ABSTRACT
Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis (RA), but the relationship between the two phenomena remains unclear. We explored whether and how cholinergic dysfunction accelerates protein citrullination and consequently drives the development of RA. Cholinergic function and protein citrullination levels in patients with RA and collagen-induced arthritis (CIA) mice were collected. In both neuron-macrophage coculture system and CIA mice, the effect of cholinergic dysfunction on protein citrullination and expression of peptidylarginine deiminases (PADs) was assessed by immunofluorescence. The key transcription factors for PAD4 expression were predicted and validated. Cholinergic dysfunction in the patients with RA and CIA mice negatively correlated with the degree of protein citrullination in synovial tissues. The cholinergic or alpha7 nicotinic acetylcholine receptor (α7nAChR) deactivation and activation resulted in the promotion and reduction of protein citrullination in vitro and in vivo, respectively. Especially, the activation deficiency of α7nAChR induced the earlier onset and aggravation of CIA. Furthermore, deactivation of α7nAChR increased the expression of PAD4 and specificity protein-3 (SP3) in vitro and in vivo. Our results suggest that cholinergic dysfunction-induced deficient α7nAChR activation, which induces the expression of SP3 and its downstream molecule PAD4, accelerating protein citrullination and the development of RA.
ABSTRACT
【Objective】 To investigate the correlation of serum exosomal microRNA-301a (miR-301a) with cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy. 【Methods】 A total of 211 patients with diabetic nephropathy treated with peritoneal dialysis from June 2019 to June 2020 in the First Hospital Affiliated of Hebei North University were selected as study subjects. Serum exosomal miR-301a was detected by real-time fluorescence quantitative polymerase chain reaction. The patients were divided into high miR-301a group and low miR-301a group based on the median of miR-301a; the clinical data of the two groups were compared. The correlation of miR-301a with high-sensitivity C-reactive protein (hs-CRP) was analyzed by Spearman. Linear regression was applied to analyze the factors associated with the effect of miR-301a. The patients were followed up for two years. Kaplan-Meier and Log-Rank were conducted to compare the cumulative incidence of cardiovascular and cerebrovascular events between the two groups, and COX regression and restricted cubic spline were used to analyze the level-effect relationship between miR-301a and cardiovascular and cerebrovascular events after peritoneal dialysis. 【Results】 Thirty-seven cases (17.54%) of cardiovascular and cerebrovascular events occurred during follow-up. The hs-CRP level and dialysis duration were lower in low miR-301a group than in high miR-301a group (P<0.05). There was a positive correlation between miR-301a and hs-CRP (rs=0.237, P=0.001). Linear regression analysis showed that hs-CRP was independently associated with miR-301a (P<0.05). The cumulative cardiovascular and cerebrovascular event rate in low miR-301a group was 3.70% (4/108), which was lower than that in high miR-301a group [32.04% (33/103), P<0.001]. COX regression analysis showed that high serum albumin level was an independent protective factor for cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy,while high hs-CRP level and miR-301a >1.46 were independent risk factors for cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy. Restricted cubic spline fitting COX regression analysis showed a non-linear relationship between miR-301a and cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy (P<0.05). 【Conclusion】 hs-CRP is independently associated with miR-301a in diabetic nephropathy peritoneal dialysis patients. High miR-301a level suggests a high risk of cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy.
ABSTRACT
Objective To investigate the relationship between endoscopic diagnosis and pathological nature of gastric mucosal leukoplakia. Methods Analysis of gastric mucosal leukoplakia patients’ endoscopic features and pathological characteristics of last 3 years. Results Endoscopic diagnosis was made in 116 cases of gastric mucosal leukoplakia. Samples were gray white, round, round shaped or patchy in general observation. Diameter was 0.2 ~1.6 cm, foam cell aggregation appeared in the natural layer of mucous membrane in 88 cases (75.9 %), with varying degrees of inflammatory cells infiltration. The foam cells accumulation were observed in high rate among the gastric mucosal leukoplakia cases. Conclusion Pathological changes of gastric mucosal leukoplakia is gastric xanthoma with chronic inflammation of mucosa.
ABSTRACT
Objective Studies on the application of fast track surgery ( FTS) are comparatively limited in urologic proce-dures.This randomized controlled study was to evaluate the impact of FTS on recovery after retroperitoneal laparoscopic adrenalectomy . Methods Eighty patients undergoing retroperitoneal laparoscopic adrenalectomy were randomly assigned to an FTS and a control group of equal number to receive an FTS recovery program and conventional perioperative care , respectively .Comparisons were made between the two groups in the time of the first flatus , first oral nutrition , and first mobilization , the incidence of gastrointestinal tract complica-tions, the time of drainage and transurethral catheterization , the length of postoperative hospital stay , hospitalization expenses , visual analogue scale (VAS) pain scores, and general state of the patients . Results The FTS group, in comparison with the control, showed significantly earlier time of first flatus ([20.6 ±8.3] vs [39.8 ±18.3]h, P0.05), nor in the incidence of gastrointesti-nal tract complications between the FTS and control groups ( P >0.05). Conclusion By improving the general state and accelera-ting the recovery of the patients , FTS can be applied safely and effectively in retroperitoneal laparoscopic adrenalectomy .
ABSTRACT
This study aimed to investigate infiltration related microRNAs (miRNAs) in bladder urothelial carcinoma (BUC). Twenty patients with BUC were enrolled and divided into 2 groups according to infiltration or not: infiltrating BUC group (n=12) and non-infiltrating BUC group (n=8). Gene chip was used to detect infiltration related miRNAs in the BUC samples. In other recruited 17 patients with BUC who were divided into infiltrating BUC samples (n=14) and non-infiltrating BUC samples (n=3), and in 4 BUC cell lines (EJ, 5637, T24 and BIU-87), the expression of miRNAs was assayed by using reverse transcription-polymerase chain reaction (RT-PCR). In infiltrating BUC group, as compared with non-infiltrating BUC group, there were 7 differentially expressed miRNAs: hsa-miR-29c, hsa-miR-200a, hsa-miR-378, hsa-miR-429, hsa-miR-200c and hsa-miR-141 were up-regulated, while hsa-miR-451 was down-regulated. In the BUC samples, the results of RT-PCR were consistent with those by the miRNA array. In the cancer cell lines, RT-PCR in T24 only revealed the similar expression pattern of miRNAs to that by the miRNA array. It is suggested that infiltration of BUC is related with different expression of miRNAs, which may provide a novel platform for further study on function and action mechanism of miRNAs.
ABSTRACT
Objective To figure out and verify infiltration related miRNAs in bladder urothelial carcinoma (BUC). Methods Fresh tissues (20 samples,12 were infiltrative BUC samples,8 were non-infiltrative BUC samples) were collected in liquid nitrogen.The total RNA was extracted by using Trizol reagents.RNA quality control; miRNA microarray hybridization; data analysis.Another 22 samples were collected in fresh (15 were infiltrative BUC samples,7 were non-infiltrative BUC samples) for verifying purpose.4 types of bladder cancer cell lines were used for the study.BUC cell strain; total RNA was extracted by Trizol reagents; RNA quality control; RT-PCR and analysis of the data. Results ①In infiltrative BUC group,compared with non-infiltrative BUC group,there were 7 differentially expressed miRNAs:hsa-miR29c,hsa-miR-200a,hsa-miR-378,hsa-miR-429,hsa-miR-200c and hsa-miR-141 were up-regulated; hsamiR-451 was down-regulated.②In collected samples,the result of RT-PCR was consistent with miRNA array.③In bladder cancer cell lines,only the results of T24 were consistent with miRNA array. Conclusion Infiltration of BUC might relate with different expression of miRNAs.
ABSTRACT
This study aimed to investigate infiltration related microRNAs (miRNAs) in bladder urothelial carcinoma (BUC). Twenty patients with BUC were enrolled and divided into 2 groups according to infiltration or not: infiltrating BUC group (n=12) and non-infiltrating BUC group (n=8). Gene chip was used to detect infiltration related miRNAs in the BUC samples. In other recruited 17 patients with BUC who were divided into infiltrating BUC samples (n=14) and non-infiltrating BUC samples (n=3), and in 4 BUC cell lines (EJ, 5637, T24 and BIU-87), the expression of miRNAs was assayed by using reverse transcription-polymerase chain reaction (RT-PCR). In infiltrating BUC group, as compared with non-infiltrating BUC group, there were 7 differentially expressed miRNAs: hsa-miR-29c, hsa-miR-200a, hsa-miR-378, hsa-miR-429, hsa-miR-200c and hsa-miR-141 were up-regulated, while hsa-miR-451 was down-regulated. In the BUC samples, the results of RT-PCR were consistent with those by the miRNA array. In the cancer cell lines, RT-PCR in T24 only revealed the similar expression pattern of miRNAs to that by the miRNA array. It is suggested that infiltration of BUC is related with different expression of miRNAs, which may provide a novel platform for further study on function and action mechanism of miRNAs.
Subject(s)
Humans , Carcinoma , Genetics , Cell Line, Tumor , MicroRNAs , Genetics , Urinary Bladder , Metabolism , Urinary Bladder Neoplasms , GeneticsABSTRACT
The effects of combined RNA interference (RNAi) of human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT) genes on telomerase activity in a bladder cancer cell line (BIU-87 cells) were investigated by using gene chip technology in vitro with an attempt to evaluate the role of RNAi in the gene therapy of bladder transitional cell cancer (BTCC). Three TR-specific double-stranded small interfering RNAs (siRNAs) and three TERT-specific double-stranded siRNAs were designed to target different regions of TR and TERT mRNA. The phTR-siRNA, phTERT-siRNA, and the combination of both plasmids phTR+phTERT-siRNA were transfected into BIU-87 cells. The expression of hTR and hTERT mRNA was detected by quantitative fluorescent reverse transcription-polymerase chain reaction, and a telomeric repeat amplification protocol was applied to detect telomerase activity. Growth inhibition of BIU-87 cells was measured by MTT assay. Gene chip analysis was performed to evaluate the effects of the combined RNAi of hTR+hTERT genes on telomerase activity and growth of BIU-87 cells in vitro. The results showed that the expression of hTERT and hTR mRNA was inhibited by pRNAT-hTERT-III, pRNAT-hTR-III, and pRNAT-hTR-III+hTERT-III in BIU-87 cells. The inhibition efficiency of pRNAT-hTERT-III, pRNAT-hTR-III, pRNAT-hTERT-III+pRNAT-hTR-III was 67% for TERT mRNA, 41% for TR mRNA, 57% for TR mRNA and 70% for TERT mRNA in BIU-87 cells respectively. The growth of BIU-87 cells was inhibited and telomerase activity was considerably decreased, especially in the cells treated with combined RNAi-hTR and -hTERT. Gene chip analysis revealed that 21 genes were down-regulated (ATM, BAX, BCL2, BCL2L1, BIRC5, CD44, CTNNB1, E2F1, JUN, MCAM, MTA1, MYC, NFKB1, NFKBIA, NME4, PNN, PNN, SERPINE1, THBS1, TNFRSF1A, and UCC1). The results indicated that hTR-siRNA and hTERT-siRNA, especially their combination, siRNA hTR+hTERT, specifically and effectively suppressed the expression of both hTR and hTERT mRNA and telomerase activity. Molecular biological mechanism by which combined siRNA-TR and -TERT inhibited telomerase activity and growth of BIU-87 cells in vitro may involve the down-regulation of the 21 genes.
ABSTRACT
To examine the expression of human telomere reverse transcriptase (hTERT), p53 and proliferating cell nuclear antigen (PCNA) in cystitis glandularis, 38 patients were divided into two grouips: group A (including 18 cases of papillary cystitis glandularis) and group B (including 20 subjects with normal bladder mucosa). All the cases were immunohistochemically examined by using antibodies specifically against p53 and PCNA, and hTERT was determined by in situ hybridization.hTERT was found in 6 cases (33.3%) and p53 was detected in 4 cases (22.2%) in group A, while they were not detected in group B. There were significant differences in hTERT and p53 expression between groups A and B (P＜0.05 for both). PCNA was detected in 7 cases (38.9%) in group A and 1 case (5.0%) in group B, and significant difference in PCNA expression was found between the two groups (P＜0.05). The expressions of hTERT, p53 and PCNA were significantly higher in group A than in group B, suggesting that papillary cystitis glandularis is predisposed to cancerous change, and p53, PCNA, hTERT may be related to the malignant alteration.
ABSTRACT
To investigate the effects of down-regulation of prostate androgen regulated (PAR) expression on proliferation of PC3 cells by using RNA interference (RNAi), suppression of PAR expression was achieved by transfection of PC3 cells with short hairpin RNA (shRNA) expression vectors against PAR, designated as psiRNA-PAR1, psiRNA-PAR2 and psiRNA-PAR3. The inhibitory effects were confirmed by RT-PCR. The growth features of PC3 transfectants were analyzed by cell counts, colon formation in soft agar and flow cytometry. The expression of PAR was suppressed by the three shRNA expression vectors. psiRNA-PAR1 was shown to inhibit the PAR expression most efficiently, with the inhibitory rate reaching a peak at (81.18±1.68)% 48 h after the transfection. PC3 transfectants exhibited a decreased proliferation in cell culture and a low efficiency of colon formation in soft agar. Flow cytometry revealed a G2/M arrest and induced apoptosis. Down-regulated PAR expression inhibited the growth of PC3 cells by inducing G2/M arrest and activating apoptotic pathway. As a potential proto-oncogene that triggers and/or has persistent malignant proliferation, PAR may serves as a very target for the gene therapy.