COX-2/PGE2 facilitates fracture healing by activating the Wnt/ß-catenin signaling pathway.

OBJECTIVE: The aim of this study was to explore the influence of cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2) on fracture healing by activating the Wnt/ß-catenin signaling pathway. MATERIALS AND METHODS: In this study, 36 adult Sprague-Dawley (SD) rats raised in our laboratory were selected as research objects. The rats were subjected to fracture surgery on the middle part of the right femoral shaft. Subsequently, they were randomly divided into the control group and experimental groups (including experimental group A and experimental group B). Rats in experimental group A were injected with PGE 2 or COX-2 selective inhibitor NS-398, while rats in experimental group B were injected with PGE2 (5 µmol/L). Meanwhile, rats in the control group were injected with the same amount of normal saline. After that, the transcriptional levels of PEG2, COX-2, vascular endothelial growth factor (VEGF) and ß-catenin in rats of the experimental group A, experimental group B and control group were detected via fluorescence quantitative Polymerase Chain Reaction (PCR) assay. Enzyme-linked immunosorbent assay (ELISA) and Western blotting were conducted to determine the changes in protein levels of PEG2, COX-2, VEGF and ß-catenin in rats of the experimental group A, experimental group B and control group. The expression level of VEGF in bone tissues at fracture ends of rats in the experimental group A, experimental group B and control group was observed through the hematoxylin-eosin (HE) staining. Furthermore, micro-computed tomography (CT) was employed to evaluate callus formation. RESULTS: The transcriptional and translational levels of COX-2, ß-catenin and VEGF in rats of experimental group A treated with COX-2 inhibitors were significantly down-regulated when compared with those of the control group, showing statistically significant differences (p<0.05). However, the levels of these genes were markedly elevated in the experimental group B treated with PGE2 in comparison with those in the control group, and the differences were statistically significant (p<0.05). After 6 weeks, HE staining showed that the expression level of VEGF in rats of the experimental group B was remarkably higher than that of the experimental group A (p<0.05). Micro-CT results revealed that the mean trabecular plate density (MTPD) of rats in the experimental group B (73.29±5.4) was markedly higher than the number of osteoblasts (49.6±3.9) in the experimental group A, showing a statistically significant difference (p<0.05). CONCLUSIONS: COX-2/PGE2 facilitates fracture healing by activating the Wnt/ß-catenin signaling pathway.

miR-940 promotes spinal cord injury recovery by inhibiting TLR4/NF-κB pathway-mediated inflammation.

OBJECTIVE: The aim of this study was to investigate the effects of miR-940 and Toll-like receptor 4/Nuclear Factor κB (TLR4/NF-κB) pathways on inflammatory responses and spinal cord injury (SCI). MATERIALS AND METHODS: This study first established a model of spinal cord injury in mice. The grip force measurement was used to detect the recovery of the forelimb, left forelimb and right forelimb of SCI mice. The quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of miR-940 and macrophage receptor TLR4 in SCI mice. In addition, the protein levels of TLR4 and inducible nitric oxide synthase (iNOS) in SCI mice were detected by Western blot. MiR-940 mimic was injected into the injured area of SCI mice to explore the effect of miR-940 overexpression on TLR4 and myeloperoxidase (MPO) expression as well as the protein levels of TLR4, P65 and iNOS. Furthermore, the grip strength of SCI mice with double forelimb, left forelimb and right forelimb was detected by the grip force test after miR-940 overexpression. RESULTS: Compared with the sham-operated mice, the grip strength of the forelimb, left forelimb, and right forelimb of the SCI group showed significant obstacles. Meanwhile, the expression of miR-940 was remarkably decreased in SCI mice along with significant elevation of the inflammatory response-related factors including TRL4 and iNOS. Then we injected SCI mice with miR-940 mimics into the spinal cord injury area and found that miR-940 overexpression decreased the expression levels of TLR4 and MPO. At the same time, the overexpression of miR-940 markedly decreased the protein levels of TLR4, P65, and iNOS in SCI mice. In addition, miR-940 overexpression improved the grip strength of the left and right forepaws and the simultaneous grip strength of the two claws of the SCI mice than those of the simple injury group. CONCLUSIONS: High expression of miR-940 can promote the recovery of spinal cord injury by downregulating the TLR4/NF-κB signaling pathway and inhibiting inflammation.

MicroRNA-23c inhibits articular cartilage damage recovery by regulating MSCs differentiation to chondrocytes via reducing FGF2.

OBJECTIVE: The aim of the study was to explore the role of microRNA-23c in the differentiation of marrow stromal cells (MSCs) to chondrocytes and its potential mechanism. MATERIALS AND METHODS: MSCs were first isolated from rat bone marrow for cell culture. Surface antigens of MSCs (CD29 and CD34) were identified by flow cytometry. MSCs were induced for chondrogenic differentiation in MCDM (Mesenchymal Stem Cell Chondrogenic Differentiation Medium) for 0, 3, and 7 days, respectively, followed by detection of RUNX2, microRNA-23c and FGF2 expressions by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Alcian blue staining was performed to access proteoglycan deposition in MSCs transfected with microRNA-23c mimics or inhibitor. Western blot was conducted to detect the protein expressions of ACAN and COL2A1 in MSCs. The binding condition between microRNA-23c and FGF2 was verified by dual-luciferase reporter gene assay. Finally, MSCs were co-transfected with microRNA-23c mimics and FGF2 overexpression plasmid for rescue experiments. RESULTS: On the fourth day of MSCs isolation, MSCs were in an elongated shape. Flow cytometry results showed positive expression of CD29 and negative expression of CD34, which were consistent with MSCs phenotype. QRT-PCR data elucidated that the mRNA levels of RUNX2 and FGF2 gradually increased, whereas microRNA-23c expression decreased with the prolongation of chondrogenic differentiation. Transfection of microRNA-23c mimics in MSCs remarkably elevated microRNA-23c expression. Alcian blue staining showed that microRNA-23c overexpression results in less proteoglycan deposition in MSCs than that of controls. Both mRNA and protein expressions of ACAN and COL2A1 decreased after microRNA-23c overexpression. Dual-luciferase reporter gene assay confirmed that FGF2 binds to microRNA-23c. Further Western blot results demonstrated that FGF2 expression is negatively regulated by microRNA-23c. FGF2 overexpression reversed the inhibitory effects of microRNA-23c on proteoglycan deposition, as well as expressions of ACAN and COL2A1. CONCLUSIONS: MicroRNA-23c expression decreases during chondrogenic differentiation of MSCs, which inhibits MSCs differentiation to chondrocytes by inhibiting FGF2.

[miR-30a-5p promotes the apoptosis of chondrocytes in patients with osteoarthritis by targeting protein kinase B].

Objective: To investigate the expression of miR-30a-5p in cartilage of osteoarthritis patients, and to explore its mechanism of chondrocyte apoptosis. Methods: From May 2015 to December 2016, tissue specimen of 289 patients with osteoarthritis was collected in Department of Orthopedics, Changzhou traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine.The expression of miR-30a-5p and protein kinase B(Akt) mRNA in cartilage of different patients was detected by qPCR.The apoptosis of chondrocytes was detected by Tunel method.The expression of related proteins in tissues and cells was detected by immunoblotting, and apoptosis and cell cycle were detected by flow cytometry. Results: The expression of miR-30a-5p in OA patients was significantly higher than control patients (P<0.05), but Aktwas positively related[(3.64±0.95)vs(1.03±0.31), P<0.05]. The expression of miR-30a-5p in cartilage of OA patients was negatively correlated with Akt mRNA expression (r=0.729 3, P<0.001), but it had a positive correlation with the apoptotic rate (r=0.847 5, P<0.001). miR-30a-5p targets negative regulation of Akt gene expression in SW1353 cells, and the expression of p-Akt, IkB-α, p-IkB-α, p65, p-p65 and mTOR and p-mTOR were significantly down-regulated by miR-30a-5p (P<0.05). Compared with normal SW1353 cells, the apoptosis rate of SW1353 cells which was transfected with miR-30a-5p-mimics increased by 9.65 times, G0/G1 phase cells increased by 1.37 times, S phase cells decreased by 60.94%, G2/M phase cells decreased 19.53%. Conclusion: miR-30a-5p is highly expressed in cartilage of osteoarthritis patients, and its high expression can block chondrocytes in G0/G1 phase by targeting Akt gene, and induce apoptosis of chondrocytes.

Towards water sensitive cities in Asia: an interdisciplinary journey.

Rapid urbanisation, population growth and the effects of climate change drive the need for sustainable urban water management (SUWM) in Asian cities. The complexity of this challenge calls for the integration of knowledge from different disciplines and collaborative approaches. This paper identifies key issues and sets the stage for interdisciplinary research on SUWM in Asia. It reports on the initial stages of a SUWM research programme being undertaken at Monash University, Australia, and proposes a framework to guide the process of interdisciplinary research in urban water management. Three key themes are identified: (1) Technology and Innovation, (2) Urban Planning and Design, and (3) Governance and Society. Within these themes 12 research projects are being undertaken across Indonesia, China, India and Bangladesh. This outward-looking, interdisciplinary approach guides our research in an effort to transgress single-discipline solutions and contribute on-ground impact to SUWM practices in Asia.

Transperineal end-to-end anastomotic urethroplasty for traumatic posterior urethral disruption and strictures in children.

OBJECTIVE: To report the long-term results of transperineal end-to-end anastomotic urethroplasty for post-traumatic posterior urethral stenosis in children. METHODS: From 1975 to 1996, 25 boys [aged 3 to 12 years] with post-traumatic posterior urethral stenosis or obliteration, and one boy [aged 7 years] with disrupted posterior urethra were treated with transperineal end-to-end anastomotic urethroplasty. Final follow-up assessments including voiding status, urinary continence and erectile function were performed in June 1999. RESULTS: Smooth voiding was restored in 25 boys postoperatively. one child failed an ill-prepared repair and was waiting for further intervention. Among the 25 patients, seven were lost to the final follow-up. All seven boys had a single urethroplasty for simple urethral stenosis and had been followed for 3 to 5 years postoperatively with smooth voiding. The other 18 boys, including seven with complex urethral stenosis [three with a history of failed previous urethroplasties, three with urethrorectal fistula and one with urethroperineal fistula], underwent a total of 22 end-to-end anastomotic urethroplasties [one successful primary repair, 17 successful delayed repairs and four failed repairs]. Of the 17 patients with successful delayed repair, 14 succeeded with one repair, two with two repairs and one with three repairs. The success rate per repair for simple urethral strictures was 94.7% [18 of 19], and for complex strictures 63.6% [7 of 11]. Stress incontinence was found in three cases, impotence in two. Concomitant impotence and stress incontinence were found in one of the five patients. CONCLUSION: Transperineal end-to-end anastomotic urethroplasty can achieve good long-term outcomes in children with simple post-traumatic posterior urethral stenosis. In experienced hands, good results can also be achieved for complex urethral strictures.

Glycoprotein 90K/MAC-2BP interacts with galectin-1 and mediates galectin-1-induced cell aggregation.

The glycoprotein 90K was originally described as a tumor-secreted antigen and subsequently found to have immunostimulatory activity as well as other possible functions. This protein interacts with an endogenous lectin, galectin-3, and may play a role in tumor metastasis through this interaction. Because 90K is heavily glycosylated, it may also interact with other members of the galectin family, which would contribute to the multifunctionality of 90K. To test this possibility, we studied the recognition of 90K by galectin-1, which, like galectin-3, has been associated with neoplastic transformation. In a solid-phase binding assay, human recombinant galectin-1 bound immobilized human recombinant 90K in a fashion that was inhibitable by lactose. Galectins 1 and 3 appeared to bind to separate sites on 90K because they did not affect the binding of each other. The dissociation constant of galectin-1 to 90K was on the order of 10(-7) M. Galectin-1 also induced aggregation of a human melanoma cell line, A375, in a carbohydrate-dependent manner, and this appeared to be mediated, at least in part, by 90K expressed on A375 cells, since it was inhibitable by a specific anti-90K monoclonal antibody. We conclude that 90K interacts with both galectin-1 and galectin-3 and both interactions contribute to the formation of multicell aggregates. Because both of these galectins as well as 90K are often over-expressed in neoplasm, these interactions may occur in the setting of various carcinomas and contribute to their progression and metastasis.

[The effect of FHIT gene on renal cell carcinomas].

OBJECTIVE: To study the potential role of FHIT gene in renal cell carcinomas. METHODS: Cancerous tissues derived from 23 patients with renal cell carcinomas and an established cell line were examined for the alteration of FHIT gene transcripts and structure of genomic DNA by using RT-PCR and Southern blot hybridization assays. RESULTS: Eight point three percent(2/24) of cancerous tissues exhibited aberrant transcripts. The transcript in the renal-cell cancer cell line appeared a small-size one when cDNA was amplified with outside nest primer; absent expression of transcript was shown when amplified with inside nest primer. About twenty percent(5/24) cancerous tissues showed genomic rearrangement in FHIT gene locus. CONCLUSION: FHIT gene exhibited aberrant transcripts and genomics in some of renal cell cancers. Therefore change of FHIT gene may play a role in development of renal cell carcinomas.

[Extracorporeal shock wave lithotripsy for renal calculi].

OBJECTIVE: The aim of the study was to determine the effectiveness of extracorporeal shock wave lithotripsy(ESWL) therapy for renal calculi. METHODS: 126 patients were treated with Shanghai HX-902 lithotriptor on an inpatient basis. The stone size varied from 0.5 to 2.5 cm. Some patients had a double-J stent inserted prior to treatment. RESULTS: The overall stone-free rate in 2 months was 45.6%, whereas it was 53.3%, 40.8% and 18.1% according to the stone size, < or = 0.8, 0.9-1.9 and > or = 2 cm, respectively. Complications were rare, including 1 subcapsular hematoma formation, 15 renal colics and 5 stone streets, which were managed by conservative treatment or ureteral stenting or additional ESWL and resulted in complete stone clearance. CONCLUSION: ESWL therapy is a reasonable and effective method for renal stone, percutaneous nephrolithotripsy(PCNL) or open surgery should be considered for stones larger than 2 cm.

Acute renal failure and multiple organ system failure.

We treated 23 patients with acute renal failure who had multiple organ system failure. The patients ranged in age from 16 to 66 years (mean, 36 years). All patients underwent hemodialysis. Mortality increased with the number of organ systems involved, from 22% for patients with two systems involved to 100% for patients with four systems involved. The overall mortality was 47.8%. The association of ventilatory failure and sepsis resulted in the most deaths. Early resuscitation, eradication of the primary lesion, and hemodialysis are the most important points in the treatment and prevention of multiple organ system failure in patients with acute renal failure. Thus, the problems of single and multiple organ system failure in China are similar to those in other countries.

The ontogeny of blood cells, complement, and immunoglobulins in 3- to 12-week-old 15I5-B congenic white Leghorn chickens.

Six partially developed line 15I5-B congenic White Leghorn chicken strains were used to investigate age and B haplotype effects on hematocrit, total and differential leukocyte counts, hemolytic complement, immunoglobulins, and body weights. The .C-12 line had the lowest hematocrit values throughout the study. The .15I-5 line always had the highest mean hematocrit value and differed from the .C-12 and .P-13 lines at 3, 8, and 12 weeks, as well as lines .N-15 and .6-2 at 12 weeks. Ontogenically, the total leukocyte, hemolytic complement, immunoglobulins, and body weight values increased after 3 weeks in all lines. Lymphocytes were the predominant leukocyte after 3 weeks in all lines studied. At 12 weeks, the .C-12 line had the highest total leukocytes, heterophils, lymphocytes, and monocytes among the lines studied. The .6-2 line had a significantly higher hemolytic complement level than the .P-13 or the .C-12 line at all 3 ages, and at 12 weeks it also differed from the .7-2 line. The lines were found to be comparable for immunoglobulin classes M and G. In this study the line 15I5-B congenic lines differ for the traits of hematocrit and hemolytic complement level, and the results suggest the difference may be determined by genes in the B complex.