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 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.

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.

Tumor immunotherapy,recognized as the fourth major treatment modality alongside surgery,radiotherapy,and chemotherapy,fundamentally relies on mobilizing and enhancing the body's intrinsic immune system to achieve the precise targeting and elimination of neoplastic lesions.In this therapeutic framework,macrophages derived from blood monocyte differentiation serve as critical components of the innate immune defense system and exert profound impacts within the tumor microenvironment(TME).As the dominant inflammatory cell population infiltrating the TME,tumor-associated macrophages(TAMs)not only perform a key function in immune regulation but also serve as a paradigm for the connection between inflammation and tumors.Therapeutic strategies targeting TAMs aim to reverse the immunosuppressive milieu of the TME through multifaceted regulatory mechanisms,including cellular depletion or functional reprogramming,thereby effectively impeding tumor progression.This review systematically analyzes the intricate immune regulatory mechanisms of macrophages in tumor immunotherapy and synthesizes research advancements in major therapeutic agents targeting TAMs,aiming to provide researchers in the field of tumor immunotherapy and developers of macrophage-modulating pharmaceuticals with novel theoretical insights and practical guidelines.

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.

Tumor immunotherapy,recognized as the fourth major treatment modality alongside surgery,radiotherapy,and chemotherapy,fundamentally relies on mobilizing and enhancing the body's intrinsic immune system to achieve the precise targeting and elimination of neoplastic lesions.In this therapeutic framework,macrophages derived from blood monocyte differentiation serve as critical components of the innate immune defense system and exert profound impacts within the tumor microenvironment(TME).As the dominant inflammatory cell population infiltrating the TME,tumor-associated macrophages(TAMs)not only perform a key function in immune regulation but also serve as a paradigm for the connection between inflammation and tumors.Therapeutic strategies targeting TAMs aim to reverse the immunosuppressive milieu of the TME through multifaceted regulatory mechanisms,including cellular depletion or functional reprogramming,thereby effectively impeding tumor progression.This review systematically analyzes the intricate immune regulatory mechanisms of macrophages in tumor immunotherapy and synthesizes research advancements in major therapeutic agents targeting TAMs,aiming to provide researchers in the field of tumor immunotherapy and developers of macrophage-modulating pharmaceuticals with novel theoretical insights and practical guidelines.

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 construct the evaluation index system for scientific research ability of hospital pharmacists， and provide reference for the improvement of hospital pharmacists’ scientific research ability and the formulation of relevant scientific research policies. METHODS The relevant indexes of scientific research evaluation of hospital pharmacists were extracted by literature analysis， and consultation questionnaire was designed according to Likert grade 5 scoring method. Delphi method was used to conduct two rounds of questionnaire consultation for 28 experts， and the weight of each index was determined by analytic hierarchy process. The reliability and validity of index system were analyzed by questionnaire survey. RESULTS After two rounds of expert correspondence， evaluation index system for scientific research ability of hospital pharmacists was finally determined from three core dimensions： basic scientific research ability， scientific research achievements and transformation ability， academic influence and personnel training （including 11 sub-dimensions and 34 measurement items）. The weight value of each dimension index was determined. The result of reliability and validity analysis confirmed the scientific rationality of the index system. CONCLUSIONS The established evaluation index system for scientific research ability of hospital pharmacists is innovative， comprehensive and scientific. The index system model can provide reference for the improvement of hospital pharmacists’ scientific research ability and the formulation of relevant scientific research policies.

OBJECTIVE To establish the index system of quality evaluation standard for pharmacist training in cough and wheeze pharmaceutical care （CWPC） outpatient department， and to provide a basis for the selection of CWPC pharmacist training teachers and the improvement of training plans. METHODS Based on Kirkpatrick model， using the Delphi method， a total of 15 experts from 13 tertiary hospitals in 10 provinces in China were consulted to establish the standard index system of the quality evaluation for CWPC pharmacists training. Analytic hierarchy process （AHP） was adopted to determine the weights of each indicator and quantify the index system according to the weights of indicators at all levels. RESULTS The coefficient of expert authority was 0.810， the judgment coefficient was 0.727， and the familiarity coefficient was 0.893. The Kendall coordination coefficient of each index was 0.308-0.687. The P values of χ2 test were all less than 0.05， which indicated that the degree of coordination of the experts was high. After two rounds of correspondence， Kirkpatrick model-based index system of quality evaluation standard for CWPC pharmacist training was determined. The index system included 4 first-level indexes （participants’ reaction layer， learning gain layer， behavior improvement layer， training outcome layer）， 12 second-level indexes （such as training needs， teaching methods， theoretical knowledge， practical skills， job abilities， patient benefits， etc.） and 44 third-level indexes （such as clear training objectives， core system of CWPC， special device operation and evaluation， professional knowledge related to the treatment of cough and wheeze patients， promoting the construction of CWPC， improving patient compliance）. CONCLUSIONS The constructed index system of quality evaluation standard for CWPC pharmacist training has a certain level of authority and scientificity， and provides a scientific theoretical basis for quality evaluation of CWPC pharmacist training.