GBP2 enhances paclitaxel sensitivity in triple­negative breast cancer by promoting autophagy in combination with ATG2 and inhibiting the PI3K/AKT/mTOR pathway.

Chemoresistance is a major challenge in treating triple­negative breast cancer (TNBC); chemotherapy remains the primary approach. The present study aimed to elucidate the role of guanylate­binding protein 2 (GBP2) in activating autophagy in TNBC and its impact on the sensitivity of TNBC cells to paclitaxel (PTX). Transfection with lentivirus was performed to establish TNBC cell lines with stable, high GBP2 expression. The mRNA and protein levels of GBP2 expression were evaluated utilizing reverse transcription­quantitative PCR and western blotting, respectively. Autophagy in TNBC cells was evaluated using immunoblotting, transmission electron microscopy and fluorescence microscopy. The PI3K/AKT/mTOR pathway proteins and their phosphorylation were detected by immunoblotting, and fluorescence co­localization analysis was performed to evaluate the association between GBP2 and autophagy­related protein 2 (ATG2). BALB/c NUDE mice were subcutaneously injected with GBP2 wild­type/overexpressing MDA­MB­231 cells. Low GBP2 expression was detected in TNBC, which was associated with a poor prognosis. Overexpression of GBP2 suppressed cell growth, and especially enhanced autophagy in TNBC. Forced expression of GBP2 significantly increased the PTX sensitivity of TNBC cells, and the addition of autophagy inhibitors reversed this effect. GBP2 serves as a prognostic marker and exerts a notable inhibitory impact on TNBC. It functions as a critical regulator of activated autophagy by co­acting with ATG2 and inhibiting the PI3K/AKT/mTOR pathway, which contributes to increasing sensitivity of TNBC cells to PTX. Therefore, GBP2 is a promising therapeutic target for enhancing TNBC treatment.

Can serum ferritin serve as a biomarker for the prognosis of gynecological malignant tumors? A retrospective cohort study.

PURPOSE: It is widely accepted that there is a strong relationship between iron levels and cancer. This study aimed to investigate the relationship between serum ferritin levels and the severity and prognosis of gynecological malignant tumors. METHODS: This retrospective study included patients with gynecological malignant tumors at Sir Run Run Shaw Hospital in the Department of Obstetrics and Gynecology from January 2013 to June 2019. Patients were grouped according to their serum ferritin level: low (< 13 µg/L), normal (13-150 µg/L), and high (> 150 µg/L). Correlation analyses were performed between serum ferritin level and other factors. Cox univariable and multivariable analysis and Kaplan-Meier survival curves were used to assess the impact of ferritin on survival in patients with gynecologic tumors. RESULTS: The 402 total patients were divided into a low (n= 37), normal (n= 182), and high (n= 183) ferritin level group. Correlation analyses were performed that WBC, MCV, CRP, CA125, and CA153 were significantly positively correlated with serum ferritin level. The Kaplan-Meier survival curves revealed that of the three groups analyzed, the high serum ferritin level group had a significantly shorter survival time versus the normal and low serum ferritin level groups (log-rank P= 0.003). Univariable Cox regression analysis identified that patients with high serum ferritin levels had a significant correlation with risk of death compared to the patients with lower and normal serum ferritin levels. Serum ferritin was not found to be significant (HR = 0.792, 95% CI: 0.351-1.787, P= 0.574) in the multivariable Cox analysis. CONCLUSION: Although this study did not find serum ferritin to be a significant independent prognosis indicator in gynecological malignant tumors, this study did identify that gynecological malignant tumor patients with high serum ferritin levels have significantly less survival time than patients with low or normal serum ferritin levels.

Application of the 'online assessment + FOBT at home' to improve participation and the efficacy of opportunistic screening for colorectal cancer: a retrospective cohort study.

BACKGROUND: Colorectal cancer (CRC) screening faces two major challenges: insufficient screening coverage and poor adherence. A smartphone applet named "Early Screening Assistant (ESA)" was developed to create an online risk-assessment and fecal occult blood test (FOBT) at home. This retrospective study was designed to evaluate whether the new CRC screening strategy can improve the colonoscopy participation rate (PR) and lesion detection rate (DR). METHODS: In total, 6194 individuals who accepted normal health examinations and CRC screening based on the ESA from June 2020 to May 2022 were assigned to the ESA group. Accordingly, 7923 inhabitants who only accepted normal health examinations were assigned to the control group. The colonoscopy PR and neoplastic lesion DR were then compared between the two groups. RESULTS: Overall, a higher proportion of subjects in the ESA group (285 of 6194 [4.6%]) completed colonoscopy than in the control group (126 of 7923, [1.6%]), p < 0.01). The neoplastic lesion DR also significantly increased in the ESA group (76 of 6194 [1.22%]) compared with the control group (15 of 7923 [0.19%]) (p < 0.01). The adjusted diagnostic sensitivity and specificity of the "Online assessment + FOBT at home" were 41.5% and 62.6% for neoplastic lesions, respectively. CONCLUSIONS: This retrospective cohort study confirmed that the new CRC screening strategy based on the "Online assessment + FOBT at home" can improve colonoscopy participation and the neoplastic lesion detection rate and may represent a promising screening strategy for CRC. TRIAL REGISTRATION: This study was registered in China Clinical Trial Registry ( https://www.chictr.org.cn ) on 29/09/2022. REGISTRATION NUMBER: ChiCTR2200064186.

Candida causes recurrent vulvovaginal candidiasis by forming morphologically disparate biofilms on the human vaginal epithelium.

Background: Recurrent vulvovaginal candidiasis (RVVC) is a recalcitrant medical condition that affects many women of reproductive age. The importance of biofilm formation by Candida in RVVC has been recently questioned. This study aimed to elucidate the fundamental growth modes of Candida in the vagina of patients with RVVC or sporadic vulvovaginal candidiasis (VVC) and to assess their roles in the persistence of RVVC. Methods: Vaginal tissues were sampled from twelve patients clinically and microbiologically diagnosed as RVVC or VVC at a post-antifungal-treatment and asymptomatic period. High-resolution scanning electron microscopy, fluorescence in situ hybridization in combination with Candida-specific 18S rRNA probes and viable fungal burden were used to qualitatively and quantitatively evaluate Candida growth in the human vagina. The presence of Candida biofilm extracellular polymeric substances was examined using confocal laser scanning microscopy and biopsy sections pre-stained with Concanavalin A. Histopathological analysis was carried out on infected vaginal tissues stained with hematoxylin and eosin. Lastly, the susceptibility of epithelium-associated Candida biofilms to fluconazole at the peak serum concentration was evaluated. Results: Candida species grew on the vaginal epithelium of RVVC patients as morphologically disparate biofilms including monolayers, microcolonies, and macro-colonies, in addition to sporadic adherent cells. Candida biofilm growth on the vaginal epithelium was associated with mild lymphocytic infiltration of the vaginal mucosa. These epithelium-based Candida biofilms presented an important characteristic contributing to the persistence of RVVC that is the high tolerance to fluconazole. Conclusions: In summary, our study provides direct evidence to support the presence of Candida biofilms in RVVC and an important role of biofilm formation in disease persistence.

The Sonic Hedgehog signaling pathway regulates autophagy and migration in ovarian cancer.

BACKGROUND: The Sonic Hedgehog (SHH) signaling pathway plays an important role in various types of human cancers including ovarian cancer; however, its function and underlying mechanism in ovarian cancer are still not entirely understood. METHODS: We detected the expressions of SHH and SQSTM1 in borderline ovarian tumor tissues, epithelial ovarian cancer (EOC) tissues and benign ovarian tumor tissues. Cyclopamine (Cyp, a well-known inhibitor of SHH signaling pathway) and chloroquine (CQ, the pharmaceutical inhibitor of autophagy) were used in vivo and in vitro (autophagic flux, CCK-8 assay, wound healing assay, transwell assay, tumor xenograft model). The mechanism of action was explored through Quantitative RT-PCR and Western Blot. RESULTS: We found up-regulation of SHH and accumulation of SQSTM1/P62 in epithelial ovarian cancer. Cyp induced autophagy through the PI3K/AKT signaling pathway. Moreover, low-dose Cyp and chloroquine (CQ) significantly promoted the migratory ability of SKOV3 cells. CONCLUSIONS: Our findings suggest that inhibition of the SHH pathway and autophagy may be a potential and effective therapy for the treatment of ovarian cancer.

Functionalized MoS2-nanoparticles for transdermal drug delivery of atenolol.

Molybdenum disulfide (MoS2) has excellent photothermal conversion abilities, an ultra-high specific surface area, and has been extensively explored for use in biomedicine. However, the high toxicity associated with MoS2 has limited its biological applications for in vivo photothermal therapy and drug delivery systems. Herein, we have developed cationic hydroxyethyl cellulose (JR400) surface-modified MoS2 nanoparticles (NPs) that are responsive to near-infrared (NIR) laser irradiation as a transdermal drug delivery system (TDDS). Herein, we confirmed the preparation of hexagonal phase MoS2 with robust surface modification with JR400. The flower-like morphology of the NPs had an average diameter of 355 ± 69.3 nm limiting the absorption of the NPs through the stratum corneum. With the ability to efficiently load 90.4 ± 0.3% of the model drug atenolol (ATE), where 1 g of JR400-MoS2 NPs was able to load 3.6 g ATE, we assayed the controlled release capacity in vitro skin penetration studies. These JR400-MoS2 NPs showed further enhancement under NIR stimulation, with a 2.3-fold increase in ATE skin penetration. Furthermore, we verified in vivo that these JR400-MoS2 NPs do not cause skin irritation suggesting that they are promising new TDDS candidates for small molecule drugs.

miRNA­328 overexpression confers cisplatin resistance in non­small cell lung cancer via targeting of PTEN.

Chemotherapy resistance, the molecular mechanism of which is complex and has not been fully understood, poses a major challenge in the treatment of patients with non­small cell lung cancer (NSCLC). The dysregulation of microRNAs (miRs) has been reported to serve a pivotal role in the development of cancer and drug resistance. In the present study, reverse transcription­quantitative polymerase chain reaction analysis revealed a significant increase in miR­328 and a significant decrease in phosphatase and tensin homolog (PTEN) mRNA expression levels within tumor tissues from patients with cisplatin­resistant NSCLC compared with those of cisplatin­sensitive NSCLC patients. In addition, there was a negative correlation between PTEN mRNA and the miR­328 expression levels. In addition, higher miR­328 expression levels, and lower PTEN mRNA and protein expression levels, were detected in cisplatin­resistant A549 (A549rCDDP) cells when compared with in their parental cells. A549rCDDP cells demonstrated significantly higher cell viability compared with A549 cells following treatment with all concentrations of cisplatin tested (2, 4, 6 and 8 µM). Additionally, transfection of miR­328 inhibitor significantly increased PTEN mRNA and protein expression levels. Furthermore, the present study predicted and confirmed PTEN, a well­known tumor suppressor, as a direct target of miR­328 in NSCLC cells via the online tool MiRanda and a dual luciferase assay, respectively. Cell viability assay and flow cytometry analysis demonstrated that inhibition of miR­328 also induced cellular apoptosis and decreased cell proliferation in A549rCDDP cells treated with cisplatin. In conclusion, these results suggested that abnormal expression of miR­328 may contribute to cisplatin resistance in NSCLC, and may be considered to be a novel therapeutic target and indicator for the treatment and prognosis of patients with NSCLC treated with cisplatin­based chemotherapy.

PI3K/AKT/Afadin signaling pathway contributes to pathological vascularization in glioblastomas.

Glioblastomas are brain tumors with extensive vascularization that are associated with tumor malignancy. The phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway is activated in endothelial cell tumors, although its exact function in glioblastoma neovascularization is poorly characterized. The present study identified that endothelial cells derived from human glioblastomas exhibit increased permeability and motility compared with normal brain vascular endothelial cells. Furthermore, the phosphorylation of AKT was significantly induced in glioblastoma-derived endothelial cells and glioblastoma vessels. To the best of our knowledge, the present study demonstrated for the first time that the cell-cell adhesion junction protein Afadin is phosphorylated and re-localized in glioblastoma-derived endothelial cells, and the phosphorylation and re-localization of Afadin is PI3K/AKT pathway-dependent. AKT-mediated phosphorylation and re-localization of Afadin may be critically involved in the modulation of brain endothelial permeability and migration. Therapies targeting the PI3K/AKT/Afadin pathway may therefore be beneficial for reducing the angiogenic potential of glioblastoma.

Risk factors and coping strategies of severe community-acquired pneumonia in chemotherapy induction period of acute leukemia.

The risk factors and coping strategies of severe community-acquired pneumonia (SCAP) in chemotherapy induction period of acute leukemia were investigated. Eighty-six patients with CAP in chemotherapy induction period of acute leukemia in Dezhou Hospital from March 2014 to February 2017 were selected and divided into observation group (SCAP group, n=45) and control group (non-SCAP group, n=41) according to the acute physiology and chronic health evolution II (APACHE II) score. The blood, sputum, nasopharyngeal secretion and pleural effusion samples were collected from patients in both groups, and the samples were detected for pathogens, followed by the analysis of relevant factors. The dynamic changes in the sequential organ failure assessment (SOFA) score, procalcitonin (PCT), D-dimer (D-D) and C-reactive protein (CRP) levels in patients were observed before and after the corresponding treatment strategies were taken. The total distribution ratio of pathogens from high to low in the two groups was as follows: bacterium, virus, fungus, mycoplasma and chlamydia trachomatis; there was no significant difference between the two groups (P>0.05). Logistic regression analysis showed that the repeated infection (OR=3.315, P=0.005), multi-resistant bacterial infection (OR=1.915, P=0.008) and D-D (OR=1.936, P=0.009) were independent risk factors for SCAP (P<0.05). After different coping strategies were taken, the SOFA score, PCT, D-D and CRP levels in the two groups were significantly decreased, and they were obviously higher in observation group than those in control group (P<0.05). Repeated infection, D-D level and multi-resistant bacterial infection are the risk factors affecting the SCAP in chemotherapy induction period of acute leukemia. The coping strategies can effectively relieve the patient's condition, reduce the severity of disease and improve the survival rate of patients.

SOS1, HKT1;5, and NHX1 Synergistically Modulate Na+ Homeostasis in the Halophytic Grass Puccinellia tenuiflora.

Puccinellia tenuiflora is a typical salt-excluding halophytic grass with excellent salt tolerance. Plasma membrane Na+/H+ transporter SOS1, HKT-type protein and tonoplast Na+/H+ antiporter NHX1 are key Na+ transporters involved in plant salt tolerance. Based on our previous research, we had proposed a function model for these transporters in Na+ homeostasis according to the expression of PtSOS1 and Na+, K+ levels in P. tenuiflora responding to salt stress. Here, we analyzed the expression patterns of PtSOS1, PtHKT1;5, and PtNHX1 in P. tenuiflora under 25 and 150 mM NaCl to further validate this model by combining previous physiological characteristics. Results showed that the expressions of PtSOS1 and PtHKT1;5 in roots were significantly induced and peaked at 6 h under both 25 and 150 mM NaCl. Compared to the control, the expression of PtSOS1 significantly increased by 5.8-folds, while that of PtHKT1;5 increased only by 1.2-folds in roots under 25 mM NaCl; on the contrary, the expression of PtSOS1 increased by 1.4-folds, whereas that of PtHKT1;5 increased by 2.2-folds in roots under 150 mM NaCl. In addition, PtNHX1 was induced instantaneously under 25 mM NaCl, while its expression was much higher and more persistent in shoots under 150 mM NaCl. These results provide stronger evidences for the previous hypothesis and extend the model which highlights that SOS1, HKT1;5, and NHX1 synergistically regulate Na+ homeostasis by controlling Na+ transport systems at the whole-plant level under both lower and higher salt conditions. Under mild salinity, PtNHX1 in shoots compartmentalized Na+ into vacuole slowly, and vacuole potential capacity for sequestering Na+ would enhance Na+ loading into the xylem of roots by PtSOS1 through feedback regulation; and consequently, Na+ could be transported from roots to shoots by transpiration stream for osmotic adjustment. While under severe salinity, Na+ was rapidly sequestrated into vacuoles of mesophyll cells by PtNHX1 and the vacuole capacity became saturated for sequestering more Na+, which in turn regulated long-distance Na+ transport from roots to shoots. As a result, the expression of PtHKT1;5 was strongly induced so that the excessive Na+ was unloaded from xylem into xylem parenchyma cells by PtHKT1;5.

[Clinical study of titanium mesh in conjunction with concentrate growth factors to rebuild severe bone defect of anterior maxilla].

PURPOSE: To evaluate the effect of overlaying titanium mesh with concentrate growth factors(CGF) for rebuilding severe buccal bone defect of anterior maxilla when used in association with dental implantation. METHODS: Twenty patients with severe buccal bone defect of maxilla were selected. A total of 25 dental implants were placed, including 5 cases in bilateral central incisor area and 15 cases in unilateral central incisor area. After implantation, the defects were treated with Bio-oss and Bio-guid in conjunction with fixation of titanium mesh and then CGF technology was used. Two-stage surgery was carried out after 6 months of submerged healing, and permanent prosthesis was used 3 months after temporary restoration. The repairs of the defect were observed at the second stage surgery. The height of margin bone around implants and the thickness of bone at implants lingual side were measured, at the time of the second stage operation, and 3, 6, 12, 18 months after permanent restoration. The differences were analyzed by SPSS 19.0 software package with multi-sample nonparametric test and Fierdman test. RESULTS: At the time of second operation, the bone plate at lingual side was completely reconstructed, and new bone was formed at the top of implants. Clinical measurements showed that the averaged thickness of bone at lingual side was (2.69±0.154) mm at that time. Three, 6, 12, 18 months after restoration, the values were (2.67±0.152) mm, (2.66±0.153) mm, (2.65±0.153) mm, (2.65±0.151) mm, respectively. Implant-abutment junction was used as a base line to assess vertical bone absorption, the marginal bone of implant neck at lingual side was all inferior to the base line, the distance was (0.02±0.048) mm, (0.69±0.085) mm，(0.87±0.019) mm, (0.87±0.013) mm, respectively. Statistical analysis showed the thickness of bone of labial side decreased significantly over time after permanent restoration (P<0.01). Likewise, the height of marginal bone was also decreased significantly (P<0.01). However, the difference between them at 12 months and 18 months was not statistically significant (P>0.05). CONCLUSIONS: The results indicate that bone augmentation at maxilla can be achieved using titanium mesh in conjunction with CGF. The height and thickness of newly formed bone at the implant neck margin will be stabilized after 1 year. This method is worthy of wide clinical application.

[Effect of chloride ion on corrosion of two commonly used dental alloys].

OBJECTIVE: To investigate the eff ect of chloride concentration on the corrosion of Co-Cr alloy and pure Ti in a simulated oral environment. METHODS: The electrochemical corrosion tests of pure Ti and Co-Cr alloy were carried out in neutral artificial saliva solutions with different NaCl concentrations (0.9%, 2.0%, and 3.0%). Th e morphologies of corroded surface for pure Ti and Co-Cr alloy were observed by scanning electron microscope (SEM). RESULTS: Th e changes in the self-corrosion potentials (Ecorr) for pure Ti and Co-Cr alloy in three kinds of artificial saliva solutions was not obvious. However, the self-corrosion current densities (Icorr) of pure Ti were much lower than those of Co-Cr. The Icorr of Co-Cr alloy increased in a concentration-dependent manner of NaCl, whereas the breakdown potential (Eb) of Co-Cr alloy decreased in a concentration-dependent manner. Th e potential ranged for the breakdown of oxide film (Ev) was shortened in a concentration-dependent manner of NaCl. There was no obvious difference in the Icorr of pure Ti with different concentrations of NaCl. The breakdown potential was not seen according to the polarization curves. CONCLUSION: In a certain range, the increase of the concentration of Cl- leads to accelerate the corrosion behavior of Co-Cr alloy, but it does not affect pure Ti.

[Comparison of the occlusion effect of three kinds of desensitizers on dentinal tubules].

OBJECTIVE: To explore the effects of three dentine desensitizers on the surface morphology of freshly exposed dentin and to evaluate their occlusion effects on dentinal tubules using scanning electron microscope (SEM). METHODS: A total of 16 isolated human premolar samples, which were prepared to expose dentine, were randomly divided into a control group (n=4), a Hybrid Coat group (n=4), a Prime & Bond NT group (n=4), and a anti-sensitivity toothpaste group (n=4). After treatment with dentine desensitizers, one half of the samples in each group were vertically cleaved. Finally, the surfaces and cross sections of the samples were observed by SEM. RESULTS: The exposed tubule was almost occluded in the Hybrid Coat group and the Prime & Bond NT group, while only the majority of tubules could be sealed in the anti-sensitivity toothpaste group. The cross-section images showed that sediments were visible in all groups except the control group. CONCLUSION: Hybrid Coat and Prime & Bond NT are able to effectively seal tubules, while the effect of anti-sensitivity toothpaste is slightly poor.