AIM: To investigate the effect of advanced glycosylation end products on the expression of receptor for advanced glycosylation end products in human monocyte-derived dendritic cells. METHODS: Monocytes were purified (over 98%) using anti-CD14+ microbeads. After 8 d culture in RPMI-1640 medium containing rhGM-CSF (100 ?g/L) and rhIL-4 (50 ?g/L), immature MDCs were derived, then exposed to AGE-BSA (0 or 200 mg/L) for 24 h. Expression of RAGE was semi-quantified by RT-PCR and Western blotting. At the same time, supernatants were collected. IFN-? and IL-12 were analyzed by ELISA. RESULTS: mRNA and protein of RAGE incubated by 200 mg/L AGE-BSA was higher than that in control at 24 h. Treatment of DCs with AGE-BSA resulted in about two-fold increase in the expression of RAGE (P

AIM: The purpose of this study is to investigate the mechanisms related to oxidized low-density lipoprotein(ox-LDL) and dendritic cells(DCs) in the process of atherosclerosis.METHODS: Human DCs were prepared from human CD14~+ peripheral blood monocytes using rhGM-CSF((100 ?g/L)) and rhIL-4(40 ?g/L).Cells were incubated with(100 mg/L) native or oxidized LDL for 72 h.The formation of foam cells was investigated by electron microscopy and oil red O staining.Phenotypic and immune functional assays were used with FACS,FITC-dextran phagocytosis,allogeneic mixed T lymphocytes reaction and secretion of Th1/Th2(IL-12/IL-2) cytokines were also conduced.RESULTS: DCs treated with ox-LDL,but not native LDL were induced into foam cells after cultured for 72 h.Compared with native LDL,ox-LDL-treated DCs were less potent in FITC-dextran phagocytosis.ox-LDL promoted allogeneic T cells proliferation.Moreover,ox-LDL upregulated CD80(72.4? 9.6 vs 89.5?10.1,P

Some kidney stones are caused by single gene mutations, and monogenic kidney stone diseases associated with purine metabolic disorder mainly including adenine phosphoribosyltransferase(APRT) deficiency, hypoxanthine-guanine phosphoribosyltransferase(HPRT)deficiency, hereditary xanthinuria(HX), and some diseases caused by gene mutations such as PRS1, SLC22A12, SLC2A9 and ABCG2. Such diseases can lead to abnormal metabolism of purine and uric acid, and then form 2, 8-dihydroxyadenine stones, uric acid stones or xanthine stones. This kind of diseases are rare, the genotype and phenotype of different types of monogenic diseases related to purine metabolism have their own characteristics and are not widely recognized. At present, the main treatment is medical therapy. Gene sequencing will make the diagnosis and find more disease-related genes or mutations. Gene editing, such as CRISPR/Cas9 technology, makes it possible to cure monogenic kidney stone diseases associated with purine metabolism disorder in the future.

Objective:To investigate the expression of miR-527 in bladder cancer (BC) and its effect on the proliferation, migration and invasion of bladder cancer cells.Methods:From February 2018 to June 2019, the immortalized human bladder epithelial cell line SV-HUC-1 and human bladder cancer cell lines T24, UM-UC-3, 5637 and RT-112 were cultured in vitro. Real time quantitative PCR (qRT-PCR) was used to detect the expression of miR-527 in BC bladder cancer tissues and adjacent normal bladder tissues, human bladder cancer cell lines and human bladder epithelial immortalized cell lines. MiR-527 mimics, miR-527 inhibitor, ENO1 overexpression plasmid, ENO1 siRNA and corresponding negative control were transfected into bladder cancer cell line. CCK8 test, clone formation test and Transwell test were used to study the cell proliferation, migration and invasion. Luciferase reporter gene assay was used to verify the target gene of miR-527. Western blotting was used to analyze the regulation of miR-527 on target gene expression.Result:Compared with normal bladder tissue, the expression of miR-527 in bladder cancer was significantly lower (1.723±1.070 vs. 1.148±0.760, P<0.05). The relative expression of miR-527 in T24 (0.540±0.082), UM-UC-3 (0.230±0.053), 5637(0.463±0.085) and RT-112 (0.310±0.056) were significantly lower than those in SV-HUC-1 cells (0.987±0.111) with statistical significance ( P<0.05). Compared with the negative control (NC) group, CCK8 assay results showed that the cell viability was significantly decreased after transfection of miR-527 mimics into UM-UC-3 cells ( P<0.05). The clone formation test showed that the number of cell clones in UM-UC-3 cells transfected with miR-527 mimics was significantly lower than that in the control group (157.00±15.52 vs 57.33±15.50, P<0.05). Compared with the control group, the cell activity of T24 cells transfected with miR-527 inhibitor was significantly increased ( P<0.05). Compared with the control group, the number of cell clone formation was significantly increased (76.67±9.07 vs. 141.70±10.50, P<0.05). According to the prediction of targetscan database, ENO1 was the target gene of mir-527. Luciferase reporter gene experiment showed that the luciferase activity of mir-527 mimics group was significantly lower than that of control group (0.99±0.02 vs. 0.47±0.10, P<0.05), while the luciferase activity of miR-527 mimics group was significantly lower than that of control group (0.99 ± 0.02 vs. 0.47 ± 0.10, P<0.05), without statistical significance (1.03±0.04 vs. 0.96±0.05, P>0.05). Western blot analysis showed that the expression of ENO1 in miR-527 mimics group was significantly lower than that in NC mimics group (1.09±0.17 vs. 0.31±0.13, P<0.05), and the expression of ENO1 in miR-527 inhibitor group and NC inhibitor group were significantly increased (0.97±0.09 vs. 2.17±0.15, P<0.05). Compared with miR-527 mimics group, transfection of miR-527 mimics+ ENO1 overexpression plasmid could reduce the inhibitory effect of miR-527 mimics on proliferation, migration and invasion of bladder cancer cell line ( P<0.05). Compared with miR-527 inhibitor group, transfection of miR-527 inhibitor+ ENO1 siRNA could weaken the inhibit effect of miR-527 on the proliferation, migration and invasion of bladder cancer cell lines ( P<0.05). Conclusion:miR-527 is low expressed in BC and can be used as a tumor suppressor gene to inhibit the proliferation, migration and invasion of BC cells.

Primary hyperoxaluria (PH) is a rare autosomal recessive hereditary disease, characterized by calcium oxalate kidney stone and nephrocalcinosis caused by defects in enzymes of liver glyoxylate metabolism. Up to now, treatment options for PH are limited. Although medication treatment and liver transplantation can slow down the progression and mitigate the symptoms, the evidence for them turned out to be weak. In recent years, breakthroughs in biotechnology provide novel promising directions for drug development. Small interfering RNA drugs, such as lumasiran and nedosiran, selectively reduce hepatic expression of glycolate oxidase and lactate dehydrogenase respectively, reducing hepatic oxalate production and urinary oxalate levels in PH patients. Gene-editing, such as CRISPR/Cas9, will be a potential treatment method of PH. This review encompasses recent developments in the gene therapy of PH.

Primary hyperoxaluria type 3 (PH3) is a rare monogenic nephrolithiasis caused by HOGA1 gene mutations. With the advancement of technology of genetic testing, the mutation site of PH3 patients can be clearly located, and the characteristics of genotype, phenotype, genotype-phenotype correlations are also gradually recognized. With the development of gene therapy, novel gene editing techniques and RNA interference treatments offer hope for the future of PH3 treatment. In this paper, the characteristics of genotype and phenotype, genotype-phenotype correlations of PH3 will be summarized and its future treatment will be prospected.

Renal abscess caused by fish bone ingestion is extremely rare and has not been reported in the literature. A male patient presented with a 1-week history of flank pain and a 2-day history of fever. Urinary ultrasound and CT scan showed an irregular hypodense lesion in the left kidney and blurred thickening of the descending colon wall. Three-dimensional CT reconstruction images revealed a needle-like foreign body, which perforated from the descending colonic lumen to the left kidney. The patient had accidentally eaten fish bone one week prior. On the basis of clinical data, the diagnosis of renal abscess caused by foreign body was suspected. Accordingly, laparotomy was performed, the abscess was drained, and the colon was repaired. The foreign body was confirmed to be fish bone. The postoperative condition of the patient was uneventful, and the patient remained well in the 3 months' follow-up without any further complaints.

Objective:To investigate the clinical value of prospective ECG-gated high-pitch protocol scanning of third generation DSCT in the diagnosis of pediatric congenital heart disease (CHD).Methods:A total of 243 children with confirmed CHD who were expected to undergo surgical treatment were prospectively collected and randomly divided evenly into 3 groups, with first group for prospective ECG-gated high-pitch scanning in third generation DSCT (Flash 3rd), second group for prospective ECG-gated high-pitch scanning in second generation DSCT (Flash 2nd) and third group for prospective sequential scanning in third generation DSCT (Sequence 3rd). The SD value and SNR of aortic root and pulmonary artery of each child were recorded. The 5-point system is adopted with subjective scoring. Based on the result of operation, the diagnosis accuracy in 3 groups was analyzed. Results:The E values in Flash 3rd, Flash 2nd and Sequence 3rd group were 0.24 (0.19, 0.27), 0.11 (0.10, 0.14) and 0.44 (0.39, 0.48) mSv ( H=207.04, P<0.05), respectively. Subjective scores of group Flash 3rd and Sequence 3rd were significantly higher than that of group Flash 2nd [4 (4, 4) vs. 4(3, 4) vs. 3(3, 3), H=124.05, P<0.05] and no difference between these two groups. SD value of aortic root and pulmonary artery of group Flash 3rd and Sequence 3rd were significantly lower than that of group Flash 2nd( H= -40.27-33.38, P<0.05). SNR of aortic root and pulmonary artery of group Flash 3rd was significantly higher than that of group Flash 2nd and Sequence 3rd ( H=-0.90-51.42, P<0.05). Diagnosis accuracy of intracardiac malformation for group Flash 2nd was significantly lower than that of Flash 3rd and Sequence 3rd (77.7%, 90.9%, 88.9%, K=9.36, P<0.05), and there was no significant difference between the latter two groups. There was no difference in diagnosis accuracy of extracardiac malformation among 3 groups (88.6%, 94.8%, 92.2%, K=3.11, P=0.21). Conclusions:The prospective ECG-gated high-pitch scanning in third generation DSCT can take into account radiation dose and image quality, which has important clinical value in the diagnosis of CHD.

Glutamate receptor-like (GLR) is an important class of Ca²⁺ channel proteins, playing important roles in plant growth and development as well as in response to biotic and abiotic stresses. In this paper, we performed genome-wide identification of banana GLR gene family based on banana genomic data. Moreover, we analyzed the basic physicochemical properties, gene structure, conserved motifs, promoter cis-acting elements, evolutionary relationships, and used real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) to verify the expression patterns of some GLR family members under low temperature of 4 ℃ and different hormone treatments. The results showed that there were 19 MaGLR family members in Musa acuminata, 16 MbGLR family members in Musa balbisiana and 14 MiGLR family members in Musa itinerans. Most of the members were stable proteins and had signal peptides, all of them had 3-6 transmembrane structures. Prediction of subcellular localization indicated that all of them were localized on the plasma membrane and irregularly distributed on the chromosome. Phylogenetic analysis revealed that banana GLRs could be divided into 3 subclades. The results of promoter cis-acting elements and transcription factor binding site prediction showed that there were multiple hormone- and stress-related response elements and 18 TFBS in banana GLR. RT-qPCR analysis showed that MaGLR1.1 and MaGLR3.5 responded positively to low temperature stress and were significantly expressed in abscisic acid/methyl jasmonate treatments. In conclusion, the results of this study suggest that GLR, a highly conserved family of ion channels, may play an important role in the growth and development process and stress resistance of banana.
Musa/metabolism* ; Phylogeny ; Abscisic Acid/metabolism* ; Temperature ; Stress, Physiological/genetics* ; Hormones/metabolism* ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism* ; Gene Expression Profiling