Overexpression of miR-150 alleviates mechanical stress-accelerated the apoptosis of chondrocytes via targeting GRP94.

The article "Overexpression of miR-150 alleviates mechanical stress-accelerated the apoptosis of chondrocytes via targeting GRP94, by Z.-Q. Zhang, C.-S. Wang, P. Yang, K.-Z. Wang, published in Eur Rev Med Pharmacol Sci 2019; 23 (18): 7775-7785-DOI: 10.26355/eurrev_201909_18987-PMID: 31599403" has been withdrawn from the authors. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18987.

The crosstalk between IGF-1R and ER-α in the proliferation and anti-inflammation of nucleus pulposus cells.

OBJECTIVE: Insulin-like growth factors-1 receptor (IGF-1R) and estrogen receptor (ER) are reported to co-express and engage in crosstalk involving in the synergistic effect of various aspects. It is unknown whether this crosstalk exists in the nucleus pulposus (NP) cells. We aimed to investigate the interaction between IGF-1R and ER-α in regulating NP cell proliferation and inflammation response under IGF-1 stimulation. PATIENTS AND METHODS: We analyzed the IGF-1, IGF-1R, and ER-α in different degenerated degree human NP tissues. NP cells were cultured with IGF-1 protein with or without the inhibitor of IGF-1R or ER-α to investigate their effects on the proliferation and inflammation response. In addition, we also upregulated the IFG-1R and ER-α expression by plasmid transfection to investigate the impact on each other. The content of IGF-1, IFG-1R, and ER-α was analyzed by enzyme-linked immunosorbent assay (ELISA). The proliferative cell rate was determined by flow cytometry. Additionally, intracellular collagen-II, p16, PCNA, IL-1ß, IL-6, TNF-α, and MMP-13 expression were also detected. RESULTS: We found IGF-1, IFG-1R, and ER-α content were decreased in higher degenerated NP tissues. IGF-1 protein treatment upregulated the IFG-1R and ER-α expression and promoted NP cell proliferation, collagen-II, and PCNA expression. However, the suppression of IGF-1R (or ER-α) weakened the IGF-1 induced collagen-II expression, proliferation, and anti-inflammation effects on NP cells, decreased ER-α (or IGF-1R) expression, and partly reversed the protective effect of NP cells caused by IGF-1 Similarly, the upregulation of one of IGF-1R and ER-α may increase the other as well. CONCLUSIONS: There is an interaction between IGF-1R and ER-α acts synergistically to promote the proliferation and suppress inflammation in NP cells.

LncRNA Bmncr alleviates the progression of osteoporosis by inhibiting RANML-induced osteoclast differentiation.

OBJECTIVE: To elucidate whether long non-coding RNA (lncRNA) Bmncr could inhibit RANML-induced osteoclast differentiation, thus alleviating the progression of osteoporosis. MATERIALS AND METHODS: Expression level of lncRNA Bmncr at different stages of osteoclast differentiation was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). After Bmncr overexpression or knockdown in RAW 264.7 cells, expression levels of osteoclast-related genes were detected. Bone marrow mesenchymal stem cells (BMMs) isolated from rats undergoing ovariectomy (OVX) were induced with RANKL (50 ng/mL) and M-CSF (50 ng/mL) for 120 h. TRAP staining was conducted to count the number of TRAP-positive osteoclasts containing more than three nuclei. Bone resorption area of bone fragments was quantitatively analyzed. Osteoporosis model in mice was established. Mice were subjected to MicroCT analyses for recording BMD and BV/TV. The expression level of lncRNA Bmncr in the marrow and spleen of osteoporosis mice was examined. RESULTS: LncRNA Bmncr was lowly expressed in the marrow and spleen of osteoporosis mice. Besides, Bmncr expression gradually downregulated during RANKL-induced in vitro osteoclast differentiation, reaching the lowest level at 72 h. The overexpression of Bmncr reduced the amount of osteoclasts, inhibited bone resorption capacity, and downregulated expression levels of Atp6v0d2, Acp5, Ctr, and Mmp9. Conversely, Bmncr knockdown obtained the opposite trends. CONCLUSIONS: LncRNA Bmncr inhibits RANKL-induced osteoclast differentiation, thus alleviating the progression of osteoporosis.

Overexpression of miR-150 alleviates mechanical stress-accelerated the apoptosis of chondrocytes via targeting GRP94.

OBJECTIVE: A previous study reported that glucose-regulated protein 94 (GRP94) is involved in mechanical stress-induced chondrocyte apoptosis; however, the underlying molecular mechanisms remain unknown. The present study aimed to investigate the post-transcriptional regulatory mechanism of microRNAs (miRs) in mechanical stress-induced chondrocyte apoptosis by targeting GRP94. MATERIALS AND METHODS: Annexin V-fluorescein isothiocyanate/propidium iodide (PI) staining was conducted to evaluate the apoptosis of chondrocytes. The mRNA and protein expression levels were measured by reverse transcription-quantitative polymerase chain reaction and Western blotting, respectively. The targeted genes were predicted using a bioinformatics tool and further investigated via a luciferase reporter assay. RESULTS: The results demonstrated that cyclic loading led to significant increases in GRP94 expression in chondrocytes; however, the expression levels of miR-150 were downregulated. Bioinformatics analysis and a luciferase reporter assay indicated that GRP94 was a direct target of miR-150, as the expression of GRP94 was dysregulated following transfection with miR-150 mimics or inhibitors. In addition, mechanical stress-induced chondrocyte apoptosis was suppressed by transfection with miR-150 mimics, while the protective effects of miR-150 mimics in this process were inhibited by GRP94 overexpression. CONCLUSIONS: MiR-150 upregulation suppressed mechanical stress-induced chondrocyte apoptosis; the underlying molecular mechanism may be mediated, at least partially, via the inhibition of GRP94 expression.

MST1 down-regulation in decreasing apoptosis of aortic dissection smooth muscle cell apoptosis.

OBJECTIVE: Elevated apoptosis of vascular smooth muscle cell (VSMC) is correlated with the occurrence of aortic dissection (AD). Mammalian ste20-like protein kinase 1 (MST1) is one important component of Hippo-YAP signal pathway for activation and cell apoptosis facilitation. Whether MST1 plays a role in AD pathogenesis is unclear yet. This study established an AD rat model to investigate the role of MST1 in regulating VSMC apoptosis and AD pathogenesis. MATERIALS AND METHODS: Cell apoptosis was compared between AD vascular tissues and normal rats, in addition to Caspase-3 activity, and expression of MST1, p-LATS1, p-YAP1, YAP1. In vitro cultured VSMCs from AD rats were treated with siRNA-MST1 to test apoptotic rate and Caspase-3 activity. AD model rats were treated with pGLVU6/GFP-MST1 for comparing MST1, p-LATS1, p-YAP1, and YAP1 expression, along with Caspase-3 activity, cell apoptosis, AD formation rate, diameter, and length. RESULTS: Compared to control group, AD rats had elevated vascular cell apoptosis, Caspase-3 activity, expressions of MST1, p-LATS1, and p-YAP1, plus lower YAP1 expression. siRNA interference of MST1 significantly inhibited apoptosis of in vitro cultured VSMC. shRNA lentivirus targeting MST1 pGLVU6/GFP-MST1 remarkably decreased expression of MST1, p-LATS1, and p-YAP1 in AD rat vascular tissues, increased YAP1 expression, decreased VSMC apoptosis, AD formation rate, AD diameter/length. CONCLUSIONS: MST1 up-regulation plays a role in facilitating VSMC apoptosis and AD pathogenesis. Down-regulation of MST1 decreased VSMC apoptosis and AD formation.

L1 Peptide-Conjugated Fibrin Hydrogels Promote Salivary Gland Regeneration.

Hyposalivation contributes to dental caries, periodontitis, and microbial infections. Additionally, it impairs activities of daily living (e.g., speaking, chewing, and swallowing). Treatments for hyposalivation are currently limited to medications (e.g., the muscarinic receptor agonists pilocarpine and cevimeline) that induce saliva secretion from residual acinar cells and the use of saliva substitutes. However, given that these therapies provide only temporary relief, the development of alternative treatments to restore gland function is essential. Previous studies demonstrated that laminin 1 (L1) is critical for intact salivary cell cluster formation and organization. However, the full L1 sequence is not suitable for clinical applications, as each protein domain may contribute to unwanted effects, such as degradation, tumorigenesis, and immune responses that, when compounded, outweigh the potential benefits provided by their sum. Although the L1 peptides YIGSR and A99 linked to fibrin hydrogels (FHs) promote intact salivary epithelial formation in vitro, little is known about their role during salivary gland regeneration in vivo. Therefore, the goal of this study was to demonstrate whether L1 peptides conjugated to FHs promote tissue regeneration in a wound-healing model of mouse submandibular glands (mSMGs). Our results suggest that YIGSR-A99 peptides, chemically conjugated to FHs and applied to wounded mSMGs in vivo, formed new organized salivary tissue. In contrast, wounded mSMGs treated with FHs alone or in the absence of a scaffold showed disorganized collagen formation and poor tissue healing. Together these studies indicate that damaged salivary gland tissue can grow and differentiate when treated with FHs containing L1 peptides.

The impact of smoking on incident type 2 diabetes in a cohort with hepatitis B but not hepatitis C infection.

Smoking may be a risk factor for diabetes, and it has been suggested that viral hepatitis may predispose to diabetes. We studied diabetes and smoking histories in people with viral hepatitis. From 1997 to 2004, we studied the risk of incident diabetes in a community cohort with hyperendemic HBV and HCV infection in southern Taiwan. The cohort involved 3539 people (40-70 years old) without diabetes. Four hundred and twenty-three individuals developed diabetes. Those who were ≥65 years old, frequently consumed alcohol, had a BMI ≥25, had <9 years of education, were anti-HCV+ or smoked ≥1 pack per day were more likely to develop diabetes (P < 0.05). A cumulative hazard function test showed that the higher the smoking levels, the greater the cumulative incidence rate of diabetes in HBsAg+ participants only (P = 0.03 by log-rank test). A multiple Cox proportional hazards model analysis in different hepatitis statuses showed smoking levels were strong predictors of diabetes with a dose-response relationship for type 2 diabetes in those with HBsAg+ : hazard ratio (HR) = 3.8, (95% CI: 1.2, 12.3) for light smokers (<1 pack per day) and HR = 4.4 (95% CI: 1.5, 13.3) for heavy smokers (≥1 pack per day). Increasing BMI was a common predictor in all people. Smoking is a strong predictor for diabetes with a dose-response relationship in HBsAg+ individuals and a mild predictor for seronegative individuals but not significant in anti-HCV+ individuals.

Ulnar Osteotomy with 2-Pin Unilateral Gradual Distraction for Treatment of Chronic Monteggia Fracture: A Case Report

Missed Monteggia fracture leading to chronic radial head dislocation is a known complication. The surgical treatment options remain challenging. The aim of treatment is to reduce the radial head and to maintain the stability of the elbow in all ranges of motion. A few surgical techniques have been described with complications. We report the case of a 13 years old boy with chronic radial head dislocation as a result of an unrecognised Monteggia fracture-dislocation for eight years. We successfully reduced the radial head and corrected the cubital valgus from 45 degrees to 10 degrees with a proximal ulna osteotomy and gradual distraction with 2-pin Monotube external fixator. The correction was uneventful with good functional outcome.

Developing immunological methods for detecting Macrobrachium rosenbergii nodavirus and extra small virus using a recombinant protein preparation.

Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) have been identified as the causative agents for white tail disease (WTD) of M. rosenbergii. In this study, the gene sequences encoding MrNV and XSV capsid proteins were separately ligated into the pGEX-4T-3 expression vector and transformed into Escherichia coli. After induction, glutathione-S-transferase (GST)-tagged MrNV and XSV fusion proteins were obtained with molecular masses of 68 and 43 kDa, respectively. Specific polyclonal antibodies for MrNV and XSV against viral recombinant proteins and infected prawn tissues were verified using Western blotting. According to immunodot blot assay results, the detection sensitivities of antibodies were approximately 5 ng µL(-1) for both recombinant proteins GST-MrNV and GST-XSV. In additional, MrNV and XSV were detected at dilution levels of 1:2560 and 1:640 in the infected prawn tissues, respectively. No cross-reactions with white spot syndrome virus or grouper nervous necrosis virus were observed using immunodot blot assays. MrNV and XSV in infected muscle tissues were detected using immunohistochemistry. Although the detection limit of the immunodot blot assay was lower than that of nested reverse transcription polymerase chain reaction, these polyclonal antibodies can be useful for confirming MrNV and XSV infections in field tests.

Microarray based analysis of gene expression patterns in pancreatic neuroendocrine tumors.

OBJECTIVE: Pancreatic neuroendocrine tumors (PanNETs) are a small subgroup of tumors with a variety of biological behaviors. MATERIALS AND METHODS: We sought to identify the specially expressed genes and characterize significant pathways in PanNETs compared with non-neoplastic samples. Gene expression profile GSE43795 was obtained from Gene Expression Omnibus database, which included 6 PanNETs and 5 non-neoplastic samples. The differentially expressed genes (DEGs) were identified using Limma package. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were used to enrich the functions and pathways of DEGs. Transcription factors (TFs) and tumor-associated genes (TAGs) were also identified. Finally, a protein-protein interaction (PPI) network was constructed, and hub proteins and functional module were screened out. RESULTS: Total of 821 DEGs (421 down-regulated, 400 up-regulated) were selected. GO and KEGG enrichment analyses showed that up-regulated DEGs were related to several pathways, including type 2 diabetes mellitus, Ca2+ signaling pathway, long-term potentiation, and long-term depression pathways. Down-regulated DEGs were enriched in several pathways, such as pancreatic secretion, protein digestion and absorption, and metabolic pathway. Interferon-stimulated gene protein 15 (ISG15), somatostatin (SST), and synaptosomal-associated protein 25 kDa (SNAP25) were identified as hub proteins. CONCLUSIONS: The genes involved in type 2 diabetes mellitus pathway may play important roles in the development of PanNETs. SNAP25, SST, and ISG15 may be used as potential targets for treatment of PanNETs.

Giant sacral chondrosarcoma in an elderly male : a case report.

UNLABELLED: Primary sacral tumours are rare, therefore experience of managing their associated complications are very limited. Effective surgical treatment of pelvic chondrosarcoma remains a major challenge for orthopaedic surgeons, due to the complex anatomic structure of the pelvis, the lack of defined compartment borders, the close vicinity to vital structures, and the risk of jeopardizing pelvic structural stability. We report a rare case of a giant sacral chondrosarcoma (100cm x 80cm) in an elderly male who successfully underwent tumour resection with good functional outcome and recovery. Long term follow up is essential in view of the possibility of local tumour recurrence. KEY WORDS: Giant Chondrosarcoma, Sacrum, Surgery, Elderly Male.

Viral susceptibility, transfection and growth of SPB--a fish neural progenitor cell line from the brain of snubnose pompano, Trachinotus blochii (Lacépède).

This study investigates the susceptibilities of the SPB cell line to fish viruses including giant seaperch iridovirus (GSIV-K1), red sea bream iridovirus (RSIV-Ku), grouper nervous necrosis virus (GNNV-K1), chum salmon reovirus (CSV) and eel herpesvirus (HVA). GSIV-K1, RSIV-Ku and CSV replicated well in SPB cells, with a significant cytopathic effect and virus production. However, the cells were HVA and GNNV refractory. To examine the ability of SPB cells to stably express foreign protein, expression vectors encoding GNNV B1 and B2 fused to enhanced green fluorescent protein (EGFP) and GSIV ORF35L fused to DsRed were constructed and introduced by transfection into SPB cells. Stable transfectants displayed different morphologies compared with SPB and with each other. EGFP-B1 was predominantly localized in the nuclei, EFPF-B2 was distributed throughout the cytoplasm and nucleus, and granular 35L-DsRed was localized with secreted vesicles. The expression of EFPF-B2 in SPB cells produced blebs on the surface, but the cells showing stable expression of EGFP, EGFP-B1 or 35L-DsRed showed normal morphologies. Results show the SPB cells and the transfected cells grow well at temperatures between 20 and 35 °C and with serum-dependent growth. SPB cells are suitable for studies on foreign protein expression and virology.

Shoulder impingement, an uncommon complication of distal clavicle fracture treated arthroscopically: a case report.

UNLABELLED: Clavicle fracture is commonly treated conservatively. However uncommon complication can arise causing impingement. We report a patient who sustained distal clavicle fracture and was treated conservatively. However he developed persistent shoulder pain that affected his daily life. Shoulder impingement was diagnosed and arthroscopic subacromioclavicular decompression was done. Following early physiotherapy the early recovery was good with full range of motion of the shoulder. KEY WORDS: Clavicle fracture, shoulder impingement, arthroscopic subacromioclavicular decompression.

Overexpression of CXCL1 and its receptor CXCR2 promote tumor invasion in gastric cancer.

BACKGROUND: The chemokine (C-X-C motif) ligand 1 (CXCL1) and its receptor CXCR2 are associated with metastasis potential. Our studies were designed to clarify the CXCL1 and CXCR2 expression patterns and to explore their potential role in gastric cancer. DESIGN: The expression of CXCL1 was determined in primary gastric cancer specimens using quantitative PCR, immunohistochemistry, and western blotting. To investigate the functional significance of CXCL1 expression, a CXCL1 expression vector and short hairpin RNA targeting the CXCL1 or CXCR2 were transfected into gastric cancer cell lines to examine the biological outcomes of these cells. RESULTS: The expression of CXCL1 and CXCR2 was higher in gastric cancer tissues compared with adjacent noncancerous tissues. The upregulation of CXCL1 correlated significantly with tumor progression, advanced stage of gastric cancer patients, and was one of the independent prognostic factors for patient's survival. Furthermore, cancer cells expressing CXCL1 stably exhibited an increase in their migration and invasion ability, whereas CXCL1 or CXCR2 depletion significantly reduced the migration and invasion ability of each cell line. CONCLUSIONS: These results provide strong evidence that CXCL1 plays an important role in gastric cancer progression and migration and suggest that CXCL1 is a promising marker for the detection and prognosis of gastric cancer.

Generalized convulsion resulted in hyperammonemia during treatment with tranexamic acid for hemoptysis.

BACKGROUND: Tranexamic acid is commonly used to treat various kinds of bleeding disorders. It has been shown to cause severe convulsions in animal experiments. AIMS: We report a patient who experienced a single convulsive seizure that resulted in transient hyperammonemia during treatment with tranexamic acid. CASE REPORT: A 68-year-old man was admitted and received tranexamic acid for persistent hemoptysis. After 5 days of admission, clonic convulsions that progressed to generalized seizures were noted following the intravenous administration of the tranexamic acid. Elevated ammonia level (233 µmol/l) was found. No further seizures occurred after immediate discontinuation of the drug. No other cause of seizures was found. The ammonia level on the following day normalized even without any treatment for the hyperammonemia. CONCLUSIONS: This case highlights that generalized convulsion is a very rare, but serious adverse effect of tranexamic acid. Generalized convulsion should be considered as a potential cause of transient hyperammonemia.

Establishment and characterization of a fibroblast cell line derived from the dorsal fin of red sea bream, Pagrus major (Temminck & Schlegel).

The establishment and partial characterization of a continuous cell line from the dorsal fin of red sea bream, Pagrus major, are described. The cell line, designated RSBF-2, has been subcultured for more than 100 passages since its initiation in November 2000. It was optimally maintained at 28 degrees C in Leibovitz L-15 medium with 10% foetal bovine serum. Propagation of RSBF-2 cells was serum dependent and exhibited low plating efficiency (<1.7%). Aside from long-term cryopreservation, the cells could also be kept at 4 degrees C for 72 days. The distribution of the chromosome number was 38-98 with a mode of 48. The RSBF-2 cell line was susceptible to red sea bream iridovirus but only produced a few rounded and refractory cells. Virus-inoculated RSBF-2 cells were then subcultured to generate a persistently infected cell line. RSBF-2 was also very sensitive to the extracellular products of Photobacterium damselae ssp. piscicida and produced significant fluorescent signals after transfection with pEGFP-C3. Analysis of mitochondrial cytochrome b gene sequences revealed 99% identity between the cell line and Pagrus major.

Intraperitoneal bladder rupture presenting as acute bloody ascites and oliguric acute renal failure in an alcoholic liver cirrhosis patient.

Intraperitoneal bladder rupture is a rare cause of acute abdomen with bloody ascites. We report herein the case of a patient who had alcoholic liver cirrhosis and multiple liver nodules, and experienced acute bloody ascites and oliguric acute renal failure in association with intraperitoneal bladder rupture. A 33-year-old male suffered from acute abdominal pain and oliguria following consumption of a large amount of alcohol and after blunt abdominal trauma. He was also found to have acute renal failure and newly onset bloody ascites that rapidly subsided following transurethral catheter drainage. Computed tomography cystography revealed intraperitoneal extravasation of contrast from the dome of the bladder, suggestive of intraperitoneal bladder rupture. The patient received surgical repair and was discharged with full recovery. This case shows that it is important for physicians to be aware of the possibility of intraperitoneal bladder rupture after alcohol consumption accompanied with abdominal blunt trauma. In particular, it has diagnostic complications for underlying liver tumors.

Hepatic migration of a catheter fragment followed by disconnection of a totally implantable venous access port.

Totally implantable venous access ports (TIVAPs) are frequently used in oncology patients who require long-term courses of chemotherapy. We report a silent, but potentially hazardous complication of catheter fracture and hepatic migration in a 64-year-old male. The patient presented with a painful, rapid swelling of subcutaneous tissue around the port area during a saline flush. A chest radiograph showed that the disconnected catheter had separated from the port and was no longer in its original location. A chest CT scan revealed that the disconnected catheter was found to be embolized to the right atrium, inferior vena cava and right hepatic vein. The patient was treated successfully with percutaneous transfemoral retrieval of the catheter under fluoroscopic guidance. To our knowledge, there have been no previous reports of migration of the fractured catheter of a TIVAP into the right hepatic vein. This case highlights that the integrity of TIVAPs should be ascertained before chemotherapeutic drugs are administered.