Morusin inhibits breast cancer-induced osteolysis by decreasing phosphatidylinositol 3-kinase (PI3K)-mTOR signalling.

Bone metastases caused by breast cancer pose a major challenge to the successful treatment of breast cancer patients. Many researchers have suggested that herbal medicines are extremely effective at preventing and treating cancer-associated osteolysis. Previous studies have revealed that Morusin (MOR) is cytotoxic to many cancer cells ex vivo. Nevertheless, how MOR contributes to osteolysis induced by breast cancer is still unknown, and the potential mechanism of action against osteolysis is worthy of further study. The protective effect and molecular mechanism of MOR in inhibiting breast cancer cell-induced osteolysis were verified by experiments and network pharmacology. Cell function was assessed by cell proliferation, osteoclast (OC) formation, bone resorption, and phalloidin staining. Tumour growth was examined by micro-CT scanning in vivo. To identify potential MOR treatments, the active ingredient-target pathway of breast cancer was screened using network pharmacology and molecular docking approaches. This study is the first to report that MOR can prevent osteolysis induced by breast cancer cells. Specifically, our results revealed that MOR inhibits RANKL-induced osteoclastogenesis and restrains the proliferation, invasion and migration of MDA-MB-231 breast cells through restraining the PI3K/AKT/MTOR signalling pathway. Notably, MOR prevented bone loss caused by breast cancer cell-induced osteolysis in vivo, indicating that MOR inhibited the development of OCs and the resorption of bone, which are essential for cancer cell-associated bone distraction. This study showed that MOR treatment inhibited osteolysis induced by breast cancer in vivo. MOR inhibited OC differentiation and bone resorption ex vivo and in vivo and might be a potential drug candidate for treating breast cancer-induced osteolysis.

Predictive Model Construction And Prevention Strategies Of Surgical Site Infection In Laparoscopic Appendectomy.

Objective: To construct a nomogram model for predicting the occurrence of the laparoscopic appendectomy surgical site infection (LASSI) and explore prevention strategies. Methods: A total of 995 patients who underwent laparoscopic appendectomy in Shanxi Bethune Hospital from October 2017 to August 2022 were selected. According to whether there was incision infection within 30 days after operation, the patients were divided into the LASSI (97 cases) and non-LASSI (898 cases) group. The following clinicopathological data from these two groups of patients were collected: gender, age, body mass index, ect. The subjects were randomly divided into training group and verification group according to the 7:3 ratio. Univariate and multivariate analysis was used to screen the related influencing factors and construct a nomogram model to predict the occurrence of LASSI. Rreceiver operating characteristic (ROC) curves and the calibration curve were used to evaluate the predictive value of the model. For patients with LASSI, a more effective preventive measure was explored. Results: Multivariate logistic regression analysis showed that operation time >1h (OR: 1.891; 95% CI: 1.07 to 3.36; P = .029), perforated and gangrenous appendix (OR: 4.078; 95% CI: 1.84 to 9.86; P = .001), free intraperitoneal fluid (OR: 2.836; 95% CI: 1.57 to 5.35; P = .001), BMI>30 kg/m2 (OR: 2.828; 95% CI:1.54 to 5.12; P = .001), diabetes mellitus (DM) (OR: 2.795; 95% CI: 1.54 to 5.28; P = .001) were the independent prognostic factors of LASSI. The prediction nomogram model showed satisfactory performance in predicting the occurrence of LASSI, ROC curve area value of the training and verification groups were respectively 0.753 (95 % CI: 0.688 ~ 0.818) and 0.772 (95 % CI: 0.691-0.852). In the event of LASSI, we took out appendix specimens in sections and sterilized surgical site, which effectively prevented it. Conclusion: This study evaluated the risk factors related to the occurrence of LASSI and established a prediction model for LASSI. The prediction model provides a convenient and fast risk assessment tool for clinicians to predict the occurrence of LASSI. Combined with the newly discovered prevention strategy of segmental removal of appendix and incision disinfection, it can effectively avoid the occurrence of LASSI and potentially reduce the hospitalization time and costs.

Spinosin inhibits activated hepatic stellate cell to attenuate liver fibrosis by targeting Nur77/ASK1/p38 MAPK signaling pathway.

AIM: Liver fibrosis remains a great challenge in the world. Spinosin (SPI), a natural flavonoid-C-glycoside, possesses various pharmacological activities including anti-inflammatory and anti-myocardial fibrosis effects. In this study, we investigate whether SPI can be a potential lead for the treatment of liver fibrosis and explore whether the orphan nuclear receptor Nur77, a negative regulator of liver fibrosis development, plays a critical role in SPI's action. METHODS: A dual luciferase reporter system of α-SMA was established to evaluate the effect of SPI on hepatic stellate cell (HSC) activation in LX2 and HSC-T6 cells. A mouse model of CCl4-induced liver fibrosis was used to test the efficacy of SPI against liver fibrosis. The expression levels of Nur77, inflammatory cytokines and collagen were determined by Western blotting and qPCR. Potential kinase pathways involved were also analyzed. The affinity of Nur77 with SPI was documented by fluorescence titration. RESULTS: SPI can strongly suppress TGF-ß1-mediated activation of both LX2 and HSC-T6 cells in a dose-dependent manner. SPI increases the expression of Nur77 and reduces TGF-ß1-mediated phosphorylation levels of ASK1 and p38 MAPK, which can be reversed by knocking out of Nur77. SPI strongly inhibits collagen deposition (COLA1) and reduces inflammatory cytokines (IL-6 and IL-1ß), which is followed by improved liver function in the CCl4-induced mouse model. SPI can directly bind to R515 and R563 in the Nur77-LBD pocket with a Kd of 2.14 µM. CONCLUSION: Spinosin is the major pharmacological active component of Ziziphus jujuba Mill. var. spinosa which has been frequently prescribed in traditional Chinese medicine. We demonstrate here for the first time that spinosin is a new therapeutic lead for treatment of liver fibrosis by targeting Nur77 and blocking the ASK1/p38 MAPK signaling pathway.

[Effects of Short-Term Nitrogen and Phosphorus Addition on Soil Respiration Components in a Subalpine Grassland of Qilian Mountains].

In order to investigate the effects of short-term nitrogen and phosphorus addition on soil respiration and its components in a subalpine grassland located on the Qilian Mountains, a random block design of nitrogen[10 g·(m2·a)-1, N], phosphorus[5 g·(m2·a)-1, P], nitrogen and phosphorus addition[10 g·(m2·a)-1N and 5 g·(m2·a)-1P, NP], the control (CK), and complete control (CK') was conducted from June to August 2019, and total soil respiration and its component respiration rates were measured. The results showed that nitrogen addition reduced soil total respiration and heterotrophic respiration rates at a lower rate than P addition[-16.71% vs. -19.20%; -4.41% vs. -13.05%], but the rate of decrease in autotrophic respiration was higher than that of P addition (-25.03% vs. -23.36%); N and P mixed application had no significant effect on soil total respiration rate. The total soil respiration rate and its components were significantly exponentially correlated with soil temperature, and the temperature sensitivity of soil respiration rate was decreased by nitrogen addition (Q10:-5.64%-0.00%). P increased Q10 (3.38%-6.98%), and N and P reduced autotrophic respiration rate but increased heterotrophic respiration rate Q10 (16.86%) and decreased total soil respiration rate Q10 (-2.63%- -2.02%). Soil pH, soil total nitrogen, and root phosphorus content were significantly correlated with autotrophic respiration rate (P<0.05) but not with heterotrophic respiration rate, and root nitrogen content was significantly negatively correlated with heterotrophic respiration rate (P<0.05). In general, autotrophic respiration rate was more sensitive to N addition, whereas heterotrophic respiration rate was more sensitive to P addition. Both N and P addition significantly reduced soil total respiration rate, whereas N and P mixture did not significantly affect soil total respiration rate. These results can provide a scientific basis for the accurate assessment of soil carbon emission in subalpine grassland.

Network-Based Elaboration of the Efficacy of the Dachangshu (BL25) and Tianshu (ST25) Points in the Treatment of Functional Constipation in Children through Inflammation, Adipocytokine, or Leptin Pathways.

Constipation commonly occurs during childhood, and more than 95% of cases are classified as functional constipation. If not effectively treated, 20% of patients with childhood constipation can continue to exhibit symptoms into adulthood, which seriously affects their mental health and quality of life. The main feature of acupuncture or acupoint stimulation, a special branch of traditional Chinese medicine, is the selection of different acupoints for different diseases, and many worthy guidelines have been established for matching acupoints. The back-shu and front-mu point combination adheres to an important acupoint compatibility law that has been used since its proposal 2,500 years ago but has not yet been verified by the modern evidence-based experiments. This study focused on the back-shu and front-mu point combination using the Dachangshu (BL25) and Tianshu (ST25) points as examples to explore possible research methods for network acupoint-based stimulation based on existing evidence and to elucidate the mechanisms induced by BL25 and ST25 in the treatment of functional constipation in children (FCC). The study found that BL25 and ST25 have 20 common targets, namely, AQP8, DRD2, VIP, TAC1, IL6R, TNF, FOS, KIT, CHAT, HTR3A, GAS8, SOD3, TRPV1, MPO, CALCA, IL1B, P2RX7, NPY2R, IL10RA, and TPH1, and these targets may provide a strategy for the combined usage of BL25 and ST25. In addition, BL25 and ST25 can affect FCC treatment through inflammation-relatedTh17-cell differentiation, the NF-kappa B signaling pathway, and the Toll-like receptor signaling pathway. Adipocytokines or leptin may also comprise the mechanism through which BL25 and ST25 regulate FCC. In addition, BL25 and ST25 regulate FCC through 13 core targets, namely, NFKBIA, RELA, TNF, IKBKB, IRAK1, TLR4, MYD88, TNFRSF1A, IL1R1, TLR2, IL1B, TRAF6, and TNFRSF1B. In short, this study provides new ideas and methods for studying the mechanism of acupuncture points.

Nitrogen effects on grassland biomass production and biodiversity are stronger than those of phosphorus.

Human-induced nitrogen (N) and phosphorus (P) enrichment have profound effects on grassland net primary production (NPP) and species richness. However, a comprehensive understanding of the relative contribution of N vs. P addition and their interaction on grassland NPP increase and species loss remains elusive. We compiled data from 80 field manipulative studies and conducted a meta-analysis (2107 observations world-wide) to evaluate the individual and combined effects of N and P addition on grassland NPP and species richness. We found that both N addition and P addition significantly enhanced grassland above-ground NPP (ANPP; 33.2% and 14.2%, respectively), but did not affect total NPP, below-ground NPP (BNPP), and species evenness. Species richness significantly decreased with N addition (11.7%; by decreasing forbs) probably due to strong decreased soil pH, but not with P addition. The combined effects of N and P addition were generally stronger than the individual effects of N or P addition, and we found the synergistic effects on ANPP, and additive effects on total NPP, BNPP, species richness, and evenness within the combinations of N and P addition. In addition, N and P addition effects were strongly affected by moderator variables (e.g. climate and fertilization type, duration and amount of fertilizer addition). These results demonstrate a higher relative contribution of N than P addition to grassland NPP increase and species loss, although the effects varied across climate and fertilization types. The existing data also reveals that more long-term (≥5 years) experimental studies that combine N and P and test multifactor effects in different climate zones (particularly in boreal grasslands) are needed to provide a more solid basis for forecasting grassland community response and C sequestration response to nutrient enrichment at the global scale.

Examining the Effector Mechanisms of the Feishu Acupoint (BL13) in the Treatment of Pneumonia Based on Systematic Acupuncture and Moxibustion Research.

BACKGROUND: Pneumonia is a serious global health problem. In traditional Chinese medicine, acupuncture or moxibustion is used to directly stimulate select acupoints on the surface of the human body and produce physical stimulation to further stimulate regulatory functions in the body, strengthening bodily resistance, eliminating disease, and adjusting the viscera. However, this Chinese medicine knowledge does not include the specific mechanisms of action or targets of acupoints. Therefore, an in-depth research is needed. METHODS: An acupoint-element database was constructed, and the target elements of the Feishu point were screened. The UniProt-Swiss-Prot sublibrary was used to obtain correct gene name information. The National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database and GEO2R were used to analyze differentially expressed genes in pneumonia. The STRING database was used to analyze interactions, construct a network of the Feishu point efficacy system in pneumonia, and elucidate the mechanisms of action. RESULTS: The Feishu point comprises 34 elements in total. The protein interaction analysis has 38 nodes and 115 edges. The Feishu point efficacy system-pneumonia system network shows that cytokine signaling in the immune system, signaling by interleukins (ILs), IL-4 and IL-13 signaling, and the immune system may be related to immunity and inflammation. The Feishu point efficacy system regulating pneumonia showed that FCER2, IL4R, FASLG, TGFB1, IL6R, STAT6, IL1B, CASP3, IL5RA, IL2RB, MYD88, SQSTM1, IL12RB1, IFNGR1, ADAM17, and CDH1 are the main targets. CONCLUSION: From the perspective of systematic acupuncture and moxibustion, the Feishu point regulates cytokine signaling in the immune system, signaling by ILs, IL-4 and IL-13 signaling, and the immune system by targeting FCER2, IL4R, FASLG, TGFB1, IL6R, STAT6, IL1B, CASP3, IL5RA, IL2RB, MYD88, SQSTM1, IL12RB1, IFNGR1, ADAM17, and CDH1, thereby regulating pneumonia.

α-Tomatine, a novel early-stage autophagy inhibitor, inhibits autophagy to enhance apoptosis via Beclin-1 in Skov3 cells.

Targeting the autophagy process is considered to be a promising new strategy for drug treatment of ovarian cancer. α-Tomatine, a steroidal alkaloid extracted, is mainly isolated from leaves, roots and immature green tomatoes. α-Tomatine has biological activities such as anticancer, antioxidative and anti-inflammatory. The study aimed to explore the effects of α-tomatine on proliferation, apoptosis and autophagy and the underlying mechanisms in ovarian cancer Skov3 cells. After treatment with different concentrations of α-tomatine (0, 0.75, 1 and 1.5 µM) in Skov3 cells for 24 h, proliferation was determined by the CCK-8 assay, and apoptosis was detected by flow cytometric analysis. Autophagy in cells was determined by the number of fluorescent spots using confocal fluorescence microscopy after mRFP-GFP-LC3 transfection. The relationship between autophagy and apoptosis was proved by Beclin-1 overexpression. The protein expression levels were tested by western blotting. The results demonstrated that α-tomatine effectively repressed proliferation, exerted a proapoptotic effect and inhibited early-stage autophagy in Skov3 cells in a dose- and time-dependent manner. Additionally, Beclin-1 overexpression significantly suppressed α-tomatine-treated apoptosis in Skov3 cells, indicating that α-tomatine inhibits autophagy to induce apoptosis. We also found α-tomatine inhibited the protein expression levels of PI3K/Akt/mTOR signaling pathway. However, the autophagy inhibition of α-tomatine could be reversed obviously by Beclin-1 overexpression. Taken together, α-tomatine inhibited autophagy through Beclin-1. Our study suggests that α-tomatine, as a novel early-stage autophagy inhibitor, might be a potential drug for further treatment of ovarian cancer by inhibiting proliferation and promoting apoptosis.

[Effect of acupuncture on pattern visual evoked potential of cerebral visual impairment in children aged 3-10 years].

OBJECTIVE: To observe clinical effect of acupuncture combined with conventional visual stimulation on cerebral visual impairment (CVI) in children aged 3-10 years and influence on the pattern visual evoked potential (P-VEP). METHODS: A total of 60 cases of children aged 3-10 years with CVI were randomly divided into an observation group and a control group, 30 cases in each group. The children in the control group received conventional visual stimulation therapy, 1 month as a course of treatment. On the basis of the control group, the children in the observation group was treated with acupuncture at Baihui (GV 20), Jingming (BL 1), Taiyang (EX-HN 5), Sibai (ST 2), etc. 3 times a week, and the treatment was given 4 weeks continuously as a course. Both groups received 3 courses of treatment. The visual acuity and P-VEP improvement were compared between the two groups before and after treatment. RESULTS: After treatment, the incubation period (P100-L) of the two groups was shorter than before treatment, and the amplitude (P100-A) was higher than before treatment (P<0.05); and the degree of above changes in the observation group was lager than the control group (P<0.05). The percentage of best corrected visual acuity of 0.6-0.8 in the observation group after treatment and follow-up 1 year after treatment was higher than before treatment (P<0.05), and was higher than the control group (P<0.05). CONCLUSION: Acupuncture combined with conventional visual stimulation can improve the incubation period (P100-L) and amplitude (P100-A) of P-VEP in children with CVI, and improve the best corrected visual acuity in children, the clinical effect is better than the conventional visual stimulation alone.

P2X7 receptor mediates activation of microglial cells in prostate of chemically irritated rats.

PURPOSE: Evidence shows that adenosine triphosphate (ATP) is involved in the transmission of multiple chronic pain via P2X7 receptor. This study was to investigate the P2X7 and microglial cells in the chronic prostatitis pain. MATERIALS AND METHODS: Rats were divided into control group and chronic prostatitis group (n = 24 per group). A chronic prostatitis animal model was established by injecting complete Freund's adjuvant (CFA) to the prostate of rats, and the thermal withdrawal latency (TWL) was detected on days 0, 4, 12 and 24 (n = 6 at each time point in each group). Animals were sacrificed and the pathological examination of the prostate, detection of mRNA expression of P2X7 and ionized calcium binding adaptor molecule 1 (IBA-1) and measurement of content of tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) in the dorsal horn of L5-S2 spinal cord were performed on days 0, 4, 12 and 24. In addition, the content of TNF-α and IL-1ß in the dorsal horn of L5-S2 spinal cord was measured after intrathecal injection of inhibitors of microglial cells and/or P2X7 for 5 days. RESULTS: The chronic prostatitis was confirmed by pathological examination. The expression of P2X7 and IBA-1 and the content of TNF-α and IL-1ß in rats with chronic prostatitis were significantly higher than those in the control group. On day 4, the expressions of pro-inflammatory cytokines became to increase, reaching a maximal level on day 12 and started to reduce on day 24, but remained higher than that in the control group. Following suppression of microglial cells and P2X7 receptor, the secretion of TNF-α and IL-1ß was markedly reduced. CONCLUSION: In chronic prostatitis pain, the microglial cells and P2X7 receptor are activated resulting in the increased expression of TNF-α and IL-1ß in the L5-S2 spinal cord, which might attribute to the maintenance and intensification of pain in chronic prostatitis.

P2X7 receptor mediates activation of microglial cells in prostate of chemically irritated rats

Purpose Evidence shows that adenosine triphosphate (ATP) is involved in the transmission of multiple chronic pain via P2X7 receptor. This study was to investigate the P2X7 and microglial cells in the chronic prostatitis pain. Materials and Methods Rats were divided into control group and chronic prostatitis group (n = 24 per group). A chronic prostatitis animal model was established by injecting complete Freund's adjuvant (CFA) to the prostate of rats, and the thermal withdrawal latency (TWL) was detected on days 0, 4, 12 and 24 (n = 6 at each time point in each group). Animals were sacrificed and the pathological examination of the prostate, detection of mRNA expression of P2X7 and ionized calcium binding adaptor molecule 1 (IBA-1) and measurement of content of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the dorsal horn of L5-S2 spinal cord were performed on days 0, 4, 12 and 24. In addition, the content of TNF-α and IL-1β in the dorsal horn of L5-S2 spinal cord was measured after intrathecal injection of inhibitors of microglial cells and/or P2X7 for 5 days. Results The chronic prostatitis was confirmed by pathological examination. The expression of P2X7 and IBA-1 and the content of TNF-α and IL-1β in rats with chronic prostatitis were significantly higher than those in the control group. On day 4, the expressions of pro-inflammatory cytokines became to increase, reaching a maximal level on day 12 and started to reduce on day 24, but remained higher than that in the control group. Following suppression of microglial cells and P2X7 receptor, the secretion of TNF-α and IL-1β was markedly reduced. Conclusion In chronic prostatitis pain, the microglial cells and P2X7 receptor are activated resulting in the increased expression of TNF-α and IL-1β in the L5-S2 spinal cord, which might attribute to the maintenance and intensification of pain in chronic prostatitis. .