Mammary epithelial cell-derived exosomal miR-155-inhibitor played a key role in the treatment of mastitis via down-regulation of TLRs/NF-κB signaling pathway to inhibit inflammatory response.

Mastitis is a common disorder in women capable of altering the normal physiological function of the mammary gland. It has been reported that mammary epithelial cells (MECs) could be involved in treating mastitis by regulating the inflammatory response and miR-155 might participate in this process. However, the effects of MECs-derived exosomal miR-155-inhibitor in treating mastitis and the regarding mechanism are still unknown. In our study, mouse mammary epithelial cells (HC11) were applied to study the role of MECs-derived exosomal miR-155-inhibitor in the treatment of mastitis and explore the mechanism. Results in our study showed that specific markers including CD63 and Apo-A1 were expressed in blank exosomes and exosomes containing miR-155-inhibitor isolated from transfected HC11 cells. Results of immunofluorescence showed that the blank exosomes and exosomes (containing miR-155-inhibitor) labeled with PKH26 were absorbed in HC11 cells. The level of miR-155 was decreased obviously in Engineered exosomes with miR-155-inhibitor and HC11 cells Transfected with exosome containing miR-155-inhibitor. The level of miR-155 was increased and cell apoptosis was promoted obviously in HC11 cells induced by LPS, however, they were decreased obviously after transfecting with an exosome containing miR-155-inhibitor. The level of TLR2, TLR4, TLR6, NF-κB, TNF-α, and IL-1ß was increased obviously in LPS-induced HC11 cells, however, they were decreased obviously after transfecting with an exosome containing miR-155-inhibitor. The change in IL-10 level is opposite to the above genes. Taken together, exosomal miR-155-inhibitor could decrease the apoptosis of MECs and inhibit the inflammatory response to treat mastitis by down-regulation in the TLRs/NF-κB signaling pathway, which might be a new therapeutic target for mastitis.

Tooth loss as a risk factor for dementia: systematic review and meta-analysis of 21 observational studies.

BACKGROUND: Tooth loss is suggested to be associated with an increased risk of dementia in many studies. But the relationship between tooth loss and dementia is not yet fully understood. This systematic review and meta-analysis aimed to determine the relative effect of tooth loss on dementia risk. METHODS: An electronic search of PubMed, Scopus, Embase, and Web of Knowledge was conducted in March 2018 to identify relevant observational studies with the English language restriction. Studies were included if they assessed the relationship between tooth loss and risk of dementia. Study quality was detected by the modified Downs and Black scale. Odds risks (ORs) were pooled using a random-effects model in the crude model. RESULTS: The literature search initially yielded 1574 articles, and 21 observational studies published between 1994 and 2017 were finally included for the analyses. The crude results with random-effects model showed that patients with multiple tooth loss had higher incidence of dementia (OR 2.62, 95% CI 1.90-3.61, P < 0.001, I2 = 90.40%). The association remained noted when only adjusted results were pooled from 18 studies (OR 1.55, 95% CI 1.41-1.70, P = 0.13, I2 = 28.00%). Meta-regression analysis showed that study design explained about 16.52% of heterogeneity in the crude model. The overall quality rating scores of studies ranged from 11 to 16. CONCLUSIONS: Findings from this review evidenced that tooth loss is positively associated with an increased risk of dementia in adults. Future well-designed longitudinal researches examining the direct and indirect relationship between tooth loss and dementia risk are encouraged.

Late-onset cystic brain necrosis after radiotherapy for nasopharyngeal carcinoma.

BACKGROUND: Cystic brain radionecrosis (CBRN) is a late-onset devastating complication after radiotherapy for head and neck neoplasms, especially for nasopharyngeal carcinoma (NPC). To our knowledge, it has scarcely been reported. METHODS: We retrospectively reviewed all available medical records of NPC patients with CBRN who were treated with surgical intervention. RESULTS: Sixteen patients were identified in this study and the mean latency of CBRN was 9.2 ± 0.9 years. The total irradiation dose of the nasopharynx ranged from 60 to 78 Gy. Cyst-like lesions were observed and there were slightly enhancements on the cyst wall in five patients on patients' brain MRI. All the included patients underwent surgical resection of the cystic necrotic lesion thought temporal approach. Specimens from surgery revealed reactive gliosis and immunopositive cytokines including TNF-α, IL-6 and HIF-2α. Only one patient experienced recurrence and received reoperation after surgery. All the other patients made a good recovery and no operation-related mortality was observed. CONCLUSIONS: CBRN is a delayed but irreversible neurological sequel in irradiated NPC patients. Post-radiotherapy follow-up is quite necessary for those with high risk of CBRN. Proper treatment is needed for early CBRN patients to suppress inflammation in the brain. Timely neurosurgery may benefit patients with late-stage CBRN by alleviating increased intracranial pressure and inflammatory responses.

Radiation-induced brachial plexus injury after radiotherapy for nasopharyngeal carcinoma.

OBJECTIVE: Radiation-induced brachial plexus injury is a devastating complication that occurs after radiotherapy in the vicinity of the brachial plexus. Nasopharyngeal carcinoma, the most common type of cancer in Guangdong Province, is primarily treated with radiotherapy with subsequent side effects. However, radiation-induced brachial plexus injury is rarely reported in nasopharyngeal carcinoma. To draw attention to this correlation, we analyzed the clinical characteristics including the imaging findings of 10 patients suffering from radiation-induced brachial plexus injury for nasopharyngeal carcinoma. METHODS: We considered the patients' medical histories, analyzed their clinical characteristics, and monitored the long-term efficacy of treatment. RESULTS: The total irradiation dose of the nasopharynx ranged from 66.6 to 74 Gy, and that of the supraclavicular fossa ranged from 60 to 70 Gy. The mean latency was 8.2 ± 5.5 years. Seven patients initially complained of bilateral weakness, and three patients complained of isolated pain. The injuries of eight patients reached Grade 3 or worse. Magnetic resonance imaging showed a low signal on T1-weighted images and a high signal on short tau inversion recovery sequences in all cases. Swollen nerve fibers were clearly displayed in magnetic resonance diffusion tensor imaging. Electromyography showed myokymia in three patients. With conservative therapy, only one patient was temporarily relieved of pain, while the conditions of others were not ameliorated. CONCLUSIONS: Radiation-induced brachial plexus injury is a late but catastrophic complication in patients with nasopharyngeal carcinoma. Clinicians should be aware of radiation-induced brachial plexus injury when deciding on treatment and should give them regular follow-up post radiotherapy.

Minimal invasive and computer-assisted total knee replacement compared with the minimal invasive technique: a prospective, randomized trial with short-term outcomes.

INTRODUCTION: Up to now, no prospective, randomized comparisons between minimal invasive and computer-assisted total knee arthroplasty (MICA-TKA), and minimal invasive technique (MI-TKA) has been documented to evaluate not only clinical, but also radiologic results of the MICA-TKA. This prospective, randomized study was performed to compare the short-term results of MICA-TKA with minimal invasive technique MI-TKA for 6-month follow-up. PATIENTS AND METHODS: We reported the clinical and radiological results of 80 subjects who had cruciate-substituting, TKA-implanted primary total knee arthroplasties using either minimal invasive and computer-assisted technique (40 patients Group I) or minimal invasive technique (40 patients, Group II). Tourniquet time, length of skin incision, and total blood loss were compared. Knee society scores (KSSs), knee society functional scores (KSFSs), range of motion (ROM), and radiographic results were assessed and reported preoperatively and at 6-month follow-up. RESULTS: The accuracy of the implantations in relation to the coronal mechanical axis in Group I was superior to that of Group II (P < 0.05). The femoral rotational profile revealed the prosthesis in Group I that was implanted with significantly less internal rotation than in Group II. The average blood loss in patients of Group I was significantly reduced as compared to patients of Group II. No significant difference was detected in terms of tourniquet time or length of skin incision. Clinical results, with regard to ROMs and KSSs, as well as KSFSs were equally good in both the groups. CONCLUSIONS: Better alignment and similarity of good clinical results at short-term follow-up may provide subjects who receive MICA-TKA with long-term endurance of their implants. Further studies on longer-term outcomes and functional improvements are required to validate these possibilities.

Double-bundle versus single-bundle anterior cruciate ligament reconstructions: a prospective, randomized study with 2-year follow-up.

This prospective, randomized study was conducted to compare the short-term results of arthroscopic double-bundle with single-bundle anterior cruciate ligament (ACL) reconstruction. One hundred and eight patients with a symptomatic ACL rupture were randomized to either double-bundle (Group DB) or single-bundle (Group SB) ACL reconstruction. Follow-up was conducted at 6, 12, 18 and 24 months postoperatively. At the 24-month follow-up, 94 of the 108 patients (87%) were available for evaluation. The rotational stability, as evaluated by pivot shift test, was significantly superior in the Group DB to that in the Group SB. No significant difference with regard to ACL revisions, total flexion work, mean peak flexion torque and extension work between the groups was detected. There was no significant difference between the groups in terms of the Tegner activity score, the knee injury and osteoarthritis outcome score, the Lysholm functional score, anterior knee pain or mobility, subjective knee function. In addition, no significant difference in laxity on the Lachman test or the KT-1000 maximum manual force test was investigated. All the results were significantly more satisfactory at each follow-up period than preoperatively, in both groups. Both SB- and DB-ACL reconstruction resulted in satisfactory subjective outcome and objective stability. Both these techniques can therefore be considered as suitable alternatives for ACL reconstruction. Moreover, as it seems to be according to the pivot shift test, the risk for the development of degenerative changes of the knee joint in a long run could be smaller in the Group DB.

Arthroscopic versus mini-open rotator cuff repair: a prospective, randomized study with 24-month follow-up.

This prospective, randomized study was performed to evaluate the results of mini-open and arthroscopic rotator cuff repair in a comparative case series of patients followed for 24 months. A total of 125 patients were randomized to mini-open (Group I) or arthroscopic (Group II) rotator cuff repair at the time of surgical intervention. The University of California Los Angeles (UCLA) score, the American Shoulder and Elbow Surgeons (ASES) index, and muscle strength were measured to evaluate the clinical results, while magnetic resonance arthrography was used at 24-month follow-up to investigate the postoperative rotator cuff integrity. Fifty-three patients in Group I and 55 patients in Group II were available for evaluation at 24-month follow-up. At 24-month follow-up, the UCLA score, the ASES index, and muscle strength were statistically significantly increased in both groups postoperatively, while no significant difference was detected between the 2 groups. Intact rotator cuffs were investigated in 42 patients in Group I and 35 in Group II, and there was a significant difference in postoperative structural integrity between the two groups (P < 0.05). When analysis was limited to the patients with full-thickness tear, the muscle strength of the shoulder was significantly better in Group II, and the retearing rate was significantly higher in Group II. Based on the results obtained from this study, it can be indicated that arthroscopic and mini-open rotator cuff repair displayed substantially equal outcomes, except for higher retearing rate in the arthroscopic repair group. While for patients with full-thickness tear, arthroscopic rotator cuff repair displayed better shoulder strength and significantly higher retearing rate as compared to mini-open rotator cuff repair at 24-month follow-up.

Minimally invasive and computer-assisted total knee arthroplasty versus conventional technique: a prospective, randomized study.

We conducted a prospective, randomized study to compare the short-term results of minimally invasive and computer-assisted total knee arthroplasty (MICA-TKA) with those of conventional total knee arthroplasty (C-TKA) for 12-month follow-up. A total of 87 subjects who met the inclusion and exclusion criteria of the study were prospectively randomized consecutively into two groups: the C-TKA group (Group A, n = 44) and the MICA-TKA technique (Group B, n = 43). All the operations were performed by the same senior surgeon. Before surgery and at follow-up, patients were evaluated by the same observer. Tourniquet time as well as total blood loss was compared. Knee Society scores (KSSs), Knee Society functional scores (KSFSs), range of motion (ROM), and radiographic results were assessed and reported preoperatively and at 12-month follow-up. Of these patients, 82 (Group A 42; Group B 40) were available for 12-month evaluation. The two groups were found to be similar in terms of coronal mechanical axis. Similarly, the femoral rotational profile revealed that the prosthesis in Group A was implanted with similar internal rotation to Group B. The average blood loss in patients of Group B was significantly reduced as compared to patients of Group A. No significant difference was detected in terms of tourniquet time. Clinical results in Group B, with regard to ROMs and KSSs, as well as KSFSs were significantly superior to that in Group A. Based on the results obtained from this study, it is demonstrated that MICA-TKA leads to a similarly accurate restoration of leg alignment and component orientation compared to the C-TKA. Moreover, MICA-TKA produces superior clinical results to that of C-TKA. However, there is clearly a need for additional high-quality clinical trials with long-term follow-up to confirm the clinical benefits of MICA-TKA.

Mini-midvastus versus mini-medial parapatellar approach in total knee arthroplasty: a prospective, randomized study.

INTRODUCTION: To date, no English literature has evaluated the short-term results of the mini-medial parapatellar approach compared with the mini-midvastus approach. This prospective, randomized study was performed to compare the short-term results of total knee arthroplasty using either a mini-midvastus or a mini-medial parapatellar approach. PATIENTS AND METHODS: We reported the clinical and radiological results of 89 patients who had primary total knee arthroplasties with minimally invasive techniques using either a mini-midvastus or a mini-medial parapatellar approach. The mini-midvastus approach was used on 45 patients (group I) and a mini-medial parapatellar approach on 44 patients (group II). Skin incision length, tourniquet time, incidence of lateral retinacular release, total blood loss, straight leg raising time, visual analogy scale score, alignment of the knee, component position, and complication of each group were examined. Knee Society scores, range of motion were compared at 7 days, 6 weeks, 3 months, and 6 months postoperatively. RESULTS: The mean tourniquet time was 68 min in group I, significantly longer than 56 min for group II. However, comparisons of postoperative knee scores and function scores between both approaches did not yield a significant difference in outcome. No significant difference was found with respect to total blood loss, visual analogy scale score, straight-leg-raising test, range of motion or radiographic findings. CONCLUSION: Based on these results, we believe that the early results are similar between mini-midvastus and mini-medial parapatellar approach, ultimately the selection of the surgical approach will depend on the surgeon's experience and preference.

Clinical and radiographic results of Bryan cervical total disc replacement: 4-year outcomes in a prospective study.

INTRODUCTION: Early results have indicated that the Bryan cervical total disc replacement (TDR) favorably compares to anterior cervical decompression and fusion, while it is associated with fewer complications and higher levels of satisfaction. In this study, we sought to prospectively report the midterm outcomes of the Bryan TDR. PATIENTS AND METHODS: A total of 20 patients had performed their 4-year follow-up visit and had been assessed clinically and radiologically. Clinical outcomes (JOA, VAS, NDI, SF-36) and ROM measurements were investigated preoperatively and at 1 and 6 months, and 1, 2 and 4 years after operation. Complications were also investigated. Occurrences of heterotopic ossifications (HOs) and adjacent-level degeneration (ALD) radiographic changes were detected from 4-year follow-up X-rays. RESULTS: The mean JOA score, VAS score for arm and neck, NDI score and SF-36 score for PCS and MCS were reduced significantly at each postoperative time point when compared with the preoperative condition. The range of movement of the cervical spine, functional spinal unit, treated segment and the adjacent segment temporarily decreased at the early assessment, but all recovered to preoperative levels over a 6-month to 4-year time period. HO was evident in 6 of the 23 operated segments, which did not restrict the movement of the prosthesis. No obvious ALD was found on MRI. There were no cases of prosthesis migration, subsidence, loosening or wear. CONCLUSION: The midterm outcomes demonstrated that the Bryan TDR maintains favorable clinical and radiological results, with preservation of movement and satisfactory clinical outcome. There were no serious complications or cases of prosthetic wear or failure. The long-term benefits are yet to be examined.

Lnc-ANRIL modulates the immune response associated with NF-κB pathway in LPS-stimulated bovine mammary epithelial cells.

BACKGROUND: The antisense noncoding RNA in the INK4 locus (ANRIL) has been confirmed related to multiple disease progression, but the role and exact mechanisms of lnc-ANRIL in lipopolysaccharide (LPS)-induced inflammation of bovine mammary epithelial cells (MAC-T) remain unclear. AIMS: This manuscript focused on expounding the functional role of lnc-ANRIL through experiments performed in MAC-T. METHODS: At the in vitro level, we established a Bovine mammary epithelial cell (BMEC) cell model of mastitis by LPS treatment. Transfection of siRNA was examined by immunofluorescence localization and RT-qPCR. CCK8, clonogenic assay and EdU were used to detect the proliferation ability of the cells. Cell cycle and apoptosis were detected by flow cytometry and Western blot. The levels of inflammatory factors and oxidative stress markers were detected by ELISA kits. RESULTS: Cell Counting Kit-8, colony formation, and 5-ethynyl-20-deoxyuridine were adopted and the data illustrated that LPS could significantly suppress the cell proliferation, while knockdown of lnc-ANRIL expression obviously promoted MAC-T cell proliferation compared with LPS or LPS + si-NC group. Flow cytometry analysis demonstrated that lnc-ANRIL could induce MAC-T cell apoptosis. In addition, downregulation of lnc-ANRIL affected LPS-induced immune response by regulating inflammatory factor expressions and modulating the nuclear factor kappa B (NF-κB) axis in MAC-T cells. CONCLUSION: Our results suggest that lnc-ANRIL is involved in the regulation of cell proliferation, cell cycle, and cell apoptosis of MAC-T cells, and plays an important role in the inflammatory and immune response of MAC-T cells through the regulation of the NF-κB pathway, proposing new therapeutic strategies for the treatment of innate immune response-related disease such as bovine mastitis.

Taurine attenuates lipopolysaccharide-induced disfunction in mouse mammary epithelial cells.

The intent of this study was to evaluate the active defense reaction of mouse mammary epithelial cells and the cytoprotective and anti-inflammatory properties of taurine to lipopolysaccharide (LPS)-induced disfunction in mouse mammary epithelial cells. (1) Primary cultured mouse mammary epithelial cells were stimulated with LPS for 24 h (final concentration=0, 5, 10, 20 µg/mL). Western blotting demonstrated a significant decrease in the secretion of ß-casein in the 20 µg/mL LPS treatment group (P<0.05), while nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), lactoferrin (LF) and N-acetyl-ß-D-glucosaminidase (NAGase) were all significantly increased following LPS treatment (P<0.01). Furthermore, cell survival was significantly inhibited after treatment with 20 µg/mL LPS; however, neither 5 µg/mL nor 10 µg/mL LPS had any effect on cell survival. Therefore, a level of 10 µg/mL LPS was selected to test the protective effect of taurine on mouse mammary epithelial cells. (2) Primary cultured mouse mammary epithelial cells were treated with 0, 5, 15 or 45 mmol/L taurine for 3 h, followed by 10 µg/mL LPS for 24 h. Taurine significantly attenuated the LPS-induced increase in NAGase activity, NO concentrations and the level of TNF-α, IL-1ß, IL-6 and LF. Taurine at 45 mmol/L markedly increased ß-casein secretion in response to LPS-induced disfunction. This study demonstrated that the addition of taurine to a culture medium significantly inhibited the LPS-induced release of inflammatory factors and increased ß-casein secretion from mammary epithelial cells, thereby providing a possible explanation for the protective effect proposed for taurine in the prevention of LPS-induced disfunction in mammary epithelial cells.

Metabolic Landscape of Bronchoalveolar Lavage Fluid in Coronavirus Disease 2019 at Single Cell Resolution.

Abnormal function of immune cells is one of the key mechanisms leading to severe clinical symptoms in coronavirus disease 2019 patients, and metabolic pathways can destroy the function of the immune system by affecting innate and adaptive immune responses. However, the metabolic characteristics of the immune cells of the SARS-CoV-2 infected organs in situ remaining elusive. We reanalyzed the metabolic-related gene profiles in single-cell RNA sequencing data, drew the metabolic landscape in bronchoalveolar lavage fluid immune cells, and elucidated the metabolic remodeling mechanism that might lead to the progression of COVID-19 and the cytokine storm. Enhanced glycolysis is the most important common metabolic feature of all immune cells in COVID-19 patients. CCL2+ T cells, Group 2 macrophages with high SPP1 expression and myeloid dendritic cells are among the main contributors to the cytokine storm produced by infected lung tissue. Two metabolic analysis methods, including Compass, showed that glycolysis, fatty acid metabolism, bile acid synthesis and purine and pyrimidine metabolism levels of CCL2+ T cells, Group 2 macrophages and myeloid dendritic cells were upregulated and correlated with cytokine storms of COVID-19 patients. This might be the key metabolic regulatory factor for immune cells to produce large quantities of cytokines.

Dectin1 activation of ß-(1-3)/(1-6)-D-glucan produces an anti-mastitis effect in rats.

OBJECTIVE: A lipopolysaccharide (LPS)-induced mastitis model in rats was used to study the protective effect of ß-glucan. MATERIALS AND METHODS: On gestation day 10, ß-glucan was administered to rats daily by gavage at either 0 (control), 0.1 mg/kg BW, 1 mg/kg BW and 10 mg/kg BW until parturition. LPS or pyrogen-free physiological saline was inoculated into the mammary gland 72 h after parturition and the rats were euthanized at 12 h post-infection. RESULTS: LPS increased dectin1 and toll-like receptor 4 (TLR4) mRNA and protein expression significantly in mammary tissues. Tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, N-acetyl-ß-D-glucosaminidase (NAGase), inducible nitric oxide synthase (iNOs) in mammary tissues and serum, and myeloperoxidase (MPO) in mammary tissues were significantly increased 12 h after LPS infusion. ß-glucan administration could enhance dectin1 mRNA and protein expression while downregulating the expression of TLR4. Level of TNF-α, IL-1ß in mammary tissues and serum, MPO, NAGase and iNOs activity in mammary tissues was decreased, but the level of IL-2 in serum was increased by ß-glucan. CONCLUSION: The results indicate that ß-glucan may protect mammary tissue against LPS-induced rat acute mastitis.

The policy effect of green finance in promoting industrial transformation and upgrading efficiency in China: analysis from the perspective of government regulation and public environmental demands.

Industrial transformation and upgrading is an important step for China to carry out cleaner production and achieve the goal of carbon neutrality. With the help of green finance, we can optimize the allocation of financial resources and promote the optimization and upgrading of industrial structure. In order to explore the impact efficiency and policy effect of green finance on industrial transformation and upgrading in China, through the establishment of VAR model and super-efficiency DEA model, the relationship between green finance and industrial transformation and upgrading and its impact on efficiency are studied. In addition, the Tobit regression model is used to empirically test the impact of public environmental demands, government regulations, and their interaction terms on the efficiency of green finance in promoting industrial transformation and upgrading. The results show that the overall efficiency of green finance in promoting industrial transformation and upgrading is high, but it shows a downward trend, which may be due to the unbalanced development of the green financial system, information asymmetry, and the absence of enterprises from regulation. The impact coefficient of input-oriented government regulation is significantly negative, which indicates that increasing the investment in pollution control may encourage polluting enterprises' emission behavior to obtain benefits. The impact of performance-oriented government regulation and public environmental demands on efficiency is not significant, which indicates that the supervision and binding force of enterprises are insufficient. Public environmental demands and input government regulations have significant synergistic governance advantages, indicating that the government should enhance the regulatory intensity of differentiated policies. The study provides a reference for the government to enhance the intensity of policy regulation and establish a diversified environmental governance system.

miR-142-5p regulates lipopolysaccharide-induced bovine epithelial cell proliferation and apoptosis via targeting BAG5.

Bovine mastitis is a threat to the health of the dairy cow. MicroRNAs (miRs) serve an important role in the progression of bovine mastitis, regulating immune and defense responses. The present study aimed to investigate the possible effects and mechanisms of bovine mastitis underlying miR-142-5p and Bcl-2 associated athanogene 5 (BAG5) in in vitro lipopolysaccharide (LPS)-induced models. Reverse transcription-quantitative PCR and western blotting were performed to determine mRNA and protein expression levels, respectively. ELISAs were conducted to assess the levels of cytokines and an immunofluorescence assay was performed to determine the expression of BAG5. Cell Counting Kit-8, clone formation and 5-ethynyl-2'-deoxyuridine assays were conducted to determine cell viability and proliferation of bovine mammary epithelial MAC-T cells, respectively. Flow cytometry was performed to measure MAC-T cell cycle distribution and apoptosis, and a luciferase assay was conducted to verify whether BAG5 was a target of miR-142-5p. The results indicated that miR-142-5p was upregulated in MAC-T cells treated with LPS compared with the control group. miR-142-5p mimics transfection significantly activated the cytokines TNF-α, IL-1ß, IL-6 and IL-8, and significantly increased the expression levels of NF-κB signaling pathway-related proteins in LPS-treated cells. The luciferase activity of MAC-T cells treated with miR-142-5p mimics and BAG5 3'untranslated region wild type decreased, compared with mutant type. By contrast, BAG5 overexpression significantly downregulated the levels of cytokines, including TNF-α, IL-1ß, IL-6 and IL-8, in LPS-treated cells. BAG5 overexpression significantly promoted cell proliferation and viability, decreased apoptosis, and regulated Caspase-3, Caspase-9, Bcl-2 and Bax expression in LPS-treated MAC-T cells, which was significantly reversed by transfection with miR-142-5p mimics. In conclusion, the results of the present study suggested that miR-142-5p may promote the progression of bovine mastitis via targeting BAG5. Therefore, the present study provided the foundations for future investigations.

Sevoflurane Inhibited Osteosarcoma Cell Proliferation And Invasion Via Targeting miR-203/WNT2B/Wnt/ß-Catenin Axis.

BACKGROUND: Osteosarcoma is one of the most common primary bone cancers with predominant occurrence in children and adolescents. This study aimed to determine the effects of sevoflurane treatment on the osteosarcoma progression and to explore the underlying molecular mechanisms. MATERIALS AND METHODS: The mRNA and protein expression levels were determined by qPCR and Western blot, respectively. Osteosarcoma cell proliferation, apoptosis and invasion were determined by MTT, caspase-3 activity, colony formation and Transwell invasion assays, respectively. The interaction between miR-203 and WNT2B 3' untranslated region was confirmed by luciferase reporter assay. RESULTS: Sevoflurane treatment for 6 hrs concentration-dependently suppressed cell viability, increased caspase-3 activity and up-regulated miR-203 expression in both U2OS and MG63 cells. MiR-203 overexpression suppressed cell viability, increased caspase-3 activity and suppressed cell growth and invasion of osteosarcoma cells. In addition, miR-203 knockdown attenuated the tumor-suppressive effects of sevoflurane treatment on osteosarcoma cells. Mechanistic studies showed that miR-203 repressed the expression of WNT2B in U2OS cells, and inhibition of miR-203 attenuated the suppressive effects of sevoflurane on WNT2B expression. More importantly, WNT2B overexpression attenuated the effects of sevoflurane treatment on cell viability, caspase-3 activity, cell growth and invasion of U2OS cells. MiR-203 overexpression suppressed Wnt/ß-catenin signalling. Similarly, sevoflurane suppressed the activity of Wnt/ß-catenin signalling, which was partially reversed by miR-203 knockdown and WTN2B overexpression. CONCLUSION: Our data showed the tumor-suppressive effects of sevoflurane on osteosarcoma cells, and mechanistic studies revealed that sevoflurane inhibited osteosarcoma cell proliferation and invasion partly via targeting the miR-203/WNT2B/Wnt/ß-catenin axis.

Evaluation of the changes of immune cells during lipopolysaccharide-induced mastitis in rats.

The aim of this study was to evaluate in rats, changes in peripheral blood immune cells and mammary tissue after an intramammary infusion of lipopolysaccharide (LPS). The results of the study showed that infusion of LPS induced a rapid migration of neutrophils (PMNs) from the blood to mammary alveoli, increased the activity of myeloperoxidase (MPO) and the concentration of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) in mammary tissues, decreased the activity of myeloperoxidase in serum and reduced the CD4+/CD8+ ratio. This is the first report of changes in peripheral blood immune cells and mammary tissue in rat mastitis.

Graphene quantum dots conjugated neuroprotective peptide improve learning and memory capability.

Alzheimer disease (AD) is a neurodegenerative disorder and the most common form of dementia. Histopathologically is characterized by the presence extracellular neuritic plaques and with a large number of neurons lost. In this paper, we design a new nanomaterial, graphene quantum dots (GQDs) conjugated neuroprotective peptide glycine-proline-glutamate (GQDG) and administer it to APP/PS1 transgenic mice. The in vitro assays including ThT and CD proved that GQDs and GQDG could inhibit the aggregation of Aß1-42 fibrils. Morris water maze was performed to exanimate learning and memory capacity of APP/PS1 transgenic mice. The surface area of Aß plaque deposits reduced in the GQDG group compared to the Tg Ctrl groups. Furthermore, newly generated neuronal precursor cell and neuron were test by immunohistochemical. Besides, neurons were impregnated by DiI using gene gun to show dendritic spine. Results indicated enhancement of learning and memory capacity and increased amounts of dendritic spine were observed. Inflammation factors and amyloid-ß (Aß) were tested with suspension array and ELISA, respectively. Several pro-inflammatory cytokines (IL-1α, IL-1ß, IL-6, IL-33, IL-17α, MIP-1ß and TNF-α) had decreased in GQDG group compared with Control group. Reversely, anti-inflammatory cytokines (IL-4, IL-10) had increased in GQDG group compared with Control group. Thus, we demonstrate that the GQDG is a promising drug in treatment of neurodegenerative diseases such as AD.

PI3K/AKT/mTOR/p70S6K Pathway Is Involved in Aß25-35-Induced Autophagy.

Disruption or deregulation of the autophagy system has been implicated in neurodegenerative disorders such as Alzheimer's disease (AD). Aß plays an important role in this autophagic system. In many cases, autophagy is regulated by the phosphatidylinositol 3-phosphate kinase/AKT/mammalian target of rapamycin/p70 ribosomal protein S6 kinase (PI3K/AKT/mTOR/p70S6K) signaling pathway. However, whether this signaling pathway is involved in Aß-induced autophagy in neuronal cells is not known. Here, we studied whether Aß25-35 induces autophagy in HT22 cells and C57 mice and investigated whether PI3K is involved in the autophagy induction. We found that Aß25-35 inhibited HT22 cell viability in a dose- and time-dependent manner. Aß25-35 induced autophagosome formation, the conversion of microtubule-associated protein light chain 3 (LC3), and the suppression of the mTOR pathway both in vitro and in vivo. Furthermore, Aß25-35 impaired the learning abilities of C57 mice. Our study suggests that Aß25-35 induces autophagy and the PI3K/AKT/mTOR/p70S6K pathway is involved in the process, which improves our understanding of the pathogenesis of AD and provides an additional model for AD research.