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

Objective To investigate the effects of Ras-related protein Rab-2B(RAB2B)on the biological behaviors of pancreatic cancer cells and elucidate its underlying mechanism.Methods PANC-1 cells,which exhibit relatively high RAB2B expression,and BXPC-3 cells,which display relatively low RAB2B expression,were selected from five pancreatic cancer cell lines.RAB2B-siRNA and pcDNA3.1-RAB2B plasmids were transfected into PANC-1 and BXPC-3 cells using a cell transfection technique.The CCK-8 assay was employed to evaluate the prolif-erative capacity of pancreatic cancer cells following RAB2B intervention.Wound healing and Transwell chamber assays were utilized to assess the migratory and invasive capabilities of pancreatic cancer cells.Additionally,the mRNA and protein expression levels of RAB2B,NF-κB,and Fibronectin 1(FN1)were analyzed by qRT-PCR and Western blot(WB),respectively.Results RAB2B mRNA and protein expression levels were significantly down-regulated in PANC-1 cells following transfection(P<0.05).CCK-8 assay results demonstrated that the proliferative capacity of PANC-1 cells was markedly reduced(P<0.05),and the wound-healing ability was substantially impaired(P<0.01)upon RAB2B knockdown.Transwell assays revealed a significant decrease in cell migration(P<0.01),while Western blot analysis indicated that the expression levels of phosphorylated p65 and FN1 were notably diminished(P<0.01).Conversely,overexpression of RAB2B reversed these aforementioned alterations.Conclusions Knockdown of RAB2B in PANC-1 cells significantly suppresses cell proliferation and migration,whereas overexpression of RAB2B in BXPC-3 cells markedly promotes these processes.This effect is likely mediated through the activation of the NF-κB signaling pathway and the subsequent regulation of FN1 expression.

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.

Objective To investigate the effects of Ras-related protein Rab-2B(RAB2B)on the biological behaviors of pancreatic cancer cells and elucidate its underlying mechanism.Methods PANC-1 cells,which exhibit relatively high RAB2B expression,and BXPC-3 cells,which display relatively low RAB2B expression,were selected from five pancreatic cancer cell lines.RAB2B-siRNA and pcDNA3.1-RAB2B plasmids were transfected into PANC-1 and BXPC-3 cells using a cell transfection technique.The CCK-8 assay was employed to evaluate the prolif-erative capacity of pancreatic cancer cells following RAB2B intervention.Wound healing and Transwell chamber assays were utilized to assess the migratory and invasive capabilities of pancreatic cancer cells.Additionally,the mRNA and protein expression levels of RAB2B,NF-κB,and Fibronectin 1(FN1)were analyzed by qRT-PCR and Western blot(WB),respectively.Results RAB2B mRNA and protein expression levels were significantly down-regulated in PANC-1 cells following transfection(P<0.05).CCK-8 assay results demonstrated that the proliferative capacity of PANC-1 cells was markedly reduced(P<0.05),and the wound-healing ability was substantially impaired(P<0.01)upon RAB2B knockdown.Transwell assays revealed a significant decrease in cell migration(P<0.01),while Western blot analysis indicated that the expression levels of phosphorylated p65 and FN1 were notably diminished(P<0.01).Conversely,overexpression of RAB2B reversed these aforementioned alterations.Conclusions Knockdown of RAB2B in PANC-1 cells significantly suppresses cell proliferation and migration,whereas overexpression of RAB2B in BXPC-3 cells markedly promotes these processes.This effect is likely mediated through the activation of the NF-κB signaling pathway and the subsequent regulation of FN1 expression.

Objective To observe serum interferon regulatory factor-5(IRF5)and suppressors of cytokine signaling-3(SOCS-3)levels in patients with colorectal cancer,and investigate predictive value of their levels on the occurrence of postoperative intestinal obstruction in patients.Methods A total of 100 colorectal cancer patients who attended the Cangzhou People's Hospital from January 2022 to March 2023 were selected as the study subjects.According to whether the patient experienced intestinal obstruction after surgery,they were divided into the intestinal obstruction group(n=14)and the control group(n=86).Preoperative serum levels of IRF5 and SOCS-3 were detected by enzyme-linked immunosorbent assay(ELISA).Factors influencing the occurrence of postoperative intestinal obstruction in colorectal cancer patients were analyzed by Logistic regression.Receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of serum IRF5 and SOCS-3 levels on the occurrence of postoperative intestinal obstruction in colorectal cancer patients.Results Serum IRF5 levels in the intestinal infarction group(0.68±0.09ng/ml)were lower,but serum SOCS-3 levels in the intestinal infarction group(97.23±15.94pg/ml)were higher than in the control group(0.89±0.12ng/ml,75.03±12.46pg/ml),with significant differences(t=6.257,5.937,all P＜0.05).SOCS-3 was an independent risk factor(OR=2.197,95％CI:1.167～4.138,P＜0.05),and 1RF5 was an independent protective factor(OR=0.823,95％CI:0.705～0.961,P＜0.05)affecting the occurrence of postoperative bowel obstruction in patients with colorectal cancer.The areas under the curve(AUC)of serum IRF5,SOCS-3,and the combination of the two for the diagnosis of postoperative intestinal obstruction in colorectal cancer patients were 0.852(95％CI:0.767～0.915),0.817(95％CI:0.727～0.887)and 0.953(95％CI:0.891～0.985),respectively.The combined diagnostic efficacy of the serum IRF5,SOCS-3 was higher than the efficacy of the test alone,with significant differences(Z=1.967,2.034,P=0.049,0.042).Conclusion Serum IRF5 levels are reduced,but SOCS-3 levels are increased in patients with colorectal cancer.Both have a predictive value for the development of postoperative bowel obstruction in patients with colorectal cancer.

Objective To investigate the changes in the serum levels of S100β，soluble intercellular adhesion molecule-1（sICAM-1），oxidized low-density lipoprotein（ox-LDL），and soluble cluster of differentiation 40 ligand（sCD40L）in patients with acute partial anterior circulation infarction（PACI）with cerebral microbleeds（CMBs），as well as the association of each index with early neurological deterioration（END）and post-stroke cognitive impairment（PSCI）after non-thrombolytic therapy. Methods A total of 236 patients with acute PACI and CMBs who were admitted to our hospital from January 2020 to June 2022 were enrolled as subjects. The onset of END and PSCI within 6 months after treatment was recorded，and the association of the serum levels of S100β，sICAM-1，ox-LDL，and sCD40L with the onset of END and PSCI was analyzed. A logistic stepwise regression analysis was used to investigate the risk factors for END and PSCI，and the receiver operating characteristic（ROC）curve was plotted to analyze the early prediction efficiency of each index. Results The degree of carotid stenosis，the severity of CMBs，NIHSS score on admission，and the serum levels of Hcy，S100β protein，sICAM-1，ox-LDL，and sCD40L on admission were independent risk factors for END（P<0.05）. END，NIHSS score after 7 days of treatment，and the serum levels of S100β protein，sICAM-1，ox-LDL，and sCD40L after 7 days of treatment were independent risk factors for PSCI（P<0.05）. Common risk factors such as NIHSS score，degree of carotid stenosis，and severity of CMBs combined with serum S100β protein，sICAM-1，ox-LDL，and sCD40L had an area under the ROC curve of 0.887 in predicting END in the early stage of non-thrombolytic therapy and 0.912 in predicting END within 6 months after treatment，with a significantly greater AUC than each risk factor alone. Conclusion In PACI patients with CMBs，the onset of END and PSCI in the early stage of non-thrombolytic therapy are closely associated with brain tissue injury，vascular endothelial injury，oxidative stress，and vascular inflammatory response. Dynamic monitoring of the serum levels of S100β protein，sICAM‑1，ox-LDL，sCD40L combined with common risk factors such as NIHSS score and the severity of CMBs has important clinical significance in predicting the onset of END and PSCI in the early stage，guiding clinical diagnosis and treatment，and improving the prognosis of patients.

Objective:To evaluate the difference in safety and immunogenicity of live rotavirus vaccine (oral) and measles, mumps and rubella (MMR) vaccine immunized alone or in combination.Methods:This study recruited 1 752 children aged 8-9 months who had not been vaccinated with live rotavirus vaccine (oral) or MMR vaccine after birth. The subjects were divided into three groups: study group (652 subjects, immunized with live rotavirus vaccine and MMR vaccine), control group 1 (723 subjects, immunized with live rotavirus vaccine) and control group 2 (377 subjects, immunized with MMR vaccine). Local and systemic adverse reactions within 30 d after vaccination were recorded. Serum samples were collected before and 35-42 d after immunization for analyzing the changes in antibodies.Results:Immunization alone or in combination with live rotavirus vaccine (oral) and MMR vaccine achieved similar results in the positive rates and concentrations of antibodies against rotavirus, measles and rubella viruses ( P>0.05). Moreover, the positive rates and the concentrations of the three antibodies were increased after vaccination. Compared with the control group 2, the concentration of antibody against mumps virus in the study group was increased ( P<0.05), but no significant difference in the positive rate of antibody against mumps virus was found between the two groups ( P>0.05). The positive rate and the concentration of antibody against mumps virus were increased after combined immunization or immunization with MMR vaccine alone. The overall incidence of fever and diarrhea was 1.54% (27/1 752) and 0.63% (11/1 752). No other abnormal reactions, incidental reactions or adverse reactions of any clinical significance were observed. Conclusions:Live rotavirus vaccine (oral) and MMR vaccine immunized alone or in combination showed good immunogenicity and safety.

Objective:To investigate the immunogenicity and safety of a boost dose of measles, mumps, and rubella combined vaccine (MMR) for children 4 to 6 years old.Methods:Children, aged 4 to 6 years old, had vaccinated with 1 dose of measles and rubella combined vaccine(MR) at the age of 8 months and 1 dose of MMR vaccine at 18-months, were recruited in Shanxi, Inner Mongolia, and Beijing, respectively. All children were assigned into 4, 5 and 6-year-old group. The children who met inclusion and exclusion criteria were vaccinated with 1 dose MMR vaccine, and were collected blood samples before vaccination and 35 to 42 d after the vaccination. During the study period, adverse events were collected at 30 min, 1 d, 2 d, 3 d, 4-12 d, and 13 to 42 days after vaccination. Serum was tested for IgG antibodies against measles, mumps and rubella. Geometric mean concentrations (GMC) of measles, mumps, and rubella antibodies were compared among groups by analysis of variance or non-parametric test. Seropositive rates and adverse event rates were compared among groups by Chi-square test or Fisher exact test.Results:A total of 500 children were included in immunogenicity analysis and 535 children were included in safety analysis. The overall adverse event rate was 20.37%, the most of severity for adverse events was mild. The rates of local and systemic adverse events were 0.37% and 20.00%, respectively. Symptoms of local adverse events were redness. The main systemic adverse events were fever, followed by cough, rash and runny nose. Received a dose of MMR vaccine for booster immunization, the seropositive rates of measles antibody, mumps antibody and rubella antibody were above 99% for all 3 age groups, and there was no significant difference between groups. There were significant differences in mumps antibody GMC among groups ( P=0.042), but no significant differences in measles and rubella antibodies GMC. Conclusion:The immunogenicity and safety of a boosted MMR vaccintion in children aged 4, 5 and 6 years were all similar good.

Objective:To assess the antibody persistence 3-5 years following vaccination of measles and rubella combined live-attenuated vaccine (MR) at 8 months of age and measles, mumps and rubella combined attenuated live vaccine (MMR) at 18 months of age.Methods:In 2016, 18-month-old children who were vaccinated with one dose of MR vaccine at the age of 8 months were recruited in Hebei Province as group 1; 4-, 5- and 6-year-old children who were vaccinated with one dose of MR vaccine at the age of 8 months and one dose of MMR vaccine at 18 months of age were recruited in Shanxi, Inner Mongolia and Beijing as group 2, group 3 and group 4, respectively. Serum samples were collected to detect IgG antibodies against measles, mumps and rubella by ELISA. Geometric mean concentrations (GMCs) of measles, mumps, and rubella antibodies were compared among groups by analysis of variance or non-parametric test. Seropositive rates were compared among groups by Chi-square test or Fisher′s exact test. Results:A total of 650 children were included in this study. Seropositive rates of measles, mumps and rubella antibodies 30 d after vaccination of 150 18-month-old children with one dose of MMR vaccine were 100%, 91.33% and 100%, respectively, and the GMCs were 1 846.87 mIU/ml, 299.91 IU/ml and 111.33 IU/ml, respectively. Seropositive rates of measles, mumps and rubella antibodies 3-5 years after vaccination one dose of MR vaccine at 8 months of age and one dose of MMR vaccine at 18 months of age were above 94%, 79% and 71%, respectively, and the GMCs were above 830 mIU/ml, 240 IU/ml and 31 IU/ml. No significant difference in the seropositive rates of the three antibodies was observed among groups 2, 3 and 4 ( P>0.05). There was no significant difference in the GMCs of measles or mumps antibodies among the three groups ( P>0.05), but the differences in the GMCs of rubella antibodies were statistically significant ( P=0.034). Conclusions:Measles, mumps and rubella antibodies persisted for 3-5 years without significant decrease after vaccination one dose of MR vaccine at 8 months of age and one dose of MMR vaccine at 18 months of age.

Objective To evaluate the safety of a Chinese thimerosal-free trivalent split influenza virus vaccine after being marketed in a large population. Methods Through the information management system of adverse event following immunization (AEFI), the adverse events in healthy people aged 6 months and above who were vaccinated with split influenza virus vaccine in Hubei Province from October to December 2015 were collected. The data was analyzed by descriptive methodology. Results From October 1, 2015 to June 30, 2016, among the 227 920 people in Hubei Province who were vaccinated with split influenza virus vaccine, the common adverse reactions were mainly fever, redness, irritability, pain and itching. Four cases of AEFI were passively observed and reported in the system, with a reporting rate of 1.76/100 000, among which 3 cases were anaphylactic rash and 1 case was optic neuritis. Conclusion The Chinese thimerosal-free trivalent split influenza virus vaccine used in Hubei Province had a good safety record and is suitable for the general vaccination of people without vaccination contraindications.