Different expressions of aquaporin water channels and macrophages infiltration in human cervix remodeling during pregnancy.

Despite aquaporin water channels (AQPs) play a critical role in maintaining water homeostasis in female reproductive tract and prompt a gradual increase in water content in cervical edema as pregnancy progressed, their relationship with macrophage infiltration and collagen content in human cervical remodeling need to be further investigated. This is the first study to examine the expression and localization of AQP3, AQP4, AQP5, AQP8, and macrophages simultaneously in human cervical ripening. The immunoreactivity of these AQPs was 2.6 to 6-fold higher on gestational weeks 26 (GD26W) than that on GD6W and GD15W, but AQP4 expression on GD39W dropped a similar extent on GD15W, other AQPs continued to rise on GD39W. The AQP3, AQP4, and AQP5 intensity seemed more abundant in cervical stroma than in the perivascular area on GD26W; the distribution of AQP3, AQP5, and AQP8 in cervical stroma was equivalent to that in the perivascular area on GD39W. Macrophage numbers were 1.7-fold higher in subepithelium region and 3.0-fold higher in center area on GD26W than that on GD15W; such numbers remained elevated on GD39W. The electron micrographs showed that cervical extensibility increased significantly on GD26W and GD39W accompanied with increased macrophage infiltration, cervical water content, and much more space among collagen fibers. These findings suggest that the upregulation of AQPs expression in human cervix is closely related to enhanced macrophage infiltration during pregnancy; there may be a positive feedback mechanism between them to lead the increase of water content and the degradation of collagen.

Increased uterine NLRP3 inflammasome and leucocyte infiltration in a rat model of preeclampsia.

The disruption of the inflammatory microenvironment in the uterus affects pregnancy outcome. However, the exact quantification and distribution of leukocyte subpopulations in the uterus in preeclampsia (PE) have not been clearly characterized. Inflammasomes promote the release of proinflammatory cytokines interleukin (IL)-ß and IL-18. A higher expression of NLRP3 inflammasome in placentas contributes to excessive inflammation in PE. However, related studies on the uterus are scarce. We aimed to investigate changes in the infiltration of leukocyte subpopulations in decidual and uterine tissues, and explore the role of activation of uterine NLRP3 inflammasomes in PE. Decidual tissues were collected from normotensive pregnant women and preeclamptic women. A PE-like model was established via administration of lipopolysaccharide to normal pregnant rats. Uterine and decidual tissues were collected from all experimental groups. It was found that the number of leukocytes was significantly elevated in decidual and uterine tissues in PE patients compared to normal controls. The leukocytes (predominantly macrophages and NK cells) particularly infiltrated into the decidua and uterine decidua in PE-like rats, and these were sparse in the myometrium. The NLRP3 immunoreactivity in the uterus was extremely little in control rats, its immunoreactivity and caspase-1 immunoreactivity were significantly elevated in the PE-like rats; the mRNA expression results also indicated an upward trend in the activation of NLRP3 inflammasomes. These results support that leucocyte infiltration in the decidua and uterine deciduas, and the activation of NLRP3 inflammasome in the uterus, which participate in the pathogenesis, are responsible for the excessive inflammation at the maternal-fetal interface during PE.

Circ_0008956 contributes to IL-1ß-induced osteoarthritis progression via miR-149-5p/NAMPT axis.

Circular RNAs (circRNAs) have been identified to involve in the pathophysiology of osteoarthritis (OA). Herein, this study aimed to investigate the role and mechanisms underlying circ_0008956 in the process of OA. The expression of circ_0008956 and microRNA (miR)-149-5p and Nicotinamide phosphoribosyl transferase 1 (NAMPT) was detected using quantitative real-time polymerase chain reaction and Western blot assays. Cell viability, apoptosis, cell cycle and extracellular matrix (ECM) degradation were analyzed using cell counting kit-8, flow cytometry, and Western blot assays, respectively. The binding interaction between miR-149-5p and circ_0008956 or NAMPT was confirmed using dual-luciferase reporter assay. Circ_0008956 was highly expressed in OA cartilage tissues and interleukin (IL)-1ß mediated chondrocytes. Knockdown of circ_0008956 promoted cell viability, cell cycle, suppressed cell apoptosis, and increased type II collagen and aggracan expression in IL-1ß-treated chondrocytes. MiR-149-5p was verified to be a target of circ_0008956, inhibition of miR-149-5p reversed the protective effects of circ_0008956 knockdown on IL-1ß-stimulated chondrocytes. NAMPT was a target of miR-149-5p, miR-149-5p attenuated IL-1ß-induced growth arrest and ECM degradation in chondrocytes, which was abolished by NAMPT overexpression. Importantly, circ_0008956 served as a sponge for miR-149-5p to up-regulate NAMPT expression in chondrocytes. Circ_0008956 contributed to IL-1ß-induced growth arrest and ECM degradation in chondrocytes via miR-149-5p/NAMPT axis, suggesting a new insight into the pathogenesis of OA and a promising therapeutic target for OA treatment.

Effects of Nogo-A and its receptor on the repair of sciatic nerve injury in rats.

Regeneration of injured peripheral nerves is an extremely complex process. Nogo-A (neurite outgrowth inhibitor-A) inhibits axonal regeneration by interacting with Nogo receptor in the myelin sheath of the central nervous system (CNS). The aim of this study was to investigate the effects of Nogo-A and its receptor on the repair of sciatic nerve injury in rats. Sprague-Dawley rats (n=96) were randomly divided into 4 groups: control group (control), sciatic nerve transection group (model), immediate repair group (immediate repair), and delayed repair group (delayed repair). The rats were euthanized 1 week and 6 weeks after operation. The injured end tissues of the spinal cord and sciatic nerve were obtained. The protein expressions of Nogo-A and Nogo-66 receptor (NgR) were detected by immunohistochemistry. The protein expressions of Nogo-A, NgR, and Ras homolog family member A (RhoA) were detected by western blot. At 1 week after operation, the pathological changes in the immediate repaired group were less, and the protein expressions of Nogo-A, NgR, and RhoA in the spinal cord and sciatic nerve tissues were decreased (P<0.05) compared with the model group. After 6 weeks, the pathological changes in the immediate repair group and the delayed repair group were alleviated and the protein expressions decreased (P<0.05). The situation of the immediate repair group was better than that of the delayed repair group. Our data suggest that the expression of Nogo-A and its receptor increased after sciatic nerve injury, indicating that Nogo-A and its receptor play an inhibitory role in the repair process of sciatic nerve injury in rats.

Effects of Nogo-A and its receptor on the repair of sciatic nerve injury in rats

Regeneration of injured peripheral nerves is an extremely complex process. Nogo-A (neurite outgrowth inhibitor-A) inhibits axonal regeneration by interacting with Nogo receptor in the myelin sheath of the central nervous system (CNS). The aim of this study was to investigate the effects of Nogo-A and its receptor on the repair of sciatic nerve injury in rats. Sprague-Dawley rats (n=96) were randomly divided into 4 groups: control group (control), sciatic nerve transection group (model), immediate repair group (immediate repair), and delayed repair group (delayed repair). The rats were euthanized 1 week and 6 weeks after operation. The injured end tissues of the spinal cord and sciatic nerve were obtained. The protein expressions of Nogo-A and Nogo-66 receptor (NgR) were detected by immunohistochemistry. The protein expressions of Nogo-A, NgR, and Ras homolog family member A (RhoA) were detected by western blot. At 1 week after operation, the pathological changes in the immediate repaired group were less, and the protein expressions of Nogo-A, NgR, and RhoA in the spinal cord and sciatic nerve tissues were decreased (P<0.05) compared with the model group. After 6 weeks, the pathological changes in the immediate repair group and the delayed repair group were alleviated and the protein expressions decreased (P<0.05). The situation of the immediate repair group was better than that of the delayed repair group. Our data suggest that the expression of Nogo-A and its receptor increased after sciatic nerve injury, indicating that Nogo-A and its receptor play an inhibitory role in the repair process of sciatic nerve injury in rats.

MiR-296 Promotes Osteoblast Differentiation by Upregulating Cbfal.

We aim to identify the potential role of miR-296, which is a class of endogenous non-coding RNAs in regulating osteoblast differentiation. Human osteoblast cell line, hFOB 1.19 cells, were transfected with miR-296 overexpression or inhibition plasmid to upregulate or downregulate miR-296 expression, and then co-transfected with anti-Cbfal antibody to knockdown Cbfal. Alizarin red staining, alkaline phosphatase (ALP) activity, and enzyme-linked immunosorbent assay assays were performed to evaluate the extent of osteoblast differentiation. MTT assays were applied to measure cell proliferation. Wound healing and transwell assays were used to determine the ability of osteoblast migration and invasion. Flow cytometry assay was used to measure cell apoptosis and cell cycle change. The mRNA and protein expression of Cbfal, OSX, and Col-1 were confirmed by RT-qPCR and western blot respectively. We found that miR-296 overexpression contributed to a significant enhancement of matrix mineralization, ALP activity, and osteocalcin expression comparing with the negative control, whereas knockdown of miR-296 caused an opposite result. Meanwhile, the treatment of miR-296 promotes osteoblast migration, invasion, and proliferation, while it inhibits cell apoptosis. Compared with the negative control groups, the cells transfected with miR-296 also presented a much higher expression of Cbfal, OSX, and Col-1. Further experiments confirmed that the positive regulatory role of miR-296 in osteoblast differentiation is achieved by upregulating Cbfal expression. In conclusion, miR-296 promotes osteoblast differentiation by upregulating Cbfal expression in hFOB 1.19 cells, which may be used as a potential therapeutic target for the treatment of bone diseases in future.

Screening of NogoA/NTR-related differential genes in rat sciatic nerve injury signal pathway.

AIM: To screen the differential genes in NogoA/NTR-related pathways that associate with sciatic nerve injury. RESULTS: There was no difference in the expression of NogoA, NTR and Ntrk2. Differential genes existed in 11 differential pathways that include NogoA, NTR and Ntrk2. Pathways closely related to sciatic nerve injury are MAPK, endophagocytosis, apoptosis, neurotrophin signaling and inflammatory mediators. NTRK1, FASLG, LDLR ADRB1 and HTR2A in model rats were downregulated compared with control rats, IL1R1, CSF1R, BCL2L1 and HRH1 in model rats were upregulated compared with control rats. CONCLUSION: MAPK, endophagocytic, apoptotic, neurotrophic factor and inflammatory mediators of ductal mediators may be involved in the sciatic nerve injury in rats. The differentially expressed genes in these pathways may play important roles in sciatic nerve injury.

Successful surgical repair of central slip rupture in finger extensor tendon.

UNLABELLED: The aim of the present study was to develop a novel effective surgical method to repair boutonniere deformity resulting from central slip rupture of finger extensor tendon. Currently, there are not standard methods available to treat boutonniere deformity. PATIENTS AND METHODS: In the current study, 8 patients with boutonniere deformity on the left hand were included. All patients had a central slip rupture of the ring fingers. Autologous palmaris longus tendon was used to surgically repair the central slip rupture. RESULTS: We successfully used the autologous palmaris longus tendons in the effective treatment of central slip rupture of the ring fingers resulting in boutonniere deformity on the left hands. CONCLUSION: To our knowledge, our study was the first illustration for a successful surgical repair of boutonniere deformity which resulted from central slip rupture, using autologous palmaris longus tendon. This method provides an alternative approach for the effective treatment of boutonniere deformity.