Determination of the signaling proteins responsible for the antioxidant potential of the Yishenhuoxue formula for treating asthenospermia in rats.

Asthenospermia is a predominant cause of male infertility, and antioxidant supplements can be effective in treating asthenospermia. We demonstrate the antioxidant potential of traditional Chinese medicine, the Yishenhuoxue (YSHX) formula, in treating polyglycosides of Tripterygium wilfordii (GTW)-induced asthenospermia in rats. Fifty male rats were randomly divided into the normal, model, and treatment groups. HE staining was used to evaluate the improvement of spermatogenic function of rats, and TBA reaction, qRT-PCR, Western Blot and other methods were used to determine the changes of oxidative stress indicators and to evaluate the improvement of antioxidant capacity of rats by YSHX. Comparison with the model group showed significant improvement in pathological damage caused by GTW to seminiferous tubules. MDA and NO content in rat testes decreased, especially in middle- and high-dosage groups. No significant changes were observed in SOD and CAT activity or mRNA expression. GSH-Px activity and GSH mRNA expression were significantly higher in the low-dosage group than in the model group. Compared to the model group, GR activity was significantly lower in the middle and high dosage groups, while the mRNA expression was higher. The PKC-beta level increased, while p-ERK1/2, NF-κB, and the ratio of p-ERK1/2*(ERK1/2)-1 decreased significantly in the treatment groups. Therefore, YSHX can alleviate GTW-induced testicular damage, enhance GSH-Px activity, regulate GSH redox cycling, and mitigate oxidative stress injury. Furthermore, YSHX can promote PKC-beta expression and inhibit the phosphorylation of ERK1/2 and NF-κB. Using YSHX may be an effective way to increase sperm motility via the PKC-ERK1/2-NF-ĸB axis.

Opsonization Inveigles Macrophages Engulfing Carrier-Free Bilirubin/JPH203 Nanoparticles to Suppress Inflammation for Osteoarthritis Therapy.

Osteoarthritis (OA) is a chronic inflammatory disease characterized by cartilage destruction, synovitis, and osteophyte formation. Disease-modifying treatments for OA are currently lacking. Because inflammation mediated by an imbalance of M1/M2 macrophages in the synovial cavities contributes to OA progression, regulating the M1 to M2 polarization of macrophages can be a potential therapeutic strategy. Basing on the inherent immune mechanism and pathological environment of OA, an immunoglobulin G-conjugated bilirubin/JPH203 self-assembled nanoparticle (IgG/BRJ) is developed, and its therapeutic potential for OA is evaluated. After intra-articular administration, IgG conjugation facilitates the recognition and engulfment of nanoparticles by the M1 macrophages. The internalized nanoparticles disassemble in response to the increased oxidative stress, and the released bilirubin (BR) and JPH203 scavenge reactive oxygen species (ROS), inhibit the nuclear factor kappa-B pathway, and suppress the activated mammalian target of rapamycin pathway, result in the repolarization of macrophages and enhance M2/M1 ratios. Suppression of the inflammatory environment by IgG/BRJ promotes cartilage protection and repair in an OA rat model, thereby improving therapeutic outcomes. This strategy of opsonization involving M1 macrophages to engulf carrier-free BR/JPH203 nanoparticles to suppress inflammation for OA therapy holds great potential for OA intervention and treatment.

Ultraviolet B radiation-induced JPH203-loaded keratinocyte extracellular vesicles exert etiological interventions for psoriasis therapy.

Psoriasis is a multifactorial immuno-inflammatory skin disease, characterized by keratinocyte hyperproliferation and aberrant immune activation. Although the pathogenesis is complex, the interactions among inflammation, Th17-mediated immune activation, and keratinocyte hyperplasia are considered to play a crucial role in the occurrence and development of psoriasis. Therefore, pharmacological interventions on the "inflammation-Th17-keratinocyte" vicious cycle may be a potential strategy for psoriasis treatment. In this study, JPH203 (a specific inhibitor of LAT1, which engulfs leucine to activate mTOR signaling)-loaded, ultraviolet B (UVB) radiation-induced, keratinocyte-derived extracellular vesicles (J@EV) were prepared for psoriasis therapy. The EVs led to increased interleukin 1 receptor antagonist (IL-1RA) content due to UVB irradiation, therefore not only acting as a carrier for JPH203 but also functioning through inhibiting the IL-1-mediated inflammation cascade. J@EV effectively restrained the proliferation of inflamed keratinocytes via suppressing mTOR-signaling and NF-κB pathway in vitro. In an imiquimod-induced psoriatic model, J@EV significantly ameliorated the related symptoms as well as suppressed the over-activated immune reaction, evidenced by the decreased keratinocyte hyperplasia, Th17 expansion, and IL17 release. This study shows that J@EV exerts therapeutic efficacy for psoriasis by suppressing LAT1-mTOR involved keratinocyte hyperproliferation and Th17 expansion, as well as inhibiting IL-1-NF-κB mediated inflammation, representing a novel and promising strategy for psoriasis therapy.

Calycosin protects against chronic prostatitis in rats via inhibition of the p38MAPK/NF-κB pathway.

Currently, the effect and molecular mechanism of calycosin, the main active ingredient of Qinshi Simiao San, which can alleviate chronic prostatitis (CP), on CP remain unclear. This study aimed to elucidate the potential mechanism of action of calycosin in CP in a rat CP model. The prostate tissue morphology was evaluated based on hematoxylin-eosin staining. Enzyme-linked immunosorbent assay was conducted to evaluate inflammatory cytokine and immune factor levels (secretory immunoglobulin A [SIgA]; immunoglobulin G [IgG]) in prostate tissues and serum. Additionally, representative biomarkers of oxidative stress, including malondialdehyde, superoxide dismutase, and catalase were detected using detection kits, and reactive oxygen species release was evaluated using immunofluorescence staining. Furthermore, the p38 mitogen-activated protein kinase (p38MAPK)/NF-kappaB (NF-κB) signaling pathway was analyzed by western blotting. The results showed that calycosin substantially ameliorated the pathological damage to prostate tissues of the CP rats. Moreover, calycosin significantly downregulated interleukin (IL)-1ß, IL-6, and tumor necrosis factor-alpha, IgG, and SIgA levels. Furthermore, we found that calycosin considerably suppressed oxidative stress and inhibited the activation of the p38MAPK/NF-κB signaling pathway in rats with CP. In summary, our findings revealed that calycosin protects against CP in rats by inhibiting the p38MAPK/NF-κB pathway.

Protein tyrosine phosphatase nonreceptor type 2 exerts antimetastatic functions in pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is a highly invasive tumor with a dismal prognosis. Recent studies have demonstrated PTPN2 (protein tyrosine phosphatase nonreceptor type 2) as a potential target for cancer therapy. However, the functions of PTPN2 in PDAC progression remain poorly understood. In this study, we found PTPN2 expression was downregulated in PDAC tissues, and decreased PTPN2 expression was associated with unfavorable prognosis. Functional studies indicated that PTPN2 knockdown promoted the migration and invasion abilities of PDAC cells in vitro, and the liver metastasis in vivo through epithelial-mesenchymal transition process. Mechanistically, MMP-1 was identified as a downstream target of PTPN2 via RNA-seq data and was responsible for the enhanced metastasis of PDAC cells upon PTPN2 knockdown. Moreover, according to chromatin immunoprecipitation and electrophoretic mobility shift assay, PTPN2 depletion transcriptionally activated MMP-1 via regulating the interaction of p-STAT3 with its distal promoter. This study, for the first time, demonstrated that PTPN2 inhibited PDAC metastasis, and presented a novel PTPN2/p-STAT3/MMP-1 axis in PDAC progression.

Glutamine-Based Metabolism Normalization and Oxidative Stress Alleviation by Self-Assembled Bilirubin/V9302 Nanoparticles for Psoriasis Treatment.

Psoriasis is an immune-mediated chronic inflammatory skin disorder characterized by epidermal hyperplasia and infiltration of inflammatory cells. Even though the pathogenesis remains unclear, T helper 17 (Th17) cells-mediated inflammation and keratinocyte-involved proliferation are considered to play key roles during the occurrence and the development of psoriasis. Therefore, suppressing the infiltration/function of Th17 and the abnormal hyperplasia of keratinocytes can be a rational strategy for ameliorating and treating psoriasis. In this study, a self-assembly nanoparticle (BVn) is developed with bilirubin (an endogenous antioxidant) and V9302 (a blocker of ASCT2, an amino acid transporter mediating glutamine influx for providing energy and activating mammalian target of rapamycin [mTOR] pathway) to intervene the local metabolism and alleviate oxidative stress for psoriasis treatment. BVn effectively suppresses inflammatory keratinocyte proliferation and scavenges excess reactive oxygen species (ROS). In the in vivo psoriasis mouse model, BVn shows increased permeation and delayed retention in the psoriatic lesion and reverses the psoriasis-related symptoms, evidenced by the normalized keratinocyte condition and decreased Th17 infiltration/activation. Mechanism study indicates that BVn not only cut off the energy supply but also suppressed cell proliferation or lymph cell expansion by deactivating mTOR pathway, besides alleviated oxidative stress. BVn-based glutamine metabolism modulation strategy offers a promising strategy for psoriasis therapy.

Early Identification of High-Risk Factors for Upper Gastrointestinal Bleeding.

Objective: To identify simple and accurate pre-endoscopy risk factors for early identification of high-risk upper gastrointestinal bleeding. Methods: Patients who were admitted to Suzhou Hospital of Integrated Traditional Chinese and Western Medicine from January 1, 2016, to December 31, 2019, due to upper gastrointestinal bleeding were retrieved, and the detailed clinical data of the above patients were collected. Patients with a definite diagnosis of bleeding from esophageal/and gastric varices were assigned to the high-risk group. Patients with bleeding not caused by varices were divided into a high-risk and a low-risk group according to the Forrest grading and scoring standard (high-risk group Forrest Ia-IIb, low-risk group Forrest IIc-III). Univariate analysis, t-test, chi-square test, binary logistic regression, ROC curve (Receiver-operating characteristic curve), etc. were employed for analysis in order to identify some simple and accurate risk factors for high-risk upper digestion tract bleeding before endoscopy. Results: A total of 916 patients were collected. Three risk factors among the screened risk factors (1) hemoglobin ≤ 85 g/L, (2) vomiting red blood, and (3) "red bloody stool" were analyzed by ROC curve analysis. The specificities of each factor were 78.4%, 94.5%, and 96.7%, respectively, and the sensitivities were 71.8%, 55.9%, and 23.1%, respectively. We also derived a risk prediction scoring system for the three factors that meet the high risk such as (1) hemoglobin ≤ 83 g/L, (2)vomiting red blood, and (3) "red bloody stool." The area under the ROC curve (AUROC), sensitivity, and specificity were 0.877, 0.904, and 0.746. Conclusion: Hemoglobin ≤ 85 g/L, vomiting red blood, and red bloody stool were included in a simple scoring standard for predicting high-risk UGIB patients before endoscopy. The new risk prediction scoring system requires only three indicators and has the advantages of high accuracy, short time-consuming, and easy application.

Upregulation of exonuclease 1 caused by homology-dependent repair confers cisplatin resistance to gastric cancer cells.

The homology-dependent repair (HDR) pathway is involved in deoxyribonucleic acid (DNA) damage response (DDR), which is crucial to cancer cell survival after treatment with DNA damage agents, including cisplatin (CDDP). Here, we explored the interactions between exonuclease 1 (EXO1), a core gene in the HDR pathway, and CDDP resistance in gastric cancer (GC). Using bioinformatics analysis, we identified the HDR pathway as the most amplified pathway in DDR in GC. In addition, EXO1 was the core gene in the HDR pathway and showed the most significant amplification in GC. The amplification of EXO1 resulted in higher EXO1 expression in cancerous tissues, with malignant prognostic effects. Moreover, we upregulated or downregulated EXO1 in GC cells to examine its effects on the cell malignant phenotype and CDDP resistance in vitro and in vivo. The depletion of EXO1 inhibited cell proliferatory, migratory, and invasive activities, and provided apoptosis resistance to GC cells. EXO1 expression was elevated in CDDP-resistant cells. Ectopic expression of EXO1 increased the resistance of GC cells to CDDP, while downregulation of EXO1 increased the sensitivity of GC cells. Taken together, our study indicates that the HDR pathway is an important player in CDDP resistance in GC through the regulation of EXO1.

Cu(OAc)2-Triggered Cascade Reaction of Malonate-Tethered Acyl Oximes with Indoles, Indole-2-alcohols, and Indole-2-carboxamides.

A Cu(OAc)2-promoted cascade reaction of malonate-tetherd acyl oximes with indoles, indole-2-alcohols, or indole-2-carboxmides provides facile access to polysubstituted 3-pyrrolin-2-ones. The reaction features the generation of two adjacent electrophilic centers at the same time as cyclization to lactam. The subsequent double addition with nucleophiles followed by oxidation realizes the difunctionalization of the imine sp2-carbon and the adjacent α-sp3-carbon.

[Comparison of major bioactive components from leaves of Chrysanthemum morifolium].

Leaves of Chrysanthemum morifolium were potential medicinal resource. The present study aims to estimate the main bioactive components: total flavonoids (TF), galuteolin (GA), quercitrin (QU), chlorogenic acid (CA) and 3 ,5-O-caffeoylquinic acid ( CQ), which were considered to be the main effective components, in leaves of C. morfolium cultivars in China. The TF content was estimated hy UV-VIS spectrophotometry, while GA, QU, CA, and CQ were quantitatively determined by HPLC. The highest TF content (7. 13% w/w) was found in cultivar Wan Cong (Shexian county). Cultivar Da Bo ( Bozhou county) had the highest GA content (33. 45 mg - g-1); Cultivar Hong Xin (Sheyang county) contained the highest QU content (29.25 mg · g(-1)); Cultivar Chang Ban (Sheyang county) had the highest CA content (13.14 mg ·(-1)). The maximum CQ content (7.35 mg · g(-1)) was observed in culti- r Da Yang ( Tongxiang county). Different cultivars of C. morfolium had significant difference in components, but the leaf and capitulum of C. morifolium. were found to possess similar chemical compositions. The high content of bioactive components in several cultivars suggested the potential utilization of C. morifolium leaves.

[Relationship between seedling grade and plant growth, yield and quality of medicinal Chrysanthemum morifolium].

OBJECTIVE: To provide the basis for standardization cultivation of medicinal Chrysanthemum morifolium, the relationship between the seedling grading and plant growth, yield and quality were studied. METHOD: The morphological index of the seedlings was measured and the method of principal component and correlation analysis were used to determine the grading index, and step-wise cluster analysis was applied for clustering analysis. Pot experiments were used to measure the indicators of plant growth and development, yield and quality. RESULT: The height and ground diameter were determined for the quality indicators of the seedlings grading, and the standard quality grading of seedlings of Ch. morifolium was initially set up. The ground diameter of the class I and II were larger than that of the class III, and the number of branches of class I and II was more than that of the class III, on the contrary, the plant height of the class III was higher than that of the class I and II. The shape and appearance of the plant had no effect on the intrinsic quality. Flower center diameter and tubular floret number of the class I and II were significantly larger than those of the class III, so as the yield. The seedling grading had no obvious effect on the internal quality of medicinal material. CONCLUSION: Seedlings of the class I and class II were suitable for transplanting.

[Rapid identification system for seedlings of medicinal Chrysanthemum morifolium].

OBJECTIVE: To achieve the rapid identification for seedlings of medicinal Chrysanthemum morifolium, the discriminant equation was established and the software for rapid identification was designed. METHOD: Leaf structure of medicinal Chrysanthemum of 12 cultivars was analyzed to establish the discriminant equation based on variance analysis and discriminant analysis. On this basis, the identification program and software (based on the python language) were designed. RESULT: Through the analysis of variance and multiple comparisons for the 11 leaf parameter index data of 12 different cultivars, it was found that that the leaf parameters were significant different from each other and reached significant levels. The discriminant equation and the rapid identification software were set up based on the analysis of various indicators. CONCLUSION: The rapid identification system of seedlings of medicinal Chrysanthemum could be achieved through the establishment of discriminant equation combined with computer technology.

[Comparison of botanical characteristics of cultivated Chrysanthemum morifolium cv. 'Hangju' of different origins].

OBJECTIVE: To compare botanical characteristics of cultivated Chrysanthemum morifolium cv. 'Hangju' of different origins in order to provide the basis for introduction and cultivation of Ch. morifolium cv. 'Hangju'. METHOD: The characteristics of plants, leaves and capitulum of Ch. morifolium cv. 'Hangju' were measured, and the obtained data were analyzed and compared. RESULT: The range of plant height was 60.87-99.47 cm, number of branches 2.76-5.20, leave length 4.90-8.40 cm, leave width 3.25-5.38 cm, aspect ratio of leave 1.35-1.83, number of leave split 1.92-3.08. Numbers of capitulum were 21.92-53.12, diameter of capitulum 3.41-5.48 cm, lays of ray florets 3.28-7.16, number of ray florets 55.32-114.60, ray florets length 1.58-2.37 cm, ray florets width 0.50-0.69 cm, aspect ratio of ray florets 2.90-3.99, diameter of tubular flower 1.10-1.58 cm. CONCLUSION: The botanical characteristics of cultivated Ch. morifolium cv. 'Hangju' were distinguished from different origins. With the cultivation environment change, the botanical characteristics of the cultivars are changed.

[Numerical analysis of morphological variation of germplasm resources of Chrysanthemum morifolium for medicine].

OBJECTIVE: Botanical characters of germplasm resources of Chrysanthemum morifolium for medicine were observed and compared, which could offer reference for its genetic improvement and germplasm resources protection. METHOD: Based on the random blocks field experiments design, twenty-six morphological traits were observed. The morphological differences among germplasm resources were compared by principal component analysis and cluster analysis. RESULT: The coefficient of variation values for 17 of 26 traits indicated a high level of variation (above 20%). Six principal components which accounted for 77.14% of total variance were extracted from the principal component analysis. The 29 germplasm resources could be divided into two clusters. CONCLUSION: There were large morphological variation among germplasm resources on Ch. morifolium for medicine.