miR-223-3p mediates the diabetic kidney disease progression by targeting IL6ST/STAT3 pathway.

Diabetic kidney disease (DKD), the most pervasive complication in diabetic patients, has become a major health threat to the aging population. Our previous miRNA profiling identified hsa-miR-223-3p as a dysregulated miRNA in the DKD samples, which may serve as a biomarker for DKD diagnosis. However, the specific mechanism of miR-223-3p in the pathogenesis of DKD remains to be elucidated. In this study, we first verified that miR-223-3p level was significantly decreased in the in vitro cell model and in vivo db/db DKD model, accompanied with endothelial cell damage. Importantly, inhibiting the expression of miR-223-3p exacerbated high-glucose induced damages in Human Umbilical Vein Endothelial Cells (HUVECs) and Human Renal Glomerular Endothelial Cells (HRGECs), while miR-223-3p overexpression showed the opposite effect. We further demonstrated that miR-223-3p associated with IL6T mRNA and attenuated the progression of DKD by suppressing the downstream STAT3 activation, indicative of the implication of miR-223-3p/IL6T/STAT3 axis in the pathogenesis of DKD.

Identifying key genes for diabetic kidney disease by bioinformatics analysis.

BACKGROUND: There are no reliable molecular targets for early diagnosis and effective treatment in the clinical management of diabetic kidney disease (DKD). To identify novel gene factors underlying the progression of DKD. METHODS: The public transcriptomic datasets of the alloxan-induced DKD model and the streptozotocin-induced DKD model were retrieved to perform an integrative bioinformatic analysis of differentially expressed genes (DEGs) shared by two experimental animal models. The dominant biological processes and pathways associated with DEGs were identified through enrichment analysis. The expression changes of the key DEGs were validated in the classic db/db DKD mouse model. RESULTS: The downregulated and upregulated genes in DKD models were uncovered from GSE139317 and GSE131221 microarray datasets. Enrichment analysis revealed that metabolic process, extracellular exosomes, and hydrolase activity are shared biological processes and molecular activity is altered in the DEGs. Importantly, Hmgcs2, angptl4, and Slco1a1 displayed a consistent expression pattern across the two DKD models. In the classic db/db DKD mice, Hmgcs2 and angptl4 were also found to be upregulated while Slco1a1 was downregulated in comparison to the control animals. CONCLUSIONS: In summary, we identified the common biological processes and molecular activity being altered in two DKD experimental models, as well as the novel gene factors (Hmgcs2, Angptl4, and Slco1a1) which may be implicated in DKD. Future works are warranted to decipher the biological role of these genes in the pathogenesis of DKD.

Study on the mitochondrial apoptosis pathways of small cell lung cancer H446 cells induced by Trichinella spiralis muscle larvae ESPs.

Trichinella spiralis (T.spiralis) muscle-larva (ML) excretory-secretory proteins (ESPs) contain antitumour-active substances. ESPs have been shown to inhibit tumour growth. To explore the effects of these proteins on small cell lung cancer cells and the possible mechanisms of their antineoplastic action, H446 SCLC cells were co-cultured with different concentrations of T. spiralis ML ESPs for 12, 24 and 48 h. Our results showed that T. spiralis ML ESPs significantly inhibited H446 cell proliferation, which was dose-and time-dependent. The results of flow cytometry testing indicate a clear apoptosis trend in H446 cells co-cultured with ESPs for 24 h. Reverse transcription polymerase chain reaction and Western blotting results showed increased expression of pro-apoptosis genes Bax, Cyt-C, Apaf-1, caspase-9 and caspase-3, compared with the negative control group, and decreased the expression of anti-apoptosis genes Bcl-2 and Livin. Our results suggest that T. spiralis ML ESPs can induce apoptosis in H446 cells through a mitochondrial pathway, which may be a mechanism of antineoplastic action in T. spiralis ML ESPs.

[LC-MS/MS Analysis on the Components of Excretory-secretory Protein of Trichinella spiralis Muscle Larvae].

Objective: To analyze the components of excretory-secretory protein（ESP） of Trichinella spiralis muscle larvae, and search for the anti-tumor protein components. Methods: The Trichinella spiralis muscle larvae were collected, and ESP was prepared. The ESP was separated in 15% SDS-PAGE. Proteins extracted from the protein bands were lysed with trypsin, and analyzed by LC-MS/MS. The identified proteins were classified by Gene Ontology（GO） according to cell component, molecular function, and biological processes. Results: SDS-PAGE revealed clear protein bands at Mr 10 000-142 000. A total of 162 proteins were analyzed with LC-MS/MS, of which 63 were identified, 34 were putative proteins, and 65 were unidentified proteins. Six anti-tumor relevant proteins were revealed, which were tropomyosin, histone H2A, cleavage and polyadenylation specificity factor subunit 2, serine proteinase inhibitor Kazal-type 4, Armadillo segment polarity protein and eukaryotic initiation factor 4A. The GO enrichment analysis showed that the identified proteins possessed 54 different types of molecular functions, and participated in cell structure and 382 biological processes. Conclusion: The ESP of Trichinella spiralis muscle larvae has complex protein components, many with unknown identities. Six anti-tumor relevant proteins were determined from the 63 identified proteins.

[Effect of the excretory/scretory proteins from Trichinella spiralis on apoptosis of NCI-H446 small-cell lung cancer cells].

OBJECTIVE: To investigate the effect of excretory/secretory proteins from Trichinella spiralis on apoptosis of NCI-H446 small-cell lung cancer cells. METHODS: Trichinella spiralis muscle stage larvae (5 x 10(6)/ml) were cultured in culture media for 24 h, the excretory/secretory proteins were collected from the supernatant of culture media. NCI-H446 small-cell lung cancer cells (No. A05) were randomly divided into three groups: experiment group (A), standard control group (apoptosis group, B), and control group (C). NCI-H446 cells in groups A and B were cultured with 0.3 mg/ml T. spiralis excretory/secretory proteins, and 6.4 microg/ml cisplatin for 24 h, respectively. NCI-H446 cells of group C were cultured for 24 h without any treatment. The expression of Bcl-2, Fas and Fasl mRNA was detected by RT-PCR. C-myc protein expression level was examined by Western blotting and immunofluorescence assay. RESULTS: The level of Bcl-2 mRNA was lowest in group A(0.575 +/- 0.047) , Bcl-2 mRNA level in group C (0.975 +/- 0.069) was higher than that of group B (0.850 +/- 0.073) (P<0.05). Fas mRNA level was highest in group A (0.975 +/- 0.115), followed by group B (0.817 +/- 0.121) and group C(0.769 +/- 0.061) (P<0.05). The level of Fasl mRNA in groups A, B, and C was 0.669 +/- 0.051, 0.787 +/- 0.124, and 0.875 +/- 0.125, respectively (P<0.05). Fas/Fasl mRNA ratio in groups A, B, and C was 1.475, 1.038, and 0.878. Western blotting showed that the expression of C-myc protein in group C (1.172 +/- 0.026) was highest, followed by group B (1.074 +/- 0.069) and A (0.566 +/- 0.054) (P<0.05). Immunofluorescence test indicated that the C-mye protein was found in the cytoplasm and the nucleus 24 h after treated with 0.3 mg/ml T. spiralis excretory/secretory proteins and 6.4 p.g/ml cisplatin. CONCLUSION: Trichinella spiralis excretory/secretory proteins may inhibit apoptosis of NCI-H446 small-cell lung cancer cells by reducing the apoptosis protein C-myc and Bcl-2 mRNA levels, and causing the increase of Fas/Fasl mRNA ratio.

Sodium glucose cotransporter2 inhibitors for type 1 diabetes mellitus: A meta-analysis of randomized controlled trials.

AIMS: Sodium glucose cotransporter2 (SGLT2) inhibitors are controversial in the treatment of type 1 diabetes mellitus (T1DM). This study is a systematic evaluation of the safety of SGLT2 inhibitors usage in T1DM. METHODS: Comprehensive literature search in six databases from inception to September 2022. Randomized controlled trials (RCTs) of T1DM treated with SGLT2 inhibitor vs. placebo were included. Data were extracted from the literature that met the inclusion criteria. After quality evaluation by the Cochrane risk bias assessment tool, meta-analysis was performed using Revman 5.4 and Stata 17.1. RESULTS: The study consisted of 16 RCTs with 7192 patients. The results indicated that SGLT2inhibitors reduce glycated hemoglobin (HbA1c, Mean difference (MD)- 0.29%, P < 0.05), fasting plasma glucose (FPG, MD-0.85 mmol/L, P < 0.05), mean amplitude of glucose excursions (MAGE, 15.75 mg/dL, P < 0.05), body weight (MD-3.49 kg, P < 0.05), and total insulin dosage (MD-7.14 IU/day, P < 0.05). Furthermore, cautious SGLT2 inhibitors did not induce the risk of hypoglycemia (RR1.00, P = 0.86), urinary tract infections (RR1.02, P = 0.085), and diarrhea (RR1.34, P = 0.523). CONCLUSION: Based on this meta-analysis, SGLT22 inhibitors reduce insulin dosage without increasing the risk of hypoglycemia and diabetic ketoacidosis for type 1 diabetes mellitus in 1month.

Type 1 diabetes mellitus induced by PD-1 inhibitors in China: a report of two cases.

The immune-related adverse events associated with immunotherapy may affect endocrine glands and other tissues. Two Chinese patients with malignancies were treated with programmed cell death-1 (PD-1) inhibitors (nivolumab and pembrolizumab) and followed up with biochemical tests over 1 year. After PD-1 treatment for 6 to 10 months, the patients developed symptoms of diabetes, ketoacidosis, and insulin secretion failure. Type 1 diabetes mellitus was confirmed by the characteristic fluctuation of blood glucose that was controlled with multiple daily insulin injections. Neither patient's insulin depletion status was reversed in subsequent years. To decrease the life-threatening complications of diabetic hyperosmolar syndrome and ketoacidosis caused by type 1 diabetes mellitus, it is necessary to monitor the blood glucose and hemoglobin A1c levels. Islet ß-cell autoantibodies and human leukocyte antigen genes can provide additional information in select cases.

Free-radical oxidation and superoxide dismutase activity in synovial fluid of patients with temporomandibular disorders.

AIMS: To measure the activity of oxygen free radicals and the level of antioxidant enzyme superoxide dismutase (SOD) in the synovial fluid (SF) of the temporomandibular joints (TMJs) of patients with temporomandibular disorders (TMD). METHODS: Thirty-two patients were divided into 3 subgroups: anterior disc displacement with reduction, anterior disc displacement without reduction, and osteoarthrosis. Six healthy volunteers served as controls. A pumping procedure was used to take SF from the superior TMJ space. The concentration of lipid peroxidation products was assessed by means of the thiobarbituric acid reaction, and the level of SOD was assessed with spectrophotometry. RESULTS: SF levels of lipid peroxides (LPO) in the control and patient groups were 0.010 (+/- 0.004) x 10(-3) nmol/mg protein and 0.61 (+/- 0.121) x 10(-3) nmol/mg protein, respectively. SF SOD levels were 4.61 (+/- 1.30) NU/mg protein and 9.83 (+/- 2.66) NU/mg protein, respectively. Both the concentration of LPO and the level of SOD activity were significantly higher in the TMD patients than in the normal control subjects (P < .001 and P < .01, respectively). There were no significant differences in the level of LPO or SOD between the 3 subgroups. CONCLUSION: These results demonstrate oxygen free radicals and antioxidant enzymes may be connected in the pathogenesis of TMD.