LncRNA AGAP2 antisense RNA 1 stabilized by insulin-like growth factor 2 mRNA binding protein 3 promotes macrophage M2 polarization in clear cell renal cell carcinoma through regulation of the microRNA-9-5p/THBS2/PI3K-Akt pathway.

BACKGROUND: Increasing evidence highlights the potential role of long non-coding RNAs (lncRNAs) in the biological behaviors of renal cell carcinoma (RCC). Here, we explored the mechanism of AGAP2-AS1 in the occurrence and development of clear cell RCC (ccRCC) involving IGF2BP3/miR-9-5p/THBS2. METHODS: The expressions of AGAP2-AS1, IGF2BP3, miR-9-5p, and THBS2 and their relationship were analyzed by bioinformatics. The targeting relationship between AGAP2-AS1 and miR-9-5p and between miR-9-5p and THBS2 was evaluated with their effect on cell biological behaviors and macrophage polarization assayed. Finally, we tested the effect of AGAP2-AS1 on ccRCC tumor formation in xenograft tumors. RESULTS: IGF2BP3 could stabilize AGAP2-AS1 through m6A modification. AGAP2-AS1 was highly expressed in ccRCC tissues and cells. The lentivirus-mediated intervention of AGAP2-AS1 induced malignant behaviors of ccRCC cells and led to M2 polarization of macrophages. In addition, THBS2 promoted M2 polarization of macrophages by activating the PI3K/AKT signaling pathway. AGAP2-AS1 could directly bind with miR-9-5p and promote the expression of THBS2 downstream of miR-9-5p. These results were further verified by in vivo experiments. CONCLUSION: AGAP2-AS1 stabilized by IGF2BP3 competitively binds to miR-9-5p to up-regulate THBS2, activating the PI3K/AKT signaling pathway and inducing macrophage M2 polarization, thus facilitating the development of RCC.

[Applying Serum Vitamin D Metabolites in the Assessment of Renal Impairment in Diabetic Kidney Disease of Type 2 Diabetes Mellitus Patients: A Retrospective Study].

Objective: Total 25(OH)D (t-25[OH]D), a marker traditionally used in the assessment of vitamin D (VitD) in the human body, includes 25(OH)D 2, 25(OH)D 3, and C 3-epimers-25(OH)D 3(C 3-epi). In this study, we analyzed the relationship between serum VitD metabolites and renal impairment in patients with diabetic kidney disease (DKD). Methods: We covered, in the study, 339 subjects, including 114 otherwise healthy controls (HC), 74 type 2 diabetes mellitus (DM) patients with no glomerular filtration dysfunction, and 151 DKD patients. According to the results of combined evaluation of estimated glomerular filtration rate (eGFR) and urine albumin-to-creatinine ratio (UACR), the DKD patients were further divided into four subgroups, stage 2 subgroup of patients of DM combined with stage-2 chronic kidney disease (CKD2), stage 3 subgroup of patients of DM combined with CKD3, stage 4 subgroup of patients of DM combined with CKD4, and stage 5 subgroup of patients of DM combined with CKD5. The levels of 25(OH)D 2, 25(OH)D 3, and C 3-epi were measured by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and the activity level of 25(OH) 3 (AVitD 3), t-25(OH)D concentration, 25(OH)D 2/25(OH)D 3 ratio, C 3-epi/t-25(OH)D ratio, and C 3-epi/AVitD 3 ratio were calculated. Results: The levels of 25(OH)D 3, t-25(OH)D, and AVitD 3 were lower in the DKD group than those in the DM and HC groups (all P<0.05). C 3-epi/t-25(OH)D ratio and C 3-epi/AVitD 3 ratio were higher in the DKD group than those in the HC group (all P<0.05). The levels of 25(OH)D 3, t-25(OH)D, AVitD 3, and C 3-epi were lower in the stage 5 subgroup than those in the stage 2 and stage 3 subgroups (all P<0.05). The levels of 25(OH)D 3, t-25(OH)D, and C 3-epi were lower in the stage 4 subgroup than those in the stage 3 subgroup (all P<0.05). The 25(OH)D 3, t-25(OH)D, and AVitD 3 levels were lower in the stage 4 subgroup than those in the stage 2 subgroup (all P<0.05). Conclusions: UPLC-MS/MS can be used to perform accurate evaluation of VitD nutritional status in DKD patients. DKD patients have decreased levels of serum t-25(OH)D, 25(OH)D 3, and AVitD 3, all of which progressively decrease along with the rise in CKD staging. The trend of C 3-epi and 25(OH)D 3 changes were not consistent.

KMT2B promotes the growth of renal cell carcinoma via upregulation of SNHG12 expression and promotion of CEP55 transcription.

BACKGROUND: This study aims to clarify the mechanistic action of long non-coding RNA (lncRNA) SNHG12 in the development of renal cell carcinoma (RCC), which may be associated with promoter methylation modification by KMT2B and the regulation of the E2F1/CEP55 axis. METHODS: TCGA and GEO databases were used to predict the involvement of SNHG12 in RCC. Knockdown of SNHG12/E2F1/CEP55 was performed. Next, SNHG12 expression and other mRNAs were quantified by RT-qPCR. Subsequently, CCK-8 was used to detect cell proliferation. Wound healing assay and Transwell assay were used to detect cell migration and invasion, respectively. The in vitro angiogenesis of human umbilical vein endothelial cells (HUVECs) was explored by matrigel-based capillary-like tube formation assay. ChIP assay was used to detect H3K4me3 in SNHG12 promoter region. The binding of E2F1 to CEP55 promoter region was analyzed with ChIP and dual luciferase reporter assays. RIP assay was used to detect the binding of SNHG12 to E2F1. Finally, the effect of SNHG12 on the tumor formation and angiogenesis of RCC was assessed in nude mouse xenograft model. RESULTS: SNHG12 was highly expressed in RCC tissues and cells, and it was related to the poor prognosis of RCC patients. SNHG12 knockdown significantly inhibited RCC cell proliferation, migration, and invasion and HUVEC angiogenesis. KMT2B up-regulated SNHG12 expression through modifying H3K4me3 in its promoter region. In addition, SNHG12 promoted CEP55 expression by recruiting the transcription factor E2F1. Knockdown of SNHG12 blocked E2F1 recruitment and down-regulated the expression of CEP55, thereby inhibiting tumor formation and angiogenesis in nude mice. CONCLUSION: The evidence provided by our study highlighted the involvement of KMT2B in up-regulation of lncRNA as well as the transcription of CEP55, resulting in the promotion of angiogenesis and growth of RCC.

Correlation between vitamin D metabolites and rheumatoid arthritis with osteoporosis by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

INTRODUCTION: Our aim is to study the correlation between vitamin D metabolites and osteoporosis in rheumatoid arthritis (RA) by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). At the same time, other influencing factors and serum biomarkers of osteoporosis in patients with RA were studied. MATERIALS AND METHODS: Patients with RA admitted from January 2020 to December 2020 were selected at our hospital. The subjects were divided into the normal bone mineral density (BMD), osteopenia, and osteoporosis groups. The differences of vitamin D (VD) metabolites among groups were compared. The Pearson correlation coefficient was used to analyze the relationship between BMD and various parameters. The relationship between BMD and influencing factors was studied by a multiple linear regression equation. RESULTS: A total of 287 patients with RA were included. RA patients with 25-hydroxy vitamin D [25(OH)D] deficiency accounted for 43.63% and 25(OH)D insufficient levels accounted for 31.37%. There were 31 cases (10.80%) in the normal BMD group, 161 cases (56.10%) in the osteopenia group, and 95 cases (33.10%) in the osteoporosis group. The BMD of L1-4 (T- score) was negatively correlated with age (P < 0.05), course of disease (P < 0.05), and erythrocyte sedimentation rate (ESR) (P < 0.05), and positively correlated with 25(OH)D3 (P < 0.05). The multiple linear regression model results showed that age and 25(OH)D3 were independent predictors of BMD; this explained 22.11% of the total variation. CONCLUSIONS: VD deficiency and insufficient are common in RA patients. RA patients can be appropriately supplemented with VD. VD3 may be a better choice.

Circular RNA circ_001842 plays an oncogenic role in renal cell carcinoma by disrupting microRNA-502-5p-mediated inhibition of SLC39A14.

Renal cell carcinoma (RCC) is a common urologic malignancy, and up to 30% of RCC patients present with locally advanced or metastatic disease at the time of initial diagnosis. Increasing evidence suggests that circular RNAs (circRNAs) serve as genomic regulatory molecules in various human cancers. Our initial in silico microarray-based analysis identified that circRNA circ_001842 was highly expressed in RCC. Such up-regulation of circ_001842 in RCC was experimentally validated in tissues and cell lines using RT-qPCR. Thereafter, we attempted to identify the role of circ_001842 in the pathogenesis of RCC. Through a series of gain- and loss-of function assays, cell biological functions were examined using colony formation assay, Transwell assay, annexin V-FITC/PI-labelled flow cytometry and scratch test. A high expression of circ_001842 in tissues was observed as associated with poor prognosis of RCC patients. circ_001842 was found to elevate SLC39A14 expression by binding to miR-502-5p, consequently resulting in augmented RCC cell proliferation, migration and invasion, as well as EMT in vitro and tumour growth in vivo. These observations imply the involvement of circ_001842 in RCC pathogenesis through a miR-502-5p-dependent SLC39A14 mechanism, suggesting circ_001842 is a potential target for RCC treatment.

Simultaneous multielement analysis by ICP-MS with simple whole blood sample dilution and its application to uremic patients undergoing long-term hemodialysis.

Few studies were reported on trace elements' alterations in uremic patients undergoing long-term (>24 months) hemodialysis (HD), especially by using the whole blood as the biological fluid for the measuring purpose. Our objective was to develop an improved micro-sampling inductively coupled plasma-mass spectrometry (ICP-MS) method to determine the levels of Ca, Mg, Cu, Zn, Fe, Mn, Se and Pb in uremic patients receiving long-term HD. A ICP-MS method with a modified whole blood sample preparation procedure with small volumes was established and applied for the simultaneous quantification of the various elements in uremic patients undergoing long-term HD. 124 eligible uremic patients receiving long-term HD (75 males and 49 females) and 77 healthy subjects (54 males and 23 females) were recruited and Ca, Mg, Cu, Zn, Fe, Mn, Se and Pb levels were further determined. Our results revealed that uremic patients with HD had significantly higher blood levels of Ca, Mg, Zn and Pb and lower Cu, Fe, Se and Mn concentrations than healthy controls. In conclusion, a reproducible and reliable ICP-MS method using minimal whole blood sample volume (50 µL) with a simple dilution-based preparation procedure was successfully improved, validated and applied. Uremic patients undergoing long-term HD might be at increased risk of some essential trace elements deficiency (especially for Cu, Fe and Se) or toxic trace element excess (Pb) in respect to healthy subjects. Monitoring of blood levels and supplementation of some trace elements may be indicated in uremic patients undergoing long-term HD.

The effect of soy intervention on insulin-like growth factor 1 levels: A meta-analysis of clinical trials.

A low insulin-like growth factor 1 (IGF-1) level is known to be associated with many disorders. Several studies have shown that soy consumption may influence IGF-1, but the findings remain inconclusive. In this work, we conducted a systematic review and meta-analysis to provide a more accurate estimation of the effect of soy consumption on plasma IGF-1. A comprehensive systematic search was performed in Scopus, Embase, Web of Science, and PubMed/MEDLINE databases from inception until October 2019. Eight studies fulfilled the eligibility criteria. The pooled weighted mean difference (WMD) of the eligible studies was calculated with random-effects approach. Overall, a significant increment in plasma IGF-1 was observed following soy intervention (WMD: 13.5 ng/ml, 95% CI: 5.2, 21.8, I2 = 97%). Subgroup analyses demonstrated a significantly greater increase in IGF-1, when soy was administered at a dosage of ≤40 g/day (WMD: 11.7 ng/ml, 95% CI: 10.9 to 12.6, I2 = 98%), and when the intervention duration was <12 weeks (WMD: 26.6 ng/ml, 95% CI: 9.1 to 44.1, I2 = 0.0%). In addition, soy intervention resulted in a greater increase in IGF-1 among non-healthy subjects (WMD: 36 ng/ml, 95% CI: 32.7 to 39.4, I2 = 84%) than healthy subjects (WMD: 9.8 ng/ml, 95% CI: 8.9 to 10.7, I2 = 90%). In conclusion, this study provided the first meta-analytical evidence that soy intake may increase IGF-1 levels, but the magnitude of the increase is dependent on the intervention dosage, duration, and health status of the participants.

A meta-analysis of platinum-based neoadjuvant chemotherapy versus standard neoadjuvant chemotherapy for triple-negative breast cancer.

Aim: Platinum agents are DNA damaging agents with promising activity in breast cancers, especially in triple-negative subgroup. This meta-analysis was conducted to compare the treatments of platinum-based neoadjuvant chemotherapy (NAC) and standard NAC for triple-negative breast cancers (TNBCs). Materials & methods: Diverse electronic databases were searched to identify the randomized clinical trials that directly compared the treatments of platinum-based NAC versus NAC in TNBC patients. Toxicity of platinum-based regimens was further evaluated. Results: Addition of platinum agents significantly improved the pathological complete response rates in TNBC patients compared with the standard NAC. Unfortunately, platinum-based regimens were more likely to develop higher incidence of hematologic toxicities. Conclusion: Platinum-based NAC regimens could achieve significant pathological complete response improvement with well-tolerated toxicity in TNBC patients.

MiR-27a Promotes Hemin-Induced Erythroid Differentiation of K562 Cells by Targeting CDC25B.

BACKGROUND/AIMS: MicroRNAs (miRNAs) play a crucial role in erythropoiesis. MiR-23a∼27a∼24-2 clusters have been proven to take part in erythropoiesis via some proteins. CDC25B (cell division control Cdc2 phosphostase B) is also the target of mir-27a; whether it regulates erythropoiesis and its mechanism are unknown. METHODS: To evaluate the potential role of miR-27a during erythroid differentiation, we performed miR-27a gain- and loss-of-function experiments on hemin-induced K562 cells. We detected miR-27a expression after hemin stimulation at different time points. At the same time, the γ-globin gene also was measured via real-time PCR. According to the results of the chips, we screened the target protein of miR-27a through a dual-luciferase reporter assay and identified it via Western blot analyses. To evaluate the function of CDC25B, benzidine staining and flow cytometry were employed to detect the cell differentiation and cell cycle. RESULTS: We found that miR-27a promotes hemin-induced erythroid differentiation of human K562 cells by targeting cell division cycle 25 B (CDC25B). Overexpression of miR-27a promotes the differentiation of hemin-induced K562 cells, as demonstrated by γ-globin overexpression. The inhibition of miR-27a expression suppresses erythroid differentiation, thus leading to a reduction in the γ-globin gene. CDC25B was identified as a new target of miR-27a during erythroid differentiation. Overexpression of miR-27a led to decreased CDC25B expression after hemin treatment, and CDC25B was up-regulated when miR-27a expression was inhibited. Moreover, the inhibition of CDC25B affected erythroid differentiation, as assessed by γ-globin expression. CONCLUSION: This study is the first report of the interaction between miR-27a and CDC25B, and it improves the understanding of miRNA functions during erythroid differentiation.

Total Oxidant/Antioxidant Status in Sera of Patients with Esophageal Cancer.

BACKGROUND Oxidative stress parameters such as total oxidant status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI) have been studied in breast, thyroid, and simple esophageal cancers (EC). We evaluated these parameters in patients with EC and analyzed their correlations with treatment outcomes. MATERIAL AND METHODS Serum TOS, TAS, and OSI in 92 patients with EC at different clinical stages and in 64 healthy people (controls) were measured. RESULTS Serum TOS, TAS, and OSI were significantly different between patients with EC and healthy controls (all p<0.001); however, there were no significant differences across different clinical stages (all p>0.05). These factors are not correlated with smoking or drinking history (all p>0.05). Patients with EC with higher TOS and OSI and lower TAS had better responses to chemotherapy and/or radiotherapy, but there was no significant correlation with different responses (all p>0.05). In a receiver operating characteristic curve analysis comparing patients with EC with healthy controls, the Youden indices were 0.391, 0.886, and 1, respectively. CONCLUSIONS Serum TOS, TAS, and OSI were significantly different between patients with EC and healthy controls. In patients with EC, these factors were not correlated with smoking or drinking history or with clinical stage. Patients with EC with higher TOS and OSI and lower TAS had a trend towards better outcomes but it did not reach significance. Serum TOS and OSI are potential diagnostic biomarkers that can be used to identify cases of EC.

[Clinical Significance of Serum ADA Combined with GLB, CREA, ß2-MG and HGB in Newly Diagnosed Multiple Myeloma].

OBJECTIVE: To investigate the role of serum adenosine deaminase (ADA) combined with globulin (GLB), creatinine (CREA), ß2-microglobulin (ß2-MG) and hemoglobin (HGB) in the initial screening of multiple myeloma (MM), in order to reduce missed diagnosis and misdiagnosis of MM. METHODS: A retrospective analysis was performed on 62 newly diagnosed multiple myeloma (NDMM) patients who were admitted to the Department of Hematology of the First Affiliated Hospital of Chengdu Medical College from April 2018 to December 2021, and 33 patients with benign hematologic diseases and 30 healthy subjects were selected as the control group. The expression of ADA in pan-cancer was analyzed using TCGA and GTEx databases. The general data and laboratory indicators of the subjects were collected, and the differences of ADA activity and other laboratory indicators in each group were compared. The relationship between serum ADA activity and clinical data of NDMM patients was analyzed. The changes of ADA activity before and after chemotherapy in NDMM patients and the differences of ADA activity in NDMM patients with different DS and ISS stages were compared. Multivariate logistic regression was used to analyze the risk factors of NDMM. The receiver operating characteristic(ROC) curve was used to evaluate the diagnostic efficacy of ADA and other laboratory indicators in MM. Bioinformatics method was used to analyze the co-expression networks and enrichment pathways of ADA. RESULTS: ADA level was significantly upregulated in tissues of 14 types of cancer in TCGA database, and ADA was highly expressed in 11 types of cancer in TCGA combined with GTEx databases. The serum levels of ADA, GLB, uric acid (UA), cystatin C (CysC) and ß2-MG in the NDMM group were significantly higher than those in benign hematologic disease group and healthy control group ( P < 0.05), while the levels of ALB and the value of albumin to globulin ratio (A∶G) in the NDMM group were significantly lower than those in the other two groups ( P < 0.001). There were significant differences in DS stage (P =0.036), ISS stage (P =0.019) and the levels of CREA (P =0.036), UA (P =0.034), ß2-MG (P =0.019) in NDMM patients with different ADA activity levels. After primary chemotherapy, ADA activity and ß2-MG concentration were decreased in NDMM patients ( P < 0.01). The comparison results of patients in different stages showed that ADA activity of patients in DS stage I+II was significantly lower than that of patients in DS stage III (P <0.05), and ADA activity of patiens in ISS stage I+II was significantly lower than that of patients in ISS stage III ( P < 0.01). Multivariate logistic regression analysis showed that increased GLB, increased ADA activity, increased CREA, increased ß2-MG and decreased HGB were independent risk factors for NDMM. The area under the curve (AUC) of ADA in the diagnosis of MM was 0.847, and the AUC of ADA combined with GLB, CREA, ß2-MG and HGB in the diagnosis of MM was 0.940. The results of co-expression network and enrichment pathway analysis showed that ADA bounded to 20 proteins and it was significantly associated with the metabolic pathways of purine, pyrimidine, nicotinate and nicotinamide. CONCLUSION: The detection of ADA activity in serum is of positive significance for the auxiliary diagnosis, therapeutic evaluation and monitoring the progress of NDMM patients. ADA combined with GLB, CREA, ß2-MG and HGB can improve the detection rate of MM, and reduce missed diagnosis and misdiagnosis to a certain extent.

Role of uranium toxicity and uranium-induced oxidative stress in advancing kidney injury and endothelial inflammation in rats.

OBJECTIVE: Uranium exposure may cause serious pathological injury to the body, which is attributed to oxidative stress and inflammation. However, the pathogenesis of uranium toxicity has not been clarified. Here, we evaluated the level of oxidative stress to determine the relationship between uranium exposure, nephrotoxic oxidative stress, and endothelial inflammation. METHODS: Forty male Sprague-Dawley rats were divided into three experimental groups (U-24h, U-48h, and U-72h) and one control group. The three experimental groups were intraperitoneally injected with 2.0 mg/kg uranyl acetate, and tissue and serum samples were collected after 24, 48, and 72 h, respectively, whereas the control group was intraperitoneally injected with 1.0 ml/kg normal saline and samples were collected after 24 h. Then, we observed changes in the uranium levels and oxidative stress parameters, including the total oxidative state (TOS), total antioxidant state (TAS), and oxidative stress index (OSI) in kidney tissue and serum. We also detected the markers of kidney injury, namely urea (Ure), creatine (Cre), cystatin C (CysC), and neutrophil gelatinase-associated lipocalin (NGAL). The endothelial inflammatory markers, namely C-reactive protein (CRP), lipoprotein phospholipase A2 (Lp-PLA2), and homocysteine (Hcy), were also quantified. Finally, we analyzed the relationship among these parameters. RESULTS: TOS (z = 3.949; P < 0.001), OSI (z = 5.576; P < 0.001), Ure (z = 3.559; P < 0.001), Cre (z = 3.476; P < 0.001), CysC (z = 4.052; P < 0.001), NGAL (z = 3.661; P < 0.001), and CRP (z = 5.286; P < 0.001) gradually increased after uranium exposure, whereas TAS (z = -3.823; P < 0.001), tissue U (z = -2.736; P = 0.001), Hcy (z = -2.794; P = 0.005), and Lp-PLA2 (z = -4.515; P < 0.001) gradually decreased. The serum U level showed a V-shape change (z = -1.655; P = 0.094). The uranium levels in the kidney tissue and serum were positively correlated with TOS (r = 0.440 and 0.424; P = 0.005 and 0.007) and OSI (r = 0.389 and 0.449; P = 0.013 and 0.004); however, serum U levels were negatively correlated with TAS (r = -0.349; P = 0.027). Partial correlation analysis revealed that NGAL was closely correlated to tissue U (rpartial = 0.455; P = 0.003), CysC was closely correlated to serum U (rpartial = 0.501; P = 0.001), and Lp-PLA2 was closely correlated to TOS (rpartial = 0.391; P = 0.014), TAS (rpartial = 0.569; P < 0.001), and OSI (rpartial = -0.494; P = 0.001). Pearson correlation analysis indicated that the Hcy levels were negatively correlated with tissue U (r = -0.344; P = 0.030) and positively correlated with TAS (r = 0.396; P = 0.011). CONCLUSION: The uranium-induced oxidative injury may be mainly reflected in enhanced endothelial inflammation, and the direct chemical toxicity of uranium plays an important role in the process of kidney injury, especially in renal tubular injury. In addition, CysC may be a sensitive marker reflecting the nephrotoxicity of uranium; however, Hcy is not suitable for evaluating short-term endothelial inflammation involving oxidative stress.

Tibial cortex transverse transport surgery improves wound healing in patients with severe type 2 DFUs by activating a systemic immune response: a cross-sectional study.

BACKGROUND: Tibial cortex transverse transport (TTT) surgery has become an ideal treatment for patients with type 2 severe diabetic foot ulcerations (DFUs) while conventional treatments are ineffective. Based on our clinical practice experience, the protective immune response from TTT surgery may play a role against infections to promote wound healing in patients with DFUs. Therefore, this research aimed to systematically study the specific clinical efficacy and the mechanism of TTT surgery. MATERIALS AND METHODS: Between June 2022 and September 2023, 68 patients with type 2 severe DFUs were enrolled and therapized by TTT surgery in this cross-sectional and experimental study. Major clinical outcomes including limb salvage rate and antibiotics usage rate were investigated. Ten clinical characteristics and laboratory features of glucose metabolism and kidney function were statistically analyzed. Blood samples from 6 key time points of TTT surgery were collected for label-free proteomics and clinical immune biomarker analysis. Besides, tissue samples from 3 key time points were for spatially resolved metabolomics and transcriptomics analysis, as well as applied to validate the key TTT-regulated molecules by RT-qPCR. RESULTS: Notably, 64.7% of patients did not use antibiotics during the entire TTT surgery. TTT surgery can achieve a high limb salvage rate of 92.6% in patients with unilateral or bilateral DFUs. Pathway analysis of a total of 252 differentially expressed proteins (DEPs) from the proteomic revealed that the immune response induced by TTT surgery at different stages was first comprehensively verified through multi-omics combined with immune biomarker analysis. The function of upward transport was activating the systemic immune response, and wound healing occurs with downward transport. The spatial metabolic characteristics of skin tissue from patients with DFUs indicated downregulated levels of stearoylcarnitine and the glycerophospholipid metabolism pathway in skin tissue from patients with severe DFUs. Finally, the expressions of PRNP (prion protein) to activate the immune response, PLCB3 (PLCB3, phospholipase C beta 3) and VE-cadherin to play roles in neovascularization, and PPDPF (pancreatic progenitor cell differentiation and proliferation factor), LAMC2 (laminin subunit gamma 2) and SPRR2G (small proline rich protein 2G) to facilitate the developmental process mainly keratinocyte differentiation were statistically significant in skin tissues through transcriptomic and RT-qPCR analysis. CONCLUSION: Tibial cortex transverse transport (TTT) surgery demonstrates favorable outcomes for patients with severe type 2 DFUs by activating a systemic immune response, contributing to anti-infection, ulcer recurrence, and the limb salvage rate for unilateral or bilateral DFUs. The specific clinical immune responses, candidate proteins, genes, and metabolic characteristics provide directions for in-depth mechanistic research on TTT surgery. Further research and public awareness are needed to optimize TTT surgery in patients with severe type 2 DFUs.

Estimation of safe and effective dose of vancomycin in MRSA-infected patients using serum cystatin C concentrations.

OBJECTIVE: To utilize serum cystatin C (CysC) concentration to identify the daily dosage regimen of vancomycin (Van) for the treatment of patients with methicillin-resistant Staphylococcus aureus (MRSA) infections. METHODS: Serum Van, CysC, and serum and urine creatinine (Cr) concentrations were detected in 65 MRSA-infected patients. The estimated glomerular filtration rate (eGFR), Cr clearance (CLcr) and Van clearance (CLvan) were calculated and the correlation equation between CysC and CLvan was obtained using mathematical methods. Finally, the daily dosage equation of Van was derived according to pharmacokinetic theory. RESULTS: In the test sample, serum Cr was 183.27 ± 68.34 µmol/l, CLcr was 75.56 ± 30.02 ml/min, eGFR was 70.79 ± 40.79 ml/min, and serum CysC was 1.35 ± 0.61 mg/l. There was significant correlation between eGFR and CLcr (R2 = 0.8051, p = 0.000). Bland-Altman analysis showed an agreement of 96.9% (63/65) between eGFR and CLcr. eGFR was significantly correlated with CLvan (R2 = 0.8465, p = 0.000) and the correlation was significantly higher than that between CLvan and CLcr (R2 = 0.6367, p = 0.000). CysC fits a high correlated CLvan estimating equation (R2 = 0.9211, p = 0.000): CLvan(ml/min) = 64.4026 × (CysC)-1.1488. Accordingly, the predicted equation was created for calculation of the Van dosage to achieve the appropriate target steady-state serum concentration (Css): IR (the rate of continuous infusion, g/D) = 64.4026 × (CysC)-1.1488 × Css (mg/l) × (60/1,000) × 24. CONCLUSIONS: Serum CysC is a good marker of renal function in comparison with serum Cr for the dose determination of Van. CysC can estimate the daily dose of Van, and may improve therapeutic success rates of MRSA-infected patients.

[Comparative study of antitumor activity of cantharidin and cantharis peptides].

OBJECTIVE: To prepare cantharidin and cantharidin peptides from Mylabris and compare their antitumor activity. METHODS: Cantharis peptides was prepared by bionic enzymolysis approach and cantharidin was prepared by alkaline water supersonic extraction. The inhibitory effects of both compounds on BEL-7402 cells proliferation of human liver cancer were tested by Prestoblue method. The influence of both compounds on the grown of tumor, thymus, spleen of S180 tumor-bearing mice were detected. RESULTS: Cantharis peptides and cantharidin could inhibit the proliferation of BEL-7402 cells (P < 0.05) in vivo, and the inhibitory effect of cantharis peptide was 12.54% lower than that of cantharidin. At the same time, they could inhibit the grown of S180 sarcoma (P < 0.05), and the inhibitory effect of cantharidin was higher than that of cantharis peptides (5.93%). Furthermore, cantharis peptides could't inhibit the grown of thymus and spleen. CONCLUSION: Both cantharis peptides and cantharidin have antineoplastic activity, but cantharidin peptides have no immunosuppression.

Untargeted and spatial-resolved metabolomics characterize serum and tissue-specific metabolic reprogramming in acute kidney injury.

Background: Acute kidney injury (AKI) is one of the most common clinical emergencies characterized by rapid progression, difficulty in early diagnosis, and high mortality. Currently, there are no effective AKI early diagnostic methods and treatments. Therefore, identifying new mechanisms of AKI have become urgent for development new targets for early diagnosis and treatment of AKI in the current clinical setting. Methods: In this study, systematic analysis and comparison of serum metabolic profiles of clinical AKI patients, chronic kidney disease (CKD) patients, and healthy subjects were performed using untargeted metabolomics. Moreover, the first spatial metabolomic analysis of kidney tissues in an AKI mouse model using MALDI-TOF MS technology was conducted. Differentially expressed metabolites were identified using a comprehensive, publicly available database. The metabolic data obtained were evaluated using principal component analysis, (orthogonal) partial least squares discriminant analysis, and metabolic pathway analysis to explore the unique serum metabolic profile of the patients, as well as to characterize the spatial distribution of differential metabolites in the kidneys of AKI mice. Results: Significant changes in the metabolite levels of amino acids, carnitine, and lipids were observed in the AKI and CKD groups versus the healthy population, suggesting that kidney injury may lead to abnormalities in various metabolic pathways, such as amino acids, fatty acids, and lipids. The significant difference between the AKI and CKD groups were found for the first time in these indexes including amino acid, carnitine, fatty acid, and lipid levels. Additionally, spatial metabolomics results revealed that amino acid, carnitine, organic acid, and fatty acid metabolites were more likely significantly altered in the renal cortex, while lipid metabolites were both differentially distributed in the cortex and medulla of the AKI group. Conclusion: Abnormalities in the serum metabolism of amino acids, carnitine, and lipids in patients with kidney diseases, such as AKI and CKD, are closely associated with the physiological dysfunction of kidney injury. Metabolic differences between patients with AKI and CKD were compared for the first time, showing that fatty acid oxidative inhibition was more severe in patients with AKI. Furthermore, spatial metabolomics has revealed metabolic reprogramming with tissue heterogeneity in AKI mice model. Our study provides valuable information in the molecular pathological features of AKI in the kidney tissues.

C3-epi-25(OH)D3 percentage, not level, may be a potential biomarker to reflect its pathological increase in multiple diseases: a cross-sectional case-control study.

National surveys in developed countries have examined the presence of C3-epimer of 25-hydroxyvitamin D3 [C3-epi-25(OH)D3]. However, controversy remains regarding its association with disease occurrence due to its high correlation with 25-hydroxyvitamin D3 [25(OH)D3]. This study aims to investigate whether %C3-epi-25(OH)D3 can serve as an indicator for this relationship with various diseases. A total of 3086 healthy participants and 4120 patients were included in this study. We investigated the association between C3-epi-25(OH)D3 and %C3-epi-25(OH)D3 levels with gender, age, and season; compared the performance of C3-epi-25(OH)D3 and %C3-epi-25(OH)D3 across different disease conditions; and explored the correlation between %C3-epi-25(OH)D3 and various diseases. Results indicated that C3-epi-25(OH)D3 varied significantly by gender, age, and season (z/χ2 = 3.765, 10.163, and 150.975, all P < 0.01), while only season for %C3-epi-25(OH)D3 (χ2 = 233.098, P < 0.001). In contrast to the significant decrease in C3-epi-25(OH)D3, %C3-epi-25(OH)D3 showed a significant increase in 8 out of 11 disease categories (z = 3.464 ~ 11.543, all Padj < 0.05). Similar opposite changes were also observed in most of the investigated 32 specific diseases. Moreover, an elevation in %C3-epi-25(OH)D3 was found to be significantly associated with 29 specific diseases both in univariate analysis (OR = 1.16 ~ 2.10, all P < 0.05) and after adjusting for gender, age, and season (OR = 1.15 ~ 1.50, all P < 0.05). However, after further adjustment for 25(OH)D3 levels, the association remained significant only for 15 specific diseases (OR = 1.11 ~ 1.50, all P < 0.05). Seasonal stratification analysis further supports the consistent association of %C3-epi-25(OH)D3 with disease across all or nearly all four seasons. In conclusion, %C3-epi-25(OH)D3 may better reflect the production of C3-epi-25(OH)D3 in disease conditions, thereby offering a more applicable approach to investigate its association with diseases. However, the interpretation of this relationship may be confounded by 25(OH)D3 as a potential covariate.

Analysis of non-targeted serum metabolomics in patients with chronic kidney disease and hyperuricemia.

Hyperuricemia (HUA) is a common complication of chronic kidney disease (CKD). Conversely, HUA can promote the disease progression of CKD. However, the molecular mechanism of HUA in CKD development remains unclear. In the present study, we applied ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to analyze the serum metabolite profiling of 47 HUA patients, 41 non-hyperuricemic CKD (NUA-CKD) patients, and 51 CKD and HUA (HUA-CKD) patients, and then subjected to multivariate statistical analysis, metabolic pathway analysis and diagnostic performance evaluation. Metabolic profiling of serums showed that 40 differential metabolites (fold-change threshold (FC) > 1.5 or<2/3, variable importance in projection (VIP) > 1, and p < 0.05) were screened in HUA-CKD and HUA patients, and 24 differential metabolites (FC > 1.2 or<0.83, VIP>1, and p < 0.05) were screened in HUA-CKD and NUA-CKD patients. According to the analysis of metabolic pathways, significant changes existed in three metabolic pathways (compared with the HUA group) and two metabolic pathways (compared with the HUA-CKD group) in HUA-CKD patients. Glycerophospholipid metabolism was a significant pathway in HUA-CKD. Our findings show that the metabolic disorder in HUA-CKD patients was more serious than that in NUA-CKD or HUA patients. A theoretical basis is provided for HUA to accelerate CKD progress.

Serum total oxidant/antioxidant status and trace element levels in breast cancer patients.

BACKGROUND: Oxidative stress and trace elements have been implicated in the development of breast cancer. However, how they contribute to the pathogenesis of the disease and the relationship between them remain unclear. In addition, most previous studies detecting one or a few oxidant/antioxidant markers failed to consider the overall oxidant/antioxidant status of the subjects. This study was designed to address this and to investigate the association between oxidative status and trace elements in the pathogenesis of breast cancer. METHODS: Fifty-six patients with breast carcinoma at different clinical stages, 32 patients with benign breast tumor, and 20 healthy subjects (controls) were recruited into this study. Their serum total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI), and levels of Cu, Zn, Fe, Se, Mg, and Mn were measured. RESULTS: Levels of TAS, TOS, OSI, and trace elements significantly differed between the study groups. Among subgroups of patients with different clinical stages of breast cancer, the levels of all the trace elements except Zn were similar, whereas TAS, TOS, and OSI levels were all significantly different. There were significant correlations between oxidative stress parameters and levels of trace elements in patients with breast carcinoma but not in patients with benign breast tumor or in the healthy controls. CONCLUSIONS: Disturbed oxidative stress status and trace element levels may contribute to the pathogenesis of breast tumors. TAS, TOS, and OSI may be useful biomarkers for monitoring the clinical status of breast cancer.

A mathematical model for calculating the shelf life of ascorbic acid solution under given conditions.

The objective of this paper is to calculate the shelf life of ascorbic acid solution under given conditions by using a mathematical model. An antioxidant, sodium metabisulfite, was added to the ascorbic acid solution. The kinetic parameters of the degradation reaction of ascorbic acid and sodium metabisulfite, were investigated, respectively, and then a mathematical model was developed. According to the mathematical model, the calculated shelf lives of ascorbic acid solution were 783, 835, 873, and 885 days for specifications 2, 5, 10, and 20 mL, respectively. The results showed that the obtained mathematical model can be used to calculate the shelf life of ascorbic acid solution under given conditions.