The mTORC2/Akt/NFκB Pathway-Mediated Activation of TRPC6 Participates in Adriamycin-Induced Podocyte Apoptosis.

BACKGROUND/AIMS: Although increased expression and gain function of transient receptor potential cation channel 6 (TRPC6) has been associated with the pathogenesis of some proteinuric glomerular diseases, it remains elusive how TRPC6 participates in the process of podocyte damage. METHODS: The potential signaling responsible for TRPC6 activation was investigated using immunoblot assays in an in vitro podocyte injury model induced by Adriamycin (ADR). Podocyte apoptosis was measured using FITC-conjugated Annexin V and Propidium Iodide staining. The channel activity of TRPC6 was assessed using the Ca2+ influx assay. RESULTS: Increase of TRPC6 expression was detected in ADR-treated podocytes, and TRPC6 knockdown significantly decreased ADR-induced podocytes apoptosis. Following ADR treatment, phospho-mTORSer2481 and phospho-AktSer473 was significantly increased in a time-dependent manner, whereas phospho-mTORSer2448 and phospho-p70S6KThr389 showed no change. ADR-induced apoptosis was prevented by ku0063794 (a dual mTOR complexes inhibitor), not by rapamycin (a specific mTORC1 inhibitor). Furthermore, nuclear translocation of NFκB/p65 was detected in ADR-treated podocytes, which was prevented by an Akt inhibitor triciribine. Of note, NFκB inhibitor PDTC prevented ADR-induced increase of TRPC6, and decreased ADR-induced apoptosis. We found that Akt activation and NFκB nuclear translocation was significantly inhibited by knockdown of mTORC2 protein Rictor, not by mTORC1 protein Raptor. In comparison with control, the Ca2+ influx was significantly increased in ADR-treated podocytes, which was remarkably prevented by TRPC6 knockdown. ADR-induced increase of TRPC6 channel activity was dramatically prevented by ku0063794, but not by rapamycin. Additionally, knockdown of Rictor, not Raptor, prevented ADR-induced increase of the Ca2+ influx. Moreover, the application of NFκB inhibitor PDTC also prevented the Ca2+ influx in ADR-treated podocytes. CONCLUSIONS: Our findings revealed that the mTORC2/Akt/NFκB pathway-mediated activation of TRPC6 participates in ADR-induced podocyte apoptosis.

Invalid phase values removal method for absolute phase recovery.

A novel approach is presented for more effectively removing invalid phase values in absolute phase recovery. The approach is based on a detailed study involving the types and cases of invalid phase values. Meanwhile, some commonalities of the existing removal algorithms also are thoroughly analyzed. It is well known that rough absolute phase and fringe order maps can very easily be obtained by temporal phase unwrapping techniques. After carefully analyzing the components and fringe order distribution of the rough fringe order map, the proposed method chiefly adopts an entirely new strategy to refine a pure fringe order map. The strategy consists of three parts: (1) the square of an image gradient, (2) subregion areas of the binary image, and (3) image decomposition and composition. In combination with the pure fringe order map and a removal criterion, the invalid phase values can be identified and filtered out from the rough absolute phase map. This new strategy not only gets rid of the limitations of traditional removal methods but also has a two-fold function. The paper also offers different metrics from the experiment to evaluate the quality of the final absolute phase. In contrast with other removal methods, experimental results have verified the feasibility, effectiveness, and superiority of the proposed method.

Matrine inhibits the growth of retinoblastoma cells (SO-Rb50) by decreasing proliferation and inducing apoptosis in a mitochondrial pathway.

Matrine, one of the main active components of extracts from the dry roots of Sophora flavescens, has potent anti-tumor activity in vitro and in vivo. Here, we investigated the apoptosis in matrine-treated retinoblastoma cells. The results showed that matrine could inhibit cell proliferation and induce apoptosis in a dose- and time-dependent manner. Further investigation revealed that a disruption of mitochondrial transmembrane potential and an up-regulation of reactive oxygen species in matrine-treated cells. By western blot analysis, we found that the up-regulation of cleaved Apaf-1, cleaved caspase-3, cleaved caspase-9, cleaved caspase-7, Bax/Bcl-2, varying with different concentration of matrine. These protein interactions may play a pivotal role in the regulation of apoptosis. Taken together, these results overall indicate that matrine could be used as an effective anti-tumor agent in therapy of retinoblastoma targets the caspase-dependent signaling pathway.

Development and validation of a novel scoring system based on a nomogram for predicting inadequate bowel preparation.

BACKGROUND AND AIMS: Adequate bowel preparation (BP) is crucial for the diagnosis of colorectal diseases. Identifying patients at risk of inadequate BP allows for targeted interventions and improved outcomes. We aimed to develop a model for predicting inadequate BP based on preparation-related factors. METHODS: Adult outpatients scheduled for colonoscopy between May 2022 and October 2022 were enrolled. One set (N = 913) was used to develop and internally validate the predictive model. The primary predictive model was displayed as a nomogram and then modified into a novel scoring system, which was externally validated in an independent set (N = 177). Inadequate BP was defined as a Boston Bowel Preparedness Scale (BBPS) score of less than 2 for any colonic segment. The model was evaluated by the receiver operating characteristic (ROC) curve, calibration plots, and decision curve analysis (DCA). RESULTS: Independent factors included in the prediction model were stool frequency ≤ 5 (15 points), preparation-to-colonoscopy interval ≥ 5 h (15 points), incomplete dosage (100 points), non-split dose (90 points), unrestricted diet (88 points), no additional water intake (15 points), and last stool appearance as an opaque liquid (0-80 points). The training set exhibited the following performance metrics for identifying BP failure: area under the curve (AUC) of 0.818, accuracy (ACC) of 0.818, positive likelihood ratio (PLR) of 2.397, negative likelihood ratio (NLR) of 0.162, positive predictive value (PPV) of 0.850, and negative predictive value (NPV) of 0.723. In the internal validation set, these metrics were 0.747, 0.776, 2.099, 0.278, 0.866, and 0.538, respectively. The external validation set showed values of 0.728, 0.757, 2.10, 0.247, 0.782, and 0.704, respectively, indicating strong discriminative ability. Calibration curves demonstrated close agreement, and DCA indicated superior clinical benefits at a threshold probability of 0.73 in the training cohort and 0.75 in the validation cohort for this model. CONCLUSIONS: This novel scoring system was developed from a prospective study and externally validated in an independent set based on 7 easily accessible variables, demonstrating robust performance in predicting inadequate BP.

Nicorandil protects against ischaemia-reperfusion injury in newborn rat kidney.

Ischaemia-reperfusion injury (IRI) is the predominant cause of acute kidney injury. Nevertheless, the underlying molecular mechanisms are still unclear. The current study investigated the effects of nicorandil on ATP-sensitive potassium (KATP) channels and the potential signal transduction pathway(s) in a rat kidney IRI model and in cultured tubular HK-2 cells subjected to oxygen and glucose deprivation/reoxygenation (OGD/R) injury. The standard procedure for IRI was performed in newborn rat kidneys. Pretreatment with nicorandil (10 mg/kg) 2 h prior to induction of IRI improved renal function, attenuated tubule damage, and prevented apoptosis of tubule cells, infiltration of neutrophils and macrophages, and production of inflammatory cytokines interleukin (IL)-6, IL-17 and tumour necrosis factor-α. Ischaemia-reperfusion-induced reduction of KIR6.2 was restored to normal levels by nicorandil. The activation of the phosphoinositide-3-kinase (PI3K)-Akt-nuclear factor (NF)-κB axis was detected in this rat kidney IRI model, which was blocked by nicorandil. The renoprotection of nicorandil against IRI was abolished by its inhibitor glibenclamide (1 mg/kg). Similar results were obtained in OGD/R-damaged HK-2 cells. Taken together, our findings demonstrated the specific renoprotective role of nicorandil in the newborn rat IRI kidney by decreasing the production of inflammatory cytokines, and restoring the expression of KIR6.2 potentially through the PI3K-Akt-NF-κB axis.

Role of peripheral blood mononuclear cell transportation from mother to baby in HBV intrauterine infection.

PURPOSE: We aimed to investigate the role of peripheral blood mononuclear cell transportation from mother to baby in hepatitis B virus (HBV) intrauterine infection. METHODS: Thirty HBsAg-positive pregnant women in the second trimester and their aborted fetuses were included in this study. Enzyme-linked-immunosorbent-assay was utilized to detect HBsAg in the peripheral blood of pregnant women and the femoral vein blood of their aborted fetuses. HBV-DNA in serum and peripheral blood mononuclear cells (PBMC) and GSTM1 alleles of pregnant women and their aborted fetuses were detected by nested polymerase chain reaction (PCR) and seminested PCR, respectively. We also examined the location of placenta HBsAg and HBcAb using immunohistochemical staining. The expression of placenta HBV-DNA was detected by in situ hybridization. RESULTS: For the 30 aborted fetuses, the HBV intrauterine infection rate was 43.33%. The HBV-positive rates of HBsAg in peripheral blood, serum, and PBMC were 10% (3/30), 23.33% (7/30), and 33.33% (10/30), respectively. Maternal-fetal PBMC transport was significantly positively correlated with fetal PBMC HBV-DNA (P = 0.004). Meanwhile, the rates of HBV infection gradually decreased from the maternal side to the fetus side of placenta (decidual cells > trophoblastic cells > villous mesenchymal cells > villous capillary endothelial cells). However, no significant correlation between placenta HBV infection and HBV intrauterine infection was observed (P = 0.410). CONCLUSIONS: HBV intrauterine infection was primarily due to peripheral blood mononuclear cell maternal-fetal transportation in the second trimester in pregnant women.

Enhancing bowel preparation quality and tolerability in a low health literacy population in Western China: a multicenter randomized trial.

BACKGROUND AND AIMS: Insufficient bowel preparation (BP) presents substantial challenges to the effectiveness of outpatient colonoscopy for colorectal cancer screening, particularly within populations characterized by low health literacy and poor adherence. METHODS: We conducted a prospective, randomized, blinded, endoscopic controlled study involving 474 colonoscopy outpatients aged 18-80 years hailing from a low health literacy population with convenient access to WeChat. These patients were subsequently randomized into three groups: the control group, WeChat group, and the automatic reminder group (ARG). All people were administered 3 L of polyethylene glycol. The Boston Bowel Preparation Scale (BBPS) score of 6 or a segmental score of 2 was used as the primary outcome to evaluate BP quality. Secondary outcomes included polyp detection rate (PDR) and adverse events, etc. RESULTS: Our findings revealed that both the WeChat group (n = 158) and ARG (n = 158) exhibited significantly higher rates of adequate BP compared to the control group (n = 158) (WeChat vs. control, 79.1% vs. 61.4%; ARG vs. control, 74.7% vs. 61.4%; p < 0.001). Furthermore, these educationally reinforced groups displayed improved BP compliance (p < 0.05). According to the Hospital Anxiety and Depression Scale (HADS), patients in the reinforced education groups exhibited lower overall anxiety levels (p = 0.001) and experienced fewer adverse reactions (p = 0.019). Compared to the control group, the PDR in the right hemi-colon was significantly greater in the WeChat group (11.4%) (2.5%), and a similar trend was observed in the ARG (7.6%). Additionally, individuals in the WeChat group reported higher levels of satisfaction with their colonoscopy experience (p = 0.043). In a multivariate analysis, adjusting for potential confounding factors, WeChat-based re-education ([OR] 1.496, 95% CI 1.154-1.939; p = 0.002)) emerged as a protective factor for achieving adequate BP. CONCLUSION: Enhanced education through WeChat can improve BP quality, and ARG applies equally to low health literacy populations.

Is the Occurrence or Reversal of Nonalcoholic Fatty Liver Disease Associated with Long-Term Helicobacter pylori Infection among Chinese Adults? A Cohort Study.

BACKGROUND: Previous studies have suggested a link between Helicobacter pylori (H. pylori) infection and nonalcoholic fatty liver disease (NAFLD), yet long-term follow-up studies to elucidate this association are lacking. We aimed to identify the relationship between NAFLD and H. pylori in these people. METHODS: A total of 2,934 adults between June 2013 and October 2017 were collected; among them, 675 people met the requirements. People were assessed for H. pylori infection diagnosis as detected by the carbon-13 urea breath test; they were also assessed for NAFLD diagnosis by ultrasound. RESULTS: H. pylori infection was present in 206 patients (30.5%), and 469 (69.5%) participants were classified as controls. Participants with H. pylori infection had a higher rate of incident NAFLD than those who were uninfected (37/206; 18% versus 73/469; 15.6%) (p < 0.001). Compared with the control group, the recovery rate of NAFLD in the H. pylori+ve group was low (6/206, 2.9% versus 33/469, 7.0%) (p < 0.001). Besides, the incidence of uric acid, postprandial blood glucose, TG, LDL-C, HDL-C, and fasting plasma glucose was significantly different between the two groups (p < 0.001), but no difference was found in alanine aminotransferase (ALT), liver-total protein, urea nitrogen, and cholesterol (p > 0.05). CONCLUSION: H. pylori infection was a risk factor for NAFLD and affected the occurrence or reversal of NAFLD, indicating that H. pylori infection eradication might play a role in reducing the risk of NAFLD.

Effect of Gender and Age on the Correlation between Helicobacter pylori and Colorectal Adenomatous Polyps in a Chinese Urban Population: A Single Center Study.

OBJECTIVES: To investigate whether Helicobacter pylori (H. pylori) infection increases the risk of colorectal adenomatous polyp (CAP) in the context of age and gender. METHODS: A total of 563 study subjects (male/female, 368/195) from Beijing, China, with higher nursing level who underwent colonoscopy were retrospectively collected. H. pylori and CAP were detected by carbon-13 urea breath test and colorectal colonoscopy. The correlations between the number, size, distribution, and pathological grade of CAP and H. pylori infection were analyzed. The population was further stratified by age and gender in order to examine the risk of H. pylori and CAP in the context of these variables. The influence of H. pylori on the risk of CAP was assessed by logistic regression model. RESULTS: 315 participants were diagnosed with CAP, and 207 participants were classified as healthy controls. The prevalence of H. pylori in the CAP group was significantly higher than that in the healthy control group (119/315, 37.8% versus 44/207, 21.3%) (p < 0.001). The proportion of H. pylori positive plus CAP in participants <50 years old was significantly higher than that in participants >50 years old (87/250; 34.8% versus 32/65; 49.2%) (p < 0.001). The proportion of H. pylori positive plus CAP in participants <50 years old was significantly higher than that in participants >50 years old (87/250; 34.8% versus 32/65; 49.2%) (p < 0.001). The proportion of H. pylori positive plus CAP in participants <50 years old was significantly higher than that in participants >50 years old (87/250; 34.8% versus 32/65; 49.2%) (p < 0.001). The proportion of H. pylori positive plus CAP in participants <50 years old was significantly higher than that in participants >50 years old (87/250; 34.8% versus 32/65; 49.2%) (. CONCLUSIONS: H. pylori is a major risk factor for CAP. Further studies are needed to assess the effects of H. pylori treatment or persistent infection on the occurrence or recurrence of CAP.

Activation of integrin ß1-focal adhesion kinase-RasGTP pathway plays a critical role in TGF beta1-induced podocyte injury.

The depletion of glomerular podocytes is the key mechanism of glomerulosclerosis and progressive renal failure. Transforming growth factor-ß (TGFß) is a central mediator of signaling networks that control a diverse set of cellular processes, such as cell proliferation, differentiation, and apoptosis. Though many key events in TGFß1 signaling have been documented at cellular and molecular level in podocytes, the complete effects of TGFß1 on podocyte integrity are still elusive. In this study, the function of adhesion protein integrin ß1, focal adhesion kinase (FAK), and a small GTPase Ras was explored in TGFß1-induced podocyte injury. In cultured mouse podocyte, caspase 3-positive cells were counted by flow cytometry to evaluate podocyte damage at different time points after TGFß1 treatment. Immunoblotting assay showed that integrin ß1, FAK, Src kinase, and an adaptor protein Grb2 were activated rapidly after TGFß1 stimulation. Active Ras Pull-Down assay revealed that the active Ras (GTP-bound Ras) level was upregulated in TGFß1-treated cell. Immunoprecipitation results displayed that TGFß1 enhanced the complex formation of integrin ß1, FAK and Src kinase, as well as FAK, Grb2 and Ras. The FAK inhibitor TAE226 and the specific knockdown of Grb2 remarkably alleviated TGFß1-induced podocyte apoptosis. The activation of p38MAPK and Erk1/2, and the nuclear translocation of NFκB(p65) were increased evidently in TGFß1-treated cell, which could be dramatically prohibited by the application of the p38MAPK inhibitor SB202190 and the Ras inhibitor FPT Inhibitor III. The Src kinase inhibitor PP2 obviously prevented the activation of FAK and Ras, as well as the translocation of NFκB(p65) from cytoplasm to nuclei. The PP2, FPT Inhibitor III, and SB202190 significantly decreased TGFß1-induced podocyte apoptosis. Taken together, these data demonstrated that the activation of integrin ß1/Src/FAK and Grb2/RasGTP should be responsible for TGFß1-induced podocyte damage through the p38MAPK and Erk1/2-mediated nuclear translocation of NFκB(p65).

Effect of carbamylated erythropoietin on major histocompatibility complex expression and neural differentiation of human neural stem cells.

The expression of major histocompatibility complex (MHC) on human neural stem cells (hNSCs) is tightly related to the fate of these cells in transplantation, therefore strategies to relieve rejection and promote graft survival are necessary to be applied. This study investigated the effect of carbamylated erythropoietin (CEPO) on MHC expression and differentiation of hNSCs with or without IFN-gamma incubation. Results showed that low levels of MHC molecules were expressed on hNSCs and increased by IFN-gamma. CEPO enhanced MHC-I antigens in both proliferative and differentiated hNSCs, but decreased MHC-II antigens in differentiated hNSCs and those cells exposed to IFN-gamma. Furthermore, CEPO promoted neural differentiation of hNSCs and outgrowth of neurites. Western blot analysis revealed activation of Stat3, Stat5 and Akt during these processes. These results suggest that CEPO may have immunoregulatory function in hNSCs besides its neuroprotection.

Ag(I) and Cu(II) discrete and polymeric complexes based on single- and double-armed oxadiazole-bridging organic clips.

Four new oxadiazole-bridging ligands (L1-L4) were designed and synthesized by the reaction of 2,5-bis(2-hydroxyphenyl)-1,3,4-oxadiazole with isonicotinoyl chloride and nicotinoyl chloride, respectively. L1 and L3 are unsymmetric single-armed ligands (4- or 3-pyridinecarboxylate arm), and L2 and L4 are symmetric double-armed ligands (4- or 3-pyridinecarboxylate arms). Nine new complexes, [Ag(L1)]PF6.CH3OH (1), [Ag(L1)]ClO4.CH3OH (2), Cu(L2)(NO3)2.2(CH2Cl2) (3), [Cu(L2)2](ClO4)2.2(CH2CCl2) (4), Cu(L2)Cl2 (5), [Cu4(L3)2(H2O)2](L3)4(ClO4)4 (6), [Ag(L4)(C2H5OH)]ClO4 (7), [Ag(L4)(C2H5OH)]BF4 (8), and [Ag(L4)(CH3OH)]SO3CF3 (9), were isolated from the solution reactions based on these four new ligands, respectively. L1, L2, and L3 act as convergent ligands and bind metal ions into discrete molecular complexes. In contrast, L4 exhibits a divergent spacer to link metal ions into one-dimensional coordination polymers. New coordination compounds were fully characterized by infrared spectroscopy, elemental analysis, and single-crystal X-ray diffraction. In addition, the luminescent and electrical conductive properties of these new compounds were investigated.