An exploration of facilitators and barriers to patient navigator core functions with breast cancer patients: Implications for the development of a human-centered mHealth app.

Objectives: To understand the barriers to core functions and workflow among patient navigators (PN) who navigate people diagnosed with breast cancer (BC). To identify how a mobile health (mHealth) app could assist PNs in providing care to BC patients. Methods: This qualitative research study used purposive sampling to recruit stakeholders (N = 33) from January to August 2021. We conducted individual semi-structured interviews with PNs (n = 11), oncology care providers (n = 12), and BC patients (n = 10). We used conventional content analysis to analyze the interview data. Results: Participants identified the following sociotechnical systems barriers in PN workflows that negatively impact BC patient care: 1) resources, 2) insurance coverage, 3) communication challenges, and 4) impact of logistical tasks. Participants identified the user experience, app features, and interoperability customizations to enhance PNs' provision of patient care as important design elements to include in a mHealth app. Conclusion: Feedback from stakeholders provided valuable insights into key design considerations, functions, and content areas for developing a mHealth app for PN use in BC care delivery. Innovation: This is one of the first studies to incorporate the human-centered design and sociotechnical systems frameworks to understand barriers to PN workflow and provision of BC patient care across the cancer care continuum.

Human Disturbance Increases Health Risks to Golden Snub-Nosed Monkeys and the Transfer Risk of Pathogenic Antibiotic-Resistant Bacteria from Golden Snub-Nosed Monkeys to Humans.

From the perspective of interactions in the human-animal-ecosystem, the study and control of pathogenic bacteria that can cause disease in animals and humans is the core content of "One Health". In order to test the effect of human disturbance (HD) on the health risk of pathogenic antibiotic-resistant bacteria (PARBs) to wild animals and transfer risk of the PARBs from wild animals to humans, golden snub-nosed monkeys (Rhinopithecus roxellana) were used as sentinel animals. Metagenomic analysis was used to analyze the characteristics of PARBs in the gut microbiota of golden snub-nosed monkeys. Then, the total contribution of antibiotic resistance genes (ARGs) and virulence factors (VFs) of the PARBs were used to assess the health risk of PARBs to golden snub-nosed monkeys, and the antimicrobial drug resistance and bacterial infectious disease of PARBs were determined to assess the transfer risk of PARBs from golden snub-nosed monkeys to humans. There were 18 and 5 kinds of PARBs in the gut microbiota of golden snub-nosed monkeys under HD (HD group) and wild habitat environments (W group), respectively. The total health risks of PARBs to the W group and the HD group were -28.5 × 10-3 and 125.8 × 10-3, respectively. There were 12 and 16 kinds of KEGG pathways of human diseases in the PARBs of the W group and the HD group, respectively, and the gene numbers of KEGG pathways in the HD group were higher than those in the W group. HD increased the pathogenicity of PARBs to golden snub-nosed monkeys, and the PARBs in golden snub-nosed monkeys exhibited resistance to lincosamide, aminoglycoside, and streptogramin antibiotics. If these PARBs transfer from golden snub-nosed monkeys to humans, then humans may acquire symptoms of pathogens including Tubercle bacillus, Staphylococcus, Streptococcus, Yersinia, Pertussis, and Vibrio cholera.

[Molecular mechanism of ligustilide attenuating OGD/R injury in PC12 cells by inhibiting ferroptosis].

The aim of this study is to explore the mechanism of ligustilide, the main active constituent of essential oils of traditional Chinese medicine Angelicae Sinensis Radix, on alleviating oxygen-glucose deprivation/reperfusion(OGD/R) injury in PC12 cells from the perspective of ferroptosis. OGD/R was induced in vitro, and 12 h after ligustilide addition during reperfusion, cell viability was detected by cell counting kit-8(CCK-8) assay. DCFH-DA staining was used to detect the level of intracellular reactive oxygen species(ROS). Western blot was employed to detect the expression of ferroptosis-related proteins, glutathione peroxidase 4(GPX4), transferrin receptor 1(TFR1), and solute carrier family 7 member 11(SLC7A11), and ferritinophagy-related proteins, nuclear receptor coactivator 4(NCOA4), ferritin heavy chain 1(FTH1), and microtubule-associated protein 1 light chain 3(LC3). The fluorescence intensity of LC3 protein was analyzed by immunofluorescence staining. The content of glutathione(GSH), malondialdehyde(MDA), and Fe was detected by chemiluminescent immunoassay. The effect of ligustilide on ferroptosis was observed by overexpression of NCOA4 gene. The results showed that ligustilide increased the viability of PC12 cells damaged by OGD/R, inhibited the release of ROS, reduced the content of Fe and MDA and the expression of TFR1, NCOA4, and LC3, and improved the content of GSH and the expression of GPX4, SLC7A11, and FTH1 compared with OGD/R group. After overexpression of the key protein NCOA4 in ferritinophagy, the inhibitory effect of ligustilide on ferroptosis was partially reversed, indicating that ligustilide may alleviate OGD/R injury of PC12 cells by blocking ferritinophagy and then inhibiting ferroptosis. The mechanism by which ligustilide reduced OGD/R injury in PC12 cells is that it suppressed the ferroptosis involved in ferritinophagy.

Does radiation therapy affect adhesion of tricalcium silicate cements to root dentin?

Abstract Objective This study aimed to analyze the effect of irradiation on the push-out bond strength of mineral trioxide aggregate (MTA) and Biodentine to radicular dentin. Methodology A total of 60 extracted mature human teeth with single root canals were categorized into two groups (irradiated and non-irradiated) (n=30). Each group was further divided into two sub-groups based on cements used (Biodentine and MTA). Then, a cumulative radiation dose of 60 Gy was divided into 30 fractions (two Gy for every fraction) and administered for five successive days per week over six weeks. Obturation was then performed using MTA and Biodentine. Afterwards, 1.5 mm thick horizontal sections were procured from the middle one-third of all the specimens and then subjected to push-out bond test. Results were analyzed using one-way analysis of variance with post-hoc Tukey's test. Results The bond strength of Biodentine and MTA to irradiated teeth was lower than non-irradiated teeth. Highest push-out bond strength was observed in non-irradiated specimens filled with Biodentine (p=0), followed by irradiated specimens filled with Biodentine (p=0); non-irradiated specimens filled with MTA (p=0); and irradiated specimens filled with MTA (p=0.9). Conclusion The push-out bond strength of Biodentine and MTA to root canal dentin decreased significantly post irradiation.

[Development and clinical application of a new transverse process retractor based on computer-aided design and 3D printing technology].

OBJECTIVE: To manufacture a new type of transverse process retractor by using computer-aided design(CAD) combined with 3D printing technology and investigate its clinical application effect. METHODS: A new type of transverse protrusion retractor was developed by CAD combined with 3D printing technology. From September 2018 to September 2019, the new transverse process retractor was applied in clinic. Sixty patients with lumbar single segment lesions who needed treatment by pedicle screw fixation, bone grafting and interbody fusion were divided into new transverse process retractor group and control group, with 30 cases in each group. There were 14 males and 16 females in new type transverse process retractor group, the age was (68.0±4.3) years old on average; lesion segment of 8 cases were L3,4, 9 cases were L4,5, 13 cases were L5S1;5 cases of lumbar disc herniation, 20 cases of lumbar spinal stenosis, 5 cases of degenerative lumbar spondylolisthesis;new transverse process retractor was used to pedicle screw placement. While there were 15 males and 15 females in control group, with an average age of (69.2±4.5) years old;lesion segment of 8 cases were L3,4, 10 cases were L4,5, 12 cases were L5S1;5 cases of lumbar disc herniation, 21 cases of lumbar spinal stenosis, 4 cases of degenerative lumbar spondylolisthesis;the traditional lamina retractor was used for soft tissue pulling and finished pedicle screw placement by freehand. The length of surgical incision, the time required for inserting a single screw, fluoroscopy times, the times of adjusting the positioning needle or screw in insertion process, and the visual analogue scale (VAS) of surgical incision 72 hours after operation were compared between two groups. RESULTS: Using CAD and 3D printing technology, a new type of transverse protrusion retractor was developed quickly. The length of surgical incision, the time required for inserting a single screw, fluoroscopy time, and the times of adjusting the positioning needle or screw in insertion process in new transverse process retractor group were less than those in control group(P<0.05). There was no significant difference in VAS of lumbar incision pain at 72 hours after operation between two groups(P>0.05). CONCLUSION: Using CAD combined with 3D printing technology to develop a new transverse protrusion retractor has the advantages of convenient design, short development cycle and low cost. It provides a new idea for the research and development of new medical devices. The new transverse process retractor has the advantages of easy operation, reliable fixation, less damage to paravertebral muscle, convenient pedicle screw placement, reducing fluoroscopy time and so on.

Germline Mutational Landscape in Chinese Patients With Advanced Breast Cancer.

Background: Genetic testing for breast cancer (BC) patients may shift the paradigm towards more personalized management and treatment strategies. While gene alterations may be ethnic-specific in breast cancer, our understanding of genetic epidemiology of BC remains mainly driven by data from Caucasian populations and further limited to selected handful of genes. Methods: We collected whole blood samples from 356 BC patients at metastatic first line BC and primary stage IV disease at Beijing Cancer Hospital between Jan. 2013 to Dec. 2019. A comprehensive 600-gene cancer panel was used to detect germline variants in the covered genes with a median 300x sequencing depth. Variants were classified into pathogenic, likely pathogenic, variant of uncertain significance, likely benign and benign groups according to the ACMG/AMP Standards and Guidelines. Pathogenic and likely pathogenic variants were considered as deleterious mutations. Results: The median age of 356 BC patients was 49 years (range, 21-87 years) at the first diagnosis of BC. Deleterious germline mutations across 48 cancer-related genes were identified in 21.6% (77/356) of the patients. The most prevalent mutations were BRCA1/2 mutations (7.0%), followed by ATM and RAD50 mutations (1.4% each). In addition, patients with family history were more likely to carry BRCA1 mutations (P=0.04). Moreover, patients with triple-negative breast cancer (TNBC) were more likely to harbor BRCA1 mutations than those with HR+ or HER2+ breast cancer (P=0.006). While there was no significant survival difference observed in BRCA1/2 carriers relative to non-carriers, patients with DNA damage repair (DDR) gene mutations (mostly frequently BRCA, ATM, RAD50) had worse disease-free survival (P=0.02). Conclusions: The most prevalent germline mutations in a large cohort of Chinese patients with advanced BC were BRCA1/2 mutations, followed by ATM and RAD50 mutations. In total, approximately 16.0% (57/356) of patients carry deleterious mutations in DDR pathway. Patients with breast or ovarian cancer family history were more likely to carry BRCA1/2 mutations, and ones with DDR mutations had worse survival. These findings suggest that DDR mutations are prevalent in Chinese BC patients who may potentially benefit from treatment with Poly (ADP-ribose) polymerase inhibitors.

Lipoxin A4 protects primary spinal cord neurons from Erastin-induced ferroptosis by activating the Akt/Nrf2/HO-1 signaling pathway.

Ferroptosis is an iron-dependent programmed cell death, which participates in the pathogenesis of spinal cord injury (SCI). Our previous study has revealed that Lipoxin A4 (LXA4) exerts a protective role in SCI. Here, we investigated whether LXA4 can protect SCI through inhibiting neuronal ferroptosis. We treated primary spinal cord neurons with Erastin (ferroptosis activator) to induce ferroptosis. Erastin treatment reduced cell viability and enhanced cell death of primary spinal cord neurons, which was rescued by ferrostatin-1 (ferroptosis inhibitor). Moreover, Erastin repressed glutathione peroxidase 4 (GPX4) expression and the levels of glutathione and cysteine in primary spinal cord neurons. Erastin also enhanced the expression of ferroptosis biomarkers (PTGS2 and ACSL4) and the levels of reactive oxygen species (ROS) in primary spinal cord neurons. The influence conferred by Erastin was effectively abolished by LXA4 treatment. Furthermore, LXA4 enhanced the protein expression of p-AKT, nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and haem-oxygenase-1 (HO-1) in primary spinal cord neurons. LXA4-mediated inhibition of ferroptosis of primary spinal cord neurons was prohibited by LY294002 (AKT inhibitor), brusatol (Nrf2 inhibitor) or zinc protoporphyrin (HO-1 inhibitor). In conclusion, this work demonstrated that LXA4 exerted a neuroprotective effect in Erastin-induced ferroptosis of primary spinal cord neurons by activating the Akt/Nrf2/HO-1 signaling pathway. Thus, this work provides novel insights into the mechanisms of action of LXA4 in ferroptosis of primary spinal cord neurons and indicates that LXA4 may be a potential therapeutic agent for SCI.

Wireless Microfluidic Sensor for Metal Ion Detection in Water.

In the present work, a wireless microfluidic sensor based on low-temperature cofired ceramic (LTCC) technology for real-time detection of metal ions in water is proposed. The wireless sensor is composed of a planar spiral inductor and parallel plate capacitor (LC) resonant antenna, which integrates with the microchannel in the LTCC substrate between the capacitor plates. Aqueous solutions of Pb(NO3)2, Cd(NO3)2, Mg(NO3)2, Ca(NO3)2, NaNO3, and KNO3 with concentrations of 0-100 mM were tested with the sensors. The metal ion and its concentration in water can be tested by the amplitude of the reflection coefficient (S 11) and the resonance frequency (f r) of the wireless microfluidic sensor. The metal ion species can be distinguished from the wireless response behavior of the sensor. The detection limit of the sensor for the selected metal ionic solutions could reach as low as 5 µM. The normalized sensitivity of the sensor is 0.47%, which is higher than that of the reported liquid microfluidic sensors based on microwave resonators. The wireless microfluidic sensor of this study is promising for rapid and convenient detection of heavy metal ion pollutants in the industrial wastewater.

[Experimental study of individualized 3D printing-guided template combined with thoracolumbar pedicle screw placement for the treatment of ankylosing spondylitis].

OBJECTIVE: To evaluate accuracy and safety of individualized 3D printing guided template for thoracolumbar pedicle screw placement in patients with ankylosing spondylitis. METHODS: From January 2016 to September 2019, thoracolumbar spine three-dimensional CT data of 8 patients with ankylosing spondylitis were included, Mimics 17.0 and ideaMaker computer software were applied to design thoracolumbar pedicle screw guided template of patients with AS, physical model of all patients (T10-L2)were printed by 3D printer, 2 parts in each patient, and divided into guide-plate-assisted screw group (experimental group) and free-hand nail group (control group). Thoracolumbar pedicle screws of both groups were placed by the same spinal surgeon. The accuracy of pedicle screw placement between two groups were evaluated according to results of postoperative CT, the accuracy of the fixation of thoracolumbar pedicle screw was divided into 4 grades, grade 0 and 1 screws were acceptable nails, grade 2 and 3 screws were unacceptable nails. The diameter and length of pedicle screws, the distance between entry point and posterior median line designed by preoperative 3D printing were compared with actual use in operation. RESULTS: Twenty three blocks of thoracolumbar 3D printing screw of ankylosing spondylitis guided templates were designed and printed in guide-plate-assisted screw group, 46 screws were inserted and 44 screws were accepted. The time of implanting a screw into thoracolumbar pedicle was (4.20±1.15) min, the frequency of X-ray was (5.00±1.25) times and the average adjustment times of screw and Kirschner needle during screw placement was (1.76±1.32) times. In the control group, 46 nails were placed by traditional surgical method and 30 screws were accepted. The time of implanting a screw into thoracolumbar pedicle was (14.67±2.23) min, the frequency of X-ray fluoroscopy was (14.46±2.21) times and the average times of Kirschner needle adjustment was (4.76±3.39) times. The success rates between experimental group and control group were 95.65%(44 / 46) and 56.22%(30 / 46) respectively, and had statistical difference (χ2=13.538, P<0.05). There was no significant difference in diameter, length of pedicle screws and the distance of posterior median line between virtual designed by 3D printing before operation and actual situation in opertaion (P>0.05). The operation time of inserting a single screw, the times of X-ray fluoroscopy, and the average times of adjustment screw and Kirschner needle in experimental group were significant less than those in control group(P<0.01). CONCLUSION: The personalized guide template assisted the thoracolumbar fixation designed by 3D printing could significantly improve safety, accuracy and efficiency of surgery, especially suitable for thoracolumbar vertebral bodies requiring posterior pedicle screw fixation for fracture or dislocation with AS.

[3D printing technology assisted the preoperative planning and application value in adult kyphoscoliosis deformity].

OBJECTIVE: To explore the application value of 3D printing technology in preoperative surgery plan and intraoperative auxiliary operation for adult kyphoscoliosis deformity. METHODS: The clinical data of 12 adult patients with kyphoscoliosis deformity treated from September 2017 to January 2019 were retrospectively analyzed. There were 3 males and 9 females, aged from 21 to 63 years old with an average of (47.67±13.32) years old. Among them, 4 cases were congenital kyphoscoliosis, 2 cases were old tuberculosis thoracolumbar kyphosis ; 2 cases were idiopathic kyphoscoliosis, 4 cases were degenerative kyphoscoliosis. The CT scan data of the patient's spine was imported into Mimics17.0 software to establish the three dimensional model of the spine, and the spine model was produced by 3D printer. Using the spine model simulated operation, preoperative surgery program planning and formulated a precise surgery, and further analysed postoperative imaging parameters improvement. All the patients were followed up for more than 1 year. Before and after operation and at the last follow-up, the scoliosis Cobb angle, maximum kyphosis Cobb angle, and coronal plane balance (distance between C 7 plumbline and center sacral vertical line, C7PL-CSVL), sagittal plane balance (sagittal vertical axis, SVA), pelvic parameters and other related imaging parameters were measured to further evaluate its orthopedic effect. RESULTS: Twelve patients with spine deformity were treated with different osteotomy and internal fixation fusion methods under the guidance of a 1∶1 spine model (pedicle screw placement of 4 patients with severe deformity were assisted by pedicle screw guide plates), nail placement and osteotomy have good effects, no major tissue damage such as blood vessels, nerves and spinal cord during and after surgery, no complications such as cerebrospinal fluid leakage and infection. Preoperative Cobb angle of scoliosis was (56.5±22.5) °, Cobb angle of kyphosis was (65.2±19.5) °, C7 PL-CSVL was (45.8±16.9) mm, SVA was (48.7±25.4) mm. Postoperative at 4 weeks, Cobb angle of scoliosis was (20.8±11.5) °, and Cobb angle of kyphosis was (22.0±6.6) °, with correction rates of (65.1±9.7)% and (64.6± 10.6)%, respectively ; C7 PL-CSVL was (22.3±8.9) mm, and SVA was (23.3±13.1) mm, all of which were significantly improved compared with preoperative results. The mean follow-up time was (18.5±7.9) months in 12 patients. At the last follow-up, the Cobb angles of scoliosis and kyphosis were (22.2±10.8) ° and (23.6±7.7) °, respectively, C7 PL-CSVL was (23.5±10.8) mm, and SVA was (24.7±12.5) mm. The results were statistically significant compared preoperative (P<0.05). There was no significant difference at the postoperative at 4 weeks and the last follow-up (P>0.05). CONCLUSION: The 3D print model can visually and clearly show the vertebral morphology and structure of adult kyphoscolisis and its spatial relationship with the adjacent vertebrae, blood vessels, and nerves, which provides a good and intuitive stereoscopic anatomical structure observation for the individualization of the surgical plan. Pre-simulation of operations to determine the internal fixation, fusion segment and osteotomy orthopedic way, may to provide a reference for actual clinical surgery, and can improve the accuracy and safety of surgery.

Lipoxin A4 protects against spinal cord injury via regulating Akt/nuclear factor (erythroid-derived 2)-like 2/heme oxygenase-1 signaling.

Spinal cord injury (SCI) is a devastating physical trauma worldwide. The mechanisms of SCI are still not clear and the effective treatment is limited. Lipoxin A4 (LXA4) possesses anti-inflammatory and neuroprotective effects. The present study was designed to further evaluate the molecular mechanisms of LXA4-induced protective effects in a rat model of SCI. We found that LXA4 increased Basso, Beattie and Bresnahan (BBB) scores, increased mechanical paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) to a radiant heat, reduced the lesion volume, decreased Bax mRNA expression and increased Bcl-2 expression after SCI. The phosphorylation of Akt and protein expression of Nrf2 and HO-1 were reduced after SCI. LXA4 treatment significantly inhibited the reduction of Akt phosphorylation and Nrf2 and HO-1 protein expression. Injection of LY294002 notably inhibited the phosphorylation of Akt, and the expression of total Akt and Nrf2 and HO-1 after SCI in LXA4-treated rats. LY294002 prohibited LXA4-induced effects after SCI. shNrf2 injection markedly decreased both Nrf2 and HO-1 expression in LXA4-treated rats after SCI. ZnPP notably decreased HO-1 expression but did not markedly affect Nrf2 expression. shNrf2 and ZnPP prohibited LXA4-induced increase of BBB scores, and PWT and PWL, decrease of lesion volume of spinal cord, reduction of Bax expression and increase of Bcl-2 expression. The results indicate that LXA4 protects against SCI through Akt/Nrf2/HO-1 signaling. The data provide novel insights into the mechanisms of LXA4-mediated neuprotective effects against SCI and suggest that LXA4 may be a potential therapeutic agent for SCI and its associated complications.

MicroRNA-137 Affects Proliferation and Migration of Placenta Trophoblast Cells in Preeclampsia by Targeting ERRα.

OBJECTIVE: To investigate the effects of microRNA-137 (miRNA-137) in proliferation and migration of placenta trophoblast cells of preeclampsia and the targeting gene of miRNA-137. METHODS: A total of 134 cases of puerperants were divided into normal pregnancy (n = 50), mild preeclampsia (n = 38), and severe preeclampsia groups (n = 46). MiRNA-137, estrogen-related receptor α (ERRα), and wingless INT (WNT)11 messenger RNAs (mRNAs) were measured in placental tissue and trophoblast cells after transfection, and ERRα protein in placental tissues was detected by immunohistochemistry. The target genes of miRNA-137, trophoblast cell proliferation, migration, and invasion abilities were detected. Both ERRα and WNT11 proteins in the trophoblast cells were measured after transfection. RESULT: Relative expressions of miRNA-137 were higher, and positive expression rates and relative expression levels of ERRα protein were lower in mild and severe preeclampsia and early- and late-onset preeclampsia than in normal pregnancy group (all P < .05). MiRNA-137 in the placental tissues was negatively correlated with ERRα protein ( P < .05). Luciferase reporter gene assay analysis showed that ERRα was a direct target gene of miRNA-137. Absorbance values, relative scratch-covered areas, cell membrane permeable rate, ERRα, and WNT11 mRNA and protein relative expressions were significantly lower, while cells at G1/G0 phase were higher in miRNA-137 mimic group than those in the blank, negative control, and miRNA-137 inhibitor group. CONCLUSION: MiRNA-137 significantly reduced the proliferation and migration of placenta trophoblast cells of preeclampsia by targeting ERRα, which might be a potential target for gene therapy.

Critical role of regulator of calcineurin 1 in spinal cord injury.

Spinal cord injury (SCI) is a severe clinical problem worldwide. The pathogenesis of SCI is complicated and much is unknown. The current study was designed to investigate the possible role of regulator of calcineurin 1 (RCAN1) in SCI and to explore the possible molecular mechanisms. Rats were injected with LVshRNAi-RCAN1 and then contusion-induced SCI was established. We found that RCAN1 was significantly increased in spinal cord of rats with SCI. Knockdown of RCAN1 markedly facilitated the structural and functional recovery in the spinal cord, as illustrated by decrease of lesion volume and increase of Basso, Beattie, and Bresnahan (BBB) and combined behavioral score (CBS) scores. Downregulation of RCAN1 suppressed the increase of pro-inflammatory cytokines, including IL-1ß and TNF-α, and inhibited the increase of TUNEL-positive cell numbers and caspases 3 and 9 activities. The decrease of oxygen consumption rate and increase of expression of glucose-regulated protein 78 (GRP78) and phosphorylation of protein kinase RNA-like endoplasmic reticulum (ER) kinase (PERK) in rats with SCI were inhibited by LVshRNAi-RCAN1. Moreover, knockdown of RCAN1 ameliorated oxidative stress in rats with SCI, as evidenced by decrease of TBA reactive substances (TBARS) and GSSG content and increase of glutathione (GSH) level. These results suggested that RCAN1 played an important role in SCI through regulation of various pathological processes. Overall, the data provide novel insights into the role of RCAN1 in SCI and novel therapeutic targets of the treatment of injury in the spinal cord.

Costunolide, an active sesquiterpene lactone, induced apoptosis via ROS-mediated ER stress and JNK pathway in human U2OS cells.

Costunolide, an active sesquiterpene lactone, is derived from many herbal medicines and it exhibits a broad spectrum of bioactivities such as anti-inflammatory, potential anti-tumor activity. Herein we assessed the anti-cancer effects of costunolide on U2OS cells and explored the underlying molecular mechanisms. The experiment data show that Costunolide exhibited significant anti-tumor activity by apoptosis related assays including Annexin V-FITC/PI flow cytometric analysis and 4,6-diamino-2-phenyl indole (DAPI) staining morphological analysis. Furthermore, we found Costunolide induced the loss of mitochondrial transmembrane potential, down-regulated Bcl-2/Bax ratio, encouraged Cyt-c release and caspase activation. All those effects are contributed by reactive oxygen species (ROS) generation and ER stress-induced mitochondrial dysfunction which are also responsible for c-Jun N-terminal kinase (JNK) activation. After the treatment of JNK inhibitor SP600125, it obviously reversed costunolide-induced apoptosis. Given N-acetyl-l-cysteine (NAC) effectively blocked the activation of JNK. Taken together, our results demonstrate that costunolide induces apoptosis in human U2OS cells through ROS generation and p38 MAPK/JNK activation.

Isoquercetin ameliorates tunicamycin-induced apoptosis in rat dorsal root ganglion neurons via suppressing ROS-dependent endoplasmic reticulum stress.

BACKGROUND: Recent studies showed that Isoquercetin (Iso), a novel extracts of plants and fruits could protect neuronal cells from neurotoxicity and neuro-inflammation. However, its role in acute spinal cord injury (ASCI) have not been elucidated. In the present study, we investigated whether Iso could prevent Tunicamycin (TUN)-induced rat dorsal root ganglion (DRG) neurons from apoptosis and endoplasmic reticulum (ER) stress. METHODS: DRG neurons were pre-treated with different doses of Iso for 24h (h) and then were stimulated with TUN (0.75µg/ml) for 24h. The cytotoxic effects and apoptosis were detected by MTT assay and TUNEL staining. The protein and mRNA expression levels were detected by Real Time PCR and Western blot, respectively. The localization of cleaved caspase-12 was evaluated by immunofluorescent staining and Western blot. The activation of caspase were measured by colorimetric assays and Western blot. Lactate Dehydrogenase (LDH) and Malondialdehyde (MDA) leakage were detected by the LDH or MDA Detection Kit PLUS. RESULTS: Iso protected TUN-associated DRG neurons from being damaged by cytotoxicity and apoptosis in a dose-dependent manner. Increased LDH and MDA leakage and proportion of TUNEL-positive cells, activation of caspase-3 and -9, increased Bcl-2 Assaciated X protein (Bax)/B cell lymphoma/lewkmia-2 (Bcl-2) ratio and mRNA levels of p53 Upregulated Modulator Of Apoptosis (PUMA) and DP5, and mitochondrial Cytochrome c release. Additionally, Iso down-regulated mRNA levels of ER stress genes, such as glucose-regulated protein 78 (GRP78), GRP94, C/EBP homologous protein (CHOP), and cleaved caspase-12 in TUN-induced DRG neurons. Moreover, Iso blocked the activation of three key branches of unfolded protein response in DRG neurons, including phosphorylation of pancreatic ER stress kinase (PERK), eukaryotic initiation factor 2 alpha (eIF2α), inositol-requiring enzyme 1α (IRE1α), and transcription factor 6 (ATF6). CONCLUSIONS: Collectively, Iso prevented TUN-induced DRG neurons apoptosis by inhibiting mitochondrial and ER stress-associated apoptotic signaling cascades, suggesting that Iso possessed neuro-protective ability which could be beneficial in the prevention of ASCI.

Report: Bacterial susceptibility and resistance analysis of traumatic osteomyelitis.

With the rapid development of industry, agriculture and transportation, the high energy trauma happened accordingly, thus greatly increased the incidence of traumatic osteomyelitis. The clinical traumatic osteomyelitis was mainly the local bone tissue inflammation caused by bacteria infection as trauma or iatrogenic causes. The delaying recovery could cause bone defection or bone nonunion. The purpose of this paper was to contribute new reference for the clinical prevention and treatment through tremendous of disease-causing bacteria susceptibility and resistance analysis of osteomyelitis.

MiR-374a promotes the proliferation of osteosarcoma cell proliferation by targeting Axin2.

BACKGROUND: MicroRNA-374a (miR-374a) has been implicated in several cancers. However, its role in osteosarcoma (OS) remains unclear. Thus the aim of this study was to investigate its expression and role in progression of OS. METHODS: Quantitative real-time PCR (qRT-PCR) was performed to detect the expression of miR-374a in OS cell lines and tissues. To further understand its role, we restored expression of miR-374a in MG63 cell line by transfection with miR-374a mimics or inhibitors. Effects of miR-374a on cell proliferation on targets were also determined. RESULTS: In the present study, our results showed that miR-374a was significantly up-regulated in both OS cell lines and OS tissues. Over expression of miR-374a markedly accelerated proliferation of OS cells, while its inhibition significantly suppressed cell proliferation. Moreover, Axin2 was identified to be a functional downstream target of miR-374a, and decreased expression of Axin2 could promote OS cell proliferation. CONCLUSION: Our study suggested that miR-374a functions as an oncogene in OS, and the miR-374a/Axin2 axis might represent a potential therapeutic target for OS intervention.

Association between XRCC3 Thr241Met polymorphism and risk of osteosarcoma in a Chinese population.

The aim of this study was to investigate whether XRCC3 Thr241Met polymorphism could affect the development of osteosarcoma in a Chinese population. A total of 152 osteosarcoma patients and 304 health control subjects were included in our study. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay was applied to assess the XRCC3 Thr241Met gene polymorphism. By conditional logistic regression analysis, we found that TT genotype of XRCC3 Thr241Met was associated with increased risk of osteosarcoma in codominant model (OR = 2.53, 95% CI = 1.28-5.39). Moreover, XRCC3 Thr241Met gene polymorphism was correlated with an elevated increased risk of osteosarcoma in dominant (OR = 1.55, 95% CI = 1.03-2.34) and recessive models (OR = 2.30, 95% CI = 1.16-4.56). In conclusion, we found that XRCC3 Thr241Met gene polymorphism was associated with increased risk of osteosarcoma in codominant, dominant and recessive models.

Rocaglamide-A Potentiates Osteoblast Differentiation by Inhibiting NF-κB Signaling.

Rheumatoid arthritis is a chronic inflammatory disease that leads to bone and cartilage erosion. The inhibition of osteoblast differentiation by the inflammatory factor TNF-α is critical for the pathogenesis of rheumatoid arthritis. To modulate TNF-α mediated inhibition of osteoblast differentiation is required to improve therapeutic efficacy of rheumatoid arthritis. Here, we explored the potential role of rocaglamide-A, a component of Aglaia plant, in osteoblast differentiation. Rocaglamide-A prevented TNF-α mediated inhibition of osteoblast differentiation, and promoted osteoblast differentiation directly, in both C2C12 and primary mesenchymal stromal cells. Mechanistically, Rocaglamide-A inhibited the phosphorylation of NF-κB component p65 protein and the accumulation of p65 in nucleus, which resulted in the diminished NF-κB responsible transcriptional activity. Oppositely, overexpression of p65 reversed rocaglamide-A's protective effects on osteoblast differentiation. Collectively, rocaglamide-A protected and stimulated osteoblast differentiation via blocking NF-κB pathway. It suggests that rocaglamide-A may be a good candidate to develop as therapeutic drug for rheumatoid arthritis associated bone loss diseases.

Elevated serum levels of Interleukin-37 are associated with inflammatory cytokines and disease activity in rheumatoid arthritis.

Interleukin-37 (IL-37) is closely associated with several inflammatory diseases. However, the role of IL-37 in the pathogenesis of rheumatoid arthritis (RA) remains unclear. The aim of this study was to assess the associations between serum levels of IL-37 and disease activity, inflammatory cytokines, and bone loss in patients with RA. Serum cytokines levels were examined by Enzyme-linked immunosorbent assay (ELISA). Radiographic bone erosion was assessed using the van der Heijde-modified Sharp score and bone mineral density (BMD) was measured using DXA. Serum IL-37 levels in RA patients were significantly higher than those in HCs (p < 0.001), and were significantly positively correlated with clinical parameters of disease activity and serum levels of IL-17 and IL-23. In addition, serum IL-37 levels were significantly higher in patients with stage IV of radiographic bone erosion than those with stage III and stage I-II, and they were significantly higher in those with osteopenia and osteoporosis than in those with normal BMD. Our results suggest that serum IL-37 levels were increased in patients with RA and were positively associated with disease activity, IL-17/IL-23 and bone loss in RA, suggesting that IL-37 may play a critical role in the pathogenesis of RA.