PPP1R3A inhibits osteogenesis and negatively regulates intracellular calcium levels in calcific tendinopathy.

Calcific tendinopathy (CT) is defined by the progressive accumulation of calcium crystals in tendonic regions that results in severe pain in patients. The etiology of CT is not fully elucidated. In this study, we elucidate the role of PPP1R3A in CT. A significant decrease in PPP1R3A expression was observed in CT patient tissues, which was further confirmed in tissues from a CT-induced rat model. Overexpression of PPP1R3A ex vivo reduced the expression of osteo/chondrogenic markers OCN and Sox9, improved tendon tissue architecture, and reduced intracellular Ca2+ levels. Overexpression of SERCA2 and knockdown of Piezo1 decreased expression of osteo/chondrogenic markers and intracellular calcium in PPP1R3A-knockdown tendon cells. Lastly, PPP1R3A expression was regulated at the posttranscriptional level by binding of HuR. Collectively, the present study indicates that PPP1R3A plays an important role in regulating calcium homeostasis in tendon cells via Piezo1/SERCA2, rendering it a promising target for therapeutic interventions of CT.

Systematic analysis of the roles of c-di-GMP signaling in Xanthomonas oryzae pv. oryzae virulence.

Cyclic di-guanosine monophosphate (c-di-GMP) is a ubiquitous second messenger, i.e. essential to bacterial adaptation to environments. Cellular c-di-GMP level is regulated by the diguanylate cyclases and the phosphodiesterases, and the signal transduction depends on its receptors. In Xanthomonas oryzae pv. oryzae strain PXO99A, 37 genes were predicted to encode GGDEF, EAL, GGDEF/EAL, HD-GYP, FleQ, MshE, PilZ, CuxR, Clp, and YajQ proteins that may be involved in c-di-GMP turnover or function as c-di-GMP receptors. Although the functions of some of these genes have been studied, but the rest have not been extensively studied. Here, we deleted these 37 genes from PXO99A and analyzed the virulence, motility, biofilm, and EPS production of these mutants. Our results show that most of these genes are required for PXO99A virulence, motility, biofilm formation, or exopolysaccharide production. Although some of them have been reported in previous studies, we found four novel genes (gedpX8, gdpX11, pliZX4, and yajQ) are implicated in X. oryzae pv. oryzae virulence. Our data demonstrate that c-di-GMP signaling is vital for X. oryzae pv. oryzae virulence and some virulence-related factors production, but there is no positive correlation between them in most cases. Taken together, our systematic research provides a new light to understand the c-di-GMP signaling network in X. oryzae pv. oryzae.

NUPR1 promotes the proliferation and migration of breast cancer cells by activating TFE3 transcription to induce autophagy.

Recurrence and metastasis affect the survival rate of breast cancer patients. The fundamental reason lies in the lack of understanding of the mechanism of breast cancer metastasis. In this study, the proliferation, migration and invasion abilities of breast cancer cells were evaluated. The mechanism of NUPR1/TFE3 signaling pathway on autophagy-related proteins and migration-invasion-related proteins was examined in cell model in vitro. The effects of NUPR1 on malignancy formation and metastasis were investigated in vivo. We found that NUPR1 was upregulated in breast cancer cells and tissues. NUPR1 knockdown inhibited the proliferation, migration and invasion of ZR-75-30 cells and inhibited malignancy formation and metastasis in vivo. Mechanically, NUPR1 promoted autophagy by activating of TFE3 transcription, thereby regulating breast cancer metastasis. This paper indicates that NUPR1 activates autophagy through the TFE3 signaling pathway to promote breast cancer metastasis, and provides a biological basis for the intervention of blocking distant metastasis.

The RavA/VemR two-component system plays vital regulatory roles in the motility and virulence of Xanthomonas campestris.

Xanthomonas campestris pv. campestris (Xcc) can cause black rot in cruciferous plants worldwide. Two-component systems (TCSs) are key for bacterial adaptation to various environments, including hosts. VemR is a TCS response regulator and crucial for Xcc motility and virulence. Here, we report that RavA is the cognate histidine kinase (HK) of VemR and elucidate the signalling pathway by which VemR regulates Xcc motility and virulence. Genetic analysis showed that VemR is epistatic to RavA. Using bacterial two-hybrid experiments and pull-down and phosphorylation assays, we found that RavA can interact with and phosphorylate VemR, suggesting that RavA is the cognate HK of VemR. In addition, we found that RpoN2 and FleQ are epistatic to VemR in regulating bacterial motility and virulence. In vivo and in vitro experiments demonstrated that VemR interacts with FleQ but not with RpoN2. RavA/VemR regulates the expression of the flagellin-encoding gene fliC by activating the transcription of the rpoN2-vemR-fleQ and flhF-fleN-fliA operons. In summary, our data show that the RavA/VemR TCS regulates FleQ activity and thus influences the expression of motility-related genes, thereby affecting Xcc motility and virulence. The identification of this novel signalling pathway will deepen our understanding of Xcc-plant interactions.

MiR-6924-5p-rich exosomes derived from genetically modified Scleraxis-overexpressing PDGFRα(+) BMMSCs as novel nanotherapeutics for treating osteolysis during tendon-bone healing and improving healing strength.

Osteolysis at the tendon-bone interface can impair pullout strength during tendon-bone healing and lead to surgery failure, but the effects of clinical treatments are not satisfactory. Mesenchymal stem cell (MSC)-derived exosomes have been used as potent and feasible natural nanocarriers for drug delivery and have been proven to enhance tendon-bone healing strength, indicating that MSC-derived exosomes could be a promising therapeutic strategy. In this study, we explored Scleraxis (Scx) dynamically expressed in PDGFRα(+) bone marrow-derived mesenchymal stem cells (BMMSCs) during natural tendon-bone healing. Then, we investigated the role of PDGFRα(+) BMMSCs in tendon-bone healing after Scx overexpression as well as the underlying mechanisms. Our data demonstrated that Scx-overexpressing PDGFRα(+) BMMSCs (BMMSCScx) could efficiently inhibit peritunnel osteolysis and enhance tendon-bone healing strength by preventing osteoclastogenesis in an exosomes-dependent manner. Exosomal RNA-seq revealed that the abundance of a novel miRNA, miR-6924-5p, was highest among miRNAs. miR-6924-5p could directly inhibit osteoclast formation by binding to the 3'-untranslated regions (3'UTRs) of OCSTAMP and CXCL12. Inhibition of miR-6924-5p expression reversed the prevention of osteoclastogenic differentiation by BMMSCScx derived exosomes (BMMSCScx-exos). Local injection of BMMSCScx-exos or miR-6924-5p dramatically reduced osteoclast formation and improved tendon-bone healing strength. Furthermore, delivery of miR-6924-5p efficiently inhibited the osteoclastogenesis of human monocytes. In brief, our study demonstrates that BMMSCScx-exos or miR-6924-5p could serve as a potential therapy for the treatment of osteolysis during tendon-bone healing and improve the outcome.

Three-dimensional printed talar prosthesis with biological function for giant cell tumor of the talus: A case report and review of the literature.

BACKGROUND: Giant cell tumors (GCT) are most commonly seen in the distal femur. These tumors are uncommon in the small bones of the hand and feet, and a very few cases have been reported. A giant cell tumor of the talus is rarely seen clinically and could be a challenge to physicians. CASE SUMMARY: We report a rare case of GCT of the talus in one patient who underwent a new reconstructive surgery technique using a three-dimensional (3D) printing talar prosthesis. The prosthesis shape was designed by tomographic image processing and segmentation using technology to match the intact side by mirror symmetry with 3D post-processing technologies. The patient recovered nearly full range of motion of the ankle after 6 mo. The visual analogue scale and American Orthopaedic Foot and Ankle Society scores were 1 and 89 points, respectively. CONCLUSION: We demonstrated that 3D printing of a talar prosthesis is a beneficial option for GCT of the talus.

Three dimension printing talar prostheses for total replacement in talar necrosis and collapse.

BACKGROUND: Reconstructing bone structures and stabilizing adjacent joints are clinical challenges in treating talar necrosis and collapse (TNC). 3D printing technology has been demonstrated to improve the accuracy of talar replacement. This study aimed to evaluate anatomical talar replacement and the clinical results. METHODS: Nine patients with TNC were enrolled between 2016 and 2020. The prosthetic shape and size were designed by CT post-processing and mirror symmetry technology. The clinical outcomes included radiographic parameters of the forefoot, hindfoot, and ankle alignment, ankle activity, recurrent pain, and peri-operative complications. RESULTS: After a mean follow-up of 23.17 ± 6.65 months, degenerative arthritis and prosthetic dislocation and other complications were not observed on plain radiographs. Each 3D-printed talar prosthesis was placed in the original anatomical position. The parameters which have significant changes pre-operative and post-operative are as follows: talar height, 27.59 ± 5.99 mm and 34.56 ± 3.54 mm (95% CI - 13.05 to - 0.87, t = 2.94, P = 0.032) and Meary's angle, 11.73 ± 4.79° and 4.45 ± 1.82° (95% CI 1.29~22.44, t = 2.89, P = 0.034). The AOFAS hindfoot score improved from 26.33 ± 6.62 to 79.67 ± 3.14 at the final follow-up (95% CI 43.36~63.30, t = 13.75, P = 0.000). The VAS score decreased from 6.33 ± 1.03 to 0.83 ± 0.75 (95% CI 4.40~6.60, t = 12.84, P = 0.000). The post-operative satisfaction scores regarding pain relief, activities of daily living, and return to recreational activities were good to excellent, and the change of activity range was statistically significant. CONCLUSIONS: The 3D printing patient-specific total talar prostheses allowed anatomical reconstruction in TNC. This novel treatment with 3D-printed prostheses could serve as a reliable patient-specific alternative in TNC.

The prevalence of suicidal behaviors and their mental risk factors among young adolescents in 46 low- and middle-income countries.

BACKGROUND: To describe the prevalence of suicidal behaviors (ideation, planning, and attempt) and their associated factors in young adolescents in low- and middle-income countries (LMICs). METHODS: We used the latest data from the Global School-Based Health Survey (GSHS) for adolescents aged 12-15 years during 2009-2015. The weighted prevalence and 95% confidential intervals (CIs) of suicidal behaviors were calculated using a random-effects model. The factors associated with suicidal behaviors were examined using logistic regression analysis. RESULTS: Data from 130,488 adolescents (48.13% boys) in 46 LMICs were included in the study. Across all countries, the pooled 12-month prevalence of suicidal ideation, planning, and attempt were 14.5%, 14.6%, and 12.7%, respectively. The highest prevalence of suicidal ideation, planning, and attempt were all in Africa (16.7%, 19.3% and 17.0%), and the lowest prevalence were all in South-East Asia (8.2%, 10.5% and 7.4%). The overall prevalence of three suicidal behaviors were higher in girls (all P < 0.001). Suicidal ideation and planning were more common in the 14-15 age group than 12-13 age group (both P < 0.001). The factors associated with suicidal behaviors were being female, older age, loneliness, anxiety, a lack of close friends, and having family supportive (all P < 0.001). LIMITATIONS: The GSHS data were obtained from a self-report questionnaire and the participants included in the GSHS were adolescents in school. CONCLUSIONS: The prevalence of suicidal behaviors remains high among young adolescents in LMICs, especially in Africa. These countries should be intervention priorities.

[Inhibitory effect of siRNA on heparanase expression and invasion ability of gastric cancer cells: an in vitro experiment].

OBJECTIVE: To investigate the inhibitory effect of siRNA on heparanase expression and invasion ability gastric cancer cells. METHODS: A heparanase mRNA-targeting double-stranded siRNA was designed with the bioinformatics technology. Human gastric cancer cells of the line SGC7901 were cultured and transfected with the siRNA of the concentrations of 5, 10, 20, and 40 nmol/L respectively. Forty-eight hours later RT-PCR and Western blotting were applied to detect the mRNA and protein expression of heparanase. Millicell chamber assay was performed to detect the invasion ability of the SGC7901 cells. Blank control group and negative control group were set. RESULTS: The mRNA expression level of the cells transfected with the siRNA of the concentrations 20 nmol/L and 40 nmol/L were 0.207 +/- 0.095 and 0.200 +/- 0.085 respectively, both significantly lower than that of the control group (0.60 +/- 0.09, both P < 0.05). Western blotting showed that the protein expression of heparanase of the different siRNA subgroups were all decreased dose-dependently; and no heparanase band was seen in the 40 nmol/L subgroup. The invasion rate of the siRNA group was significantly lower than that of the control group with a mean inhibition rate of (61 +/- 36)%. CONCLUSION: RNAi inhibits the expression of heparanase and the invasion ability of human gastric cancer cells. Heparanase may be a new target in treatment of gastric cancer's metastasis.

[Loss of heterozygosity at chromosome 8p21-p23 in adenocarcinoma of gastric cardia].

Chromosome 8p21-p23 harbors tumor suppressor gene(s) implicated in multiple types of cancers. To investigate the involvement of the gene(s) in the carcinogenesis of adenocarcinoma of gastric cardia, loss of heterozygosity (LOH) for microsatellite markers at chromosome 8p21-p23 was examined. Laser capture microdissection (LCM) was used to obtain homogeneous tumor cells from 19 surgical specimens. Subsequently, genomic DNA extracted from the LCM-captured cells was amplified by multiple displacement amplification. Each tumor was assessed for allelic loss using 13 microsatellite markers. An overall LOH frequency of 63.2% (12/19) was observed and the LOH frequency for individual markers varied from 25% to 55.6%. One common deleted region of about 1.2 Mb (8p22GGAA-8p22ATCT) was defined. Our data indicated that the tumor suppressor gene at chromosome 8p22 might play an important role in the development of adenocarcinoma of gastric cardia.

Vitamin C and E supplements improve the impaired antioxidant status and decrease plasma lipid peroxides in hemodialysis patients small star, filled.

This study investigated the supplementation with vitamin C or/and E on the antioxidant system in hemodialysis patients. Thirty-eight hemodialysis patients (27 males and 11 females) with the average of 60 years old were divided into four groups: placebo (400 mg starch/time), vitamin C (400 mg/time)-, vitamin E (400 mg d,l- alpha-tocopheryl acetate/time)-, and vitamin C (400 mg/time) + E (400 mg d,l- alpha-tocopheryl acetate/time)-supplemented groups for 6-week supplementation. The patients orally received three capsules of placebo or antioxidant(s) three times a week after finishing hemodialysis. Thirty-six healthy subjects (22 males and 14 females) with the average of 58 years old were recruited as the control group. Hemodialysis patients significantly decreased plasma vitamin C by 32%, erythrocyte glutathione by 26%, and plasma total antioxidant status by 9%, but increased plasma lipid peroxide levels by 102% compared with the control group at the baseline. The levels of plasma vitamin C and total antioxidant status significantly decreased by 24% and 18%, respectively, from the post-dialysate compared with those from the pre-dialysate. At week 6, vitamin C + E-supplemented group significantly increased plasma vitamin C and E, erythrocyte glutathione, and plasma antioxidant status, and inhibited plasma lipid peroxides compared with placebo group. Additionally, vitamin C + E-supplemented group had higher plasma vitamin C, vitamin E, and total antioxidant status, and lower plasma lipid peroxides than placebo group even at least 2 weeks after the termination of the supplements. Therefore, antioxidant vitamin supplements could improve antioxidant status and decrease lipid peroxides of hemodialysis patients.