Validating the role of PTGIS gene in colorectal cancer by bioinformatics analysis and in vitro experiments.

Prostaglandin I2 synthase (PTGIS) is a member of the cytochrome P450 family. Studies have revealed that differential expression of the PTGIS gene is closely related to the pathological and physiological processes of many diseases, including breast cancer, oral squamous cell carcinoma, and head and neck cancer. However, the mechanism of action of the PTGIS gene in colorectal cancer is not fully understood. This study explored the role of PTGIS in colorectal cancer through comprehensive bioinformatics analysis and in vitro experiments, and found that the expression of PTGIS gene in colorectal cancer tissue was significantly lower than that in normal colorectal tissue (P < 0.05), and high expression of PTGIS gene was associated with poor prognosis in patients (P < 0.05). The KEGG results showed that PTGIS-related genes were mainly enriched in metabolic pathways, arachidonic acid metabolism, steroid biosynthesis, and cancer pathways. The expression of PTGIS may be related to immune infiltration. Cell experiments showed that PTGIS was expressed at a lower level in cancer. Overexpression of PTGIS inhibited apoptosis and promoted proliferation, invasion, and migration ability of SW480 colorectal cancer cells. Analysis of the PTGIS gene in this study provides a theoretical basis for further exploring the pathogenesis of colorectal cancer and finding more accurate new targets for early screening and treatment of the cancer.

Cornel Iridoid Glycoside and Its Effective Component Regulate ATPase Vps4A/JNK to Alleviate Autophagy Deficit with Autophagosome Accumulation.

Improving autophagy-lysosome fusion has been considered a key method in the treatment of Alzheimer's disease (AD). Cornel iridoid glycoside (CIG) is extracted from Cornus officinalis and has been shown to promote the clearance of tau oligomers via the autophagy pathway. However, the mechanisms of CIG on autophagy deficits are not understood. Here, we found autophagy deficit and tau aggregation in the brains of P301S tau transgenic mice and MAPT cells edited using CRISPR-Cas9 technology. CIG decreased tau aggregation and alleviated autophagic markers involving the JNK/Beclin-1 signaling pathway which demonstrated CIG that might enhance lysosome formation by upregulating ATPase Vps4A expression. Knocking down VPS4A increased autophagosome accumulation and attenuated the effect of CIG on p62. In addition, CIG had no effect on tau oligomers but still inhibited the level of tau monomer in VPS4A knockout cells. The effective component (Sweroside, SWE) of CIG attenuated tau oligomers accumulation and increased Vps4A level but not CHMP2B. SWE could not change the level of tau oligomers in VPS4A knockout cells. In conclusion, CIG suppressed autophagosome accumulation by regulating the ATPase Vps4A/JNK. SWE is a core of active factors of CIG in Vps4A regulation. These findings suggest CIG may be a potential drug in AD treatment.

Cornel Iridoid Glycoside Regulates Modification of Tau and Alleviates Synaptic Abnormalities in Aged P301S Mice.

Alzheimer's disease (AD), also defined as a tauopathology, is a common neurodegenerative disease. Hyper-phosphorylation, cleavage or truncation, and aggregation of tau contribute to AD. Thus, targeting the post-translational modifications on tau may be a therapeutic strategy to treat AD. This study understood how cornel iridoid glycoside (CIG) affects tau post-translational modifications and synaptic abnormalities. The 10-month old P301S tau transgenic mice were given CIG at 100 and 200 mg/kg every day orally for 1 month. Hyperphosphorylated and truncated tau, synapse-associated proteins and glutamatergic receptors were all detected using Western blotting. The interactions between Morroniside (MOR) or Loganin (LOG) and tau were detected using Autodock and Surface Plasmon Resonance (SPR). The effects of CIG on the aggregation of tau were investigated using a cell-free system. CIG attenuated tau hyperphosphorylation at Thr205, Ser212, Ser262, Thr231 and Ser235 (AT180), but had no effect on tau truncation in the brains of 10-month old P301S mice. Binding free energies and interactions revealed that MOR and LOG bound with tau. We also found that CIG upregulated synapse-associated proteins such as PSD-95, syntaxin1A and synaptotagmin. In addition, CIG restored N-methyl-D-aspartic acid receptor and glutamate receptor levels. CIG improves post-translational modification of tau as well as synaptic abnormalities. The data presented here reveal that CIG may be used in the treatment of AD.

Essential Oil from Carpesium abrotanoides L. Induces Apoptosis via Activating Mitochondrial Pathway in Hepatocellular Carcinoma Cells.

The effects of essential oil from Carpesium abrotanoides L. (CAEO) on the proliferation and apoptosis of human hepatic cancer cells were investigated in this study. MTT assays indicated that CAEO inhibited the proliferation of HCC cells with the IC50 values ranging from 41.28±3.06 to 130.36±20.79 µg/mL. Moreover, many obviously nuclear morphological changes of apoptotic cells in CAEO-treated HepG2 cells were detected by Hoechst 33258 staining and fluorescence microscopy. Flow cytometry was used to detect cell apoptosis and cell cycle, and noticeable findings showed that CAEO arrested cell-cycle at S and G2/M phases. The decreased Bcl-2/Bax protein ratio and the activation of caspase-3, caspase-9 were also detected by Western blotting. All results suggested that CAEO is a potential agent to fight against liver cancer, and the mitochondria-mediated intrinsic apoptotic pathway could be involved in CAEO-mediated apoptosis of human liver carcinoma cells.

Essential oil from Siegesbeckia pubescens induces apoptosis through the mitochondrial pathway in human HepG2 cells.

Siegesbeckia pubescens (SP) has been used as a traditional medicine for the treatment of and inflammatory diseases. However, the activities of SP against hepatocellular carcinoma and the related mechanisms remain unclear. The present study aimed to examine the effects of the essential oil of SP (SPEO) on the proliferation of hepatocellular carcinoma cells and the possible mechanisms. The growth inhibition of HepG2 cells was analyzed by MTT assay. Hoechst 33258 and fluorescence microscopy were utilized to observe the nuclear morphological changes of apoptotic cells. Flow cytometry was used to detect cell apoptosis and cell cycle. The expressions of the target proteins were detected by Western blotting. The results showed that SPEO obviously inhibited the proliferation of HepG2 cells in a dose-dependent manner. SPEO activated a series of apoptotic proteins in HepG2 cells, increasing expression levels of Bax, caspase-3 and caspase-9, and decreasing the bcl-2 expression level. SPEO displayed promising anti-hepatocellular carcinoma activities in vitro, partly by inducing apoptosis in HepG2 cells through activating the mitochondrial pathway.

[Studies on the genetic variation of two mitochondrial DNA molecules of Schistosoma japonicum].

OBJECTIVE: To study the variation of Schistosoma japonicum through two mitochondrial DNA molecules. METHODS: Genomic DNA was isolated with kit, and the mitochondrial NADH dehydrogenase 1 (ND1) and cytochrome c oxidase I (COI) gene fragments were amplified by polymerase chain reaction (PCR) and sequenced. The gene trees were constructed and the acquired data were analyzed with the help of bioinformatics. RESULTS: The gene trees showed that the Taiwan isolate and the mainland isolates can be divided in two groups: a group from the hilly region (Yunnan and Sichuan), another group from the lake region (Hunan, Jiangxi and Anhui); isolates from Hubei are at a different position on the gene trees. CONCLUSION: There are variations among the geographic isolates of Schistosoma japonicum in China, nevertheless, they have close kinship.

Essential Oil from Siegesbeckia pubescens Induces Apoptosis through the Mitochondrial Pathway in Human HepG2 Cells / 华中科技大学学报（医学）（英德文版）

Siegesbeckia pubescens (SP) has been used as a traditional medicine for the treatment of and inflammatory diseases.However,the activities of SP against hepatocellular carcinoma and the related mechanisms remain unclear.The present study aimed to examine the effects of the essential oil of SP (SPEO) on the proliferation of hepatocellular carcinoma cells and the possible mechanisms.The growth inhibition of HepG2 cells was analyzed by MTT assay.Hoechst 33258 and fluorescence microscopy were utilized to observe the nuclear morphological changes of apoptotic cells.Flow cytometry was used to detect cell apoptosis and cell cycle.The expressions of the target proteins were detected by Western blotting.The results showed that SPEO obviously inhibited the proliferation of HepG2 cells in a dose-dependent manner.SPEO activated a series of apoptotic proteins in HepG2 cells,increasing expression levels of Bax,caspase-3 and caspase-9,and decreasing the bcl-2 expression level.SPEO displayed promising anti-hepatocellular carcinoma activities in vitro,partly by inducing apoptosis in HepG2 cells through activating the mitochondrial pathway.

[The effect of CTLA4Ig-modified dendritic cells on proliferation and cytotoxicity of lymphocytes in-vitro].

AIM: To study the effect of dendritic cells (DCs) modified by CTLA4Ig on T-cell proliferation and the cytotoxic T lymphocyte killer activity. METHODS: The expressing vector pG/CTLA4Ig was transfected into DCs via lipofectamine reagent mediation. CTLA4Ig fusion protein secreted by DCs in the culture supernatant was confirmed by sandwich ELISA and SDS-PAGE. Peripheral blood mononuclear cells (PBMC) from C57BL/6 mice (as reaction cells) and DCs modified or unmodified by CTLA4Ig (as stimulation cells) were co-cultured for 6 days. MTT colorimetry was used to detect lymphocyte proliferation. Lactate dehydrogenase release method and sandwich ELISA assay was used to examine the cytotoxic activity and T-cell apoptosis. RESULTS: CTLA4Ig fusion protein and DCs modified by CTLA4Ig could significantly inhibit lymphocyte proliferation response and cytotoxic activity of specific cytotoxic T lymphocytes, and induce T-cell apoptosis, while unmodified-DCs could markedly induce lymphocyte proliferation response. CONCLUSION: The DCs of stably expressing CTLA4Ig fusion protein could not only markedly inhibit T-cell proliferation and cytotoxic activity of CTL, but also induce T-cell apoptosis.