decreased the body weight gain of mice from 15.4 ± 1.7 to 12.2 ± 1.8 g in a concentration-dependent manner. Analysis of short-chain fatty acids (SCFAs) indicated that GPP increased the levels of acetate, propionate, butyrate, and total SCFAs in the cecum contents of normal mice. Furthermore, GPP improved the species richness and abundance in the gut microbiota but reduced the Firmicutes/Bacteroidetes ratio from 0.8021 to 0.3873. This work provides a basis for incorporating GPP into diet to prevent or mitigate the occurrence of obesity via gut microbiota regulation.

Phosphoproteomic analysis of FAC overload-triggered human hepatic cells reveals G2/M phase arrest.

Hepatic iron overload is a universal phenomenon in patients with myelodysplastic syndromes (MDS) who undergo bone marrow transplantation and may experience the toxicity of peri- and post-bone marrow transplantation. To clarify the mechanisms of iron overload-triggered liver injury, we determined the effects of iron overload on changes in protein phosphorylation in human hepatocyte cell line HH4 in vitro by using a phosphoproteomics approach. The hepatocytes were exposed to high concentrations of ferric ammonium citrate (FAC) to build up an iron overload model in vitro. Changes in protein phosphorylation initiated by iron overloading were studied by 2D-LC/MS. We identified 335 differentially expressed phosphorylated proteins under the condition of excess hepatocyte iron, 11% of which were related to cell cycle progression. The results of phosphoproteomics showed that iron overload induced 10.9 times increase in Thr 14/Tyr 15-phosphorylated Cdk1 in HH4 cells. Flow cytometry analysis revealed that FAC-treated HH4 cells showed significant G2/M phase arrest. Our subsequent RT-PCR and Western blot experiments indicated that FAC-induced G2/M phase arrest was related to the activation of p53-p21-Cdk1, p53-14-3-3 sigma-Cdk1, and 14-3-3 gamma pathway. Our findings demonstrate the first evidence that iron overload causes G2/M arrest in HH4 hepatocytes.

Positive Psychology 2.0 in a Foreign Language Classroom: Students' Emotional Experience in English Classroom Interaction in China.

The second wave of positive psychology (PP 2.0) focuses on the way positive and negative psychology complement each other in social contexts. It offers a balanced interactive model that aims at enhancing the optimal learning outcome through the interplay of positive and negative emotions. Building on a large qualitative study of students' and teachers' experiences in EFL classrooms in China, this paper argues that adopting the principles of PP 2.0 could deepen our understanding of learners' emotional experience in SLA. Using one illustrative case, it shows the dynamic and complexity of students' shifting emotions as they interact in the classroom over a span of 2 months. One major finding is that the students' positive emotions could transcend negative emotions and influence their engagement in classroom interaction. This study contributes to the existing research into emotional experiences of classroom interaction that integrates the observable, reflective, and participatory. It draws on interrelated sets of data, including a student and teacher profile questionnaire, classroom observation and recording, student and teacher reflective journals documenting their classroom interaction experiences, and stimulated recall interviews based on recordings and reflective journals. The study in the first place has implications for English teachers and teacher trainers in China and abroad as well as researchers interested in the role of emotional experience in English language learning and teaching.

First Report of Anthracnose on Bletilla striata Caused by Colletotrichum fructicola in China.

Bletilla striata (Thunb.) Rchb. f. (Orchidaceae) is a traditional Chinese medicinal plant. In April 2018 and 2019, a leaf spot disease was observed on ∼20% of B. striata plants in two fields (∼1.4 h) in Guilin, Guangxi Province, China (Fig.1 A). Small, circular, brown spots were initially observed on the leaf surfaces, which progressively expanded into large, sunken, dark brown, necrotic areas. As the disease progressed, lesions merged into large, irregular spots, ultimately resulting in abscission. To determine the causal agent, small pieces (5 mm x 5 mm) were collected from the infected leaf tissues (n = 18), surface sterilized in 1% NaOCl for 2 min, and rinsed three times with sterile water. Then, the tissues were placed on potato dextrose agar (PDA) with chloramphenicol (0.1 g/L) and incubated under 12 h photoperiod at 26°C for 3 days. Seventeen isolates were obtained, of which twelve isolates with similar morphological characteristics were obtained from the germinated spores on PDA. Seven-day-old colonies on PDA appeared cottony, pale white to pale gray from above, and grayish-green from below. Conidia of strain BJ-101.3 were hyaline, aseptate, straight, and cylindrical, with rounded ends (Fig.1 E-G), measuring 11.3 to 15.9 µm × 4.0 to 6.4 µm (n = 50). Appressoria were brown to dark brown, with different shapes and a smooth edge (Fig.1 H-I), measuring 6.3 to 10.0 µm × 4.1 to 8.0 µm (n = 50). Morphological features were similar to C. gloeosporioides species complex (Weir et al. 2012, Fuentes-Aragón et al. 2018). For molecular identification, DNA was extracted from two isolates BJ-101.3 and BJ-101.13, following the CTAB method (Guo et al. 2000). The internal transcribed spacer (ITS) region, partial actin (ACT), calmodulin (CAL), chitin synthase (CHS-1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), manganese superoxide dismutase (SOD2), beta-tubulin (TUB2), glutamine synthetase (GS), and Apn2-Mat1-2 intergenic spacer and partial mating-type (ApMat) genes were amplified by PCR and sequenced (Weir et al. 2012, Silva et al. 2012, Vieira et al. 2017). The obtained sequences were deposited in GenBank (MW386818, MW386819, MW403508 to MW403519, and MW888410 to MW888413). BLASTN analysis of the obtained sequences showed 99% identity with those of C. fructicola (JX010165，JX010033, FJ917508, FJ907426, JX009866, JX010095, JX010327, JX010405, JQ807838) (Weir et al. 2012, Liu et al. 2015). A phylogenetic tree based on the concatenated sequences confirmed the isolates as C. fructicola (Fig.2). Furthermore, pathogenicity tests were conducted on six 1.5-year-old B. striata plants. Healthy leaves on the plants were inoculated with the conidial suspensions (106 conidia/mL; 10 µL) of the strains BJ-101.3 and BJ101.13. The conidial suspension of each isolate was inoculated onto at least three leaves. Another three plants inoculated with sterile water served as the control. All plants were covered with transparent plastic bags and incubated in a greenhouse at 26°C for 14 days with a 12 h photoperiod. Nine days post-inoculation, the inoculated leaves showed leaf spot symptoms, while the control plants remained symptomless (Fig.1 B-C). The experiments repeated three times showed similar results. Finally, C. fructicola was consistently reisolated from the infected leaves and confirmed by morphology and sequencing, fulfilling Koch's postulates. The outcome of this study will help in developing effective management measures against anthracnose of B. striata.

SNAI1 interacts with HDAC1 to control TGF­ß2­induced epithelial­mesenchymal transition in human lens epithelial cells.

The opacity of the lens capsule after cataract surgery is caused by epithelial­to­mesenchymal transition (EMT) of lens epithelial cells. Snail family transcriptional repressor 1 (SNAI1) is a transcriptional repressor that recruits multiple chromatin enzymes including lysine­specific histone demethylase 1A, histone deacetylase (HDAC) 1/2, polycomb repressive complex 2, euchromatic histone lysine methyltransferase 2 and suppressor of variegation 3­9 homolog 1 to the E­cadherin promoter, thereby suppressing E­cadherin expression. However, the functional relationship between SNAI1 and HDAC in the induction of EMT in human lens epithelial cells (HLECs) is still unclear. Therefore, the objective of the present study was to explore the possible functional relationship between SNAI1 and HDAC1 in the induction of EMT in HLECs. In the present study, SNAI1 was found to be increased in HLECs during transforming growth factor­ß2 (TGF­ß2)­induced EMT. Knockdown of SNAI1 by siRNA reversed TGF­ß2­induced downregulation of E­cadherin and upregulation of α­Smooth Muscle Actin. Furthermore, SNAI1 was found to be associated with HDAC1 in the E­cadherin promoter in TGF­ß2­treated HLECs. Inhibition of HDAC by trichostatin A and suberoylanilide hydroxamic acid could prevent TGF­ß2­induced EMT in HLECs. Collectively, SNAI1 interacted with HDAC1 to repress E­cadherin in the TGF­ß2­induced EMT in HLECs, suggesting that HDAC inhibitors may have potential therapeutic value for the prevention of EMT in HLECs.

[Role of lipophagy in the regulation of lipid metabolism and the molecular mechanism].

Recent studies have discovered a selective autophagy-lipophagy, which can selectively identify and degrade lipids and plays an important role in regulating cellular lipid metabolism and maintaining intracellular lipid homeostasis. The process of lipophagy can be directly or indirectly regulated by genes, enzymes, transcriptional regulators and other factors. This review examines the role of lipophagy in reducing liver lipid content, regulating pancreatic lipid metabolism, and regulating adipose tissue differentiation, and summarizes the findings of the molecules (Rab GTPase, enzymes, ion channels, transcription factors, small molecular substances) involved in the regulation of lipophagy, which points to new directions for the treatment of diseases caused by lipid accumulation.

Addressing adverse childhood experiences in diverse racial and ethnic settings: a review.

PURPOSE OF REVIEW: This article reviews the current literature on adverse childhood experiences (ACEs) with particular focus on racial and ethnic factors. RECENT FINDINGS: Limited research exists with respect to ACEs and certain racial and ethnic groups. In this article, we highlight the unique challenges specific populations may face in their lives. Further research on ACEs in these and other racial and ethnic groups is needed to help clinicians contextualize their cases in the clinical setting. SUMMARY: As the US population continues to diversify, it is pivotal that clinicians and researchers understand some of the challenges faced by different racial and ethnic populations and demonstrate more sensitivity to patient needs. Clinicians who are sensitive to an individual's history of ACEs would be expected to have a better understanding of his/her patient's predilection to specific mental health issues.

Scientometric analysis of published papers in global ophthalmology in the past ten years.

AIM: To investigate the published papers of ophthalmology in past ten years and explore the development of ophthalmology. METHORDS: The data of this study retrieved from Science Citation Index Expanded and downloaded online in November 2017, including all the papers with publication year from 2007-2016 were analyzed. The papers were based on the Web of Science category and the journals were based on the Journal Citation Report category. RESULTS: The number of ophthalmology papers increased from 7450 to 9089 during 2007 to 2017. The average rate increased 2.2% annually. USA accounts for one thirds of the total and two thirds of the highly cited papers. In Asia, China, Japan and South Korea were in Top 10 by the number of ophthalmology papers. UK, Germany, Japan and Australia also had great impact in global ophthalmology. The hot spots included endothelial growth factor, optical coherence tomography and open-angle glaucoma. CONCLUSION: USA is in the leading position in global ophthalmology. Part of Asian countries play an important role in the development of ophthalmology, but the impact needs to be improved.

Expression of transcription factors Slug in the lens epithelial cells undergoing epithelial-mesenchymal transition induced by connective tissue growth factor.

AIM: To investigate the expression of transcription factors Slug in human lens epithelial cells (HLECs) undergoing epithelial-mesenchymal transition (EMT) induced by connective tissue growth factor (CTGF). METHODS: HLECs were treated with CTGF of different concentrations (20, 50 and 100 ng/mL) or without CTGF (control) for 24h. The morphological changes of HLECs were analysed by microscopy. The expression and cellular localization of Slug was evaluated by immumo-fluorescence. Expressions of Slug, E-cadherin and alpha smooth muscle actin (α-SMA) were further determined by Western blot analysis. RESULTS: HLECs showed spidle fibrolasts-like characteristics and loosely connected each other after CTGF treatment. The immuno-fluorescence staining indicated that Slug was localized in the nuclei and its expression was induced by CTGF. The relative expressions of Slug protein were 1.64±0.11, 1.96 ±0.03, 3.12 ±0.10, and 4.08±0.14, respectively, in response to control group and treatment with CTGF of 20, 50 and 100 ng/mL (F=443.86, P<0.01). The increased Slug protein levels were correlated well with up-expression of α-SMA (0.78±0.05, 0.85±0.06, 2.17±0.15, 2.86±0.10; F=449.85, P<0.01) and down-expression of E-cadherin (2.50±0.11, 1.79±0.26, 1.05±0.14, 0.63±0.08; F=101.55, P<0.01). CONCLUSION: Transcription factor Slug may be involved in EMT of HLECs induced by CTGF in vitro.

Artemisinin resistance at the China-Myanmar border and association with mutations in the K13 propeller gene.

Artemisinin resistance in Plasmodium falciparum parasites in Southeast Asia is a major concern for malaria control. Its emergence at the China-Myanmar border, where there have been more than 3 decades of artemisinin use, has yet to be investigated. Here, we comprehensively evaluated the potential emergence of artemisinin resistance and antimalarial drug resistance status in P. falciparum using data and parasites from three previous efficacy studies in this region. These efficacy studies of dihydroartemisinin-piperaquine combination and artesunate monotherapy of uncomplicated falciparum malaria in 248 P. falciparum patients showed an overall 28-day adequate clinical and parasitological response of >95% and day 3 parasite-positive rates of 6.3 to 23.1%. Comparison of the 57 K13 sequences (24 and 33 from day 3 parasite-positive and -negative cases, respectively) identified nine point mutations in 38 (66.7%) samples, of which F446I (49.1%) and an N-terminal NN insertion (86.0%) were predominant. K13 propeller mutations collectively, the F446I mutation alone, and the NN insertion all were significantly associated with day 3 parasite positivity. Increased ring-stage survival determined using the ring-stage survival assay (RSA) was highly associated with the K13 mutant genotype. Day 3 parasite-positive isolates had ∼10 times higher ring survival rates than day 3 parasite-negative isolates. Divergent K13 mutations suggested independent evolution of artemisinin resistance. Taken together, this study confirmed multidrug resistance and emergence of artemisinin resistance in P. falciparum at the China-Myanmar border. RSA and K13 mutations are useful phenotypic and molecular markers for monitoring artemisinin resistance.

[Clinical evaluation of the consistency between two diagnostic criteria for diabetic peripheral neuropathy].

OBJECTIVE: To evaluate the consistency between the clinical diagnostic criteria and the ascertained diagnostic criteria for diabetic peripheral neuropathy (DPN) in the Preventive and Treatment Guidelines of Diabetes in China (2013) and explore an economic, convenient, and accurate approach to DPN diagnosis. METHODS: The patients with type 2 diabetes admitted in our department from April to June, 2014 were examined for nerve conduction velocity, 10 g nylon silk, vibration threshold value, sense of temperature and pain, and ankle reflex. The sensitivity, specificity, positive predictive value, negative predictive value, Youden index, and Kappa value were calculated to assess the diagnostic power of the two diagnostic criteria. RESULTS: Of the 151 patients enrolled, 106 (70.2%) had a diagnosis of DPN consistent with the ascertained diagnostic criteria, as compared to 86 (56.95%) who were diagnosed according to the clinical diagnostic criteria; the latter patients accounted for 81.13% of former cases. The sensitivity, specificity, positive predictive value, negative predictive value, Youden index, and Kappa value of the clinical diagnostic criteria were 80.19%, 97.78%, 98.84%, 67.69%, 77.97%, and 0.69, respectively, which were highly consistent with those of the ascertained diagnostic criteria; the sensitivity to compression showed a poor consistency between the two diagnostic criteria. In the 5 screening tests, the combined test of temperature sensation, vibration perception, and ankle reflex showed the highest AUC value among their different combinations. CONCLUSION: The clinical diagnostic criteria for DPN show good consistency with the ascertained diagnostic criteria, and for patients with clinical symptoms or with only one positive sign, combination of the two diagnostic criteria can achieve the maximum diagnostic power.

[Study on mechanism of traditional Chinese medicines reducing phlegm and resolving masses in treatment of goiter].

OBJECTIVE: To discuss the mechanism of traditional Chinese medicines reducing phlegm and resolving masses in treatment of iodine deficiency-induced goiter by observing the expression of growth factors and the balance-regulating mechanism of proliferation and apoptosis. METHOD: 180 four-week-old Wistar rats were selected to establish the iodine deficiency model. After the modeling, the rats were randomly divided into six groups: the normal control group, the model control group, the iodine group, the phlegm compound group, the L-T4 group and the phlegm compound and L-T4 group. At the 21st day and 77th day after administration, 15 rats in each group were killed to collect specimens. Doses were calculated and adjusted according to body surface area and body weight. TT3, TT4 radioimmunoassay, TSH, immunoradiometric method were adopted. Fas, FasL and PCNA protein expressions are detected using immunohistochemical methods. RESULT: Compared with the normal group and the model group, the expressions of fas and FasL in the phlegm Group significantly increased, the expressions of fas and FasL in the phlegm and L-T4 group were also increased significantly. The expression of fas in the L-T4 Group was significantly lower than that of the L-T4 group and the phlegm compound and L-T4 group. Compared with the normal group, the expression of PCNA of the phlegm group and the phlegm and L-T4 group was significantly lower. Compared with the model group, the expression of PCNA of the iodine group, the phlegm groups and the phlegm and L-T4 group were significantly lower. Compared with the normal group, the expression of VEGF in the iodine group significantly decreased after treatment. Compared with the iodine group, the expression of VEGF in the phlegm group and the L-T4 group significantly reduced. Compared with the normal group, the expression of TGF-beta1 in the model group and the phlegm group significantly increased. Compared with model group, the expression of TGF-beta1 in the iodine group significantly reduced. Compared with the phlegm group, the expression of TGF-beta1 in the phlegm compound and L-T4 group was significantly reduced. CONCLUSION: Traditional Chinese medicines reducing phlegm and resolving masses can completely recover goiter by promoting apoptosis of thyroid cells, inhibiting their proliferation and the expression of growth factors and enhancing the expression of TGF-beta, without causing injury on thyroid cells.