Chlorogenic acid can improve spermatogenic dysfunction in rats with varicocele by regulating mitochondrial homeostasis and inhibiting the activation of NLRP3 inflammasomes by oxidative mitochondrial DNA and cGAS/STING pathway.

In recent years, Varicocele (VC) has been recognized as a common cause of male infertility that can be treated by surgery or drugs. How to reduce the damage of VC to testicular spermatogenic function has attracted extensive attention in recent years. Among them, overexpressed ROS and high levels of inflammation may play a key role in VC-induced testicular damage. As the key mediated innate immune pathways, cGAS-STING shaft under pathological conditions, such as in cell and tissue damage stress can be cytoplasmic DNA activation, induce the activation of NLRP3 inflammatory corpuscle, triggering downstream of the inflammatory cascade reaction. Chlorogenic acid (CGA), as a natural compound from a wide range of sources, has strong anti-inflammatory and antioxidant activities, and is a potential effective drug for the treatment of varicocele infertility. The aim of this study is to investigate the role of CGA in the spermatogenic dysfunction of the rat testis induced by VC and the potential mechanisms. The results of this study have shown that CGA gavage treatment ameliorated the pathological damage of seminiferous tubules, increased the number of sperm in the lumen, and increased the expression levels of Occludin and ZO-1, which indicated the therapeutic effect of CGA on spermatogenic dysfunction in the testis of VC rats. Meanwhile, the damage of mitochondrial structure was alleviated and the expression levels of ROS, NLRP3 and pro-inflammatory cytokines (IL-1ß, IL-6, IL-18) were significantly reduced in the testicular tissues of model rats after CGA treatment. In addition, we demonstrated for the first time the high expression status of cGAS and STING in testicular tissues of VC model rats, and this was ameliorated to varying degrees after CGA treatment. In conclusion, this study suggests that CGA can improve the spermatogenic function of the testis by reducing mitochondrial damage and inhibiting the activation of the cGAS-STING axis, inhibiting the activation of the NLRP3 inflammasome, and improving the inflammatory damage of the testis, highlighting the potential of CGA as a therapeutic agent for varicocele infertility.

[Action mechanisms of Qianlie Jindan Tablets on chronic nonbcterial prostatitis in rats: An exploration based on non-targeted urine metabolomics].

OBJECTIVE: To explore the mechanisms of Qianlie Jindan Tablets (QLJD) acting on chronic nonbacterial prostatitis (CNP) in rats based on non-targeted urine metabolomics. METHODS: According to the body mass index, we equally randomized 30 eight-week-old male SD rats into a blank control, a CNP model control and a QLJD medication group. We established the CNP model in the latter groups and, from the 4th day of modeling, treated the rats in the blank and model control groups intragastrically with normal saline and those in the QLJD medication group with QLJD suspension, qd, for 30 successive days. Then we detected the changes in the metabolites of the rats by ultra-high-performance liquid chromatography-tandem mass spectrometry, and identified the differential metabolites in different groups by multivariate statistical analysis, followed by functional annotation of the differential metabolites. RESULTS: Eight common metabolites were identified by metabolomics analysis, of which 5 were decreased in the CNP model controls and increased in the QLJD medication group, while the other 3 increased in the former and decreased in the latter group. Creatinine and genistein were important differential metabolites, and the arginine and proline metabolic pathways and isoflavone biosynthesis pathways were the main ones for QLJD acting on CNP. Compared with the blank controls, the model controls showed up-regulated arginine and proline metabolic pathways, increased production of creatinine, down-regulated isoflavone biosynthetic pathway and decreased production of genistein. The above changes in the model controls were all reversed in the QLJD medication group. CONCLUSION: QLJD acts effectively on CNP in male rats by regulating L-arginine and proline metabolic pathways, as well as the isoflavone biosynthesis pathway and naringenin metabolism.

Tissue factor/activated factor VIIa induces matrix metalloproteinase-7 expression through activation of c-Fos via ERK1/2 and p38 MAPK signaling pathways in human colon cancer cell.

PURPOSE: Increased expression of tissue factor (TF) is associated with tumor invasion and metastasis in human colorectal cancer. We have previously observed that TF/FVIIa upregulates matrix metalloproteinase-7 (MMP-7) expression at the transcriptional level in colon cancer cells. MMP-7 overexpression is believed to play an important role in tumor invasion and metastasis. The aim of this study is to elucidate the molecular mechanisms by which TF/FVIIa induced MMP-7 expression and cell invasion in vitro. METHODS: Reverse transcription polymerase chain reaction, Western blot, luciferase assay, and chromatin immunoprecipitation (ChIP) were used to determine the potential mechanism and signaling pathways by which TF/FVIIa induced MMP-7 expression and cell invasion in LoVo cells. Small interfering RNA (siRNA) and cell invasion assay was used to examine whether blocking c-Fos expression could abolish FVIIa-mediated upregulation of MMP-7 and cell invasion in vitro. RESULTS: The results showed that FVIIa induced the upregulation of MMP-7 both at the mRNA and protein levels in a time- and dose-dependent manner and increased the invasive behavior of LoVo cells. FVIIa enhanced the promoter activity of MMP-7, and the activator protein-1 (AP-1) binding site was responsible for the activation. Site mutation of the AP-1 binding site in the promoter almost completely abolished FVIIa-mediated response. Furthermore, ChIP assay confirmed that FVIIa promoted the direct binding of c-Fos with the MMP-7 promoter in vivo. FVIIa also induced the expression and nuclear accumulation of the AP-1 subunit c-Fos. siRNA-mediated knockdown of c-Fos eliminated FVIIa-stimulated MMP-7 expression and cell migration in vitro. In addition, selective mitogen-activated protein kinase (MAPK) kinase (MEK1/2) inhibitor (PD98059) and p38 MAPK inhibitor SB203580 suppressed MMP-7 upregulation induced by FVIIa. CONCLUSIONS: Our data suggest that a novel TF/FVIIa/MAPK/c-Fos/MMP-7 axis plays an important role in modulating the invasion of colon cancer cells and blockage of this pathway holds promise to treat colon cancer metastasis.

Morphological description of the third instar larva of Lasiotrichius succinctus hananoi (Sawada) (Coleoptera: Scarabaeidae: Cetoniinae: Trichiini), using scanning electron microscopy.

Cetoniinae is one of the showiest scarab groups, exhibiting bright-metallic body colors, and usually attract great attention from entomologists and amateur collectors. Larvae of Cetoniinae show dramatically diversity on morphology and living habits. Although being considered one of the best-studied groups of Scarabaeidae, larvae have been described for less than 5% species to the known Cetoniinae. In this study, the final instar larva of Lasiotrichius succinctus hananoi was described using scanning electron microscopy. The larvae are peculiar for bearing a haptomeral process dividing 10 spines into two groups: six on the left side, four on the right side, different from the previous descriptions on L. succinctus (Pallas, 1781). The morphological differences under SEM imply the further requirement of taxonomic revision in Lasiotrichius. Both advantage and disadvantage of SEM utilizing in larval descriptions were briefly discussed.

Morphological description of the white grub Melolontha incana (Coleoptera: Scarabaeidae: Melolonthinae: Melolonthini).

Melolonthinae are the largest subfamily of Scarabaeidae, considered as serious pests for their larvae attacking plant roots and tubers. The edaphic larvae are difficult to be identified because the study on larval taxonomy is far from satisfactory. In this study, multivoltine white grubs Melolontha incana (Motschulsky, 1853) were investigated using light and scanning electron microscopy, in order to provide more morphological characters for the pest identification. The white grubs are atypical for the epipharynx bearing 14 heli arranged in two rows; the mandible is furnished with a patch of minute granules; the maxilla is equipped with 18 acute stridulatory teeth arranged in line; each femur and tibiotarsus is furnished ventrally with a cluster of fossorial setae. The morphological comparisons with the other melolonthine species were provided. The adaptative relationship between the morphological feature and the multivoltine life history were briefly discussed.

Description of the third instar larva of the stag beetle Prismognathus dauricus Motschulsky, 1860 (Coleoptera: Scarabaeoidea: Lucanidae) using electron microscopy.

Lucanidae have long received great attention for their adults. However, differentiating between the larvae of stag beetles is difficult and remains unsatisfactorily resolved due to the microscopic features separating groups. In the current study, the larvae of Prismognathus dauricus Motschulsky, 1860 were investigated using light and scanning electron microscopy. The larvae of P. dauricus are atypical for the following characters: i) the epipharynx possesses anteriorly eleven protophoba arranged in a semi-round line; ii) the par stridens consists of 31 ± 0.7 (N = 10) subconical teeth arranged in a slightly curved, longitudinal row; iii) the plectrum is composed of 60 ± 1.4 (N = 10) carinae; iv) the claws of thoracic legs are apically blunt and bear four setae. Moreover, the size arrangement of the larval spiracles also provides valuable taxonomic information. Our SEM study reveals larval ultramorphological characters useful for identification of this species for the first time.

Extrahepatic synthesis of coagulation factor VII by colorectal cancer cells promotes tumor invasion and metastasis.

BACKGROUND: Blood coagulation factor VII (FVII) is physiologically synthesized in the liver and released into the blood. Binding of FVII to tissue factor (TF) is related to the metastatic potential of tumor cells, also a significant risk factor in the development of hepatic metastasis in patients with colorectal cancer (CRC). It has been found that some cancer cells can produce FVII extrahepatically. However, little is known about FVII and CRC. We therefore hypothesized that CRC cells may synthese FVII, leading to tumor invasion and metastasis. METHODS: We detected the expression of FVII protein in 55 CRC specimens by immunohistochemical staining. The FVII mRNA in 45 of 55 CRC cases, 6 colon cancer cell lines and one hepatoma cell line was measured by real-time reverse transcription-PCR (RT-PCR). Transwell invasion assays were performed to evaluate the changes of cell migration and invasion of LoVo cancer cells in vitro. We further observed the likely effectors regulated by the TF/FVIIa complex Western blotting assay. RESULTS: Extrahepatic synthesis of FVII was detected in the cytoplasm of 32 (58.2%) CRC specimens by immunohistochemistry, but not in normal mucosa. Liver metastasis (P = 0.003) and TNM staging (P = 0.005) were significantly correlated with FVII antigen expression. The positive ratios in stages I, II, III and IV were 33.3%, 40.0%, 52.4% and 87.5%, respectively. The expression of FVII mRNA in CRC with hepatic metastasis was significantly higher than CRC without hepatic metastasis (5.33 ± 2.88 vs. 1.47 ± 0.51, P = 0.03). Ectopic FVIIa induced a slight increase (1.34-fold) in the number of migrating cells, which was inhibited by the specific TF antibody. The formation of TF/FVIIa complex resulted in a marked increase in the expression of matrix metalloproteinases (MMP)-2 (3.5-fold) and MMP-9 (4.7-fold) in a time-dependent and dose-dependent manner. CONCLUSIONS: Extrahepatic synthesis of FVII by CRC cells may promote tumor invasion and metastasis. MMPs, as downstream effectors of TF/FVIIa signaling, facilitate the development of metastasis in colon cancer.