Study on the role of CCM3 gene and lead exposure induced neurotoxicity through neurovascular units.

This study aimed to determine the toxic effects of vascular CCM3 gene deficiency and lead (Pb) exposure on the nervous system. Lentiviral transfection was performed to generate a stable strain of brain microvascular endothelial cells with low CCM3 expression. MTT assay assessed the survival rate of cells exposed to Pb, determining the dose and duration of Pb exposure in vitro. Proteomic analysis was performed on the differentially expressed proteins in bEnd3 and HT22 cells and flow cytometry was used to detect cell apoptosis. Finally, urine samples from pregnant and postpartum women were subjected to ICP-MS to detect Pb levels and HPLC to detect neurotransmitter metabolites. Based on the proteomic analysis of bEnd3 (CCM3-/-) cells co-cultured with HT22 cells, it was determined that HT22 cells and CCM3 genes interfered with bEnd3 cell differential proteins,2 including apoptosis and ferroptosis pathways. Electron microscopy observation, ICP-MS iron ion loading detection, and WB determination of protein GPX4 expression confirmed that HT22 cells undergo apoptosis, while bEnd3 cells undergo multiple pathways of iron death and apoptosis regulation. Furthermore, a linear regression model showed the interaction between maternal urine Pb levels, the rs9818496 site of the CCM3 SNP in peripheral blood DNA, and the concentration of the neurotransmitter metabolite 5-HIAA in maternal urine (F=4.198, P < 0.05). bEnd3 cells with CCM3 gene deficiency can induce HT22 cell apoptosis through iron death and apoptosis pathways under Pb exposure in a combined cell culture Pb exposure model, and CCM3 gene deficiency in endothelial cells and Pb exposure interacts with neural cell HT22. Epidemiological studies on maternal and newborn infants further confirmed the interaction between urine Pb levels in mothers and the SNP rs9818496 site of the CCM3 gene in peripheral blood DNA.

Applications of single­cell omics and spatial transcriptomics technologies in gastric cancer (Review).

Gastric cancer (GC) is a prominent contributor to global cancer-related mortalities, and a deeper understanding of its molecular characteristics and tumor heterogeneity is required. Single-cell omics and spatial transcriptomics (ST) technologies have revolutionized cancer research by enabling the exploration of cellular heterogeneity and molecular landscapes at the single-cell level. In the present review, an overview of the advancements in single-cell omics and ST technologies and their applications in GC research is provided. Firstly, multiple single-cell omics and ST methods are discussed, highlighting their ability to offer unique insights into gene expression, genetic alterations, epigenomic modifications, protein expression patterns and cellular location in tissues. Furthermore, a summary is provided of key findings from previous research on single-cell omics and ST methods used in GC, which have provided valuable insights into genetic alterations, tumor diagnosis and prognosis, tumor microenvironment analysis, and treatment response. In summary, the application of single-cell omics and ST technologies has revealed the levels of cellular heterogeneity and the molecular characteristics of GC, and holds promise for improving diagnostics, personalized treatments and patient outcomes in GC.

The murein endopeptidase MepA regulated by MtrAB and MprAB participate in cell wall homeostasis.

The complete genome of Corynebacterium glutamicum contain a gene encoding murein endopeptidase MepA which maintain cell wall homeostasis by regulating peptidoglycan biosynthesis. In this study, we investigate the physiological function, localization and regulator of MepA. The result shows that mepA overexpression lead to peptidoglycan degradation and the defects in cell division. MepA-mCherry was shown to localizes exclusively at the cell cell septum. In addition, mepA overexpression increased cell permeability and reduced the resistance of cells to isoniazid, an antibiotic used to treat Mycobacterium tuberculosis infection. Furthermore, transcription analysis showed that mepA affected cell division and membrane transport pathways, and was coordinately regulated by the two-component systems MtrAB and MprAB(CgtS/R2).

Effect of tamoxifen in patients with thin endometrium who underwent frozen-thawed embryo transfer cycles: a retrospective study.

Introduction: Thin endometrium leads to an impaired implantation rate. The aim of the study is to compare the clinical outcomes of tamoxifen (TAM) and hormone replacement therapy (HRT) used in patients with thin endometrium (<7mm) in frozen-thawed embryo transfer (FET)cycles. Methods: A total of 176 FET cycles with thin endometrium were retrospectively analyzed in our center from Jan 2020 to May 2022. According to patients' own will, 112 patients were allocated to the HRT group and 64 patients chose the TAM protocol. Clinical outcomes were compared between two groups. Result: The duration of treatment was shorter in the TAM group(12.03±2.34d) than the HRT group (16.07±2.52 d), which was statistically different (p<0.05). The endometrial thickness on the transfer day of the TAM group (7.32±1.28 mm) was significantly thicker than that of the HRT group (6.85±0.89mm, p<0.05). The clinical pregnancy rate of the TAM group (50.0%) was higher than that of the HRT group (36.6%), but there was no significant difference (p >0.05). The early miscarriage rate was significantly lower in the TAM group compared with the HRT group (5.9% Vs 26.8%, adjusted OR 0.10, p<0.05), while the live birth rate was higher in the TAM group (46.9% Vs 26.8%, adjusted OR 2.24, p<0.05) compared with the HRT group. Conclusion: For patients with thin endometrium, TAM effectively improved the endometrial thickness and increased the live birth rate. TAM can be used as an alternative protocol for patients with thin endometrium.

Water quality status response to multiple anthropogenic activities in urban river.

Water quality evaluation and degrading factors identification are crucial for predicting water quality evolution trends in an urban river. However, under the coupling of multiple factors, these targets face great challenges. The water quality status response to multiple anthropogenic activities in an urban river was evaluated and predicted based on comprehensive assessment methods and random forest (RF) model. We found that the distribution of each physicochemical parameter exhibits an obvious spatial clustering. The mean pollution level and trophic status of the urban river are medium pollution (water quality index = 59.79; Nemerow's pollution index = 2.00) and light eutrophication (trophic level index = 57.30). The water quality status is sensitive to anthropogenic activities, showing the following order of TLI and NPI values: residential district > industrial district > agricultural district and downtown > suburbs > countryside. According to the redundancy analysis, constructed land (F = 15.90, p < 0.01) and domestic sewage (F = 14.20, p < 0.01) evinced as the crucial factors that aggravated the water quality pollution level. Based on the simulation results of the RF model (variation explained = 94.91%; R2 = 0.978), improving domestic sewage treatment standards is the most effective measure to improve the water quality (increased by 40.3-49.3%) in residential and industrial districts. While in a suburban district, improving the domestic sewage collection rate has more effectively (23%) than those in the residential and industrial districts. Conclusively, reducing exogenous pollution input and improving domestic sewage treatment standards are vital to urban river restoration. Clinical trial registration Not applicable.

Exposure to a real traffic environment impairs brain cognition in aged mice.

Traffic-related air pollution (TRAP) has been a common public health problem, which is associated with central nervous system dysfunction according to large-scale epidemiological studies. Current studies are mostly limited to artificial laboratory exposure environments and specific genetic mechanisms remain unclear. Therefore, we chose a real-world transportation environment to expose aged mice, transporting them from the laboratory to a 1-m-high dry platform inside the campus and tunnel, and the mice were exposed daily from 7 a.m. to 7 p.m. for 2, 4 and 12 weeks respectively. Compared with the control group (in campus), the memory function of mice in the experimental group (in tunnel) was significantly impaired in the Morris water maze test. TRAP exposure increased the number of activated microglia in the hippocampal DG, CA1, CA3 regions and dorsolateral prefrontal cortex (dPFC). And neuroinflammation and oxidative stress levels were up-regulated in both hippocampus and dPFC of aged mice. By screening the risk genes of Alzheimer's disease, we found the mRNA and protein levels of ABCA7 were down-regulated and those of PYK2 were up-regulated. The DNA methylation ratios increased in four CpG sites of abca7 promoter region and decreased in one CpG site of pyk2 promoter region, which were consistent with the altered expression of ABCA7 and PYK2. In conclusion, exposure to the real traffic environment impaired memory function and enhanced neuroinflammation and oxidative stress responses, which could be relevant to the altered expression and DNA methylation levels of ABCA7 and PYK2. Our work provides a new and promising understanding of the pathological mechanisms of cognitive impairment caused by traffic-related air pollution.

Surface Modification of Biomedical Ti and Ti Alloys: A Review on Current Advances.

Ti is widely used as a material for orthopedic implants. As rapid and effective osseointegration is a key factor for the successful application of implants, biologically inert Ti materials start to show inherent limitations, such as poor surface cell adhesion, bioactivity, and bone-growth-inducing capabilities. Surface modification can be an efficient and effective approach to addressing the biocompatibility, mechanical, and functionality issues of the various Ti implant materials. In this study, we have overviewed more than 140 papers to summarize the recent progress in the surface modification of Ti implants by physical and/or chemical modification approaches, aiming at optimizing their wear resistance, biocompatibility, and antimicrobial properties. As an advanced manufacturing technology for Ti and Ti alloys, additive manufacturing was particularly addressed in this review. We also provide an outlook for future research directions in this field as a contribution to the development of advanced Ti implants for biomedical applications.

Type 2 diabetes mellitus promotes the proliferation, metastasis, and suppresses the apoptosis in oral squamous cell carcinoma.

BACKGROUND: Our previous study revealed that patients with oral squamous cell carcinoma and concomitant type 2 diabetes mellitus presented a lower 5-year survival rate. Hyperglycemia has been increasingly recognized as a risk factor for more advanced disease and poorer prognosis in patients with oral squamous cell carcinoma. However, its role remains unclear. METHODS: The expressions of BRIP1, Ki67, E-cadherin, and cleaved caspase-3 were detected by immunohistochemistry in oral squamous cell carcinoma tissues with or without type 2 diabetes mellitus. Cell counting kit-8 assay and wound healing assay were used to determine the proliferative and migratory ability of oral squamous cell carcinoma cells cultured with or without high glucose in vitro. Flow cytometry was applied to distinguish the role of high glucose on the cell cycle and apoptosis rates. RESULTS: The expression level of Ki67 was elevated while BRIP1, E-cadherin, and cleaved caspase-3 were downregulated in patients with oral squamous cell carcinoma coexisting with diabetes. The cell proliferation and migration in oral squamous cell carcinoma cell lines were significantly enhanced by high glucose. Flow cytometric analysis suggested that high glucose predisposed cancer cells to stay at S/G2 phase and to exhibit lower apoptosis rates. CONCLUSION: Our results implicated that type 2 diabetes mellitus may play a crucial role in the development and progression of oral squamous cell carcinoma through hyperglycemia, affecting cancer cell proliferation, migration, and apoptosis. This finding might provide a new direction for the prevention and treatment of oral squamous cell carcinoma.

miR-22 eluting cardiovascular stent based on a self-healable spongy coating inhibits in-stent restenosis.

The in-stent restenosis (IRS) after the percutaneous coronary intervention contributes to the major treatment failure of stent implantation. MicroRNAs have been revealed as powerful gene medicine to regulate endothelial cells (EC) and smooth muscle cells (SMC) in response to vascular injury, providing a promising therapeutic candidate to inhibit IRS. However, the controllable loading and eluting of hydrophilic bioactive microRNAs pose a challenge to current lipophilic stent coatings. Here, we developed a microRNA eluting cardiovascular stent via the self-healing encapsulation process based on an amphipathic poly(ε-caprolactone)-poly(ethylene glycol)-poly(ε-caprolactone) (PCL-PEG-PCL, PCEC) triblock copolymer spongy network. The miR-22 was used as a model microRNA to regulate SMC. The dynamic porous coating realized the uniform and controllable loading of miR-22, reaching the highest dosage of 133 pmol cm-2. We demonstrated that the sustained release of miR-22 dramatically enhanced the contractile phenotype of SMC without interfering with the proliferation of EC, thus leading to the EC dominating growth at an EC/SMC ratio of 5.4. More importantly, the PCEC@miR-22 coated stents showed reduced inflammation, low switching of SMC phenotype, and low secretion of extracellular matrix, which significantly inhibited IRS. This work provides a simple and robust coating platform for the delivery of microRNAs on cardiovascular stent, which may extend to other combination medical devices, and facilitate practical application of bioactive agents in clinics.

[Preliminary clinical study on the effect of metformin on prognosis of patients with oral squamous cell carcinoma after surgical treatment].

PURPOSE: To investigate the effect of metformin on the prognosis of patients with oral squamous cell carcinoma after surgical treatment. METHODS: Three hundred and forty-six patients with oral squamous cell carcinoma after operation in Xiangya Hospital of Central South University from October 2015 to October 2016 were selected and divided into experimental group and control group. In the experimental group, 71 patients with oral squamous cell carcinoma received metformin after surgery. The control group included 275 patients without metformin after surgery of oral squamous cell carcinoma. The clinical follow-up results of patients in the two groups were compared. SPSS 21.0 software package was used to analyze the data. RESULTS: Compared with the control group, the recurrent rate of the experimental group was lower. The difference was more significant in male patients, patients with primary tongue tumor, patients with highly differentiated squamous cell carcinoma, patients with cervical lymph node metastasis, and patients with a history of chewing areca nut (P<0.05). CONCLUSIONS: Metformin can decrease postoperative recurrent rate and metastatic rate of oral squamous cell carcinoma after surgery.

Hair levels of heavy metals and essential elements in Chinese children with autism spectrum disorder.

BACKGROUND: Disproportional heavy metals and essential elements were reported in children with autism spectrum disorder (ASD) that is obscure in etiology. Inevitably, the association is biased by diet and environmental factors. METHODS: Fifty pairs, one with ASD and the other living together from the same special school with cerebral palsy (CP), were recruited in Hangzhou (China), aged from 2 to 11 years old (74.0 % male). All samples were divided into two subgroups: preschool-aged (2-5 years old) and school-aged (6-10 years old). Heavy metals (As, Hg, Pb) and essential elements (Al, Ca, Cu, Mg, Mn, Zn) in hair were quantified by inductively coupled plasma mass spectrometry analysis and flame atomic absorption spectroscopy. RESULTS: The children with ASD generally had lower hair levels of Mn (ASD 0.124 µg/g, CP 0.332 µg/g, P = 0.001) compared to the children with CP. After stratification for age, there were no significant differences detected in preschool-aged group. In school-aged group, the results exhibited the children with ASD had higher hair Pb (1.485 µg/g, 0.690 µg/g, P = 0.007) and Cu/Zn ratio (0.092, 0.060, P = 0.003), while hair Hg (0.254 µg/g, 0.353 µg/g, P = 0.016)、Mn (0.089 µg/g, 0.385 µg/g, P = 0.002)、Mg (17.81 µg/g, 24.53 µg/g, P = 0.014) and Zn (100.15 µg/g, 135.83 µg/g, P = 0.007) showed an opposite pattern. CONCLUSIONS: These results suggest an imbalance of Mn in Chinese children with ASD.

Metformin reduces the increased risk of oral squamous cell carcinoma recurrence in patients with type 2 diabetes mellitus: A cohort study with propensity score analyses.

BACKGROUND: To investigate the impact of type 2 diabetes mellitus (T2DM) and metformin treatment on the prognosis of oral squamous cell carcinoma (OSCC) patients received radical surgical treatment. METHODS: Eight hundred and fifty-two patients with OSCC between January 2011 and January 2015 were included in the cohort study. Propensity score analysis was used to balance the characteristics of patients with or without T2DM and those of patients with T2DM treated with or without metformin. Five-year OSCC-free survival (OFS) was used to evaluate the prognosis of OSCC patients. RESULTS: Two hundred and sixty-nine patients without T2DM and 138 patients with T2DM were selected after the propensity score matching. The 5-year OFS of patients with T2DM was significantly lower than that of those without T2DM, both before (P = 0.019) and after (P = 0.014) the propensity score matching. Forty-four metformin users of OSCC patients with T2DM and 44 patients never users were further compared after the propensity score matching. The 5-year OFS of metformin users was significantly higher than that of metformin never users both before (P = 0.005) and after (P = 0.002) the propensity score matching. CONCLUSIONS: T2DM is associated with a higher risk of OSCC recurrence that can be reduced by metformin treatment.

CD147 promotes proliferation and migration of oral cancer cells by inhibiting junctions between E-cadherin and ß-catenin.

BACKGROUND: Although association between oral squamous cell carcinoma (OSCC) with epithelial-mesenchymal transition (EMT) has been demonstrated, we found CD147, one transmembrane protein we previously studied in oral submucous fibrosis, was correlated with E-cadherin, one marker of EMT. Here, we investigated CD147 expression in the different stages of OSCC and assessed its association with epithelial-mesenchymal transition (EMT). MATERIALS AND METHODS: CD147 and E-cadherin expression in tissue microarrays containing 48 OSCC specimens and matched adjacent tissues was analysed using immunohistochemistry. CD147 was overexpressed or knocked down using exogenous cloning vector and RNA interference, respectively, in OSCC cell lines. Cell proliferation and migration were measured using the CCK8 assay and scratch test, respectively. The expression and localization of EMT-associated proteins was analysed by Western blotting and immunofluorescence. RESULTS: CD147 expression in OSCC tissues was significantly higher than that in adjacent tissues and was markedly higher in cancer tissues with metastasis (P < .05). CD147 expression showed significant negative correlation with E-cadherin expression. CD147 overexpression downregulated E-cadherin and inhibited its complex with ß-catenin and then upregulated N-cadherin and vimentin. Additionally, alterations in CD147 protein expression affected proliferation and migration ability in OSCC cells and were related to ß-catenin nuclear translocation. CONCLUSION: CD147 plays an important role in tumorigenesis and metastasis by promoting EMT progression in OSCC. It may be considered as a novel potential diagnostic and therapeutic target for OSCC.

DEC1: a potential biomarker of malignant transformation in oral leukoplakia.

The purpose of this study was to analyze the differential expression of DEC1 in oral normal mucosa (NM), oral leukoplakia (OLK) and oral squamous cell carcinoma (OSCC). Surgically excised specimens from patients with OLK (n = 47), OSCC (n = 30) and oral normal mucosa (n=11) were immunostained for DEC1. The expression of DEC1 protein was evaluated, and its association with the clinicopathological features was analyzed. The expression of DEC1 in NM, OLK and OSCC tissues increased in turn, and significant differences were observed among the groups (P < 0.0001). In terms of the association between DEC1 expression and epithelial dysplasia, DEC1 expression was lower in hyperkeratosis without dysplasia (H-OLK) than in OLK with moderate to severe dysplasia (S-OLK), and these differences were significant (p < 0.05). The expression of DEC1 in OSCC with OLK was significantly higher than that in OSCC without OLK (p < 0.01). Therefore, DEC1 could be a potential biomarker of malignant transformation in the carcinogenesis of OSCC, which may provide a new research direction for the transformation of oral potentially malignant disorders (OPMDs) into OSCC.

Functionalized biomaterials to combat biofilms.

Pathogenic microbial biofilms that readily form on implantable medical devices or human tissues have posed a great threat to worldwide healthcare. Hopes are focused on preventive strategies towards biofilms, leaving a thought-provoking question: how to tackle the problem of established biofilms? In this review, we briefly summarize the functionalized biomaterials to combat biofilms and highlight current approaches to eradicate pre-existing biofilms. We believe that all of these strategies, alone or in combination, could represent a blueprint for fighting biofilm-associated infections in the postantibiotic era.

Biodegradable phosphorylcholine copolymer for cardiovascular stent coating.

Phosphorylcholine (PC) based polymer coatings with excellent biocompatibility have shown successful commercialization in drug-eluting stents. However, poor degradability represents a challenge in the application of biodegradable stents. Herein, a biodegradable phosphorylcholine copolymer is developed based on one-step radical ring-opening polymerization (RROP). This copolymer was synthesized by copolymerization of a PC unit, degradable ester (2-methylene-1,3-dioxepane, MDO) unit and non-degradable butyl methacrylate (BMA) unit, which showed ratio controllability by changing the monomer ratio during polymerization. We demonstrated that the copolymer with the ratio of 34% MDO, 19% MPC and 47% BMA could form a stable coating by ultrasonic spray, and showed good blood compatibility, anti-adhesion properties, biodegradability, and rapamycin eluting capacity. In vivo study revealed its promising application as a biodegradable stent coating. This work provides a facile path to add biodegradability into PC based polymers for further bio-applications.

Study on the expression and function of smad family member 7 in oral submucous fibrosis and oral squamous cell carcinoma.

OBJECTIVE: To investigate the expression and function of smad family member 7 (SMAD7) in the progress of oral submucous fibrosis (OSF) and oral squamous cell carcinoma (OSCC). METHODS: Mucosa tissue microarray containing 12 normal oral mucosa samples, 69 OSF samples, 28 OSCC sample and paired adjacent tissues was used to explore the expression levels in OSF and OSCC by immunochemistry. Several online bioinformatics analysis tools were used to explore "transcriptome level" and mostly probable "functions and pathways" of SMAD7 in head and neck squamous cell carcinoma. RESULTS: SMAD7 expression was up-regulated significantly in "OSF" (P < 0.0001) and "OSCC" (P < 0.05). In the status of "OSF and OSCC with OSF", the trends of SMAD7 expression were consistent and up-regulated (P < 0.0001). Based on bioinformatics analysis results, SMAD7 was significantly higher in head and neck squamous cell carcinoma than that of normal tissues. No mutation was found in head and neck squamous cell carcinoma. Pathway analysis results showed three mostly probable functions (extracellular matrix organization, blood vessel development and laminin interactions) and two mostly probable pathways (regulation of actin cytoskeleton and ras-associated protein-1 signaling pathway) that SMAD7 participated in. CONCLUSIONS: In OSF and OSCC, SMAD7 is indicated to be a promoter, as well as a potential diagnostic biomarker.

DEC1: a potential biomarker of malignant transformation in oral leukoplakia

Abstract The purpose of this study was to analyze the differential expression of DEC1 in oral normal mucosa (NM), oral leukoplakia (OLK) and oral squamous cell carcinoma (OSCC). Surgically excised specimens from patients with OLK (n = 47), OSCC (n = 30) and oral normal mucosa (n=11) were immunostained for DEC1. The expression of DEC1 protein was evaluated, and its association with the clinicopathological features was analyzed. The expression of DEC1 in NM, OLK and OSCC tissues increased in turn, and significant differences were observed among the groups (P < 0.0001). In terms of the association between DEC1 expression and epithelial dysplasia, DEC1 expression was lower in hyperkeratosis without dysplasia (H-OLK) than in OLK with moderate to severe dysplasia (S-OLK), and these differences were significant (p < 0.05). The expression of DEC1 in OSCC with OLK was significantly higher than that in OSCC without OLK (p < 0.01). Therefore, DEC1 could be a potential biomarker of malignant transformation in the carcinogenesis of OSCC, which may provide a new research direction for the transformation of oral potentially malignant disorders (OPMDs) into OSCC.

Antibacterial and hydroxyapatite-forming coating for biomedical implants based on polypeptide-functionalized titania nanospikes.

Titanium (Ti)-based implants often suffer from detrimental bacterial adhesion and inefficient healing, so it is crucial to design a dual-functional coating that prevents bacterial infection and enhances bioactivity for a successful implant. Herein, we successfully devised a cationic polypeptide (Pep)-functionalized biomimetic nanostructure coating with superior activity, which could not only kill pathogenic bacteria rapidly and inhibit biofilm formation for up to two weeks, but also promote in situ hydroxyapatite (HAp) formation. Specifically, a titania (TiO2) nanospike coating (TNC) was fabricated by alkaline hydrothermal treatment firstly, followed by immobilization of rationally synthesized Pep via robust coordinative interactions, named TNPC. This coating was able to effectively kill (>99.9%) both Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Escherichia coli (E. coli) bacteria, while being non-toxic to murine MC3T3-E1 osteoblastic cells. Furthermore, the in vivo infection studies denoted that the adherent bacteria numbers on the TNPC implants were significantly reduced by 6 orders of magnitude than those on the pure Ti implants (p < 0.001). Importantly, in the presence of cationic amino groups and residual Ti-OH groups, substantial HAp deposition on the TNPC surface in Kokubo's simulated body fluid (SBF) occurred after 14 days. Altogether, our results support the clinical potential of this biomimetic dual-functional coating as a new approach with desirable antibacterial properties and HAp-forming ability in orthopedic and dental applications.