Needle-perc-assisted endoscopic surgery versus retrograde intrarenal surgery for the treatment of 1- to 2-cm lower-pole renal stones in patients with unfavorable infundibulopelvic anatomy: a matched-pair analysis.

PURPOSE: To compare the safety and efficacy of needle-perc-assisted endoscopic surgery (NAES) and retrograde intrarenal surgery (RIRS) for the treatment of 1- to 2-cm lower-pole stones (LPS) in patients with complex infundibulopelvic anatomy. METHODS: Between June 2020 and July 2022, 32 patients with 1- to 2-cm LPS and unfavorable lower-pole anatomy for flexible ureteroscopy were treated with NAES. The outcomes of these patients were compared with patients who underwent RIRS using matched-pair analysis (1:1 scenario). The matching parameters such as age, gender, body mass index, stone size, hardness, and pelvicalyceal anatomy characteristics including infundibular pelvic angle, infundibular length, and width were recorded. Data were analyzed using the Student's t-test, Mann-Whitney U test, and Fisher's exact test. RESULTS: The two groups had similar baseline characteristics and lower-pole anatomy. The stone burden was comparable between both groups. NASE achieved a significantly better initial stone-free rate (SFR) than RIRS (87.5% vs 62.5%, p = 0.04). The auxiliary rates for the NAES and RIRS groups were 12.5% and 31.3%, respectively (p = 0.13). Finally, the SFR after 1 month follow-up period was still higher for the NAES group than RIRS group (93.8% versus 81.3%), but the difference was not statistically significant (p = 0.26). Concerning the operation duration, overall complication rates, and postoperative hospital stay, there were no differences between two groups. CONCLUSION: Compared to RIRS for treating 1- to 2-cm LPS in patients with unfavorable infundibulopelvic anatomy for flexible ureteroscopy, NAES was safe and effective with higher SFR and similar complication rate.

Sr-HA-graft-Poly(γ-benzyl-l-glutamate) Nanocomposite Microcarriers: Controllable Sr2+ Release for Accelerating Osteogenenisis and Bony Nonunion Repair.

The microcarrier system offers an attractive method for cellular amplification and phenotype enhancement in the field of bone tissue engineering. However, it remains a challenge to fabricate porous microcarriers with osteoinductive activity for speedy and high-quality osseointegration in regeneration of serious complication of bone fracture, like nonunion. Here, we present a facile method for the first time manufacture microcarriers with osteogenic effects and properties based on well controlled and long-term Sr2+ release. At first, strontium-substituted hydroxyapatite was prepared (Sr-HA) and a novel Sr-HA-graft-poly(γ-benzyl-l-glutamate) (Sr-HA-PBLG) nanocomposite was synthesized. Then, the microcarriers with highly interconnected macropores were fabricated by a double emulsion method, which allowed cells to adhere and proliferate and secrete extracellular matrix. Besides, the microcarriers with a relatively uniform diameter of 271.5 ± 45.0 µm are feasible for injection. The Sr-HA-PBLG microcarriers efficiently promoted osteogenic gene expression in vitro. With injection of the Sr-HA-PBLG microcarriers loading adipose derived stem cells (ADSCs) into the nonunion sites, bone regeneration was observed at 8 weeks after injection in a mice model.

Role of c-Jun N-terminal kinase in the osteogenic and adipogenic differentiation of human adipose-derived mesenchymal stem cells.

Although previous studies have characterized the osteogenic potential of adipose-derived mesenchymal stem cells (AMSCs) in vitro and in vivo, the molecular mechanism involved remains to be fully determined. Previously, we demonstrated that the ERK pathway plays an important role in osteogenesis and regulation of the balance between osteogenesis and adipogenesis. Here, we explored the possible role of JNKs in osteogenesis and adipogenesis of AMSCs. JNK activation in osteo-induced AMSCs was initiated at 15 min, peaked at 30 min, and declined from 45 min to basal levels. Inhibition of the JNK signaling pathway using SP600125 blocked osteogenic differentiation in a dose-dependent manner, which was revealed by an ALP activity assay, extracellular calcium deposition detection, and expression of osteogenesis-relative genes (Runx2, ALP, and OCN) via RT-PCR and real-time PCR. However, blockage of JNK did not induce a switch between osteogenesis and adipogenesis of AMSCs in the presence of dexamethasone, which is different from that of blockage of ERK. Significantly, the blockage of JNK activation in adipo-induced AMSCs by SP600125 stimulated adipogenic differentiation, which was confirmed by Oil Red O staining to detect intracellular lipid droplets, and RT-PCR and real-time PCR analysis for expression of adipogenesis-relative genes (PPARγ2 and aP2). This study suggested a potential function of the JNK pathway in committing osteogenic and adipogenic differentiation of AMSCs in vitro. However, blockage of the JNK pathway is not sufficient to induce a switch from osteogenesis to adipogenesis of AMSCs.

Case fatality rates of COVID-19 during epidemic periods of variants of concern: A meta-analysis by continents.

OBJECTIVES: To calculate the case fatality rates (CFR) of COVID-19 during epidemic periods of different variants of concern (VOC) by continents. METHODS: We systematically searched five authoritative databases (Web of Science, PubMed, Embase, Cochrane Library, and MedRxiv) for epidemiological studies on the CFR of COVID-19 published between January 1, 2020, and March 31, 2023. After identifying the epidemic trends of variants, we used a random-effects model to calculate the pooled CFRs during periods of different VOCs. This meta-analysis was conducted following the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) and registered with PROSPERO (CRD42023431572). RESULTS: There were variations in the CFRs among different variants of COVID-19 (Alpha: 2.62%, Beta: 4.19%, Gamma: 3.60%, Delta: 2.01%, Omicron: 0.70%), and disparities in CFRs also existed among continents. On the whole, the CFRs of COVID-19 in Europe and Oceania were slightly lower than in other continents. There was a statistically significant association between the variant, HDI value, age distribution, coverage of full vaccination of cases, and the CFR. CONCLUSIONS: The CFRs of COVID-19 varied across the epidemic periods of different VOCs, and disparities existed among continents. The CFR value reflected combined effects of various factors within a certain context. Caution should be exercised when comparing CFRs due to disparities in testing capabilities and age distribution among countries, etc.

A Retrospective Study of 91 Patients Treated with Percutaneous Kyphoplasty for Mild Osteoporotic Vertebral Compression Fractures and a New Evaluation Scale of Shape and Filling Effect of Cement.

BACKGROUND: Percutaneous kyphoplasty (PKP) is commonly used to treat severe osteoporotic vertebral compression fractures (OVCFs) by restoring vertebral height. However, its application in mild cases is not frequently discussed. METHODS: The study retrospectively included 100 treated vertebral bodies of the 91 patients mentioned before, and efficacy was evaluated using visual analog scale (VAS) and Oswestry Disability Index (ODI) scores preoperatively, 2 days postoperatively, and at 1 and 6 months after treatment, as well as mean variation in vertebral body height. The study also examined complications such as pain recurrence, delayed vertebral fracture, and loss of vertebral height, and developed a scale to assess the shape and filling effect of cement (SFEC) and its impact on complications. RESULTS: The results showed significant reductions in mean VAS and ODI scores from pre-to post-surgery and an increase in vertebral body height. However, complications occurred in 10 patients who received treatment for 11 vertebral bodies, including pain recurrence, fractures, and loss of vertebral height. Among the 10 patients with complications, 7 (63.6%) vertebral bodies had dissatisfied SFEC scores, compared with 22 (24.7%) vertebral bodies with dissatisfied SFEC scores in 81 patients without complications (89 vertebral bodies). CONCLUSIONS: PKP is a safe and effective method for treating mild OVCFs, but attention should be paid to the shape and filling effects of cement during surgery to prevent later complications. The developed SFEC scale provides a specific and quantitative standards for evaluating the recovery status after PKP, which need further validations.

Evaluation and Analysis of the Rationality of Clinical Use of Carbapenems in Surgical Departments of a Tertiary Hospital in Southwest China.

Purpose: With the increasing frequency and intensity of carbapenem consumption, carbapenem-resistant organisms (CRO) have become a focus of anti-infection research. This study aimed to evaluate the rationality of the clinical use of carbapenems among inpatients in the surgical departments of a tertiary hospital in southwest China. Patients and methods: A point-score system was established for evaluation based on the clinical practices in surgical departments and selected carbapenem prescriptions from June 2020 to June 2021 for hepatobiliary surgery, gastrointestinal surgery, and neurosurgery in the study hospital. Prescriptions with a total score ≥ 270 were defined as rational. Descriptive statistics were used to describe the characteristics and rationality of the prescriptions. The chi-square test, Mann-Whitney U-test, and Kruskal-Wallis H-test were used to compare characteristics between rational and irrational prescriptions. Linear regression analysis was used to determine the factors affecting the rationality of carbapenem prescriptions. Results: According to 192 carbapenem prescription records, the median age of patients was 62 years [IQR, 48.0-73.0], and 20% of patients had abdominal infections, 10% had lung infections, 14% had intracranial infections, and 3% had urinary tract infections. 56% of carbapenem prescriptions were irrational. Compared with rational carbapenem prescriptions, irrational prescriptions had a higher proportion of those with inappropriate indications (49% vs 0%, p < 0.05), incorrect variety selection (15% vs 0%, p<0.05), and unreasonable assessment of etiology and efficacy (46% vs 8%, p < 0.05). Linear regression analysis suggested that the diagnosis of cholecystitis (standardized regression coefficient=0.183, p<0.05) and replaced medication (standardized regression coefficient = 0.154, p<0.05) influenced the rationality of carbapenem prescriptions. Conclusion: Our study shows that the irrational use of carbapenems deserves attention, especially in surgical departments. Interventions for carbapenem use that are based on evaluation criteria should be developed to reduce the emergence and spread of carbapenem-resistant bacteria.

Prevalence, awareness, treatment, and control of dyslipidemia in Chinese adults: a systematic review and meta-analysis.

Background: Researchers have conducted a considerable number of epidemiological studies on dyslipidemia in China over recent years. Nevertheless, a representative study to comprehensively appraise for the epidemiological status of dyslipidemia is still lacked. This meta-analysis is intended to explore the pooled prevalence, rates of awareness, treatment, and control of dyslipidemia among adults in Chinese Mainland. Materials and methods: A systematic review was performed on relevant cross-sectional studies published since January 2012 by searching six authoritative literature databases. Meta-analyses were conducted in included studies based on a random-effect model to summarize the epidemiological status of dyslipidemia in China. A potential source of heterogeneity was detected by subgroup analysis and meta-regression. Publication bias was assessed by Egger's test and funnel plots. A sensitivity analysis was conducted to examine the study quality's influence on the pooled estimate of prevalence and rates of awareness, treatment, and control. Results: Forty-one original researches with a total of 1,310,402 Chinese participants were finally included in the meta-analysis. The prevalence, rates of awareness, treatment, and control of dyslipidemia were 42.1%, 18.2%, 11.6%, and 5.4%, respectively. With a pooled prevalence estimate at 24.5%, low HDL-C was the most prevalent among various dyslipidemia types, followed by hypertriglyceridemia (TG) (15.4%), hypercholesterolemia (TC) (8.3%), and high LDL-C (7.1%). The pooled prevalence of elevated serum lipoprotein(a) [Lp(a)] was 19.4%. By gender, the prevalence of dyslipidemia was 47.3% in males and 38.8% in females. Subgroup analyses revealed that the prevalence in southern and urban areas were higher than their counterparts. Females and population in urban areas tended to possess higher rates of awareness, treatment, and control. Meta-regression analyses suggested that the year of screening influenced prevalence estimates for dyslipidemia. The impact of the study's quality on the pooled estimates is insignificant. Conclusion: Our study suggested a severe epidemic situation of dyslipidemia among adults in Chinese Mainland. More importantly, the awareness, treatment, and control rates were extremely low, revealing that dyslipidemia is a grave health issue. Consequently, we should attach more importance to the management of dyslipidemia, especially in economically underdeveloped areas. Systematic review registration: PROSPERO [CRD42022366456].

Exosome miR-4738-3p-mediated regulation of COL1A2 through the NF-κB and inflammation signaling pathway alleviates osteoarthritis low-grade inflammation symptoms.

This study aimed to elucidate the roles of microRNA (miR)-4738-3p and the collagen type I alpha 2 chain (COL1A2) gene in the pathogenesis of osteoarthritis (OA) through bioinformatics analysis and cellular assays. The GSE55235 dataset was analyzed using the weighted gene co-expression network analysis (WGCNA) method to identify gene modules associated with OA. Key overlapping genes were identified from these modules and the GSE55235-differential expressed genes (DEGs). The expression levels of selected genes were determined in C28/I2 cells using the quantitative real-time polymerase chain reaction (qRT-PCR). The interaction between miR-4738-3p and COL1A2 was examined in the context of interleukin 1 beta (IL-1ß) induction. Exosome characterization was achieved through transmission electron microscopy (TEM), western blotting (WB), and other analyses. The study also investigated the functional relevance of miR-4738-3p in OA pathology through various molecular and cellular assays. Our findings revealed that the green module exhibited a strong correlation with the OA phenotype in the GSE55235 dataset, with COL1A2 emerging as a hub gene and miR-4738-3p as its key downstream target. IL-1ß induction suggested that COL1A2 is involved in inflammation and apoptosis, while miR-4738-3p appeared to play an antagonistic role. The analysis of exosomes underscored the significance of miR-4738-3p in cellular communication, with an enhanced level of exo-miR-4738-3p antagonizing IL-1ß-induced inflammation and promoting cell survival. Conversely, a reduction in exo-miR-4738-3p led to increased cell damage. This study established a clear regulatory relationship between miR-4738-3p and COL1A2, with the nuclear factor kappa B (NF-κB) signaling pathway playing a central role in this regulation. The miR-4738-3p significantly influences the OA-associated inflammation, primarily through modulation of COL1A2 and the NF-κB pathway. Therefore, targeting miR-4738-3p offers a potential therapeutic approach for OA, with exosome miR-4738-3p presenting a promising strategy.

Risk factors for moderate-to-severe postoperative pain after percutaneous nephrolithotomy: a retrospective cohort study.

Percutaneous nephrolithotomy (PCNL) is a minimally invasive procedure for removing renal calculi, while a large number of patients experience acute moderate-to-severe pain despite the analgesia provided. This study aimed to explore the risk factors for postoperative pain after PCNL, which may provide a novel perspective to refine the enhanced recovery after surgery (ERAS) program and to improve clinical outcomes. The clinical data of 331 patients who underwent PCNL in our hospital from September 2020 to February 2021 were retrospectively analyzed. The pain intensity was assessed every 4 h until 24 h post-surgery. According to the visual analog scale (VAS) score, patients were divided into two groups: mild or no pain group (VAS score, 0-3) and moderate-to-severe pain group (VAS score, 4-10). The pre-, peri-, and post-operative data were collected and analyzed. The indicators with statistically significant differences were selected, and multivariate logistic regression analysis was employed to determine the risk factors for postoperative pain after PCNL. Among 331 patients, 221 patients had moderate-to-severe pain and the incidence rate was 66.77%. Multivariate logistic regression analysis showed that the independent risk factors for moderate-to-severe pain after PCNL were the diameter of the renal calculus (odds ratio (OR) = 6.23, 95% confidence interval (CI) 2.50-15.56, P = 0.001), the number of renal calculi (OR = 15.892, 95% CI 7.721-32.711, P < 0.01), the presence of residual calculi (OR = 1.780, 95% CI 0.897-3.533, P = 0.01), and operation time (OR = 1.033, 95% CI 1.020-1.046, P < 0.01). The diameter of the renal calculus, the number of renal calculi, the presence of residual calculi, and operation time were significant predictors of postoperative pain after PCNL.

Pneumatosis cystoides intestinalis accompanied by schistosomiasis: a case report.

Pneumatosis cystoides intestinalis (PCI) is a rare disease that most frequently occurs in the large and small intestine and has no obvious clinical symptoms. The main pathological feature is the presence of air-filled cysts in the intestinal submucosa, intermuscular wall, and subserous membrane. Conservative treatment is the first choice when no serious complications are present, whereas timely surgical treatment is needed for serious and life-threatening complications. This report presents the clinical and pathological analysis of PCI in a man in his early 90s. The patient was hospitalized because of acute abdomen and diagnosed with perforation of the sigmoid colon due to PCI with schistosomiasis after emergency surgery. Emergency partial sigmoid colon resection and permanent colostomy were performed under general anesthesia. Preoperative diagnosis of PCI is difficult because of the nonspecific clinical manifestations and endoscopic findings, and missed diagnosis and misdiagnosis easily occur. Pure PCI has no specific symptoms and does not require special treatment, and there is a lack of special treatment methods in clinical practice. However, when PCI is combined with other intestinal diseases such as schistosomiasis enteropathy, intestinal perforation is likely to occur, leading to severe acute abdomen with the need for prompt surgical treatment.

Expression Profile Analysis of Long Non-coding RNA in OVX Models-Derived BMSCs for Postmenopausal Osteoporosis by RNA Sequencing and Bioinformatics.

Osteoporosis (OP) has the characteristics of a systematically impaired bone mass, strength, and microstructure. Long non-coding RNAs (lncRNAs) are longer than 200 nt, and their functions in osteoporosis is yet not completely understood. We first harvested the bone marrow mesenchymal stem cells (BMSCs) from ovariectomy (OVX) and sham mice. Then, we systematically analyzed the differential expressions of lncRNAs and messenger RNAs (mRNAs) and constructed lncRNA-mRNA coexpression network in order to identify the function of lncRNA in osteoporosis. Totally, we screened 743 lncRNAs (461 upregulated lncRNAs and 282 downregulated lncRNAs) and 240 mRNAs (128 upregulated and 112 downregulated) with significantly differential expressions in OP compared to normal. We conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional analyses to investigate the functions and pathways of the differential expression of messenger RNAs (mRNAs), a coexpressed network of lncRNA/mRNA. Quantitative PCR (qPCR) validated that the expressions of NONMMUT096150.1, NONMMUT083450.1, and NONMMUT029743.2 were all downregulated, whereas NONMMUT026970.2, NONMMUT051734.2, NONMMUT003617.2, and NONMMUT034049.2 were all upregulated in the OVX group. NONMMUT096150.1, as a key lncRNA in OP, was identified to modulate the adipogenesis of BMSCs. Further analysis suggested that NONMMUT096150.1 might modulate the adipogenesis of BMSCs via the peroxisome proliferator-activated receptor (PPAR) signaling pathway, AMPK signaling pathway, and the lipolysis regulation in adipocyte and adipocytokine signaling pathway. Our study expands the understanding of lncRNA in the pathogenesis of OP.

Downregulation of microRNA­143 promotes osteogenic differentiation of human adipose­derived mesenchymal stem cells through the k­Ras/MEK/ERK signaling pathway.

MicroRNAs (miRNAs) are known to have regulatory roles in the osteogenic differentiation of various mesenchymal stem cells (MSCs), although their regulatory role on human adipose­derived mesenchymal stem cells (hADSCs) remains unclear. The aim of the present study was to investigate the biological function and underlying molecular mechanism of miRNAs in regulating the osteogenic differentiation of hADSCs using microarray assay. hADSCs differentiated into osteoblasts under culture with osteogenic medium, with an increase observed in calcium deposits and alkaline phosphatase activity. The mRNA levels of bone sialoprotein, osteopontin and osteocalcin increased, whereas Runt­related transcription factor­2 expression decreased during osteogenic differentiation. In addition, miR­143 was markedly downregulated during osteogenic differentiation, while miR­143 overexpression inhibited and miR­143 knockdown enhanced this process. miR­143 overexpression also blocked extracellular signal­regulated kinase 1/2 (ERK1/2) pathway activation, while miR­143 inhibition enhanced it. The promoting effects of miR­143 knockdown on the osteogenic differentiation of hADSCs were partly diminished by the mitogen­activated protein kinase (MEK) inhibitors U0126 and PD98059. Bioinformatics analysis further revealed that miR­143 targets k­Ras and directly binds to the 3'­untranslated region of its mRNA. Inhibition of miR­143 enhanced the activation of the k­Ras/MEK/ERK pathway during osteogenic differentiation, whereas miR­143 overexpression had the opposite effect. Collectively, these results demonstrated that miR­143 negatively regulates the osteogenic differentiation of hADSCs through the k­Ras/MEK/ERK pathway, providing further insight into the underlying molecular mechanisms.

LINC00511 Promotes Osteosarcoma Tumorigenesis and Invasiveness through the miR-185-3p/E2F1 Axis.

Osteosarcoma is a malignant tumor that seriously threatens human health. Numerous studies have pointed out the potential of long noncoding RNAs (lncRNAs) as new therapeutic targets for various human cancers. Therefore, we mainly investigate whether there is a new type of lncRNA pathway involved in regulating the development of osteosarcoma. The present study shows the higher expression levels of LINC00511 correlates to a shorter overall survival and disease-free survival time in patients with sarcoma. It is significantly higher in the clinical samples of osteosarcoma patients than in normal adjacent cancer tissues. We used U373 and SW1353 osteosarcoma cells to determine the effect of lncRNA on osteosarcoma proliferation and invasion by knocking down LINC00511 compared with controls. The results showed that the LINC00511 knockdown significantly suppressed osteosarcoma cell growth and metastasis. To explore the mechanisms of LINC00511 in osteosarcoma, we tested whether LINC00511 could competitively stimulate miR-185-3p and regulate E2F1 as a ceRNA. The results showed that LINC00511 knockdown induced the increased level of miR-185-3p levels; however, miR-185-3p overexpression suppressed LINC00511 levels. In addition, the results also demonstrated that LINC00511 knockdown or miR-185-3p overexpression could reduce E2F1 levels in osteosarcoma cells. The dual-luciferase reporter assay verified the direct interaction between miR-185-3p and LINC00511 or E2F1. These results may offer an explanation of how the lncRNA affects the progression of osteosarcoma, and our study shows that LINC00511 can be a novel biomarker in osteosarcoma.

Circular RNA circHIPK3 Promotes Cell Metastasis through miR-637/STAT3 Axis in Osteosarcoma.

Recent studies have suggested that circular RNAs play an important role in the progression of various cancers. However, few studies have revealed the great value of circRNAs in the diagnosis and prognosis prediction of osteosarcoma (OS). In this study, we performed experiments with the human OS cell lines and the results showed that the expression of circHIPK3 in OS cell lines was significantly upregulated compared to that in the normal cell line. In addition, the results showed that circHIPK3 could promote the migration, invasion, and growth of OS cells. Furthermore, miR-637 was identified as a target of circHIPK3, while STAT3 was targeted by miR-637. circHIPK3 could promote STAT3 expression via interacting with miR-637 in OS cells. In conclusion, our research uncovered an important role of the circHIPK3/miR-637/STAT3 pathway in the migration and invasion of OS cells and suggested that circHIPK3 may be a prognostic marker and a promising therapeutic target for OS.

Identification of circRNA-associated ceRNA network in BMSCs of OVX models for postmenopausal osteoporosis.

Circular RNAs (circRNAs) serve as competing endogenous RNAs (ceRNAs) and indirectly regulate gene expression through shared microRNAs (miRNAs). However, the potential circRNAs functioning as ceRNAs in osteoporosis remain unclear. The bone marrow mesenchymal stem cells (BMSCs) were isolated from ovariectomy (OVX) mice and controls. We systematically analyzed RNA-seq and miRNA-microarray data, miRNA-target interactions, and prominently coexpressed gene pairs to identify aberrantly expressed circRNAs, miRNAs, and messenger RNAs (mRNAs) between the OVX mice and controls. A total of 45 circRNAs, 22 miRNAs, and 548 mRNAs were significantly dysregulated (fold change > 1.5; p < 0.05). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were conducted for differentially expressed mRNAs, and subsequently a circRNA-associated ceRNA network involved in osteoporosis was constructed. We identified two ceRNA regulatory pathways in this osteoporosis mouse model-novel circRNA 0020/miR-206-3p/Nnmt and circRNA 3832/miR-3473e/Runx3, which were validated by real-time PCR. This is the first study to elucidate the circRNA-associated ceRNA network in OVX and control mice using deep RNA-seq and RNA-microarray analysis. The data further expanded the understanding of circRNA-associated ceRNA networks, and the regulatory functions of circRNAs, miRNAs and mRNAs in the pathogenesis and pathology of osteoporosis.

MiR-1-3p regulates the differentiation of mesenchymal stem cells to prevent osteoporosis by targeting secreted frizzled-related protein 1.

Osteoporosis (OP) is a systemic skeletal disorder with the characteristics of bone mass reduction and microarchitecture deterioration, resulting in bone fragility and increased fracture risk. A reduction in the osteoblast-differentiation of bone marrow mesenchymal stem cells (BMSCs) is considered as a basic pathogenesis of osteoporosis. miRNAs play a substantial role in the development and differentiation of BMSCs. In the present study, we found that miR-1-3p was significantly downregulated in the bones of Chinese osteoporotic patients (n = 29). Secreted frizzled-related protein 1 (SFRP1) was predicted as a target gene of miR-1-3p via the TargetScan and PicTar softwares and validated by dual-luciferase reporter assays. The findings revealed that the expression of SFRP1 was inversely correlated with miR-1-3p in osteoporotic patients. We induced mouse MSCs (mMSCs) to osteogenesis or adipogenesis and found that miR-1-3p was upregulated during osteogenesis but downregulated during adipogenesis. The overexpression of miR-1-3p stimulated osteogenesis and inhibited adipogenesis of mMSCs. In addition, ovariectomized (OVX) mice were tested and the function of miR-1-3p in vivo was explored. Immunohistochemistry and histomorphometric assays showed that in vivo inhibition of miR-1-3p increased the expression level of SFRP1 and reduced bone formation and bone mass. Furthermore, tartrate-resistant acid phosphatase (TRAP) staining indicated that the in vivo suppression of miR-1-3p promoted osteoclast activity, suggesting that miR-1-3p may influence bone mass by regulating bone resorption. It can be concluded that miR-1-3p plays a pivotal role in the pathogenesis of osteoporosis via targeting SFRP1 and may be a potential therapeutic target for osteoporosis.

Network and pathway-based analyses of genes associated with osteoporosis.

Osteoporosis (OP) is a disease characterized by bone mass loss, bone microstructure damage, increased bone fragility, and easy fracture. The molecular mechanism underlying OP remains unclear.In this study, we identified 217 genes associated with OP, and formed a gene set [OP-related genes gene set (OPgset)].The highly enriched GOs and pathways showed OPgset genes were significantly involved in multiple biological processes (skeletal system development, ossification, and osteoblast differentiation), and several OP-related pathways (Wnt signaling pathway, osteoclast differentiation, steroid hormone biosynthesis, and adipocytokine signaling pathway). Besides, pathway crosstalk analysis indicated three major modules, with first module consisted of pathways mainly involved in bone development-related signaling pathways, second module in Wnt-related signaling pathway and third module in metabolic pathways. Further, we calculated degree centrality of a node and selected ten key genes/proteins, including TGFB1, IL6, WNT3A, TNF, PTH, TP53, WNT1, IGF1, IL10, and SERPINE1. We analyze the K-core and construct three k-core sub-networks of OPgset genes.In summary, we for the first time explored the molecular mechanism underlying OP via network- and pathway-based methods, results from our study will improve our understanding of the pathogenesis of OP. In addition, these methods performed in this study can be used to explore pathogenesis and genes related to a specific disease.

Articular Cartilage Stem Cells Influence the Postoperative Repair of Hip Replacement by Regulating Endoplasmic Reticulum Stress in Chondrocytes via PERK Pathway.

OBJECTIVE: Endoplasmic reticulum stress (ERS) is present in chondrocytes of osteoarthritis, and the intensity of ERS is related to the degree of cartilage degeneration. In vitro intervention strategies can change the status of ERS and induce the inhibition of ERS-related pathway. Therefore, this study is designed to explore the role and molecular mechanism of cartilage stem cells (ACSCs) of ERS in chondrocytes after hip replacement. METHODS: Human cartilage cell lines C28/I2 were cultured as the control group. The ERS inducer was added into C28/I2 as ERS group. The third ERS + stem cells group was formed by adding cartilage stem cells into ERS group, and further transfection of si-PERK was defined as si-PERK + ERS + stem cells group. Cell cycle and apoptosis in the four groups were determined by flow cytometry. The protein expression of GRP78, PERK, ATF4, TMEM119, CDK4, Cyclin D, and BMP6 in chondrocytes in the four groups were investigated by western blot, and the distribution of PERK, TMEM119, and BMP6 in chondrocytes were observed by immunofluorescence assay. In addition, the transcriptional levels of Bcl2, Bax, and Caspase 3 were also determined by RT-PCR. RESULTS: In cell cycle assay, ERS increased the accumulation of cells in G0 /G1 and G2 /M, while cartilage stem cells weakened the effects. The apoptosis rates in control group, ERS, ERS + stem cells, si-PERK + ERS + stem cells were 0%, 21.3%, 18.9%, and 15.9%, respectively, and the difference of apoptosis rate between the latter three groups and control group was statistically significant (P < 0.01). Stem cells could weaken the ERS-induced cell apoptosis, especially reducing the number of cells in the late stage of apoptosis from 5.4% to 1.1%. The protein level of GRP78, PERK, ATF4, TMEM119, and BMP6 in the group of ERS, ERS + stem cells, and si-PERK + ERS + stem cells were all significantly higher than those in control group, and the group of ERS + stem cells was the highest, all of the differences were significant (P < 0.01). However, the protein level of CDK4 and Cyclin D presented an absolutely opposite trend and the difference was still significant (P < 0.05). The group of si-PERK + ERS + stem cell was lower than those in the group of ERS + stem cell but higher than those in the group of ERS (P < 0.05). The level of Caspase 3 in the latter three groups was significantly higher than those in the control group, and the group of ERS was the highest (P < 0.01). Besides, the relative level of Bcl-2/Bax in control group was 1, but the group of ERS was about 0.5, and there was significant difference (P < 0.01). The ratio of Bcl-2/Bax in the group of ERS + stem cells was more than 2 and significantly higher than those of other groups. CONCLUSION: ACSCs could reduce ERS-induced chondrocyte apoptosis by PERK and Bax/Bcl-2 signaling pathway.

MiR-409-3p Inhibits Cell Proliferation and Invasion of Osteosarcoma by Targeting Zinc-Finger E-Box-Binding Homeobox-1.

Osteosarcoma (OS) is the most common bone cancer worldwide. There is evidence that microRNA-409 (miR-409-3p) is involved in tumorigenesis and cancer progression, however, its possible role in OS requires clarification. In the present study, we evaluated the expression level, clinical significance, and mode of action of miR-409-3p in OS. The miR-409-3p levels were diminished in the OS cells and tissues compared with associated adjacent non-tumor tissues and a non-cancer osteoplastic cell line. Low miR-409-3p expression levels were associated with clinical stage and distant metastasis in patients with OS. Resumption of miR-409-3p expression attenuated OS cell proliferation and invasion. Additionally, based on informatics analyses, we predicted that zinc-finger E-box-binding homeobox-1 (ZEB1) is a possible target of miR-409-3p. This hypothesis was confirmed using luciferase reporter assays, reverse transcription-quantitative real-time polymerase chain reaction, and Western blot analyses. The findings of the current study indicated that ZEB1 was up-regulated in the OS tissues and cell lines, and that this up-regulation was inversely proportional to miR-409-3p expression levels. Furthermore, down-regulation of ZEB1 decreased OS cell invasion and proliferation, illustrating that the tumor suppressive role of miR-409-3p in OS cells may be exerted via negative regulation of ZEB1. Taken together, our observations highlight the potential role of miR-409-3p as a tumor suppressor in OS partially through down-regulation of ZEB1 and suggest that miR-409-3p has potential applications in OS treatment.

Bone regeneration using injectable poly (γ-benzyl-L-glutamate) microspheres loaded with adipose-derived stem cells in a mouse femoral non-union model.

Microspheres have gained immense popularity in bone tissue engineering because of their unique properties as injectable scaffolds for bone tissue regeneration. Herein, we evaluated the feasibility of using poly (γ-benzyl-L-glutamate) (PBLG) microspheres for bone engineering by examining the attachment, proliferation, and osteogenic differentiation of adipose-derived stem cells (ASCs) in vitro and the use of PBLG microspheres in healing non-union in vivo. Scanning electron microscopy and fluorescent 3, 30-dioctadecyloxacarbocyanine perchlorate-labeling were performed to study the attachment and growth of ASCs to the PBLG microspheres, and a DNA assay was performed to quantify cell proliferation with time. Osteogenic differentiation of ASCs cultured on the PBLG microspheres was assessed by determining alkaline phosphatase expression and extracellular calcium deposition, which was further confirmed by real-time polymerase chain reaction for osteogenesis-related genes. Femoral non-union were created in mouse models and filled with ASCs/PBLG microspheres in vivo. The results showed that ASCs attached, spread, and showed good osteogenic differentiation on the PBLG microspheres in vitro. Moreover, the ASCs/PBLG microspheres could repair the mouse femoral non-union in vivo. Thus, PBLG microspheres have good biocompatibility and cytocompatibility and are potentially useful as an injectable scaffolds for bone tissue engineering.