Objective The aim of this study was to detect the expression of miR-383-5p in bladder cancer tissues and bladder cancer 5637 cells,BIU-87 cells,TCCSUP cells and HT-1376 cells,and to explore the effects of miR-383-5p on the prolif-eration,invasion,migration and apoptosis of bladder cancer cells by targeting CIP2A.Methods The expression of miR-383-5p was detected by qRT-PCR in human bladder cancer tissues and their corresponding adjacent tissues,5637 cells,BIU-87 cells,TCCSUP cells,HT-1376 cells,human bladder transitional epithelial cells.BIU-87 cells with low miR-383-5p expression were selected for subsequent experiments.BIU-87 cells were divided into the blank group(normal culture),miR-383-5p NC group(negative control,transfected with miR-383-5p negative control),miR-383-5p mimic group(transfected with miR-383-5p mimic),and miR-383-5p mimic+pc-CIP2A group(co-transfected with miR-383-5p mimic and CIP2A overexpression plasmid pc-CIP2A).CCK-8 kit was used to detect the viability of BIU-87 cells in each group;Flow cytometry was used to detect apoptosis of BIU-87 cells;Transwell assay was used to measure cell invasion ability of BIU-87 cells;Scratch assay was used to measure cell migration ability of BIU-87 cells;Western blot was used to determine the expression of proteins related to apoptosis,invasion(MMP-2,MMP-9),and CIP2A/PP2A in BIU-87 cells;The dual luciferase assay was used to verify the targeting relationship between miR-383-5p and CIP2A in BIU-87 cells.Results The expression of miR-383-5p was low in bladder cancer tissues and bladder cancer cells.Compared with the blank group,BIU-87 cells in the miR-383-5p mimic group showed a significant increase the level of miR-383-5p(0.91±0.10 vs.1.67±0.24,P<0.01)and a significant decrease in the expression of CIP2A protein(1.32±0.17 vs.0.45±0.03,P<0.001),the cell viability,invasion,migration abilities,the expression of proteins related to invasion(MMP-2,MMP-9),and the expression of Bcl-2 protein[(100.00±4.36)% vs.(32.15±2.65)% ,(150.20±12.95)vs.(82.35±7.01),(77.91±3.63)% vs.(46.12±2.54)% ,1.02±0.11 vs.0.22±0.04,1.03±0.18 vs.0.21±0.04,1.01±0.14 vs.0.27±0.05,P<0.001];The apoptosis rate,the expression of caspase-3 and Bax proteins related to apoptosis,and PP2A expression were significantly increased[(14.02±2.29)% vs.(38.21±3.20)% ],0.81±0.11 vs.1.78±0.24,0.83±0.12 vs.1.72±0.24,0.27±0.02 vs.0.95±0.16,P<0.001].Compared with the miR-383-5p mimic group,BIU-87 cells in the miR-383-5p mimic+pc-CIP2A group significantly increased the cell viability,invasion,migration abilities,the expression of proteins related to invasion,and the expression of Bcl-2 protein[(32.15±2.65)% vs.(50.18±3.77)% ,(82.35±7.01)% vs.(116.30±13.70),(46.12±2.54)% vs.(58.43±3.15)% ,0.22±0.04 vs.0.60±0.08,0.21±0.04 vs.0.5 8±0.06,0.27±0.05 vs.0.64±0.08,P<0.05];The apoptosis rate,the expression of caspase-3,Bax,and PP2A was signifi-cantly reduced in the miR-383-5p mimic+pc-CIP2A group[(38.21±3.20)% (23.15±2.74)% ,1.78±0.24 vs.1.25±0.21,1.72±0.24 vs.1.23±0.18,0.95±0.16 vs.0.60±0.13,P<0.05].The results of dual luciferase experiments showed a corresponding tar-geting relationship between miR-383-5p and CIP2A.Conclusion Increasing the expression of miR-383-5p can inhibit the prolif-eration,invasion and migration of bladder cancer BIU-87 cells,and enhance the ability of apoptosis,which may be achieved by targe-ted regulation of CIP2A.

Objective To explore the predictive value of methylenetetrahydrofolate reductase(MTHFR)gene C677T polymorphism and serum homocysteine(Hcy)level for adverse pregnancy outcomes.Methods The clinical data of 698 pregnant women who received antenatal care and deliv-ered at Ji'an Central People's Hospital were retrospectively collected.According to the research pur-pose,the pregnant women were divided into internal study cohort(n=483)or external validation co-hort(n=215).Each cohort was further divided into adverse pregnancy group and normal pregnancy group based on whether adverse pregnancy outcomes occurred.The MTHFR gene C677T polymor-phism and serum Hcy level of all pregnant women were detected.Binary Logistic regression analysis was used to investigate the relationships of MTHFR gene C677T polymorphism,Hcy level,and other indicators with adverse pregnancy outcomes.Receiver operating characteristic(ROC)curves were plotted to analyze the predictive efficacy of different indicators when used alone and their combina-tion.Results In the internal study cohort,compared with the normal pregnancy group,the detection frequency of the CC genotype at the MTHFR gene C677T locus was lower,while the detection frequencies of the CT and TT genotypes were higher in the adverse pregnancy group(P＜0.001).The proportion of pregnant women with high Hcy levels was higher in the adverse pregnancy group than that in the normal pregnancy group(P＜0.001).Binary Logistic regression analysis showed that age,folic acid level during pregnancy,MTHFR gene C677T polymorphism,and Hcy level were all independ-ent influencing factors for adverse pregnancy outcomes(P＜0.05).The ROC curve indicated that the area under the curve(AUC)of the combined prediction model was 0.938,which was greater than that of each indicator.In the external validation cohort,the AUC of the combined prediction model was 0.917,validating the stability and clinical applicability of the model.Conclusion The TT and CT genotypes at the MTHFR gene C677T locus and high Hcy level are all independent risk factors for adverse pregnancy outcomes.The prediction model constructed by combining these factors with age and folic acid level during pregnancy has a high predictive efficacy for adverse pregnancy outcomes.

Objective:To explore patterns of traditional Chinese medicine (TCM) use among patients with inflammatory bowel disease (IBD) in Jiangsu Province, China from 2019 to 2023.Methods:Using data from the IBD health data platform of the National Healthcare Big Data (Eastern) Center, a retrospective cohort study was conducted. We performed descriptive analyses on hospitalised patients diagnosed with IBD between 2019 and 2023, who received TCM treatment.Results:The study included 11 095 case records from 4 760 patients, with TCM diagnoses primarily indicating diarrhoea and abdominal pain. Ulcerative colitis (UC) accounted for 4 782 hospitalizations (3 103 patients), while Crohn's disease (CD) accounted for 6 313 hospitalizations (1 657 patients). Patient demographics showed a trend towards younger age and a higher proportion of males. Treatment utilisation was highest in southern Jiangsu compared with the central and northern regions. In terms of disease burden, all treatment costs showed a downward trend. In terms of external TCM therapies, UC patients tend to prefer plasters and enemas, while CD patients are more inclined to use acupuncture. Regarding herbal medicine, licorice, white atractylodes, and white peony root are commonly used single herbs for IBD patients.Conclusions:The number of IBD patients treated with TCM in Jiangsu Province has steadily increased from 2019 to 2023. It is important to identify effective TCM treatment methods to reduce the burden of patients.

Objective:To explore patterns of traditional Chinese medicine (TCM) use among patients with inflammatory bowel disease (IBD) in Jiangsu Province, China from 2019 to 2023.Methods:Using data from the IBD health data platform of the National Healthcare Big Data (Eastern) Center, a retrospective cohort study was conducted. We performed descriptive analyses on hospitalised patients diagnosed with IBD between 2019 and 2023, who received TCM treatment.Results:The study included 11 095 case records from 4 760 patients, with TCM diagnoses primarily indicating diarrhoea and abdominal pain. Ulcerative colitis (UC) accounted for 4 782 hospitalizations (3 103 patients), while Crohn's disease (CD) accounted for 6 313 hospitalizations (1 657 patients). Patient demographics showed a trend towards younger age and a higher proportion of males. Treatment utilisation was highest in southern Jiangsu compared with the central and northern regions. In terms of disease burden, all treatment costs showed a downward trend. In terms of external TCM therapies, UC patients tend to prefer plasters and enemas, while CD patients are more inclined to use acupuncture. Regarding herbal medicine, licorice, white atractylodes, and white peony root are commonly used single herbs for IBD patients.Conclusions:The number of IBD patients treated with TCM in Jiangsu Province has steadily increased from 2019 to 2023. It is important to identify effective TCM treatment methods to reduce the burden of patients.

Objective The aim of this study was to detect the expression of miR-383-5p in bladder cancer tissues and bladder cancer 5637 cells,BIU-87 cells,TCCSUP cells and HT-1376 cells,and to explore the effects of miR-383-5p on the prolif-eration,invasion,migration and apoptosis of bladder cancer cells by targeting CIP2A.Methods The expression of miR-383-5p was detected by qRT-PCR in human bladder cancer tissues and their corresponding adjacent tissues,5637 cells,BIU-87 cells,TCCSUP cells,HT-1376 cells,human bladder transitional epithelial cells.BIU-87 cells with low miR-383-5p expression were selected for subsequent experiments.BIU-87 cells were divided into the blank group(normal culture),miR-383-5p NC group(negative control,transfected with miR-383-5p negative control),miR-383-5p mimic group(transfected with miR-383-5p mimic),and miR-383-5p mimic+pc-CIP2A group(co-transfected with miR-383-5p mimic and CIP2A overexpression plasmid pc-CIP2A).CCK-8 kit was used to detect the viability of BIU-87 cells in each group;Flow cytometry was used to detect apoptosis of BIU-87 cells;Transwell assay was used to measure cell invasion ability of BIU-87 cells;Scratch assay was used to measure cell migration ability of BIU-87 cells;Western blot was used to determine the expression of proteins related to apoptosis,invasion(MMP-2,MMP-9),and CIP2A/PP2A in BIU-87 cells;The dual luciferase assay was used to verify the targeting relationship between miR-383-5p and CIP2A in BIU-87 cells.Results The expression of miR-383-5p was low in bladder cancer tissues and bladder cancer cells.Compared with the blank group,BIU-87 cells in the miR-383-5p mimic group showed a significant increase the level of miR-383-5p(0.91±0.10 vs.1.67±0.24,P<0.01)and a significant decrease in the expression of CIP2A protein(1.32±0.17 vs.0.45±0.03,P<0.001),the cell viability,invasion,migration abilities,the expression of proteins related to invasion(MMP-2,MMP-9),and the expression of Bcl-2 protein[(100.00±4.36)% vs.(32.15±2.65)% ,(150.20±12.95)vs.(82.35±7.01),(77.91±3.63)% vs.(46.12±2.54)% ,1.02±0.11 vs.0.22±0.04,1.03±0.18 vs.0.21±0.04,1.01±0.14 vs.0.27±0.05,P<0.001];The apoptosis rate,the expression of caspase-3 and Bax proteins related to apoptosis,and PP2A expression were significantly increased[(14.02±2.29)% vs.(38.21±3.20)% ],0.81±0.11 vs.1.78±0.24,0.83±0.12 vs.1.72±0.24,0.27±0.02 vs.0.95±0.16,P<0.001].Compared with the miR-383-5p mimic group,BIU-87 cells in the miR-383-5p mimic+pc-CIP2A group significantly increased the cell viability,invasion,migration abilities,the expression of proteins related to invasion,and the expression of Bcl-2 protein[(32.15±2.65)% vs.(50.18±3.77)% ,(82.35±7.01)% vs.(116.30±13.70),(46.12±2.54)% vs.(58.43±3.15)% ,0.22±0.04 vs.0.60±0.08,0.21±0.04 vs.0.5 8±0.06,0.27±0.05 vs.0.64±0.08,P<0.05];The apoptosis rate,the expression of caspase-3,Bax,and PP2A was signifi-cantly reduced in the miR-383-5p mimic+pc-CIP2A group[(38.21±3.20)% (23.15±2.74)% ,1.78±0.24 vs.1.25±0.21,1.72±0.24 vs.1.23±0.18,0.95±0.16 vs.0.60±0.13,P<0.05].The results of dual luciferase experiments showed a corresponding tar-geting relationship between miR-383-5p and CIP2A.Conclusion Increasing the expression of miR-383-5p can inhibit the prolif-eration,invasion and migration of bladder cancer BIU-87 cells,and enhance the ability of apoptosis,which may be achieved by targe-ted regulation of CIP2A.

Objective:To explore the effects of microRNA-126 (miR-126) on the proliferation of human myeloid leukemia mononuclear cells (THP-1)-derived macrophages in high glucose environment and the regulatory role of miR-126 in periodontitis with diabetes.Methods:THP-1 cells were cultured in vitro and 5 μg/L phorbol-12-myristate-13-acetate was applied to induce THP-1 cells differentiating into macrophages for 48 h in low glucose culture medium (5.5 mmol/L). THP-1-derived macrophages were then cultured with low glucose, medium glucose (15 mmol/L) or high glucose (25 mmol/L) media respectively. The proliferation of THP-1-derived macrophages was detected by cell counting kit-8 (CCK-8) method and the expressions of miR-126 and proliferation-associated factors were detected by quantitative real time PCR (qRT-PCR). The miR-126 mimic or inhibitor was transfected into THP-1-derived macrophages for 72 h. The proliferation of cells was detected by CCK-8 method and the expressions of miR-126 or proliferation-associated factors were detected by qRT-PCR. Results:Increasing glucose concentration decreased the proliferation of THP-1-derived macrophages (day 7, A values in low, medium and high glucose groups were 0.369±0.014, 0.214±0.009 and 0.200±0.010, respectively, P<0.01) as well as the survival rate ( P<0.05), promoted the expression of miR-126, B-cell lymphoma-2 (Bcl-2)-associated X protein (BAX) and caspase-3 ( P<0.05), and suppressed Bcl-2, phosphoinositol-3 kinase regulatory subunit 2 (PIK3R2) expression ( P<0.05). After the miR-126 mimic was transfected in cells in low glucose medium for 72 h, compared with negative control (1.005±0.118), the expression of miR-126 significantly increased (2 980.227±170.431, P<0.05), and the proliferation of THP-1 derived macrophages decreased (negative control: 1.816±0.013, mimic group: 1.310±0.048, P<0.01), the level of BAX and caspase-3 significantly increased ( P<0.01, P<0.05), PIK3R2 and Bcl-2 significantly decreased ( P<0.05, P<0.01). After the miR-126 inhibitor was transfected in cells cultured in high glucose medium for 72 h, compared with negative control (0.723±0.133), the proliferation of inhibitor group increased (0.984±0.049, P<0.05), the level of BAX and caspase-3 significantly decreased ( P<0.01, P<0.05), PIK3R2 and Bcl-2 significantly increased ( P<0.01, P<0.05). Conclusions:High glucose condition can inhibit the proliferation of THP-1-derived macrophages and increase the expression of miR-126. MiR-126 can inhibit the proliferation of THP-1-derived macrophages in high glucose environment through up-regulating the expression of BAX and caspase-3 and down-regulating the expression of PIK3R2 and Bcl-2.

AIM:To investigate the role of caveolin-1 on epithelial-mesenchymal transition (EMT) in human bronchial epithelial (HBE) cells induced by transforming growth factor beta 1 (TGF-β1).METHODS:Immunofluorescence, real-time PCR and Western blot were applied to detect the mRNA and the protein expression of caveolin-1 in the 16HBE cells during EMT.The influence of siRNA-mediated silencing of caveolin-1 on EMT in the 16HBE cells was detected by Western blot.RESULTS:Caveolin-1 was widely present on the cell membrane of the 16HBE cells.The expression of caveolin-1 at mRNA and protein levels was significantly decreased in a time-dependent manner in the 16HBE cells compared with control group (P<0.05) after stimulation with TGF-β1.The morphologic changes of the 16HBE cells induced by TGF-β1 were promoted by caveolin-1 silencing compared with TGF-β1 group.The protein expression of E-cadherin and α-SMA induced by TGF-β1 was promoted by caveolin-1 silencing compared with TGF-β1 group (P<0.05).The phosphorylation levels of AKT and Smad3 were the highest at 30 min and increased significantly compared with control group (P<0.05) after stimulated with TGF-β1.Treatment of the 16HBE cells with TGF-β1 for 30 min after silencing caveolin-1 gene for 24 h significantly increased the phosphorylation levels of AKT and Smad3 compared with TGF-β1 group (P<0.05).CONCLUSION:TGF-β1 down-regulates the expression of caveolin-1 in the 16HBE cells.Caveolin-1 may participate in TGF-β1/Smad pathway and PI3K-AKT pathway, which are the signal transduction pathways for TGF-β1 inducing EMT.

Objective To investigate the the multi-directional differentiation potential between pluripotent of human gingival fibroblasts (HGFs) and human periodontal ligament cells (HPDLCs). Methods HPDLCs and HGFs were obtained from the primary culture. HPDLCs and HGFs at 3rd-4th passage were cultured in osteogenic, adipogenic or chondrogenic me-dium. Cells without differentiation were taken as control. Alizarin red, Alcian blue and oil red O staining were performed to detect osteogenic differentiation, chondrogenic and adipogenic differentiation in vitro, respectively. Reverse transcription polymerase chain reaction (RT-PCR) was applied to examine the expression of osteocalcin (OCN), runt-related transcription factor 2 (RUNX2) and collagen 1 (Col 1), peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and collagen 10 (Col 10). Results HPDLCs and HGFs cultured in osteogenic medium showed massive calicium nodulus at day 28, but HP-DLCs formed more calicium nodulus than those of HGFs. The expressions of OCN, RUNX2 and Col 1 were significantly high-er in HPDLCs than those in HGFs (P<0.05). In chondrogenic medium both cells were found blue deposit at day 14, and the expression of Col 10 was significantly higher in HGFs than that of HPDLCs (P<0.01). Furthermore, in adipogenic medium HGFs showed more lipid-filled droplets stained with oil red O than HPDLCs at day 21. The expression of PPARγ2 was sig-nificantly higher in HGFs than that of HPDLCs (P<0.01). Conclusion HPDLCs has the better potency of osteogenic differ-etiation than HGFs, however, HGFs has the better potency of adipogenic and chondrogenic differentiation.

OBJECTIVE:To explore the method for the scientific and standard management of clinical trial drugs. METHODS:By theory analysis and empirical analysis,the management model of clinical trial drugs in our hospital was introduced in terms of software and hardware construction of clinical trial pharmacy,the formulation of drug management system and standard operation procedure,regular quality control and drug information management platform construction,etc. RESULTS:In the experience of our hospital,it could safeguard the safety of drug use in subjects and scientificity and preciseness of drug clinical trial results through the concentrated administration trial drugs by full-time pharmacists according to national laws and regulations,management system and standard operation procedure,and regular quality control inspection by quality control group. CONCLUSIONS:Drug clinical trial institute strictly abide the requirements of Good Clinical Practice,strengthen the management of trial drugs and im-prove information management continuously,which is of important significance to construct standardized,detailed and high-effi-ciency centralized management system of clinical trial drugs.

Objective To investigate the pluripotency of human gingival fibroblasts (hGFs), and provide a novel cell source for tissue engineering. Methods With informed consent from volunteers, fresh and healthy gingiva were collected. The hGFs were obtained from the gingiva by tissue culture. The third passage of hGFs was cultured in osteogenic medium, chondrogenic medium and adipogenic medium. Cells without differentiation were taken as control. Cells were examined by al?kaline phosphatase (ALP) staining, Alizarin red staining, Alcian blue staining and oil red O staining for detecting of the abili?ty of differentiation pluripotency. Real-time polymerase chain reaction was applied to examine the expression of osteogenic marker genes ALP, runt-related transcript factor 2 (Runx2), chondrogenic marker aggrecan (AGR) and adipogenic marker peroxisome proliferator-activated receptor gamma 2 (PPARγ2). Results The hGFs cultured in osteogenic medium showed massive violet deposit at day 7 and calcium nodulus at day 28, meanwhile, the expressions of ALP and Runx2 were higher than those of control (P<0.01). In chondrogenic group cells were found blue deposit at day 14. In adipogenic group lipid-filled droplets stained with oil red O were found in cells at day 14. However, hGFs in control group had no any positive stain?ing. Furthermore, expressions of AGR and PPARγ2 were significantly higher than those of control (P<0.01). Conclusion Human gingival fibroblasts have the pluripotency of osteogenic, adipogenic and chondrogenic differentiation.