OBJECTIVE To explore the mechanism of Yiqi yangyin huazhuo tongluo formula in ameliorating diabetic kidney disease （DKD） based on the gut microbiota-lipopolysaccharide （LPS）/Caspase-11/gasdermin D （GSDMD） pathway.METHODS A DKD mouse model was established by high-fat and high-sugar diet combined with intraperitoneal injection of low-dose streptozotocin. The successfully modeled mice were randomly divided into model group， metformin group （positive control， 200 mg/kg） and Yiqi yangyin huazhuo tongluo formula group （32.4 g/kg， calculated as crude drug dosage）， with 10 mice in each group； another 10 mice were set as the control group. Mice were intragastrically administered with corresponding drug solutions or normal saline once a day for 8 consecutive weeks. After the last administration， serum glycolipid metabolism indicators and urinary renal function indicators of mice were determined； the pathological and morphological changes of renal and colonic tissues were observed； LPS contents in renal tissues， colonic contents and serum were detected. 16S rRNA high-throughput sequencing was performed for gut microbiota analysis of colonic contents. The protein expressions of Occludin and zonula occludens-1 （ZO-1） in colonic tissues as well as Caspase-11 and GSDMD in renal tissues were detected.RESULTS Compared with the control group， the model group exhibited significantly elevated levels of serum fasting blood glucose， triglyceride， total cholesterol， serum creatinine， blood urea nitrogen and microalbumin， LPS contents in each sample， as well as protein expressions of Caspase-11 and GSDMD in renal tissues （ P ＜0.05）. The protein expressions of ZO-1 and Occludin in colonic tissues， together with Chao1 and Ace indices of gut microbiota， were markedly decreased （ P ＜0.05）， and the ratio of Firmicutes to Bacteroidetes was abnormally increased. Obvious histopathological changes were observed in renal and colonic tissues； the percentage of PAS-positive area in kidney was markedly increased （ P ＜0.05）. Compared with the model group， the Yiqi yangyin huazhuo tongluo formula group showed remarkable reductions in glycolipid metabolism and renal function-related indicators（except total cholesterol）， LPS contents in various samples， and renal protein expressions of Caspase-11 and GSDMD （ P ＜0.05）. The protein expression level of ZO-1 in colonic tissues and the Ace index of gut microbiota were significantly increased （ P ＜0.05）， and the relative abundance of beneficial genera including Bifidobacterium ， Faecalibaculum and Akkermansia in the gut microbiota was elevated.CONCLUSIONS The mechanism by which Yiqi yangyin huazhuo tongluo formula ameliorates DKD may be related to reshaping gut microbiota， repairing intestinal mucosal barrier， reducing LPS translocation into blood， and further inhibiting Caspase-11/GSDMD-mediated pyroptosis in kidney.

Systemic scleroderma (SSc) is a fibrotic disease involving the skin and a variety of internal organs, which is currently thought to be mainly caused by abnormal activation of fibroblasts and excessive secretion of extracellular matrix. Growing evidence has shown signaling cascades associated with SSc fibrosis, including transforming growth factor-β/Smads, Notch, Wnt/β-catenin, rat sarcoma/extracellular regulated protein kinases, etc. MicroRNAs participate in the post-transcriptional regulation of different target genes and control the cell state, determining the changes in transcriptomes and proteomes caused by various cellular signaling cascades. This review summarizes recent research progress in the regulatory role of microRNAs in SSc fibrosis-related signaling pathways and potential therapeutic targets.

Systemic scleroderma (SSc) is a fibrotic disease involving the skin and a variety of internal organs, which is currently thought to be mainly caused by abnormal activation of fibroblasts and excessive secretion of extracellular matrix. Growing evidence has shown signaling cascades associated with SSc fibrosis, including transforming growth factor-β/Smads, Notch, Wnt/β-catenin, rat sarcoma/extracellular regulated protein kinases, etc. MicroRNAs participate in the post-transcriptional regulation of different target genes and control the cell state, determining the changes in transcriptomes and proteomes caused by various cellular signaling cascades. This review summarizes recent research progress in the regulatory role of microRNAs in SSc fibrosis-related signaling pathways and potential therapeutic targets.

Objective To investigate the protective effect of Yiqi Yangyin Huazhuo Tongluo Formula(YYHT)against high glucose-induced injury in mouse renal podocytes(MPC5 cells)and the possible mechanism.Methods Adult Wistar rats were treated with 19,38,and 76 g/kg YYHT or saline via gavage for 7 days to prepare YYHT-medicated or blank sera for treatment of MPC5 cells cultured in high glucose(30 mmol/L)prior to transfection with a miR-21a-5p inhibitor or a miR-21a-5p mimic.The changes in miR-21a-5p expressions and the mRNA levels of FoxO1,PINK1,and Parkin in the treated cells were detected with qRT-PCR,and the protein levels of nephrin,podocin,FoxO1,PINK1,and Parkin were detected with Western blotting.Autophagic activity in the cells were evaluated with MDC staining.The effect of miR-21a-5p mimic on FoxO1 transcription and the binding of miR-21a-5p to FoxO1 were examined with luciferase reporter gene assay and radioimmunoprecipitation assay.Results MPC5 cells exposed to high glucose showed significantly increased miR-21a-5p expression,lowered expressions of FoxO1,PINK1,and Parkin1 mRNAs,and reduced levels of FoxO1,PINK1,parkin,nephrin,and podocin proteins and autophagic activity.Treatment of the exposed cells with YYHT-medicated sera and miR-21a-5p inhibitor both significantly enhanced the protein expressions of nephrin and podocin,inhibited the expression of miR-21a-5p,increased the mRNA and protein expressions of FoxO1,PINK1 and Parkin,and upregulated autophagic activity of the cells.Transfection with miR-21a-5p mimic effectively inhibited the transcription of FoxO1 and promoted the binding of miR-21a-5p to FoxO1 in MPC5 cells,and these effects were obviously attenuated by treatment with YYHT-medicated sera.Conclusion YYHT-medicated sera alleviate high glucose-induced injury in MPC5 cells by regulating miR-21a-5p/FoxO1/PINK1-mediated mitochondrial autophagy.

OBJECTIVES: To investigate the protective effect of Yiqi Yangyin Huazhuo Tongluo Formula (YYHT) against high glucose-induced injury in mouse renal podocytes (MPC5 cells) and the possible mechanism. METHODS: Adult Wistar rats were treated with 19, 38, and 76 g/kg YYHT or saline via gavage for 7 days to prepare YYHT-medicated or blank sera for treatment of MPC5 cells cultured in high glucose (30 mmol/L) prior to transfection with a miR-21a-5p inhibitor or a miR-21a-5p mimic. The changes in miR-21a-5p expressions and the mRNA levels of FoxO1, PINK1, and Parkin in the treated cells were detected with qRT-PCR, and the protein levels of nephrin, podocin, FoxO1, PINK1, and Parkin were detected with Western blotting. Autophagic activity in the cells were evaluated with MDC staining. The effect of miR-21a-5p mimic on FoxO1 transcription and the binding of miR-21a-5p to FoxO1 were examined with luciferase reporter gene assay and radioimmunoprecipitation assay. RESULTS: MPC5 cells exposed to high glucose showed significantly increased miR-21a-5p expression, lowered expressions of FoxO1, PINK1, and Parkin1 mRNAs, and reduced levels of FoxO1, PINK1, parkin, nephrin, and podocin proteins and autophagic activity. Treatment of the exposed cells with YYHT-medicated sera and miR-21a-5p inhibitor both significantly enhanced the protein expressions of nephrin and podocin, inhibited the expression of miR-21a-5p, increased the mRNA and protein expressions of FoxO1, PINK1 and Parkin, and upregulated autophagic activity of the cells. Transfection with miR-21a-5p mimic effectively inhibited the transcription of FoxO1 and promoted the binding of miR-21a-5p to FoxO1 in MPC5 cells, and these effects were obviously attenuated by treatment with YYHT-medicated sera. CONCLUSIONS YYHT-medicated sera alleviate high glucose-induced injury in MPC5 cells by regulating miR-21a-5p/FoxO1/PINK1-mediated mitochondrial autophagy.
Animals ; MicroRNAs/genetics* ; Podocytes/pathology* ; Drugs, Chinese Herbal/pharmacology* ; Autophagy/drug effects* ; Rats, Wistar ; Protein Kinases/metabolism* ; Rats ; Forkhead Box Protein O1 ; Mice ; Mitochondria/drug effects* ; Ubiquitin-Protein Ligases/metabolism* ; Glucose ; Diabetic Nephropathies ; Male ; Membrane Proteins/metabolism* ; Intracellular Signaling Peptides and Proteins

OBJECTIVES: To explore the association of the expression of WW domain-containing ubiquitin E3 ligase 1 (WWP1) with immune infiltration in tumor microenvironment (TME) of ovarian cancer. METHODS: Ovarian cancer patient data from The Cancer Genome Atlas (TCGA) were used to analyze the association of WWP1 expression with patient prognosis. TISCH2 was utilized to analyze the changes in immune cell subtypes in TME of metastatic tumor and after chemotherapy. The impact of WWP1 on immune cell infiltration, somatic copy number alterations of WWP1 and evolution of immune cell subtypes was evaluated using TIMER and TIGER pseudo-time analysis. A deep learning model was used to analyze TCGA pathological images to investigate the effect of WWP1 on TME of ovarian cancer. RNA-seq analysis was conducted to identify the differentially expressed genes in WWP1-overexpressing SKOV3 cells and validate immune infiltration. Multicolor immunofluorescence assay was used to analyze the immune markers in SKOV3 and SKOV3/DDP cell xenografts in nude mice. RESULTS: The patients with high WWP1 expression levels had significantly lower overall survival rate (P=0.0012). High WWP1 expression levels and Stage IV disease were both associated with a poor prognosis (P<0.05). In metastatic ovarian cancer or after chemotherapy, the percentages of malignant tumor cells and tumor-associated fibroblasts increased in the TME, accompanied by elevated WWP1 levels. WWP1 expression level was positively correlated with pro-tumorigenic immunosuppressive cells (r=0.1323-0.3955, P<0.05) and negatively with tumor-inhibiting immune cells (r=-0.1949- -0.1333, P<0.05). Specific copy number alterations of WWP1 also influenced CD8⁺ T cell percentage and neutrophil infiltration levels in the TME. RNA-seq analysis of WWP1-overexpressing SKOV3 cells and immunofluorescence assay of the tumor-bearing mice yielded findings consistent with those of bioinformatics analysis. CONCLUSIONS WWP1 may serve as a prognostic biomarker and a potential target for immune regulation in the TME of ovarian cancer.
Female ; Ovarian Neoplasms/genetics* ; Humans ; Ubiquitin-Protein Ligases/metabolism* ; Tumor Microenvironment/immunology* ; Animals ; Mice ; Cell Line, Tumor ; Mice, Nude ; Prognosis ; Gene Expression Regulation, Neoplastic

Objective:To investigate the relationship between cancer node status identified after radical resection and patterns of recurrence in stage Ⅱ/Ⅲ gastric cancer patients for developing personalized follow-up plans in node-positive patients.Methods:A retrospective analysis was conducted on 706 stage Ⅱ/Ⅲ gastric cancer patients who underwent curative intent surgery at Henan Cancer Hospital from Sep 2015 to Sep 2018. Patients were categorized into node-positive (TDs +) and node-negative (TDs -) groups based on their cancer node status. Clinical-pathological characteristics and recurrence patterns were compared between the groups to formulate an optimized follow-up plan. Results:Among the patients, there were 112 TDs + and 594 TDs - cases. TDs - patients exhibited significantly better overall survival and disease-free survival rates at 1 and 3 years compared to the TDs + group (92.4%,72.2% vs. 79.4%,38.8%;87.9%,68.6% vs. 66.7%,35.9%, all P<0.05). The cumulative recurrence rates of peritoneal and distant metastases were higher in TDs + patients compared to TDs - patients, with statistical significance (33.0% vs. 14.5%,21.4% vs. 10.8%, χ 2=21.285,8.851 all P<0.05). TDs + patients experienced significantly earlier median time to distant metastasis compared to TDs - patients (9.0 months vs. 15.4 months , Z=-2.294 P=0.022). The risk of peritoneal metastasis recurrence was higher in the TDs + group, showing a bimodal recurrence pattern at 8.5 and 15.0 months post-surgery. TDs + patients also had a higher risk of distant metastasis recurrence, with a single peak at 6 months. Conclusion:Postoperative recurrence patterns differ between TDs + and TDs - patients, and an optimized follow-up plan can enhance early detection of recurrence.

Objective:To analyze the curative effect of sandwich cheiloplasty for the treatment of macrocheilia secondary to port-wine stain (PWS).Methods:A total of 43 patients, including 20 males and 23 females, aged 37.0 years (ranged from 4 to 69 years), who underwent sandwich cheiloplasty for the treatment of macrocheilia secondary to PWS from March 2008 to March 2022 were retrospectively enrolled. According to the location of the lesions, they were divided into two groups: upper lip group (21 cases) and lower lip group (22 cases). Pathological examinations were performed in all excised specimens. Postoperative attentions should be paid to keeping incisions locally clean, observing the blood supplying, and taking care of fluid accumulation or incision split. After discharge, the lip appearance and function were evaluated every six months in the outpatient department, and the long-term efficacy was further divided into three levels: grade Ⅰ (poor), grade Ⅱ (moderate), and grade Ⅲ (good). The comparison of grade data named long-term efficacy between the two groups was conducted by Mann-Whitney U test.Results:All 43 patients underwent sandwich cheiloplasty, of which 40 patients received one operation, whereas the other 3 received two due to recurrences. One-stage incision healing was achieved in 43 cases, whereas partial mucosal necrosis appeared in 2 cases, and slight incision dehiscence occurred in 1 case, which healed well after local dressing change. The pathological examination results of 43 excised tissue specimens all showed capillary malformations. After follow-up for 1 to 10 years, long-term efficacy evaluation was made up as follows: 36 cases were evaluated as grade Ⅲ, 7 cases as grade Ⅱ, and 0 case as grade Ⅰ. There were 18 cases of grade Ⅲ, and 3 cases of grade Ⅱ in group A, compared to 18 cases of grade Ⅲ, and 4 cases of grade Ⅱ in group B. By rank sum test, there was no significant difference in overal efficacy between the two groups ( Z=0.342, P>0.05). Conclusions:Sandwich cheiloplasty for the treatment of macrocheilia secondary to PWS effectively removes malformed vascular lesions, and consequently, it can achieve a good long-term therapeutic effect.

Objective:To establish a nomogram to predict overall survival of patients with stage Ⅰ-Ⅲ gastric cancer (GC) based on preoperative inflammatory markers.Methods:Clinicopathological and follow-up data of 1 035 patients with stage Ⅰ-Ⅲ gastric cancer operated at He'nan Cancer Hospital between May 2015 and Oct 2016 were retrospectively collected. A nomogram was established based on prognostic factors. Harrell's concordance index (C-index), receiver operating characteristic (ROC) curve, calibration curve and decision curve analysis (DCA) were used to verify the performance of the model according to differentiation, calibration and clinical utility.Results:A total of 1 035 patients were enrolled . The median follow-up time was 41.9 months; According to the optimal cutoff value, 170 were with elevated neutrophil-to-lymphocyte ratio (NLR) and 865 with a reduced　ratio; 562 in elevated platelet-to-lymphocyte ratio (PLR) vs. 473 in the reduced group; fibrinogen/albumin ratio (FAR) elevated in 108 group vs. 972 in the reduced group; 180 in the prognostic nutritional index score (PNI) elevated group and 855 in the reduced group. Two hundred and sixty-seven patients were categorized at stage Ⅰ, 334 at stage Ⅱ ,434 at stage Ⅲ. Multivariate regression analysis showed tumor location, vascular tumor thrombus, pTNM stage, FAR, PNI and NLR were independent prognostic factors (all P<0.05). The C-index of the nomogram was 0.723 (95% CI: 0.710 -0.736) and had better clinical utility than the American Joint Committee on Cancer (AJCC) 8th TNM staging system 0.693 (95% CI, 0.681 -0.705). The calibration curve of the nomogram showed that the predicted survival rate was consistent with the actual survival rate in GC patients. Compared to AJCC 8th pTNM staging system, the DCA curve indicate that the nomogram has a higher net income. Conclusion:The nomogram predicting overall survival of patients with stage Ⅰ-Ⅲ gastric cancer is established and verified , which provides better individual prediction than TNM staging system.

Objective:To analyze the influencing factors of gout patients and explore whether there is an interaction between serum uric acid and diastolic blood pressure affecting the onset of tophi.Methods:A total of 4 798 gout patients were retrospective selected in the outpatient Department of the Gout Clinical Medical Center of the Affiliated Hospital of Qingdao University from September 2016 to May 2020. It was divided into tophi group and non-tophi group to compare the differences in indicators. A logistic regression model was used to analyze the influencing factors of tophi, and an interaction model was constructed to analyze the interactions.Results:Multivariate logistic regression analysis showed significant associations between age, diastolic blood pressure, alcohol consumption history, gout family history, blood uric acid, urea nitrogen, and creatinine clearance and tophi formation. The results of blood uric acid-related interaction analysis showed a significant interaction between blood uric acid and diastolic blood pressure( Pinteraction=0.014), and the risk of developing tophi in low diastolic blood pressure and high diastolic blood pressure group increased by 34.4%( OR=1.344, 95% CI 1.105-1.635, P=0.003) and 95.4%( OR=1.954, 95% CI 1.558-2.450, P<0.001) in the high blood uric acid group compared with the low blood uric acid group. The results of diastolic blood pressure and blood uric acid subgroup analysis showed that there was no statistical difference in the risk of developing tophi in people with low uric acid levels( P=0.238), but in people with high uric acid levels, the risk of developing tophi was 67%( OR=1.670, 95% CI 1.379-2.022, P<0.001) higher than that in the low uric acid group. Conclusion:Age, diastolic blood pressure, combined alcohol consumption history and gout family history, blood uric acid, renal function are related to the occurrence of tophi. High uric acid and high diastolic blood pressure have interaction on the occurrence of tophi. Attention and proactive intervention shall be applied to this group of patients.