This study investigated the effect of multi-glycosides of Tripterygium wilfordii(GTW) on renal injury in diabetic kidney disease(DKD) rats through Nod-like receptor protein 3(NLRP3)/cysteine-aspartic acid protease-1(caspase-1)/gsdermin D(GSDMD) pyroptosis pathway and the mechanism. To be specific, a total of 40 male SD rats were randomized into the normal group(n=8) and modeling group(n=34). In the modeling group, a high-sugar and high-fat diet and one-time intraperitoneal injection of streptozotocin(STZ) were used to induce DKD in rats. After successful modeling, they were randomly classified into model group, valsartan(Diovan) group, and GTW group. Normal group and model group were given normal saline, and the valsartan group and GTW group received(ig) valsartan and GTW, respectively, for 6 weeks. Blood urea nitrogen(BUN), serum creatinine(Scr), alanine ami-notransferase(ALT), albumin(ALB), and 24 hours urinary total protein(24 h-UTP) were determined by biochemical tests. The pathological changes of renal tissue were observed based on hematoxylin and eosin(HE) staining. Serum levels of interleukin-1β(IL-1β) and interleukin-18(IL-18) were detected by enzyme-linked immunosorbent assay(ELISA). Western blot was used to detect the expression of pyroptosis pathway-related proteins in renal tissue, and RT-PCR to determine the expression of pyroptosis pathway-related genes in renal tissue. Compared with the normal group, the model group showed high levels of BUN, Scr, ALT, and 24 h-UTP and serum levels of IL-1β and IL-18(P<0.01), low level of ALB(P<0.01), severe pathological damage to kidney, and high protein and mRNA levels of NLRP3, caspase-1, and GSDMD in renal tissue(P<0.01). Compared with the model group, valsartan group and GTW group had low levels of BUN, Scr, ALT, and 24 h-UTP and serum levels of IL-1β and IL-18(P<0.01), high level of ALB(P<0.01), alleviation of the pathological damage to the kidney, and low protein and mRNA levels of NLRP3, caspase-1, and GSDMD in renal tissue(P<0.01 or P<0.05). GTW may inhibit pyroptosis by decreasing the expression of NLRP3/caspase-1/GSDMD in renal tissue, thereby relieving the inflammatory response of DKD rats and the pathological injury of kidney.
Rats ; Male ; Animals ; Diabetic Nephropathies/genetics* ; Interleukin-18/metabolism* ; Glycosides/pharmacology* ; Tripterygium ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Rats, Sprague-Dawley ; Caspase 1/metabolism* ; Pyroptosis ; Uridine Triphosphate/pharmacology* ; Kidney ; Valsartan/pharmacology* ; RNA, Messenger/metabolism* ; Diabetes Mellitus

MicroRNAs (miRNAs) are mediators of the aging process. The purpose of this work was to analyze the miRNA expression profiles of spermatozoa from men of different ages with normal fertility. Twenty-seven donors were divided into three groups by age (Group A, n = 8, age: 20-30 years; Group B, n = 10, age: 31-40 years; and Group C, n = 9, age: 41-55 years) for high-throughput sequencing analysis. Samples from 65 individuals (22, 22, and 21 in Groups A, B, and C, respectively) were used for validation by quantitative real-time polymerase chain reaction (qRT-PCR). A total of 2160 miRNAs were detected: 1223 were known, 937 were newly discovered and unnamed, of which 191 were expressed in all donors. A total of 7, 5, and 17 differentially expressed microRNAs (DEMs) were found in Group A vs B, Group B vs C, and Group A vs C comparisons, respectively. Twenty-two miRNAs were statistically correlated with age. Twelve miRNAs were identified as age-associated miRNAs, including hsa-miR-127-3p, mmu-miR-5100_L+2R-1, efu-miR-9226_L-2_1ss22GA, cgr-miR-1260_L+1, hsa-miR-652-3p_R+1, pal-miR-9993a-3p_L+2R-1, hsa-miR-7977_1ss6AG, hsa-miR-106b-3p_R-1, hsa-miR-186-5p, PC-3p-59611_111, hsa-miR-93-3p_R+1, and aeca-mir-8986a-p5_1ss1GA. There were 9165 target genes of age-associated miRNAs. Gene Ontology (GO) analysis of the target genes identified revealed enrichment of protein binding, membrane, cell cycle, and so on. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of age-related miRNAs for target genes revealed 139 enriched pathways, such as signaling pathways regulating stem cell pluripotency, metabolic pathways, and the Hippo signaling pathway. This suggests that miRNAs play a key role in male fertility changes with increasing age and provides new evidence for the study of the mechanism of age-related male fertility decline.
Humans ; Male ; Young Adult ; Adult ; Middle Aged ; MicroRNAs/genetics* ; Signal Transduction/genetics* ; Spermatozoa/metabolism* ; Gene Expression Profiling

COVID-19 has been prevalent for three years. The virulence of SARS-CoV-2 is weaken as it mutates continuously. However, elderly patients, especially those with underlying diseases, are still at high risk of developing severe infections. With the continuous study of the molecular structure and pathogenic mechanism of SARS-CoV-2, antiviral drugs for COVID-19 have been successively marketed, and these anti-SARS-CoV-2 drugs can effectively reduce the severe rate and mortality of elderly patients. This article reviews the mechanism, clinical medication regimens, drug interactions and adverse reactions of five small molecule antiviral drugs currently approved for marketing in China, so as to provide advice for the clinical rational use of anti-SARS-CoV-2 in the elderly.

Objective: To develop a nomogram model for the differential diagnosis of benign and malignant breast BI-RADS (Breast Imaging Reporting and Data System) category 4 nodules based on serum tumor specific protein 70 (SP70) and conventional laboratory indicators and validate its predictive efficacy. Methods: A case-control study design was used to retrospectively analyze the data of 429 female patients diagnosed with BI-RADS category 4 breast nodules by breast color doppler flow imaging at the First Affiliated Hospital of Nanjing Medical University from January 2021 to April 2022 with an age range of 16 to 91 years and a median age of 50 years, and the patients were divided into a training cohort (314 patients) and a validation cohort (115 patients) according to the inclusion time successively. Using postoperative pathological findings as the"gold standard", univariate and multivariate logistic regression analyses were used to identify the predictor variables used for the model. The nomogram, receiver operating characteristic (ROC) curves and calibration curves were drawn for the prediction model, and the discrimination and calibration of the model were evaluated using the consistency index (C-index) and calibration plots. Results: The postoperative pathological results showed that 286 (66.7%) were malignant nodules and 143 (33.3%) were benign nodules of 429 breast BI-RADS category 4 nodules. The serum SP70 (OR=1.227,95%CI: 1.033-1.458,P=0.020), NLR (OR=1.545,95%CI: 1.047-2.280,P=0.028), LDL-C (OR=2.215, 95%CI: 1.354-3.622, P=0.002), GLU (OR=2.050,95%CI:1.222-3.438,P=0.007), PT (OR=1.383,95%CI: 1.046-1.828,P=0.023), nodule diameter (OR=1.042, 95%CI: 1.008-1.076, P=0.015) and age (OR=1.062,95%CI: 1.011-1.116,P=0.016) were independent risk factors which could be used to distinguish benign and malignant breast BI-RADS category 4 nodules (P<0.05). The nomogram was plotted by the above seven independent variables, and the concordance index (C-index) for the training cohort and validation cohort were 0.842 (95%CI:0.786-0.898) and 0.787 (95%CI:0.687-0.886), respectively. The sensitivity and specificity of using this model to identify benign and malignant breast BI-RADS category 4 nodules in the training and validation cohort were 83.5%, 72.5% and 79.2%, 73.6%, respectively. The calibration curves showed good agreement between the predicted and actual values in the nomogram. Conclusions: This study combined serum SP70, conventional laboratory indicators and breast color doppler flow imaging to develop a nomogram model for the differential diagnosis of benign and malignant breast BI-RADS category 4 nodules. The model may have good predictive efficacy and may provide a basis for clinical treatment options, which is beneficial for guiding breast cancer screening and prevention.
Female ; Humans ; Middle Aged ; Adolescent ; Young Adult ; Adult ; Aged ; Aged, 80 and over ; Diagnosis, Differential ; Retrospective Studies ; Case-Control Studies ; Breast/pathology* ; Breast Neoplasms/pathology*

OBJECTIVE: To investigate the optimal time window for observation of catheter-induced valve injury that mimics calcified aortic valve disease in mice. METHODS: A catheter was inserted into the right common carotid artery of 8-week-old C57BL6 mice under ultrasound guidance, and aortic valve injury was induced using the guide wire.At 4, 8 and 16 weeks after modeling, the mice were subjected to ultrasound measurement of the heart short axial shortening rate, aortic valve peak velocity and aortic valve orifice area.Grain-Eosin staining was used to observe the changes in the thickness of the aortic valve, and calcium deposition in the aortic valve was assessed using Alizarin red staining.Immunofluorescence assay was performed to detect the expression of alkaline phosphatase (ALP) in the aortic valve. RESULTS: At 4, 8 and 16 weeks after modeling, valve thickness (P=0.002), calcium deposition (P < 0.0001) and the expression of osteogenic protein ALP (P=0.0016) were significantly increased, but their increments were comparable at the 3 time points of observation. CONCLUSION In mouse models of calcific aortic valve disease induced by catheter valve injury, 4 weeks after the injury appears to be the optimal time window for observation of pathophysiological changes in the aortic valves to avoid further increase of the death rate of the mice over time.
Animals ; Mice ; Aortic Valve/metabolism* ; Calcium/metabolism* ; Mice, Inbred C57BL ; Aortic Valve Stenosis/metabolism* ; Catheters ; Osteogenesis ; Cells, Cultured

ObjectiveTo explore the changes of plasma levels of soluble neuropilin-1 and superoxide dismutase （SOD） in hypertensive patients and hypertensive diabetic patients. MethodsIn this cross-sectional study， eighty-eight cases were enrolled， which were divided into hypertensive group （n=31）， hypertensive diabetic group （n=31） and control group （n=26）. The levels of neuropilin-1 and SODwere measured by ELISA. Meanwhile， the levels of serum glucose， glycosylated hemoglobin A1_C （GHbA1c） and lipid profile were detected. ResultsThe levels of total cholesterol （TC） and body mass index （BMI） were significantly higher in the hypertensive group than those in control group （P<0.05）. The levels of TC， low density lipoprotein cholesterol （LDL-C）， triglyceride （TG）， BMI， waist circumference were significantly higher in the hypertensive diabetic group than those in control group （P<0.05）. The mean plasma levels of neuropilin-1 and SOD in both hypertensive diabetic group and hypertensive group were significantly decreased compared with that in the normal group［NRP-1（ng/mL）：6.8（6.0~8.3），5.2（4.0~6.8）vs 8.9（7.7~10.0）；SOD（U/mL）：157.1±18.6，145.1±31.4 vs 168.4±23.1，P<0.05］While the mean plasma levels of neuropilin-1 and SOD in hypertensive diabetic group were significantly decreased compared withthat of the hypertensive group［NRP-1（ng/mL）：5.2（4.0~6.8）vs 6.8（6.0~8.3）；SOD（U/mL）： 145.1±31.4 vs 157.1±18.6，P<0.05］. The linear correlation analysis of three groups of study population found that neuropilin-1 and SOD showed significant positive correlation （r=0.539，0.660，0.895， P<0.05）. ConclusionsThe plasma concentration of neuropilin-1 is decreased in both hypertensive and hypertensive diabetic patients， and more decreased in hypertensive diabetic patients. Decreased neuropilin-1 concentration may be associated with the reduction of SOD activity.

OBJECTIVE: To investigate whether ginsenoside Rb1 (Rb1) can protect human umbilical vein endothelial cells (HUVECs) against high glucose-induced apoptosis and examine the underlying mechanism. METHODS: HUVECs were divided into 5 groups: control group (5.5 mmol/L glucose), high glucose (HG, 40 mmol/L) treatment group, Rb1 (50 µ mol/L) treatment group, Rb1 plus HG treatment group, and Rb1 and 3-( RESULTS: Rb1 ameliorated survival in cells in which apoptosis was induced by high glucose (P<0.05 or P<0.01). Upon the addition of Rb1, mitochondrial and intracellular reactive oxygen species generation and malondialdehyde levels were decreased (P<0.01), while the activities of antioxidant enzymes were increased (P<0.05 or P<0.01). Rb1 preserved the mitochondrial membrane potential and reduced the release of Cyt-c from the mitochondria into the cytosol (P<0.01). In addition, Rb1 upregulated mitochondrial biogenesis-associated proteins (P<0.01). Notably, the cytoprotective effects of Rb1 were correlated with SIRT3 signalling pathway activation (P<0.01). The effect of Rb1 against high glucose-induced mitochondria-related apoptosis was restrained by 3-TYP (P<0.05 or P<0.01). CONCLUSION Rb1 could protect HUVECs from high glucose-induced apoptosis by promoting mitochondrial function and suppressing oxidative stress through the SIRT3 signalling pathway.

AIM:To explore the role of Sirt1/eNOS signalling pathway in the protective effect of hydrogen sul-phide(H2S)against endothelial cell senescence induced by high glucose.METHODS:High glucose(33 mmol/L)was applied to induce senescence in primary human umbilical vein endothelial cells(HUVECs).The cell viability,the propor-tion of senescence-associated β-galactosidase(SA-β-Gal)positive cells and the plasminogen activator inhibitor 1(PAI-1) expression were detected to assess the senescence model.Mean while,Sirt1 siRNA was used to examine the effect of Sirt 1 on eNOS expression and the senescence-related parameters.RESULTS: Treatment of HUVECs with high glucose de-creased the cell viability slowly with a larger proportion of the cells stained with SA-β-Gal, and the protein expression of PAI-1 was dramatically increased.The increased cell viability,reduced SA-β-Gal positive cells and decreased protein ex-pression of PAI-1 were detected after sodium hydrosulfide(NaHS,100 μmol/L)treatment.Furthermore,NaHS treatment upregulated the protein expression of Sirt 1 and eNOS,and eventually increased the production of nitric oxide(NO).CON-CLUSION:Exogenous H2S modulates Sirt1/eNOS/NO pathway to prevent HUVECs against high glucose-induced senes-cence.

AIM:To study the value of copeptin(CPP)level for the prediction of cardiorenal syndrome (CRS)in the rats with subtotal nephrectomy(SNX)combined with myocardial infarction(MI).METHODS: Male SD rats(n=60)were divided into blank control group(Con group), renal failure group(SNX group), heart failure group (MI group)and heart failure+renal failure group(CRS group).The concentrations of CPP in the serum and urine,hemo-dynamic indexes,blood pressure and renal function indexes were measured 1~5 weeks after modeling.The predictive val-ue of CPP for CRS in the rats was evaluated by the receiver operating characteristic(ROC)curve.RESULTS:Compared with Con group,left ventricular systolic pressure(LVSP)at 9 d in CRS group was significantly decreased(P<0.05),left ventricular end-diastolic pressure(LVEDP)at 9 d in CRS group was significantly increased(P<0.05), and the differ-ence of blood pressure at each time point was not statistically significant.The levels of blood urea nitrogen(BUN)and uri-nary creatinine(Ucr)in CRS group were significantly increased at 1 and 3 weeks(P<0.05).Compared with Con group, serum CPP level was significantly increased at 1,3 and 5 weeks(P<0.05), and urine CPP level was significantly in-creased at 3 weeks in CRS group.Serum brain natriuretic peptide(BNP)level was significantly increased at 1 and 3 weeks,while urine BNP level was significantly increased at 5 weeks after modeling in CRS group(P<0.05).No correla-tion between serum or urine CPP and BNP or BUN levels at 1 week in CRS group was observed.The results of ROC curve analysis indicated that the area under the curve(AUC)of serum CPP was 0.908(95%CI:0.789~1.028),and the cut-off value was 56.59 ng/L(sensitivity 0.875,specificity 0.800).CONCLUSION:The combination of SNX and MI estab-lishes a CRS rat model with both heart and kidney injury,and serum CPP can be used as a sensitive and specific biomarker for early prediction of CRS.

Objective To understand infection status,clinical and epidemiological features of respiratory tract infection(RTI) with human Bocavirus(HBoV) in children in Wuhan area.Methods A total of 968 hospitalized children with acute RTI between January and December 2016 were recruited in the study.Sputum specimens were collected by sterile negative pressure suction method.HBoV,respiratory syncytial virus,adenovirus,A/B-type influenza virus,and parainfluenza virus Ⅰ/Ⅱ/Ⅲ were detected,the prevalence and clinical features of HBoV infection were analyzed.Results Among 968 cases,75(7.75％) were positive for HBoV.The detection rates of HBoV in boys and girls were 5.68％ and 2.07％ respectively,there was no significant difference in the detection rate of HBoV between different genders(x2 =2.083,P =0.140);53 (70.67 ％) HBoV-positive children were＜ 1 year old,63(84.00％) were＜3 years old,there was significant difference in the detection rate among all age groups(x2 =4.60,P =0.043).The detection rates in spring,summer,autumn,and winter were 9.78％,5.29％,2.86％,and 5.20％ respectively;except September and October,HBoV was found positive in the other months,detection rate was highest in March (33.33％).17 cases were simple HBoV infection and 58 were mixed infection,the main clini cal diagnosis of HBoV was bronchopneumonia(n =26,34.67％).The main clinical features of HBoV cases were fever(81.33 ％),cough(77.33 ％),and wheezing (77.33 ％).Conclusion HBoV is one of the important pathogens causing RTI in hospitalized children in Wuhan area,HBoV infection can occur throughout the year,the peak season are spring and summer,there is no difference between different genders.The most common clinical diagnosis of HBoV in hospitalized children is bronchopneumonia,the main manifestations are fever,coughing,and wheezing.