RNA-binding protein GIGYF2 orchestrates hepatic insulin resistance through STAU1/PTEN-mediated disruption of the PI3K/AKT signaling cascade.

BACKGROUND: Obesity is well-established as a significant contributor to the development of insulin resistance (IR) and diabetes, partially due to elevated plasma saturated free fatty acids like palmitic acid (PA). Grb10-interacting GYF Protein 2 (GIGYF2), an RNA-binding protein, is widely expressed in various tissues including the liver, and has been implicated in diabetes-induced cognitive impairment. Whereas, its role in obesity-related IR remains uninvestigated. METHODS: In this study, we employed palmitic acid (PA) exposure to establish an in vitro IR model in the human liver cancer cell line HepG2 with high-dose chronic PA treatment. The cells were stained with fluorescent dye 2-NBDG to evaluate cell glucose uptake. The mRNA expression levels of genes were determined by real-time qRT-PCR (RT-qPCR). Western blotting was employed to examine the protein expression levels. The RNA immunoprecipitation (RIP) was used to investigate the binding between protein and mRNA. Lentivirus-mediated gene knockdown and overexpression were employed for gene manipulation. In mice, an IR model induced by a high-fat diet (HFD) was established to validate the role and action mechanisms of GIGYF2 in the modulation of HFD-induced IR in vivo. RESULTS: In hepatocytes, high levels of PA exposure strongly trigger the occurrence of hepatic IR evidenced by reduced glucose uptake and elevated extracellular glucose content, which is remarkably accompanied by up-regulation of GIGYF2. Silencing GIGYF2 ameliorated PA-induced IR and enhanced glucose uptake. Conversely, GIGYF2 overexpression promoted IR, PTEN upregulation, and AKT inactivation. Additionally, PA-induced hepatic IR caused a notable increase in STAU1, which was prevented by depleting GIGYF2. Notably, silencing STAU1 prevented GIGYF2-induced PTEN upregulation, PI3K/AKT pathway inactivation, and IR. STAU1 was found to stabilize PTEN mRNA by binding to its 3'UTR. In liver cells, tocopherol treatment inhibits GIGYF2 expression and mitigates PA-induced IR. In the in vivo mice model, GIGYF2 knockdown and tocopherol administration alleviate high-fat diet (HFD)-induced glucose intolerance and IR, along with the suppression of STAU1/PTEN and restoration of PI3K/AKT signaling. CONCLUSIONS: Our study discloses that GIGYF2 mediates obesity-related IR by disrupting the PI3K/AKT signaling axis through the up-regulation of STAU1/PTEN. Targeting GIGYF2 may offer a potential strategy for treating obesity-related metabolic diseases, including type 2 diabetes.

Analysis of the clinical outcomes of microbial contamination caused by environmental contamination of the embryology laboratory during IVF-ET treatment cycles.

BACKGROUND: Bacterial contamination may cause loss of or damage to cultured oocytes or embryos, resulting in the lack of transplantable embryos during IVF embryo culture. However, there are few reports about IVF embryo contamination caused by embryology laboratories. In this work, we evaluated clinical pregnancy outcomes and the risk of maternal and infant complications after embryo contamination caused by environmental pollution during IVF. METHODS: The authors retrospectively analyzed 2490 IVF-ET ovulation induction therapy cycles in the Reproductive Center of Yichang Central People's Hospital from January 2015 to May 2022. According to the presence or absence of embryo culture medium contamination, the two groups were divided into an embryo contamination cycle and a nonembryo contamination cycle. The primary outcome parameters were the characteristics and progress of embryo culture medium contamination. Embryo laboratory outcomes, pregnancy outcomes, and maternal and infant complications were secondary outcome parameters. RESULTS: One case of embryo contamination originated from semen contamination. The remaining 15 cases involved environmental contamination outbreaks in embryo culture chambers, caused by Staphylococcus pasteuri. Compared with conventional uncontaminated IVF cycles, the 15 cases of contaminated embryo cycles showed no significant difference in embryo laboratory outcomes, pregnancy outcomes, or maternal and infant complications except for a slightly higher rate of fetal growth retardation. Ultimately, 11 live-born infants were successfully delivered, of which 2 were premature. The remaining 4 patients did not become pregnant after 1-2 transfers due to a lack of transferable embryos. CONCLUSION: When the embryo culture medium is contaminated due to the environmental contamination of the IVF culture room, it is feasible to perform daily rapid rinsing of the culture medium and avoid blastocyst culture as remedial treatment. However, the long-term impact on offspring needs further prospective research.

Primary diffuse large B-Cell lymphoma of the uterine cervix with severe lower urinary tract Symptoms: A rare case report and review of the literature.

Background: Diffuse large B-cell lymphoma (DLBCL) is a rare disease with a crude annual incidence rate of 3.8 cases per 100,000 people. Besides, primary cervical lymphoma is very rare, accounting for only 0.008% of cervical malignancies. (Sant et al., 2010) Although DLBCL patients often present with abnormal vaginal bleeding, it was not involved in this case. In this article, we present a rare case of primary cervical diffuse large B-cell lymphoma with urinary tract symptoms. Case: A 71-year-old woman who had been suffering from dysuria for two months came to our hospital. A pelvic examination revealed a 10 cm cervical mass, while HPV and squamous cell carcinoma (SCC) antigen tests were negative. The bulky cervical mass invaded the posterior wall of the uterus, vagina, superior rectum, bladder, and bilateral lower ureters, resulting in dysuria and dilatation of the upper ureter. Histopathological and immunohistochemical examination confirmed DLBCL and PET-CT suggested that it was stage IV. After two cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone), the large lesions were eliminated. Unfortunately, the patient suffered an untimely death unrelated to her disease before the fourth cycle of R-CHOP could begin. Conclusions: DLBCL of the cervix is a rare, but potentially curable disease if the diagnosis is made accurately, and doing so requires a high index of suspicion for cervical masses with an atypical presentation in which traditional diagnostic methods are equivocal. Obtaining adequate multilayered lesion biopsies containing both cervical epithelium and mesenchyme helps to avoid misdiagnoses. Histopathological biopsy and immunohistochemistry are the gold standards for diagnosis, and R-CHOP chemotherapy is an effective treatment.

The long coding RNA AFAP1-AS1 promotes tumor cell growth and invasion in pancreatic cancer through upregulating the IGF1R oncogene via sequestration of miR-133a.

Long non-coding RNAs (lncRNAs) have been shown to play a significant role in the progression of many cancers, including pancreatic cancer (PC). However, the biological function and regulatory mechanisms of lncRNAs in PC remains largely unclear. The aim of this study was to identify and evaluate the potential functions of lncRNAs in PC and reveal the underlying mechanisms of their effects. Screening of published microarray data (GEO accession Nos. GSE16515 and GSE32688), revealed lncRNA AFAP1-AS1 to be one of the most upregulated lncRNAs in PC tissues. High expression of AFAP1-AS1 was correlated with advanced stages, tumor size and lymph node metastasis, as well as with poorer overall survival in patients with PC. Functionally, knockdown of AFAP1-AS1 by transfection with siRNA inhibited the proliferative and invasive capacities of PaCa-2 and SW1990 PC cells, promoted apoptosis of PC cells in vitro, and impaired in-vivo tumorigenicity. In particular, it was hypothesized that AFAP1-AS1 may act as a competitive endogenous RNA (ceRNA), effectively becoming a sink for miR-133a whose expression was found to be downregulated in PC tissues and cell lines, and which was negatively correlated with the expression of AFAP1-AS1. We also found that the IGF1R oncogene which is an important regulator of MEK/ERK signaling pathway, was positively regulated by AFAP1-AS1 through ameliorating miR-133a-mediated IGF1R repression in PC tissues. Moreover, we demonstrated that knockdown of IGF1R by transfection with si-IGF1R suppressed cell proliferation, invasion and migration of PaCa-2 and SW1990 PC cells, suggesting that IGF1R may function as an oncogene in PC cells. Further investigations revealed that miR-133a reversed the biological effects of AFAP1-AS1 on PC cells. Collectively, the findings provide new evidence that AFAP1-AS1 could regulate the progression of pancreatic cancer by acting as a ceRNA, and suggest it has potential for use as both a biomarker for the early detection PC and for the development of individualized therapies for PC.

Morphologic characteristics and proliferation of rabbit corneal stromal cells onto complexes of collagen-chitosan-sodium hyaluronate under simulated microgravity.

PURPOSE: We investigated the morphologic characteristics and proliferation of rabbit corneal stromal cells (CSCs) onto scaffolds under simulated microgravity. METHODS: Rabbit CSCs were cultured under simulated microgravity (SMG) and static condition. Complexes of collagen-chitosan-sodium hyaluronate with pores were used as scaffolds. Rotational speed was set at 15, 20, and 30 rpm in the first, second, and third weeks of culture, respectively. Histology, immunofluorescence staining, atomic force microscope (AFM), and scanning electron microscope (SEM) examinations were performed. The cell proliferation was analyzed by cell counting kit-8 (CCK-8) assay. RESULTS: In the SMG group, more CSCs adhered to the carriers in 24 hours. Confocal microscopic evaluation showed aggregated cells positively immunostained with vimentin. The SEM displayed the complex network of triangular or polygonal dendritic morphology of the cell bodies with many fine and long processes, which adhered to the scaffolds tightly. After 18 days of SMG culture, keratocyte-like CSCs with rich cell interconnections not only grew on the surface, but also into the interior of scaffolds. There were degradation phenomena in scaffolds in the SMG condition. Under static condition, cells just grew on scaffolds forming a monolayer. Cells showed elongating spindle shape and developed less processes. The absorbance values of the CCK-8 assay in the SMG group were significantly higher (P < 0.01) than in the conventional group. CONCLUSIONS: The condition of SMG and porous collagen-chitosan-sodium hyaluronate scaffolds facilitate the proliferation of CSCs. Cells showed robust growing characteristics and morphologic properties of keratocytes. The techniques for microgravity culture of keratocyte-like CSCs on scaffolds can yield cell aggregates or cell sheets that are favorable to the reconstruction of tissue engineering of the corneal stromal layer.

Oncostatin M promotes proliferation of ovarian cancer cells through signal transducer and activator of transcription 3.

Oncostatin M (OSM), a pleiotropic cytokine, has been shown to have distinctive effects in different tissues. In ovarian carcinoma, it is commonly co-expressed together with its receptors but its precise role and the underlying molecular mechanism governing its activity remains unclear. The aim of the present study was to investigate the function of the recombinant human OSM (rH-OSM) in human ovarian cancer. The study demonstrated that rH-OSM promotes the proliferation of SKOV3 ovarian cancer cells. Western blot analysis showed that phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), phosphorylated-extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) and p-p38 protein levels increased in the cell lines treated with rH-OSM. Proliferation in SKOV3 cells induced by rH-OSM was suppressed by inhibitors of p-p38 or p-ERK1/2. Western blot analysis showed that p-STAT3 protein levels decreased in SKOV3 cells treated with inhibitors of p-p38 prior to treatment with rH-OSM. Also, p-STAT3 levels did not increase in cells treated with inhibitors of ERK1/2 prior to treatment with rH-OSM. Cell proliferation was moderately increased, and p-ERK1/2 and p-p38 protein expression were similarly affected in STAT3-RNAi knocked-down SKOV3 cells treated with rH-OSM compared to the control group. The data demonstrate that the growth-promoting activity of rH-OSM may be mediated through different signaling pathways. ERK1/2 and p38 proteins regulate STAT3 expression in SKOV3 cells, while STAT3 may be pivotal to the proliferation of SKOV3 cells in vitro.

[Study on thermal decompositon properties of hexafluoropropane clean gaseous fire-extinguishing agent].

The thermal decomposition properties of hexafluoropropane clean gaseous fire-extinguishing agent were studied in tubular reactor from 500 to 750 degrees C and the decomposed gas was characterized by gas chromatography(GC), Fourier transform infrared spectroscopy (FTIR) and gas chromatography-mass spectrometry (GC-MS). Hydrogen fluoride was detected after the decomposed gas was analyzed by pH testing, while pentafluoropropylene was found by GC-MS. The results showed that hydrogen fluoride eliminated from hexafluoropropane was the main reaction, while pentafluoropropylene was the primary product during hexafluoropropane decomposition under high temperature. GC and FTIR results indicated that the reaction temperatures had significant effects on the thermal decomposition of hexafluoropropane. Haxafluropropane was steady at 500 degrees C, whereas started to decompose weakly at 600 degrees C. The degree of the thermal decomposition of hexafluoropropane was enhanced with the temperature increase. And hexafluoropropane underwent intense decompositon at 750 degrees C. FTIR can be used as a new method to study extinguishing mechanism of fluorine-containing fire extinguishing agent online.

An active artificial cornea with the function of inducing new corneal tissue generation in vivo-a new approach to corneal tissue engineering.

An active artificial cornea which can perform the function of inducing new cornea generation in vivo but does not need culture cells in vitro and which has similar optical and mechanical properties to those of the human cornea was constructed. An animal keratoplasty experiment using the artificial cornea as the implant showed that the animals' corneas could keep smooth surface and clear stroma postoperatively, and that the repopulation of the host's keratocytes, the degradation of the implant and new corneal tissue generation were completed at 5-6 months after surgery. Such an artificial cornea has several advantages over other corneal equivalents constructed in the typical way of tissue engineering: in having similar mechanical and optical properties to those of the human cornea and with no exogenetic cells, it can be used universally in different implantation surgeries without immunoreaction; it is easy to prepare and process into different shapes and sizes on a large scale, and suitable for long-distance transportation and long-term storage. All these characteristics make it a new approach to cornea tissue engineering having potential in many clinical applications.

Study on biocompatibility of complexes of collagen-chitosan-sodium hyaluronate and cornea.

In this study we investigated the biocompatibility of collagen-chitosan-sodium hyaluronate (Col-Chi-NaHA) complexes and cornea tissue, and the feasibility of Col-Chi-NaHA complexes as substrates for cultivating rabbit corneal cells. Different components of Col-Chi-NaHA complexes were prepared and tested. A circular complex film with a diameter of 6 mm was inserted into rabbit stomal pocket and traced for a period of 5 months. Clinical examination was made. Rabbit limbal corneal epithelial cells, corneal endothelial cells, and keratocytes were cultured primarily on complexes. Phase contrast microscope examination was made daily. Histological, immunohistochemical, and scanning electron microscopic examinations were carried out. The complexes of 20% collagen, 10% chitosan, and 0.5% sodium hyaluronate showed rather weak corneal edema and other responses. The degradation of materials was obvious after 5 months. Corneas were transparent and translucent. Cells seeded on Col-Chi-NaHA were allowed to proliferate and partly form confluent monolayer after 9 days in culture. Cultured cells were well attached to the complexes of 20% collagen, 10% chitosan, and 0.5% sodium hyaluronate, or 10% chitosan and 0.5% sodium hyaluronate. The results showed that Col-Chi-NaHA complexes had good biocompatibility with cornea. The complexes can degrade and be absorbed in cornea. Col-Chi-NaHA complex may be a suitable substrate for cultivating corneal cells and a feasible material as a scaffold of tissue-engineered cornea.

[Research advance of corneal tissue engineering].

OBJECTIVE: To review research progress of corneal tissue engineering. METHODS: The recent articles on corneal tissue engineering focus on source and selection of corneal cells, the effects of growth factors on culture of corneal cells in vitro. The preparation and selection of three-dimensional biomaterial scaffolds and their strong and weak points were discussed. RESULTS: The corneal tissue engineering cells come from normal human corneal cells. The embryo corneal cell was excellent. Several kinds of growth factors play important roles in culture, growth and proliferation of corneal cell, and incorporated into matrix. Growth factors including basic fibroblast growth factor, keratinocyte growth factor, transforming growth factor beta 1 and epidermal growth factor was favor to corneal cell. Collagen, chitosan and glycosaninoglycans were chosen as biomaterial scaffolds. CONCLUSION: Human tissue engineering cornea can be reconstructed and transplanted. It has good tissue compatibility and can be used as human corneal equivalents.

Human T-cell leukemia virus type 1 Tax protein stimulates the interferon-responsive enhancer element via NF-kappaB activity.

Gene expression in Rat-1 fibroblast cells transformed by Tax from human T-cell leukemia virus type 1 was studied using the reverse transcriptase polymerase chain reaction differential display technique. The analysis revealed eight genes that were upregulated and one gene that was suppressed in Tax-transformed cells. Interestingly, at least four of the upregulated genes were interferon-stimulated genes. Promoter analysis of the 2',5'-oligoadenylate synthetase gene, which was activated in both Tax-transformed Rat-1 cells and primary adult T-cell leukemia cells, demonstrated that Tax indirectly activates its interferon-responsive enhancer element in a nuclear factor-kappaB pathway-dependent manner, indicating a close association of interferon signaling with the transformation by Tax.