AIM and METHODS: To investigate expression of intercellular adhesion molecule-1(ICAM-1) on human umbilical vein endothelial cells (HUVEC) induced by lipopolysaccharide (LPS) , and inhibiting role of polydatin by cellular immune fluorescent staining and laser confocal microscope scanning technology. RESULTS: Compared with basic expression of ICAM-1 on HUVEC, the ICAM-1 expression was enhanced significantly after stimulated by LPS from 8 h to 36 h, dose-dependent relation appeared between expression of ICAM-1 and LPS. ICAM-1 expression on endothelial cells treated only by polydatin had no abvious change,but inducing role of LPS to expression of ICAM-1 was inhibited significantly by polydatin pretreating endothelial cells. CONCLUSION: The expression of ICAM-1 on endothelial cells can be promoted by LPS , and polydatin can inhibit LPS-induced ICAM-1 expression

AIM: To construct the plasmid vectors containing different regions of human eNOS promoter coupled to a red fluorescent protein reporter gene, which may express in mammalian cells. METHODS: Different regions of human eNOS promoter were subcloned respectively into a red fluorescent protein vector, pDsRed1-1. These recombinant vectors, pDsF1033Red, pDsF494Red and pDsF166Red, were then transfected into NIH3T3 cell lines, followed by the observation under a fluorescent microscope. RESULTS: After identified to be right by double restriction enzyme digestion, PCR and sequencing, the vectors might be effectively expressed in NIH3T3 cells. 95 % of the red fluorescent emitted by a red fluorescent protein dispersed all over the cells, appearing at 48-60 h after transfection, reaching peak at 96-144 h, becoming the strongest in light at 144 h, gradually disappearing after 168 h and remaining little red fluorescent in 21 days. The quantity and intensity in expressions of red fluorescent protein drived by different regions of human eNOS promoter were clearly lower than by a strong promoter, p CMVIE . CONCLUSION: The red fluorescent protein reporter gene vectors containing different regions of human eNOS promoter are successfully constructed and may efficaciously express in mammalian cells, appearing not strong transcriptional activities, which provide practical and feasible tools to study functions of different regions of human eNOS promoter and roles of cis-elements in it. [

Objective To detect the efficiency of the newly developed PLGA/IO MPs in tracking tendon stem cells (TSCs) by magnetic resonance (MR) and photoacoustic (PA) imaging . Methods Both PLAG/IO MPs and TSCs were prepared and acquired according to the previous study ,and TSCs were incubated with PLGA/IO MPs for labeling .TSCs were collected for MR and PA imaging ,prussian blue staining was performed ,and the iron concentration of labeled TSCs was determined using inductively coupled plasma optical emission spectrometry ( ICP-OES ) at 3 ,7 ,14 ,21 and 28 days after labeling respectively . The rotator cuff injury model was built on the right side of SD rats by surgery and the labeled TSCs were implanted instantly . Dual-modal MR/PA imaging was performed to observe the implanted labeled TSCs at day 3 ,7 ,14 ,21 and 28 after implantation respectively . Results Along with the increase of labeling time ,both MR and PA signal of labeled TSCs decreased gradually ,and the amount of intracellular Fe loading was gradually decreased . At day 28 ,the difference of Fe concentration per cell between labeled TSCs and non-labeled TSCs was not significant (1 .45 pg Fe/cell vs 1 .17 pg Fe/cell , P ＞0 .05) . MR and PA imaging allowed a long-term tracking of labeled TSCs for 21 and 7 days respectively in the rat rotator cuff injury model . Conclusions PLGA/IO MPs are able to label TSCs for up to 21 days ,and dual-modal MR/PA imaging could be used to track the labeled TSCs in the rat rotator cuff injury model .

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are confirmed to be expressed in bladder interstitial Cajal-like cells (ICC-LCs), but little is known about their possible role in cystitis-associated bladder dysfunction. The present study aimed to determine the functional role of HCN channels in regulating bladder function under inflammatory conditions. Sixty female wild-type C57BL/6J mice and sixty female HCN1-knockout mice were randomly assigned to experimental and control groups, respectively. Cyclophosphamide (CYP)-induced cystitis models were successfully established in these mice. CYP treatment significantly enhanced HCN channel protein expression and I(h) density and significantly altered bladder HCN1 channel regulatory proteins. Carbachol (CCH) and forskolin (FSK) exerted significant effects on bladder ICC-LC [Ca²⁺]i in CYP-treated wild-type (WT) mice, and HCN1 channel ablation significantly decreased the effects of CCH and FSK on bladder ICC-LC [Ca²⁺]i in both naive and CYP-treated mice. CYP treatment significantly potentiated the spontaneous contractions and CCH (0.001-10 µM)-induced phasic contractions of detrusor strips, and HCN1 channel deletion significantly abated such effects. Finally, we demonstrated that the development of CYP-induced bladder overactivity was reversed in HCN1 -/- mice. Taken together, our results suggest that CYP-induced enhancements of HCN1 channel expression and function in bladder ICC-LCs are essential for cystitis-associated bladder hyperactivity development, indicating that the HCN1 channel may be a novel therapeutic target for managing bladder hyperactivity.
Animals ; Carbachol ; Colforsin ; Cyclophosphamide ; Cystitis ; Female ; Humans ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels* ; Mice* ; Telocytes* ; Up-Regulation* ; Urinary Bladder*

Objective To prepare PLGA microparticles loaded with Iron oxide (PLGA/IO MPs) and explore their feasibility of the rat tendon stem cells (TSCs) labeled with the particles and the multimodal imaging of Ultrasonic (US)/Photoacoustic (PA)/Magnetic resonance (MR) in vitro. Methods The PLGA/IO MPs were prepared using double emulsification,and physical and chemical properties were tested and US/PA/MRI imaging was performed.The TSCs were labeled with PLGA/IO MPs,and transmission electron microscopy (TEM) and prussian blue staining were performed to test labeling effects,then the US, PA and MRI imaging of labeled TSCs were performed. Results The diameter and Zeta potential of prepared PLGA/IO MPs were ( 801.5 ± 165.6) nm and (6.36 ± 3.36) mV [the Zeta potential of microparticles which including poly-L-Lysine(PLL) was about (3.16 ± 3.69)mV],respectively.PLGA/IO MPs could be imaged by US/PA/MRI multimodal imaging. After labeling,the PLGA/IO MPs were distributed in cytoplasm of labeled TSCs which could be imaged by US,PA,MRI simultaneously. Conclusions The TSCs can be labeled with PLGA/IO MPs effectively,and imaged by using multimodal US/PA/MRI imaging in vitro,which will lay foundation for noninvasive and multimodal tracking of transplanted TSCs in vivo.

【Objective】 To evaluate the safety and efficacy of long-term indwelling of Allium ureteral stent in the treatment of ureteral stricture. 【Methods】 The clinical data of patients who underwent endoscopic Allium ureteral stent implantation for ureteral stricture in our hospital during Aug.2020 and Dec.2022 were retrospectively analyzed, and the surgical conditions and adverse events were recorded. The data of serum creatinine, blood urea nitrogen, glomerular filtration rate (GFR) and renal pelvis width under ultrasound were compared before surgery and 1, 3, 6 and 12 months after surgery. 【Results】 A total of 52 patients with ureteral stricture of 1.1 (0.7, 2.0)cm were included. All operations were successful. The operation time was 82.5 (70, 114)min, intraoperative blood loss 20 (10, 20)mL, and postoperative hospitalization stay 1 (1, 2) day. During the follow-up of (13.2±7.8) months, 14 patients had stent displacement, 5 had stone obstruction of stent tubes, 7 had occasional hematuria after movement, 9 had intermittent lumbar and abdominal pain, and 1 had recurrent urinary tract infection. The serum creatinine, blood urea nitrogen and renal pelvis width of 1 month, 3, 6 and 12 months after surgery were significantly decreased, while GFR was significantly increased. 【Conclusion】 Long-term indwelling of Allium ureteral stent is effective in the treatment of ureteral stricture, but the high incidence of stent displacement should arouse attention.