Injectable hydrogel harnessing foreskin mesenchymal stem cell-derived extracellular vesicles for treatment of chronic diabetic skin wounds.

Chronic skin wounds are a serious complication of diabetes with a high incidence rate, which can lead to disability or even death. Previous studies have shown that mesenchymal stem cells derived extracellular vesicles (EVs) have beneficial effects on wound healing. However, the human foreskin mesenchymal stem cell (FSMSCs)-derived extracellular vesicle (FM-EV) has not yet been isolated and characterized. Furthermore, the limited supply and short lifespan of EVs also hinder their practical use. In this study, we developed an injectable dual-physical cross-linking hydrogel (PSiW) with self-healing, adhesive, and antibacterial properties, using polyvinylpyrrolidone and silicotungstic acid to load FM-EV. The EVs were evenly distributed in the hydrogel and continuously released. In vivo and vitro tests demonstrated that the synergistic effect of EVs and hydrogel could significantly promote the repair of diabetic wounds by regulating macrophage polarization, promoting angiogenesis, and improving the microenvironment. Overall, the obtained EVs-loaded hydrogels developed in this work exhibited promising applicability for the repair of chronic skin wounds in diabetes patients.

Application of symptom-based mind mapping combined with PBL teaching method in emergency trauma standardized resident training in MDT model.

Explore the feasibility and effectiveness of accepting mind mapping combined with problem-based learning (PBL) teaching method in the standardized training of emergency surgery residents in the multi-disciplinary team (MDT) model of emergency trauma. Eighty-nine doctors under training who rotated in the Department of Emergency Surgery of the First Affiliated Hospital of Anhui Medical University from January 2021 to January 2022 were selected as the study subjects, and randomly divided into a group receiving mind mapping combined with PBL teaching and a group receiving traditional lecture-based learning teaching. Mini-clinical evaluation exercise (Mini-CEX), direct observation of procedural skills (DOPS), teaching adherence, and satisfaction assessments were completed at the time of discharge from the department. There were no significant differences between the observation and control group trainees in terms of gender, age, education, and entry grades. Both groups of doctors were better able to participate in their respective teaching modes and made significant progress. The participants in the observation group had significantly higher Mini-CEX, DOPS, and teaching satisfaction scores than the control group (P < .05). Under the MDT model of emergency trauma, the combination of mind mapping and PBL teaching can improve the comprehensive clinical ability of the trainees more than participating in the traditional lecture-based learning teaching, which is worth promoting and implementing in the clinical standardized training.

Defluorinative Alkylation of Trifluoromethylbenzimidazoles Enabled by Spin-Center Shift: A Synergistic Photocatalysis/Thiol Catalysis Process with CO2•.

We describe a catalytic strategy for direct single C(sp3)-F bond alkylation of trifluoromethylbenzimidazoles under a photoinduced thiol catalysis process. The CO2 radical anion (CO2•-) proved to be the most efficient single-electron reductant to realize such a transformation. The spin-center shift of the generated radical anion intermediate is the key step in realizing C-F bond activation under mild conditions with high efficiency.

An Alternatively Spliced Variant of METTL3 Mediates Tumor Suppression in Hepatocellular Carcinoma.

Many post-transcriptional mRNA processing steps play crucial roles in tumorigenesis and the progression of cancers, such as N6-methyladenosine (m6A) modification and alternative splicing. Upregulation of methyltransferase-like 3 (METTL3), the catalytic core of the m6A methyltransferase complex, increases m6A levels and results in significant effects on the progression of hepatocellular carcinoma (HCC). However, alternative splicing of METTL3 has not been fully investigated, and the functions of its splice variants remain unclear. Here, we analyzed both our and online transcriptomic data, obtaining 13 splice variants of METTL3 in addition to canonical full-length METTL3-A in HCC cell lines and tissues. Validated by RT-qPCR and Western blotting, we found that METTL3-D, one of the splice variants expressing a truncated METTL3 protein, exhibits higher levels than METTL3-A in normal human livers but lower levels than METTL3-A in HCC tumor tissues and cell lines. Further functional assays demonstrated that METTL3-D expression decreased cellular m6A modification, inhibited the proliferation, migration, and invasion of HCC cells, and was negatively associated with the malignancy of patient tumors, exhibiting functions opposite to those of full-length METTL3-A. This study demonstrates that the METTL3-D splice variant is a tumor suppressor that could potentially be used as a target for HCC therapy.

Effect of PLC-ß1/CaM signaling pathway mediated by AT1R on the occurrence and development of hepatocellular carcinoma.

OBJECTIVE: To study the roles of AT1R, PLC-ß1, CaM and other related signal molecules in the formation and development of hepatocellular carcinoma (HCC) and their correlation. METHODS: ELISA and immunohistochemistry were used to analyze the expressions of target proteins in serum and liver tissue of HCC patients, and the correlation between AT1R, PLC-ß1 and CaM and postoperative survival status of patients was followed up and determined. CCK-8 method was used to screen the doses of Ang II and candesartan sensitive to HepG2 and HCCLM3 cells. Transwell experiment was used to observe the effects of different drugs on the migration and invasion activity of HCC cells. Meanwhile, flow cytometry and Western blot were used to detect the expression levels of AT1R, PLC-ß1 and CaM in the cells. Then PLC-ß1 siRNA was selected to transfect HCC cells, so as to further clarify the mechanism of the above signal proteins. HepG2 cells were inoculated under the hepatic capsule of mice to induce the formation of HCC in situ. Ang II and candesartan were used to stimulate HCC mice to observe the difference in liver appearance and measure the liver index. Finally, ELISA and immunofluorescence experiments were selected to analyze the levels of target proteins in mouse serum and liver tissue. RESULTS: The expression levels of target proteins in serum and liver tissue of HCC patients were significantly increased, and the postoperative survival time of patients with high expression of AT1R, PLC-ß1 or CaM was obviously shortened. Ang II and candesartan could significantly promote and inhibit the motility of HCC cells, and had different effects on the levels of AT1R, PLC-ß1 and CaM in cells. However, in hepatocellular carcinoma cells transfected with PLC-ß1 siRNA, the intervention ability of drugs was obviously weakened. Ang II could significantly promote the formation and progression of mouse HCC, while candesartan had the opposite effect. Meanwhile, medications could affect the expressions of target proteins in mouse serum and liver tissue. CONCLUSION: AT1R, PLC-ß1 and CaM may be risk factors affecting the formation and prognosis of HCC, and the PLC-ß1/CaM signaling pathway mediated by AT1R is an important way to regulate the migration and invasion activity of HCC cells.

Exploring DNA Methylation Profiles Altered in Cryptogenic Hepatocellular Carcinomas by High-Throughput Targeted DNA Methylation Sequencing: A Preliminary Study for Cryptogenic Hepatocellular Carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) includes cryptogenic hepatocellular carcinomas (CR-HCC) that lack a defined cause. Specific DNA methylation patterns and comparisons of the aberrant alterations in DNA methylation between CR-HCC and adjacent peritumor tissues (APTs) have not yet been reported. METHODS: The SureSelectXT Methyl-Seq Target Enrichment System was used to sequence targeted DNA methylation in three paired CR-HCC tissues and APTs. Gene Ontology (GO) enrichment and KEGG pathway analysis were performed to investigate the DNA methylation mechanism of CR-HCC. The mRNA expression levels of HOXB-AS3, HOXB6, HOXB3, USP18, MAP3K6, TIRAP, TNNI2, SHC3, CTTN, and TFAP2A, selected from the identified signaling pathways, were evaluated by quantitative real-time PCR (qPCR). RESULTS: A total of 1728 differentially methylated regions (DMRs) were identified in tumor tissues compared with non-tumor tissues, of which 868 DMRs were hypermethylated and 860 were hypomethylated. The DMRs were mapped within 2091 DMR-associated genes (DMGs). The mRNA expression of HOXB-AS3, HOXB3, and MAP3K6 was downregulated in CR-HCC tissues compared to the APTs. However, the mRNA expression of TIRAP, SHC3, and CTTN was upregulated in the CR-HCC tissues. Differences between the mRNA expression of HOXB6, USP18, TNNI2, and TFAP2A in the CR-HCC and APTS tissues were not statistically significant. GO analysis showed that the molecular functions of "binding", "protein binding", and "cytoskeletal protein binding" were the main categories for the hypermethylated DMGs. The hypomethylated DMGs were mostly enriched in the molecular functions "binding", "protein binding", "calcium ion binding", among others. KEGG pathway analysis showed that the hypermethylated DMGs were enriched in several pathways such as "estrogen signaling pathway", while hypomethylated DMGs were enriched in several pathways such as "proteoglycans in cancer", suggesting that epigenetic modifications play important roles in the cryptogenic hepatocarcinogenesis. CONCLUSION: These results provide useful information for future work to characterize the functions of epigenetic mechanisms on CR-HCC.

[Correlation between periodontitis and metabolic syndrome of the middle-aged and aged population in Shijingshan community of Beijing].

OBJECTIVE: To survey the metabolic status of the middle-aged and aged population with periodontitis in Shijingshan community of Beijing, and investigate the relationship between periodontitis and metabolic syndrome (MS). METHODS: The middle-aged and aged population in the community were investigated by questionnaires, periodontal clinical examinations and blood biochemical tests in 2005. A total of 903 subjects were enrolled, who were divided into two groups by severity of periodontitis. Their waist circumferences, values of high-density lipoprotein cholesterol (HDL-C), systolic blood pressure, fasting glucose, MS and its individual components (central obesity, insulin resistance, hypertriglyceridemia, low HDL-C and hypertension) were compared between the two groups. The Logistic regression model was set to analyze the relationship between periodontitis and MS. RESULTS: There was a significantly higher mean of systolic blood pressure, fasting glucose in the subjects with moderate-severe periodontitis than that with no-mild periodontitis. With severity of periodontitis increasing, the prevalence of MS, high blood glucose and low HDL-C increased significantly. After adjustment for gender, age, and smoking, the subjects with moderate-severe periodontitis were 1.524, 1.527 and 2.349 times more likely to suffer from MS, high blood glucose and low HDL-C than those with no-mild periodontitis, respectively. CONCLUSION: With severity of periodontitis increasing, the prevalence of MS, high blood glucose and low HDL-C increased significantly in the middle-aged and aged population of the community in Beijing. Severity of periodontitis is associated with MS, high blood glucose and low HDL-C.

An essential function for MKP5 in the formation of oxidized low density lipid-induced foam cells.

The uptake of oxidized low density lipoprotein (ox-LDL) by macrophages usually leads to the formation of lipid-laden macrophages known as "foam cells," and this process plays an important role in the development of atherosclerosis. Ox-LDL activates mitogen-activated protein kinase (MAP) kinases and nuclear factor (NF)-κB, and activations of p38 and NF-κB are important for the formation of foam cells. MAP kinase phosphatase (MKP) 5 is a member of the dual specificity phosphatases (DUSPs) family that can selectively dephosphorylate activated MAPKs to regulate innate and adaptive immune responses. However, the role of MKP5 in the formation of foam cells remains unknown. Here, we found that stimulation of ox-LDL induces the expression of MKP5 in macrophages. MKP5 deficiency blocked the uptake of ox-LDL and the formation of foam cells. Further analysis revealed that deletion of MKP5 reduced the ox-LDL-induced activation of NF-κB. Also, MKP5 deficiency markedly inhibited the production of TNF-α, but enhanced the levels of TGF-ß1 in ox-LDL-stimulated macrophages. Moreover, inhibition of NF-κB by p65 RNAi significantly reduced foam cell formation in macrophages from WT mice relative to MKP5-deficient mice. Thus, MKP5 has an essential role in the formation of foam cells through activation of NF-κB, and MKP5 represents a novel target for the therapeutic intervention of atherosclerosis.

[Impact on the risk of obesity due to interactions between fat mass- and obesity-associated gene rs9939609 variants and behavioral factors, in the Chinese school-aged children].

OBJECTIVE: To investigate how the interactions between fat mass- and obesity-associated (FTO) gene rs9939609 variants and daily-life related behavioral factors would influence the risk of obesity among the Chinese school-aged children. METHODS: 3503 school-aged children were selected from the Beijing Child and Adolescent Metabolic Syndrome (BCAMS) Study, and divided into obese children (n = 1229) and non-obese children (n = 2274). Venipuncture blood test, genotyping and questionnaire were performed. RESULTS: Five common factors including protein foods, tobacco & alcohol, vegetables & fruits, sedentary behavior and physical exercise in spare time were extracted with factor analysis methodology. Data from logistic regression analysis showed that taking the interaction of rs9939609 variant with protein foods as an example, the risk of interaction accounted for 19.16% when both factors existing simultaneously. Similarly, the interactions of this SNP with vegetables & fruits, sedentary behavior and physical exercise in spare time appeared to be 5.97%, 19.62% and 12.43% respectively; however there might not be interaction between tobacco, alcohol and the SNP in the Chinese children. CONCLUSION: Protein foods, vegetables & fruits, sedentary behavior and physical exercise might modify the effects of FTO rs9939609 variant on the risk of obesity in Chinese school-aged children. However, large-scale, prospective studies with detailed information on related behavioral factors would be ideal models for identifying the interactions between genes and environment.

[Levels of tumor necrosis factor-alpha and interleukin-1beta in saliva of metabolic syndrome patients].

OBJECTIVE: To investigate the association between periodontitis and the low-grade inflammation in metabolic syndrome (MS) patients. METHODS: Fifty-seven MS patients, 26 healthy controls were enrolled. Non-stimulated whole saliva was collected. The levels of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta was analyzed by radioimmunoassay and enzyme-linked immunosorbent assay, respectively. Concentration of cytokines was compared between MS patients and the healthy controls. Correlations between the cytokines and various periodontal indices, and between the cytokines level and different quantity of metabolic syndrome components were also investigated. RESULTS: Levels of TNF-alpha in saliva of MS patients [(69.30+/-21.01) ng/L] were significantly higher than that in the healthy subjects [(57.85+/-15.69) ng/L, P<0.05], and of IL-1beta in MS patients [(616.42+/-360.05) ng/L] higher than that in healthy subjects [(506.06+/-245.76) ng/L], but the difference was not statistically significant. TNF-alpha was positively correlated with bleeding index (BI). In MS patients, TNF-alpha level and IL-1beta level increased with increasing severity of periodontal disease and increasing component numbers of MS. CONCLUSIONS: Periodontal inflammation may be one of the sources of low-grade inflammation in MS patients. Both systemic and periodontal conditions may influence the level of salivary TNF-alpha and IL-1beta.

[Effects of Ginkgo biloba extract 50 preconditioning on contents of inflammation-related cytokines in myocardium of rats with ischemia-reperfusion injury].

OBJECTIVE: To investigate the effects of Ginkgo biloba extract 50 (GBE50) preconditioning on contents of inflammation-related cytokines in rats with myocardial ischemia-reperfusion. METHODS: Fifty-eight SD rats were divided into sham-operated group, untreated group, Salviae miltiorrhizae (SM) injection group and low-, medium- and high-dose GBE50 groups. After intragastric administration for 7 d, the left anterior descending (LAD) coronary artery was occluded for 30 min followed by 60-min reperfusion to induce ischemia-reperfusion injury. Myocardium histopathologic change was observed by HE staining under a light microscope; myeloperoxidase (MPO) activity in myocardium was measured by colorimetric detection; tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and IL-8 were detected by radioimmunoassay; IL-4 and IL-10 were tested by enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared with untreated group, rats in medium-dose GBE50 group had lower inflammatory reaction and MPO activity (P<0.01). GBE50 also decreased the content of IL-6 (P<0.05, P<0.01) and increased the content of IL-4 in myocardium (P<0.05, P<0.01) as compared with the untreated group. The content of TNF-alpha in myocardium in the medium-dose GBE50 group was lower and IL-10 was higher than those in the untreated group, but without significant differences. CONCLUSION: GBE50 can decrease the content of IL-6 and increase the content of IL-4 in myocardium after ischemia-reperfusion injury. It suggests that GBE50 can regulate the inflammatory reaction after ischemia-reperfusion injury via inhibiting inflammatory cytokines and promoting anti-inflammatory cytokines.

[Effect of electroacupuncture on ethology and cytokines of hippocampus in rats with dysmnesy].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on learning and memory impairment and cytokines of hippocampus in aging rats induced by D-galactose for exploring its underlying mechanism in the treatment of dysmnesy. METHODS: A total of 27 SD rats were randomly divided into control group (n = 9), model group (n = 8) and EA group (n = 10). Dysmnesy model was induced by intraperitoneal injection of D-galactose. EA (3 Hz, continuous waves, 1 mA) was applied to "Baihui" (GV 20) and "Zusanli" (ST 36) for 20 min every time and on alternate days, continuously for 21 days. Morris water maze tests were conducted to detect the rat's escape latency, percentage of swimming distance in the original platform quadrant and the total distance (SD/TD) and the percentage of swimming time in the platform quadrant after removal of the platform (spatial probe test) which were used for assessing the animals' learning and memory ability. The contents of IL-1beta, IL-6 and TNF-alpha in the hippocampus tissue were assayed by radioimmunoassay. RESULTS: Location navigation test showed that the escape latencies on the 2nd day and the 3rd day in model group were significantly longer than those of control group (P < 0.05), and the percents of SD/TD and swimming time in the platform quadrant were significantly lower than those of control group (P < 0.05, P < 0.01). In comparison with model group, the escape latency of EA group on the 3rd day was significantly shorter (P < 0.05), and the percent of SD/TD in the platform quadrant of EA group was significantly longer (P < 0.01). Compared with control group, IL-1beta and TNF-alpha contents in the hippocampus increased significantly in model group (P < 0.05, P < 0.01), while IL-6 content in model group decreased significantly (P < 0.01). In comparison with model group, the levels of IL-1beta and TNF-alpha in EA group decreased significantly (P < 0.05). CONCLUSION: EA can improve the learning-memory ability in dysmnesy rats, which may be closely associated with its effects in regulating the levels of cytokines in the hippocampus.

[Expression of E-cadherin catenin complex and invasiveness of pituitary adenoma].

OBJECTIVE: To explore the relationship between the E-cadherin catenin complex and invasiveness of pituitary adenoma. METHODS: The expression of E-cadherin catenin complex was determined by immunohistochemistry in 78 cases of human pituitary adenomas including invasive adenoma 44 cases, noninvasive adenoma 34 cases and the relativity of their expressions with hormone-producing, pituitary apoplexy and necrosis or cystoid change, tumor diameter were analyzed. RESULTS: The invasive group had a significantly lower expression of E-cad and alpha-cat than that of noninvasive group (chi-squared = 13.969, P < 0.01). There was no statistical significance for beta-cat expression between the invasive group and noninvasive group (chi-squared = 0.430, P > 0.05). Moreover, the expressions of beta-cad and alpha-cat were significantly lower in macro-adenoma group than that in micro-adenoma group (chi-squared = 5.038, P < 0.05). The expression of E-cad was significantly lower in endocrine inactive group than that in endocrine active group (chi-squared = 4.614, P < 0.05). The expression of beta-cat was significantly lower in the group with apoplexy and necrosis than that in the group without apoplexy and necrosis (chi-squared = 6.701, P < 0.05). CONCLUSIONS: The reduction of E-cad catenin complex is related to invasiveness and clinical pathological characteristics.

[Suppression of c-myc expression by interference RNA in HepG2 hepatocellular carcinoma cells].

OBJECTIVE: To study the inhibitory effect of RNA interference (RNAi) on c-myc expression in hepatocellular carcinoma cell line, HepG2. METHODS: Expression vector of c-myc gene-targeting small interference RNA (siRNA) was constructed (psilencer-c-myc) and transfected into HepG2 cells by lipofectamine, and the unloaded vector was used as control (mock). The expression of c-myc mRNA and protein was identified by quantitive PCR and Western blot. Apoptosis of the transfected cells was examined by flow cytometry and immunofluorescent microscopy. RESULTS: After HepG2 cells were transfected with psilencer-c-myc, the expression of c-myc mRNA and protein was suppressed with an inhibition rate of 67% compared with the mock-transfected cells. Apoptosis was identified in the transfected HepG2 cells. CONCLUSION: The expression of c-myc at transcriptional and translational levels in HepG2 cells transfected with siRNA is markedly inhibited, which may be associated with the induction of apoptosis.

cAMP elevators inhibit LPS-induced IL-12 p40 expression by interfering with phosphorylation of p38 MAPK in murine peritoneal macrophages.

cAMP mediated signaling may play a suppressive role in immune response. We previously found that the cAMP-elevators (CTx and 8-Br-cAMP) inhibited IL-12, IL-la, IL-6 gene expression, but increased the transcriptional levels of IL-10 and IL-1Ra in LPS-treated murine peritoneal macrophages. The present study examined a possible molecular mechanism involved in cAMP elevators-induced inhibition of IL-12 p40 expression in response to LPS. Our data demonstrated that cAMP elevators downregulated IL-12 p40 mRNA expression and IL-12 p70 production in murine peritoneal macrophages. Subsequent studies revealed that cAMP-elevators blocked phosphorylation of p38 MAPK, but did not affect the activity of NF-kappaB binding to IL-12 promoter (-136/-112). This is the first report that cAMP elevators inhibit LPS-induced IL-12 production by a mechanism that is associated, at least in part, with p38-dependent inhibition by cAMP signaling pathways.