CD73-positive pediatric urethral mesenchymal stem-like cell-derived small extracellular vesicles stimulate angiogenesis.

Introduction: Angiogenesis plays an important role in the repair of urethral injury, and stem cells and their secretomes can promote angiogenesis. We obtained pediatric urethral mesenchymal stem-like cells (PU-MSLCs) in an earlier study. This project studied the pro-angiogenic effect of PU-MSLC-derived small extracellular vesicles (PUMSLC-sEVs) and the underlying mechanisms. Materials and methods: PUMSLCs and PUMSLC-sEVs were cultivated and identified. Then, biological methods such as the ethynyl deoxyuridine (EdU) incorporation assay, Cell Counting Kit-8 (CCK-8) assay, scratch wound assay, Transwell assay, and tube formation assay were used to study the effect of PUMSLC-sEVs on the proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs). We explored whether the proangiogenic effect of PUMSLC-sEVs is related to CD73 and whether adenosine (ADO, a CD73 metabolite) promoted angiogenesis. GraphPad Prism 8 software was used for data analysis. Results: We observed that PUMSLC-sEVs significantly promoted the proliferation, migration, and tube-forming abilities of HUVECs. PUMSLC-sEVs delivered CD73 molecules to HUVECs to promote angiogenesis. The angiogenic ability of HUVECs was enhanced after treatment with extracellular ADO produced by CD73, and PUMSLC-sEVs further promoted angiogenesis by activating Adenosine Receptor A2A (A2AR). Conclusions: These observations suggest that PUMSLC-sEVs promote angiogenesis, possibly through activation of the CD73/ADO/A2AR signaling axis.

LncRNA IGFBP4-1 promotes tumor development by activating Janus kinase-signal transducer and activator of transcription pathway in bladder urothelial carcinoma: retraction.

[This retracts the article DOI: 10.7150/ijbs.46986.].

Isolation and identification of a mesenchymal stem/stromal cell-like population from pediatric urethral tissue.

Mesenchymal stem cells (MSCs) are important seed cells for cell therapy and tissue engineering because of their multidirectional differentiation potential, high proliferative capacity, low immunogenicity, and immunomodulatory ability. In this study, we successfully isolated and cultured a population of mesenchymal stem-like cells from pediatric urethra (PU-MSLCs). The cells had a spindle-shaped fibroblast-like morphology, similar to MSCs derived from other tissues. The PU-MSLCs highly expressed MSC surface markers CD29, CD73, CD90, and CD105 but were negative for leukocyte common antigen CD45, and MHC class II-encoded molecule HLA-DR. After in vitro induction, the PU-MSLCs had the potential to differentiate into adipocytes, osteocytes, and chondrocytes. The PU-MSLCs maintained a normal karyotype and showed no tumorigenicity during long-term cultivation. We thus demonstrated that the mesenchymal stem/stromal cell-like population obtained from pediatric urethra tissue is capable of self-renewal and multidirectional differentiation, has promising application prospects for cell therapy and tissue engineering, and is expected to contribute to urethral tissue reconstruction.

CD73-Positive Small Extracellular Vesicles Derived From Umbilical Cord Mesenchymal Stem Cells Promote the Proliferation and Migration of Pediatric Urethral Smooth Muscle Cells Through Adenosine Pathway.

Smooth muscle cells (SMCs) are the main functional component of urethral tissue, but are difficult to proliferate in vitro. Mesenchymal stem cells (MSCs) and mesenchymal stem cell-derived small extracellular vesicles (MSC-sEV) have been shown to promote tissue repair by regulating the proliferation and migration of different types of cells. In this study, we investigated the effect of umbilical cord mesenchymal stem cell-derived sEV (UCMSC-sEV) on the proliferation and migration of pediatric urethral smooth muscle cells (PUSMCs) and the mechanism by which sEV regulates the function of PUSMCs. We observed that UCMSC-sEV can significantly promote the proliferation and migration of PUSMCs in vitro. UCMSC-sEV exerted proliferation and migration promotion effects by carrying the CD73 to PUSMCs and catalyzing the production of adenosine. Conversely, the effect of UCMSC-sEV on the proliferation and migration of PUSMCs were no longer observed with addition of the PSB12379 as a CD73 inhibitor. It was found that the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway in PUSMCs was activated by adenosine or UCMSC-sEV intervention. In summary, UCMSC-sEV promoted proliferation and migration of PUSMCs in vitro by activating CD73/adenosine signaling axis and downstream PI3K/AKT pathway. Thus, we concluded that UCMSC-sEV may be suggested as a new solution strategy for the urethral tissue repair.

Risk factors of uterine contraction after ureteroscopy in pregnant women with renal colic.

BACKGROUND: Ureteroscopy is widely applied in pregnant women with renal colic, but such patients are easy to experience uterine contraction after surgery. There are many factors which may affect uterine contraction, this study aims to explore the risk factors of uterine contraction triggered by ureteroscopy in pregnant women with renal colic. METHODS: One hundred and one pregnant women were retrospectively analyzed, the patients were hospitalized because of severe renal colic. All patients received ureteroscopy during which double J catheters were inserted into ureters for drainage. Patients received other medical treatments individually according to their condition and uterine contractions were detected by EHG within 12 h after operation. Patients were classified as group A (uterine contraction) and group B (no uterine contraction) according to the presence or absence of continuously regular uterine contraction. Clinical characteristics were collected for further analysis, including history of childbirth, anesthesia method, application of phloroglucinol or not, operation time, Oxygen inhalation or not, pain relief or not after surgery, systemic inflammatory response syndrome (SIRS) occurred or not. A binary logistic regression analysis model was established to explore whether such clinical characteristics were relevant to uterine contraction after ureteroscopy. RESULTS: Continuously regular uterine contraction presented in 46 pregnant women within 12 h after ureteroscopy, making the incidence of uterine contraction as high as 45.54%. The presence of uterine contraction was related to the following factors(P < 0.05): history of childbirth (primipara versus multipara)(OR 6.593, 95% CI 2.231-19.490), operation time (each quarter additional) (OR 2.385, 95% CI 1.342-4.238), application of phloroglucinol (yes versus not) (OR 6.959, 95% CI 1.416-34.194), pain relief after surgery(yes versus not)(OR 6.707, 95% CI 1.978-22.738), SIRS occurred after surgery (yes versus not) (OR 0.099, 95% CI 0.014-0.713). CONCLUSION: Continuously regular uterine contraction is easy to occur within 12 h after ureteroscopy in pregnant women. SIRS occurred after surgery is a risk factor for uterine contraction; on the contrary, no history of childbirth, shorter operation time, application of phloroglucinol, pain relief after surgery are protective factors.

CRISPR-CasRx Targeting LncRNA LINC00341 Inhibits Tumor Cell Growth in vitro and in vivo.

CRISPR-CasRx technology provides a new and powerful method for studying cellular RNA in human cancer. Herein, the pattern of expression of long noncoding RNA 00341 (LINC00341) as well as its biological function in bladder cancer were studied using CRISPR-CasRx. qRT-PCR was employed to quantify the levels of expression of LINC00341 in tumor tissues along with the matched non-tumor tissues. sgRNA targeting LINC00341 or the sgRNA negative control were transiently transfected into the T24 as well as 5,637 human bladder cancer cell lines. CCK-8, ELISA as well as wound healing methods were employed to explore cell proliferation, apoptosis and migration, respectively. The tumorigenicity experiment in nude mice also performed to detect cell proliferation. The expression of p21, Bax as well as E-cadherin were assayed using western blot. The results demonstrated that LINC00341 was overexpressed in bladder cancer in contrast with the healthy tissues. The LINC00341 expression level in high-grade tumors was higher in contrast with that in low-grade tumors. The expression of linc00341 was higher relative to that of non-invasive tumors. In T24 as well as 5637-cell lines harboring LINC00341-sgRNA, inhibition of cell proliferation (in vitro and in vivo), elevated apoptosis rate and diminished migration ability. Moreover, silencing LINC00341 upregulated the expressions of p21, Bax as well as E-cadherin. Knockout of these genes could eliminate the phenotypic changes caused by sgRNA targeting LINC00341. Our data demonstrate that LINC00341 has a carcinogenic role in human bladder cancer.

LncRNA IGFBP4-1 promotes tumor development by activating Janus kinase-signal transducer and activator of transcription pathway in bladder urothelial carcinoma.

Insulin-like growth factor binding protein 4-1 (IGFBP4-1), a new long noncoding RNA (lncRNA), has been reported to contribute to tumorigenesis and has been suggested to be a poor prognostic marker in human lung cancer. However, there still lacks basic studies that investigated the biological role of IGFBP4-1 in bladder urothelial carcinoma to date. In this study, we investigated the relationship between IGFBP4-1 expression and prognosis in patients with bladder cancer. Cell proliferation, cell cycle and cell apoptosis assays were performed to assess IGFBP4-1 function by up-regulating or down-regulating IGFBP4-1 in bladder cancer cells. A xenograft mice model was used to validate the in vitro results. Blockade of Janus kinase-signal transducer and activator of transcription pathway (JAK/STAT) was used to evaluate JAK/STAT signaling activity. The results showed that IGFBP4-1 was overexpressed in bladder cancer tissues compared with that in normal bladder tissues, and its expression level was positively correlated with poor prognosis in bladder cancer patients. Overexpression of IGFBP4-1 markedly promoted cell proliferation and cell cycle progression, and inhibited cell apoptosis, while knockdown of IGFBP4-1 notably suppressed the proliferation, promoted cell apoptosis, and induced cell cycle arrest at the G0/G1 phase. Mechanistically, we revealed that IGFBP4-1 promotes the activation of the JAK/STAT pathway in bladder cancer cells. Moreover, the JAK/STAT inhibitor dramatically blocked the tumor-promoting activity of IGFBP4-1. Tumor growth in vivo was also suppressed by knocking down of IGFBP4-1. In conclusion, IGFBP4-1 promoted bladder cancer progression by activating the JAK/STAT signaling pathway. These findings suggest that IGFBP4-1 exhibits an oncogenic role in the development of human bladder cancer.

Evaluation of the clinical value of retroperitoneal laparoscopic pyeloplasty in the treatment of ureteropelvic junction obstruction in infants: A single-center experience involving 22 consecutive patients.

Retroperitoneal laparoscopic pyeloplasty (RLP) is 1 method for treating ureteropelvic junction obstruction (UPJO) in children, but reports are more common in children than in infants younger than 2 years old. The purpose of this study was to evaluate the clinical value of RLP for infants with UPJO.From January 2015 to December 2017, a retrospective analysis of 22 infants aged 2 to 24 (11.95 ±â€Š6.00) months with UPJO who were treated with RLP in our hospital was performed. During the same period, 14 infants who underwent conventional transperitoneal laparoscopic pyeloplasty (TLP) were compared with those who underwent RLP. Postoperative recovery and complications, including bleeding, infection, urinary leakage and anastomotic stenosis, postoperative resumption of oral feeding, postoperative hospitalization time and surgical success rate were evaluated. Drainage and function were assessed with isotope scan at 6 months and later during the yearly follow-up and by intravenous urography (IVU) and mercaptoacetyltriglycine (MAG3) renography.Both groups underwent successful surgery. The operative time in the RLP group was 88 to 205 (120.59 ±â€Š24.59) min, and there was no significant difference compared with the TLP group (P = .767). The estimated intraoperative blood loss was 2 to 10 (3.75 ±â€Š1.59) ml, which was not significantly different between the 2 groups (P = .386). In the RLP group, the mean postoperative resumption of oral feeding was faster than that in the TLP group (3.55 ±â€Š0.74 vs 5.50 ±â€Š0.85 hour, P < .001), and the postoperative hospitalization time was shorter in the TLP group than in the RLP group (6.59 ±â€Š0.50 vs 7.07 ±â€Š0.47 day, P = .007 < .05). Follow-up lasted from 6 months to 3 years, and there was a significant reduction in postoperative hydronephrosis in both groups (P < .05, respectively).RLP is a safe procedure for infants. This procedure is associated with relatively little trauma, a quick recovery and good cosmetic effects. RLP also has the advantages of relatively little interference with the abdominal cavity and sufficient operating space; thus, this technique is worth promoting.

Application of Ureteroscope in Emergency Treatment with Persistent Renal Colic Patients during Pregnancy.

BACKGROUND: Although the application of ureteroscopy in the treatment of ureteral calculi during pregnancy has been on the rise, for persistent renal colic patients without ultrasound-detected ureteral calculi, it may represent a clinical dilemma due to the potential risks for both mother and fetus. OBJECTIVE: The aim of the present study is to present our experience with the application of the ureteroscope in the emergency treatment of persistent renal colic patients during pregnancy. METHODS: From March 2009 to September 2014, a total of 117 pregnant women who received ureteroscopy for persistent renal colic were retrospectively analyzed. Patients were divided into three groups according to duration of the persistent renal colic: Group A (within 12 hours; 24 cases); Group B (12 to 24 hours; 76 cases); and Group C (more than 24 hours; 17 cases). The stone-free rate, complications, and other qualitative data were analyzed. RESULTS: Of the 117 patients, 31 patients who were found not to have renal or ureteral calculi received ureteroscopic double-J (DJ) stent insertion, whereas 86 patients who were found with ureteral calculi received ureteroscopic lithotripsy (URSL) and DJ stent insertion. Among them, 24 patients (27.9%) were found with ureteral calculi by ureteroscopy rather than ultrasound. In addition, 73 patients (84.9%) had complete fragmentation of calculi; 12 patients (10.3%) had a threatened abortion (the rates of threatened abortion in Groups A, B and C were 8.3% vs. 6.5% vs. 29.4%; Group C compared with Groups A and B, p<0.05), and one patient (1.2%) had urosepsis (in Group C). However, these complications were cured with conservative treatment, without postpartum infant and maternal complications. CONCLUSION: For pregnant patients with persistent renal colic/ureteral calculi and hydronephrosis, ureteroscopic DJ stent insertion and URSL are effective and safe options when conservative treatment fails, even if no urinary calculi were found by ultrasound. At the same time, for patients with persistent renal colic during pregnancy, early application of ureteroscopy may reduce the risk of preterm birth.

Biomimetic gelatin methacrylamide hydrogel scaffolds for bone tissue engineering.

Bone tissue engineering is an exciting research area that develops functional strategies for the most challenging bone related clinical issues. Among the numerous materials used in this area, biomimetic materials have been developed amazingly over the past decades. In this study, biomimetic gelatin methacrylamide (Bio-GelMA) hydrogel scaffolds have been fabricated to mimic both the physical architecture and chemical composition of the natural bone extracellular matrix (ECM) by using the thermally induced phase separation (TIPS) technique, to provide three-dimensional templates and extracellular matrix microenvironments. Adipose derived stem cells (ADSCs) also play a pivotal role in osteogenesis when co-cultured with Bio-GelMA at days 7, 14, and 21. The effects of cell-biomaterial interactions such as adhesion, proliferation, and osteogenic differentiation were systematically investigated. The results showed that Bio-GelMA significantly enhanced cell attachment and viability, alkaline phosphatase (ALP) activity, and mineral deposition, as well as mRNA expression levels of osteogenic genes of ADSCs. In a subcutaneous model, H&E staining and dual immunofluorescent staining differed significantly. More importantly, in a critical-size rat calvarial bone defect model, the results of Micro-CT, H&E staining and Masson trichrome staining confirmed that the combination of the Bio-GelMA and ADSCs could enhance osteogenesis significantly. Altogether, the observations prove that the Bio-GelMA scaffolds can act as cell carriers for ADSCs, promote greater osteogenic differentiation of ADSCs and may have great potential in future clinical applications.

Urolithiasis in pregnancy: survey in clinical epidemiology.

This study examined the association of pregnancy with urolithiasis and provided new insights into urolithiasis in pregnancy. A total of 462 subjects were studied from January 2004 to December 2009 in Foshan Maternal and Child Health Hospital, China. Among the 462 subjects, 162 cases of urolithiasis during pregnancy (UPG) were selected as the observation group, 150 cases of no urolithiasis during pregnancy (NUPG) served as pregnancy control group, and 150 cases of no pregnancy (NPG) at reproductive age who took part in physical examination were randomly assigned into non-pregnant control group. At the same time, the patients in observation group were divided into the following sub-groups: no symptomatic urinary calculus (NSUC) and symptomatic urinary calculus (SUC) groups; SUC group was further divided into surgical intervention (SI) and conservative management (CM) groups. The general information and the data of blood and urine were collected and compared among the groups. The results showed that the incidence of urinary calculi in pregnant women was lower than that in non-pregnant women, the formation of urinary stone was associated with the change of metabolism of protein and sugar in pregnant women, and the surgical intervention was a practicable alternative to treat the clinical intractable symptomatic urinary calculi in pregnancy.

Urolithiasis in Pregnancy: Survey in Clinical Epidemiology / 华中科技大学学报（医学）（英德文版）

This study examined the association of pregnancy with urolithiasis and provided new insights into urolithiasis in pregnancy.A total of 462 subjects were studied from January 2004 to December 2009 in Foshan Maternal and Child Health Hospital,China.Among the 462 subjects,162 cases of urolithiasis during pregnancy (UPG) were selected as the observation group,150 cases of no urolithiasis during pregnancy (NUPG) served as pregnancy control group,and 150 cases of no pregnancy (NPG) at reproductive age who took part in physical examination were randomly assigned into non-pregnant control group.At the same time,the patients in observation group were divided into the following sub-groups:no symptomatic urinary calculus (NSUC) and symptomatic urinary calculus (SUC) groups;SUC group was further divided into surgical intervention (SI) and conservative management (CM) groups.The general information and the data of blood and urine were collected and compared among the groups.The results showed that the incidence of urinary calculi in pregnant women was lower than that in non-pregnant women,the formation of urinary stone was associated with the change of metabolism of protein and sugar in pregnant women,and the surgical intervention was a practicable alternative to treat the clinical intractable symptomatic urinary calculi in pregnancy.