Triptolide (TP) is a major bioactive compound derived from Tripterygium wilfordii Hook. F. (TwHF) known for its medicinal properties, but it also exhibits potential toxic effects. It has been demonstrated to induce severe male reproductive toxicity, yet the precise mechanism behind this remains unclear, which limits its broad clinical application. This study aimed to investigate the mechanisms underlying testicular damage and spermatogenesis dysfunction induced by TP in mice, using both mouse models and the spermatocyte-derived cell line GC-2spd. In the present study, it was found that TP displayed significant testicular microstructure damaged and spermatogenesis defects including lower concentration and abnormal morphology by promoting ROS formation, MDA production and restraining GSH level, glutathione peroxidase 4 (GPX4) expression in vivo. Furthermore, Ferrostatin-1 (FER-1), a ferroptosis inhibitor, was found to significantly reduce the accumulation of lipid peroxidation, alleviate testicular microstructural damage, and enhance spermatogenic function in mice. Besides, notably decreased cell viability, collapsed mitochondrial membrane potential, and elevated DNA damage were observed in vitro. The above-mentioned phenomenon could be reversed by pre-treatment of FER-1, indicating that ferroptosis participated in the TP-mediated spermatogenesis dysfunction. Mechanistically, TP could enhance GPX4 ubiquitin degradation via triggering K63-linked polyubiquitination of GPX4, thereby stimulating ferroptosis in spermatocytes. Functionally, GPX4 deletion intensified ferroptosis and exacerbated DNA damage in GC-2 cells, while GPX4 overexpression mitigated ferroptosis induced by TP. Overall, these findings for the first time indicated a vital role of ferroptosis in TP induced-testicular injury and spermatogenic dysfunction through promoting GPX4 K63-linked polyubiquitination, which hopefully offers a potential therapeutic avenue for TPrelated male reproductive damage. In addition, this study also provides a theoretical foundation for the improved clinical application of TP or TwHF in the future.

BACKGROUND:The development of potent non-toxic chemotherapeutic drugs against castration resistant prostate cancer (CRPC) remains a major challenge. Corosolic acid (CA), a natural triterpenoid, has anti-cancer activity with limited side effects. However, CA anti-prostate cancer activities and mechanisms, particularly in CRPC, are not clearly understood. In this study, we investigated CA anti-tumor ability against human CRPC and its mechanism of action.METHODS:The cell apoptosis and proliferation effects were evaluated via MTT detection, colony formation assay and flow cytometry. Western blot, gene transfection and immunofluorescence assay were applied to investigate related protein expression of Endoplasmic reticulum stress. A xenograft tumor model was established to investigate the inhibitory effect of CA on castration resistant prostate cancer in vivo.RESULTS:The results showed that CA inhibited cell growth and induced apoptosis in human prostate cancer cell (PCa) line PC-3 and DU145, as well as retarded tumor growth in a xenograft model, exerting a limited toxicity to normal cells and tissues. Importantly, CA activated endoplasmic reticulum (ER) stress-associated two pro-apoptotic signaling pathways, as evidenced by increased protein levels of typical ER stress markers including IRE-1/ASK1/JNK and PERK/eIF2α/ATF4/CHOP. IRE-1, PERK or CHOP knockdown partially attenuated CA cytotoxicity against PCa cells. Meanwhile, CHOP induced expression increased Tribbles 3 (TRIB3) level, which lead to AKT inactivation and PCa cell death. CHOP silencing resulted in PCa cells sensitive to CA-induced apoptosis.CONCLUSION:Our data demonstrated, for the first time, that CA might represent a novel drug candidate for the development of an anti-CRPC therapy.

Osteosarcoma (OS) is the most frequent and high mortality primary bone tumor in the adolescent. And it is well-known for poor prognosis due to high incidence of metastasis. Raddeanin A (RA), an active component of Anemone raddeana Regel, showed potential anti-cancer activities. However, the anti-tumor effect and molecular mechanism(s) of RA on osteosarcoma are still unclear. The present research is the first in vitro and in vivo investigate systematically anticancer of RA on human osteosarcoma. Our study demonstrated that RA induced mitochondria-dependent apoptosis in osteosarcoma cell lines and markedly suppressed the metastasis of osteosarcoma cells in vitro. And, RA treatment markedly inhibits tumor growth in vivo. Further mechanism study demonstrated that RA caused a significant enhance reactive oxygen species (ROS) level to stimulate phosphorylation of JNK. Moreover, RA led to decrease of p-IκBα level in the cytosol and reduction of p65 level in the nucleus, which was associated with the inhibition of NF-κB transcriptional activity. When NF-κB signaling was inhibited by siRNA targeting p65, a significant increase in cell apoptosis activity was observed. In addition, non-toxic RA concentrations (0.25, 0.5 and 1 μM) inhibited the migration and invasion of OS by suppressing MMP-2/9 expression associated with NF-κB-dependent transcription in vitro. The silencing of p65 increased the sensitivity of the osteosarcoma cells to RA suppressed migration and invasion. These findings suggest RA induces apoptosis and inhibits metastasis in OS cells, involved in provoking ROS/JNK and inhibiting NF-κB signaling pathways. Therefore, it may be a potential anti-metastatic and anti-proliferative therapeutic agent for human osteosarcoma.

AIMS:Colorectal cancer (CRC) remains one of the most lethal human malignancies with high incidence and lack of effective therapy. Raddeanin A (RA), an active triterpenoid saponins, has been demonstrated the ability to inhibit the growth of tumor. But the therapeutic effects and mechanisms of RA in CRC remain elusive. Here, we investigated the efficacy and mechanism of RA in CRC both in vitro and in vivo. MAIN METHODS:Cell viability was investigated to evaluate cytotoxic activity by MTT method. Apoptosis induced by RA was studied using Annexin V-FITC/PI binding and JC-1 staining by flow cytometry analysis. The xenograft mouse model of CRC was used to investigate anti-tumor effects in vivo. The key proteins involved in mitochondrial apoptotic, Wnt/β-catenin and NF-κB pathway were detected by Western blotting, Immunofluorescence, and Immunohistochemistry. KEY FINDINGS:RA induced apoptosis and inhibited cell proliferation of SW480 and LOVO cells in a concentration-dependent manner. Moreover, RA efficiently inhibited tumor growth in xenograft mouse model. RA could down regulate the Wnt/β-catenin signaling to display anti-tumor effects via suppression of p-LRP6, induction of AKT inactivation, removal of GSK-3β inhibition and attenuation of β-catenin. Meanwhile, RA also suppressed the NF-κB pathway by decreasing the phosphorylation of IKBα to induce subsequently mitochondrial apoptotic pathway. SIGNIFICANCE:In summary, RA suppressed the growth and triggered the apoptosis of CRC through discontinuing Wnt/β-catenin signaling and inhibiting the NF-κB pathway. These findings suggested that RA may hold a promise as a novel therapeutic agent for CRC therapy.

Triptolide (TP), an oxygenated diterpene, has a variety of beneficial pharmacodynamic activities but its clinical applications are restricted due to severe testicular injury. This study aimed to delineate the molecular mechanisms of TP-induced testicular injury in vitro and in vivo . TP (5–50000 nmol/L) dose-dependently decreased the viability of TM4 Sertoli cells with an IC 50 value of 669.5–269.45 nmol/L at 24 h. TP (125, 250, and 500 nmol/L) dose-dependently increased the accumulation of ROS, the phosphorylation of JNK, mitochondrial dysfunction and activation of the intrinsic apoptosis pathway in TM4 cells. These processes were attenuated by co-treatment with the antioxidant N-acetyl cysteine (NAC, 1 mmol/L). Furthermore, TP treatment inhibited the translocation of Nrf2 from cytoplasm into the nucleus as well as the expression of downstream genes NAD(P)H quinone oxidoreductase1 (NQO1), catalase (CAT) and hemeoxygenase 1 (HO-1), thus abrogating Nrf2-mediated defense mechanisms against oxidative stress. Moreover, siRNA knockdown of Nrf2 significantly potentiated TP-induced apoptosis of TM4 cells. The above results from in vitro experiments were further validated in male mice after oral administration of TP (30, 60, and 120 mg·kg −1 ·d −1 , for 14 d), as evidenced by the detected indexes, including dose-dependently decreased SDH activity, increased MDA concentration, altered testicle histomorphology, elevated caspase-3 activation, apoptosis induction, increased phosphorylation of JNK, and decreased gene expression of NQO1, CAT and HO-1 as well as nuclear protein expression of Nrf2 in testicular tissue. Our results demonstrate that TP activates apoptosis of Sertoli cells and injury of the testis via the ROS/JNK-mediated mitochondrial-dependent apoptosis pathway and down-regulates Nrf2 activation.

Functional organic molecules/metal-organic frameworks composites can be obtained by in situ crystalline structure transformation from ZIF-L to ZIF-8-L under double solvent conditions. Interestingly, the as-prepared molecules/ZIF-8-L composites with the leaf-like morphology exhibit good fluorescence properties and size selectivity in fluorescent quenchers due to the molecular sieving effect of the well-defined microporous ZIF-8-L.

Scattering and backscattering properties were measured for 4 main fresh water alga species cultured in the laboratory: Microcystis aeruginosa , Chlorella vulgaris , Cyclotella meneghiniana and Cryptomonas ovata . Chlorophyll a-specific scattering and backscattering were used to represent scattering ability. M. aeruginosa had the strongest scattering and backscattering ability, followed by C. meneghiniana , and C. vulgaris had the weakest. M. aeruginosa and C. meneghiniana showed greater backscattering ability than C. vulgaris and C. ovata . The main factor affecting backscattering was the ratio of chlorophyll a to phycocyanin content. Backscattering coefficient varied with chlorophyll a content and chlorophyll a proportion to total pigment. We found a good correlation between cell size and backscattering coefficient.

In this paper, we describe the synthesis and testing of a novel β-cyclodextrin derivative, carboxymethy-hydroxypropyl-β-cyclodextrin (CM-HP-β-CD). After synthesis, we verified that carboxymethyl and hydroxypropyl groups were successfully substituted onto β-cyclodextrin to form CM-HP-β-CD. Then, we performed in vitro experiments to investigate (1) the ability of CM-HP-β-CD to bind insulin, (2) the transportation of insulin-CM-HP-β-CD complexes across a Caco-2 cell monolayer, and (3) the cytotoxicity of CM-HP-β-CD. CM-HP-β-CD showed the strongest insulin-binding ability and the best transport properties compared with the other β-cyclodextrin derivatives we evaluated. In addition, CM-HP-β-CD showed no cytotoxicity up to 200 mM (IC50>200 mM). To examine the effectiveness of the cyclodextrin complexes in delivering insulin in vivo, we administered different insulin-cyclodextrin complexes to diabetic rats. In these studies, we found that the insulin-CM-HP-β-CD complex provided a significant and sustained (5 h) reduction in the blood glucose levels of diabetic rats. Overall, our study showed that CM-HP-β-CD may be a promising protein carrier for use in oral protein drug delivery.

According to ADA theory and Kramers-Kronig relationship, the real and imaginary parts of complex refractive index were estimated by the datasets of microcystis aeruginosa algal absorption and particle size distribution. Then, the optical properties of microcystis aeruginosa were calculated based on the Mie theory. The relative deviations between modeled and measured scattering efficiency factors are 10. 08%, and 4. 8% between absorption efficiency factors. The agreements between simulation result and actual measurement show that homogenous spherical model can be used to reproduce the absorption and scattering property of microcystis aeruginosa.

铜绿微囊藻是我国内陆富营养化湖泊蓝藻水华的主要藻种.将铜绿微囊藻抽象为由细胞质和叶绿体构成的两层球形模型,通过循环迭代的方式确定两层球形模型中叶绿体的体积比例及其折射率的中心值,结合Kramers-Kronig关系最终确定两层球形模型细胞质及叶绿体的复折射率,通过Aden-Kerker理论结合粒径分布情况模拟出其吸收和散射效率光谱.对比模拟结果与实验结果发现:该模型估算得到的散射光谱与实验的散射光谱之间误差为3.4％,吸收效率误差为4.3％,在后向散射模拟实验中,相比均匀球形模型,两层球形模型表现出较好的优越性.

We have reported that intracellular glutathione S-transferases P1 (GSTP1) suppresses LPS (lipopolysaccharide)-induced excessive production of pro-inflammatory factors by inhibiting LPS-stimulated MAPKs (mitogen-activated protein kinases) as well as NF-kappaB activation. But under pathogenic circumstances, physiologic levels of GSTP1 are insufficient to stem pro-inflammatory signaling. Here we show that LPS-induced up-regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in RAW246.7 cells is significantly reduced by incubating cells with recombinant GSTP1 protein. In vivo study demonstrates that treatment of mice (i.p.) with recombinant GSTP1 protein effectively suppresses the devastating effects of acute inflammation, which includes reduction of mortality rate of endotoxic shock, alleviation of LPS-induced acute lung injury and abrogation of thioglycolate (TG)-induced peritoneal deposition of leukocytes and polymorphonuclear cells (PMNs). Meanwhile, GSTP1 prevented LPS-induced TNF-alpha, IL-1beta, MCP-1 and NO production. Further investigation by using confocal microscopy and flow cytometry shows that recombinant GSTP1 protein can be delivered into RAW246.7 cells, mouse peritoneal macrophages and HEK 293 cells suggesting that extracellular GSTP1 protein could be transported across plasma membrane and act as a cytosolic protein. In conclusion our research demonstrates a new finding that increasing cellular GSTP1 level by supplement of recombinant GSTP1 effectively suppresses the devastating effects of acute inflammation.

Dipyrithione (PTS2) possesses anti‐bacterial and anti‐fungal activity. In the present study, we found that PTS2 dose‐dependently inhibited the LPS‐induced up‐regulation of nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2) protein level in RAW264.7 cells. RT‐PCR experiments showed that PTS2 suppressed LPS‐induced iNOS but not COX‐2 expression at the mRNA level. As expected, PTS2 prevented NO secretion in RAW264.7 cells. Furthermore, PTS2 administration significantly decreased LPS‐induced mortality in mice. Mechanistically, PTS2 decreased expression and phosphorylation of STAT1, but did not interfere with the MAPK and NF‐κB pathways. In conclusion, PTS2 protects mice against endotoxic shock and inhibits LPS‐induced production of pro‐inflammatory mediators, suggesting that PTS2 could play an anti‐inflammatory role in response to LPS.

Human glutathione S-transferase P1 is a ubiquitous expressed protein which plays an important role in the detoxification and xenobiotics metabolism,and it is an endogenous protein.Our previous studies showed that GSTP1 prevented LPS-induced excessive production of pro-inflammatory factors and played an anti-inflammatory role in RAW264.7 macrophage-like cells.In the present study we investigate whether exogenous GSTP1 protein can enter into RAW264.7 macrophage-like cells and mouse peritoneal macrophages by confocal laser scanning microscopy and Flow Cytometry.The results demonstrated that GSTP1 protein can be transported across plasma membrane into RAW264.7 cells and mouse peritoneal macrophages.

Aim of the study: Shuang-Qing-Cao (SQC) is a folk Chinese medicinal formula. The therapeutic effects of inclusion complexation of SQC extract in beta-cyclodextrin (SQC-beta-CD) against lipopolysaccharide (LPS)-induced acute lung injury (ALI) were studied in mice.Materials and Methods: Two protocols were designed for administration of SQC-beta-CD (10 and 20 mg/kg body weight) or DEX (2 mg/kg). According to Protocol A we intraperitoneally injected diluent (saline with 0.5% Tween 80), SQC-beta-CD or DEX respectively into mice 30 min and 3 h after LPS challenge. Alternatively, in Protocol B we administered diluent, SQC-beta-CD or DEX 3 h before and 30 min after LPS challenge.Results: The histological results showed that SQC-beta-CD (20mg/kg) protected mice from LPS-induced ALI such as oedema, haemorrhage, blood vessel and alveolar structural damage. Furthermore, SQC-beta-CD inhibited LPS-increased pulmonary MPO activity and migration of polymorphonuclear neutrophils (PMNs) into bronchoalveolar lavage fluid (BALF). Immunohistochemical experiment demonstrated that SQC-beta-CD decreased inducible nitric oxide synthase (iNOS) expression in lung 24 h after LPS administration. Consequently, SQC-beta-CD prevented LPS-induced nitric oxide (NO) release in BALEConclusions: The results indicated that SQC-beta-CD is greatly effective in inhibiting ALI. The present study indicated that SQC-beta-CD acted as a potential therapeutic reagent for treating ALI. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

The privilege of lawyers is an important part of lawyer's rights.Due to the rapid development of criminal defense,it is urgent to build up the privilege of lawyers so as to protect the accused's legal rights and the smooth development of lawyer system.

Superoxide dismutase (SOD) activity of erythrocyte in mice and intracellular free calcium concentration ([Ca2+]i) of single human lymphocyte in mice and rat neuron were investigated before and after low level laser irradiation by use of SOD inhibiting method and Ca2+ sensitive indicator Fura-2 with Spex AR-CM-MIC Cation Measurement System. The findings suggest that intravascular low level laser irradiation (ILLLI) enhanced SOD activity of mouse erythrocyte, and the effects induced by 6.75��103 J/cm2day were better than those did by 2.70��104J/cm2day. It seemed that irradiation with optimum constants would result in the best effect on organism.