Baicalin inhibits IL-1ß-induced ferroptosis in human osteoarthritis chondrocytes by activating Nrf-2 signaling pathway.

BACKGROUND: Osteoarthritis (OA) is a common degenerative disease involving articular cartilage, in which ferroptosis of chondrocytes plays an important role. Baicalin (BAI) exerts regulatory effects in a wide range of orthopedic diseases including OA, but its effect on ferroptosis of chondrocytes (CHs) is still unclear. The purpose of this study was to determine the effect of BAI on ferroptosis in human OA chondrocytes (OACs), and to explore its possible mechanism. METHODS: CHs were treated with IL-1ß (10 ng/mL) to simulate inflammation in vitro. Immunofluorescence, quantitative RT-PCR, Western blotting and cell viability assay were performed to evaluate the impacts of BAI on Fe2+ level, mitochondrial dysfunction, ferroptosis-related proteins, oxidative stress and cytotoxicity in CHs. Additionally, siRNA was made use of to knock out nuclear factor E2-related factor 2 (Nrf2) to analyze the role played by Nrf2 in BAI-induced CH ferroptosis. RESULTS: BAI eliminated IL-1ß-induced Fe2+ accumulation, changes in mitochondrial membrane potential and ferroptosis-related protein GPX4, SLC7A11, P53 and ACSL4 levels, as well as reactive oxygen species (ROS), lipid peroxidation (LPO) and malondialdehyde (MDA) accumulation in CHs. Besides, BAI reversed IL-1ß-induced decrease of Collagen II and increase of MMP13 in CHs. Meanwhile, BAI attenuated IL-1ß-induced CH toxicity and promoted Nrf2 antioxidant system activation. When Nrf2 was knocked down by siRNA, the effects of BAI on IL-1ß-induced ferroptosis-related proteins and antioxidant stress in CHs were significantly weakened. CONCLUSIONS: This study demonstrates that IL-1ß can induce CH ferroptosis. BAI is able to inhibit IL-1ß-induced CH ferroptosis and ECM degradation, and the specific mechanism may be that it can inhibit IL-1ß-induced CH ferroptosis by activating Nrf2 antioxidant system to attenuate the accumulation of intracellular ROS and lipid ROS.

Comparison of the clinical efficacy of three cannulated screws with parallel distribution and inverted triangular distribution in the treatment of femoral neck fractures in the elderly.

The aim of the present study was to investigate the clinical efficacy of three cannulated screws with parallel distribution in comparison with the 'gold standard' of inverting three cannulated screws with triangular distribution, in the treatment of femoral neck fractures in the elderly. A total of 106 elderly patients with femoral neck fractures treated at the Department of Orthopedics of the First Affiliated Hospital of Nanjing Medical University (Jiangsu Provincial Hospital) from October 2018 to March 2020 were retrospectively analyzed and divided into groups A and B. Group A consisted of 51 patients with a mean age of 73.3±6.8 years; and group B consisted of 55 patients with a mean age of 74.5±7.3 years. Group A was treated with closed reduction + inverted triangular distribution of three cannulated screws, while group B was treated with closed reduction + parallel distribution of three cannulated screws, and the general surgical conditions, including fracture reduction quality, fracture healing, incidence of avascular necrosis of the femoral head, and functional recovery scale (FRS) score of hip joint function were assessed in both groups. All patients were followed up for an average of 14.8 months. Compared with group A, group B had significant advantages in operation time, number of times intraoperative fluoroscopy was performed, number of intraoperative guide wire adjustments, and proportion of postoperative referrals to the intensive care unit (P<0.05). There were no significant differences in fracture reduction quality, fracture healing, incidence of avascular necrosis of the femoral head, and FRS score of hip joint function between groups A and B (P>0.05). For elderly patients with femoral neck fractures, distributing three cannulated screws in parallel after closed reduction achieved similar clinical efficacy to 'gold standard' inverted triangular distribution, and had obvious advantages in operation time, with significantly reduced surgical difficulty. This procedure is therefore deemed worthy of promotion and clinical application in the primary hospitals of China.

Under flooding conditions, controlled-release fertiliser coated microplastics affect the growth and accumulation of cadmium in rice by increasing the fluidity of cadmium and interfering with metabolic pathways.

The combined pollution of microplastics (MPs) and Cd can affect plant growth and development and Cd accumulation, with most studies focusing on dryland soil. However, the effects of polyurethane (PU) controlled-release fertiliser coated MPs (PU MPs), which widely exist in rice systems, coupled with Cd on plant growth and Cd accumulation under flooding conditions are still unknown. Therefore, in the present study, in situ techniques were used to systematically study the effects of PU MPs and Cd coupling on the physiological and biochemical performance, metabolomics characteristics, rhizosphere bacterial community, and Cd bioavailability of rice in different soil types (red soil/cinnamon soil). The results showed that the effects of PU MPs on rice growth and Cd accumulation were concentration-dependent, especially in red soil. High PU concentration (1 %) inhibited rice root growth significantly (44 %). The addition of PU MPs inhibited photosynthetically active radiation, net photosynthesis, and transpiration rate of rice, mainly with low concentration (0.1 %) in red soil and high concentration (1 %) in cinnamon soil. PU MPs can enhance the expression of Cd resistance genes (cadC and copA) in soil, enhance the mobility of Cd, and affect the metabolic pathways of metabolites in the rhizosphere soil (red soil: fatty acid metabolism; cinnamon soil: amino acid degradation, heterobiodegradation, and nucleotide metabolism) to promote Cd absorption in rice. Especially in red soil, Cd accumulation in the root and aboveground parts of rice after the addition of high concentration PU (1 %) was 1.7 times and 1.3 times, respectively, that of the control (p < 0.05). Simultaneously, microorganisms can affect rice growth and Cd bioavailability by affecting functional bacteria related to carbon, iron, sulfur, and manganese. The results of the present study provide novel insights into the potential effects of PU MPs coupled with Cd on plants, rhizosphere bacterial communities, and Cd bioavailability.

[A multi-center observation of the therapeutic efficacy of Bencycloquidium bromide in the treatment of seasonal allergic rhinitis with predominant symptoms of rhinorrhea].

Objective:To observe the efficacy and safety of the M receptor antagonist Bencycloquidium bromide nasal spray in treatment of seasonal allergic rhinitis with runny nose as the main symptom. Methods:From August 2021 to September 2021, 134 patients with seasonal allergic rhinitis were enrolled in the otolaryngology Outpatient Department of Peking University Third Hospital, First Affiliated Hospital of Harbin Medical University and China-Japanese Friendship Hospital of Jilin University, including 71 males and 63 females, with a median age of 38 years. TNSS score and visual analogue scale（VAS） of total nasal symptoms were observed during 2 weeks of treatment with Bencycloquidium bromide nasal spray. Results:TNSS score decreased from （8.89±3.31） on day 0 to （3.71±2.51） on day 14（P<0.001）, VAS score of nasal symptoms decreased from （24.86±7.40） on day 0 to （6.84±5.94） on day 14（P<0.001）, VAS score of rhinorrhoea decreased from （6.88±2.06） on day 0 to （1.91±1.81） on day 14（P<0.001）. Rhinoconjunctivitis quality of life questionnaire（RQLQ） score decreased from （94.63±33.35） on day 0 to （44.95±32.28） on day 14（P<0.001）. The incidence of adverse reaction was low and no serious adverse events occurred during the whole experiment. Conclusion:Bencycloquidium bromide nasal spray has significant efficacy and good safety in the treatment of seasonal allergic rhinitis.

Pretreatment fibrinogen levels are associated with survival outcome in patients with cancer using immunotherapy as a second­line treatment.

The present study aimed to investigate the predictive value of pretreatment fibrinogen (FIB) levels in patients with cancer who received immunotherapy as a second-line treatment. A total of 61 patients with stage III-IV cancer were included. The cut-off value of FIB for predicting overall survival (OS) was determined by receiver operating characteristic curve analysis. The prognostic value of pretreatment FIB on progression-free survival (PFS) and OS was determined by univariate and multivariate analyses. Based on a cut-off point of 3.47 g/l, patients were divided into low pretreatment FIB (<3.47 g/l) and high pretreatment FIB (≥3.47 g/l) groups. A high pretreatment FIB level was more common in older patients (P=0.03). Kaplan-Meier analysis showed that patients with high pretreatment FIB levels had shorter PFS and OS times than patients with low FIB levels (P<0.05). In multivariate analysis, pretreatment FIB was an independent prognostic factor for OS [hazard ratio (HR), 6.06; 95% CI, 2.01-18.28; P<0.01] and OS from the initiation of second-line treatment (HR, 3.69; 95% CI, 1.28-10.63; P=0.02). Overall, FIB is associated with survival outcome in patients with cancer who are administered immunotherapy as a second-line treatment.

Molecular mechanism of XB130 adaptor protein mediates trastuzumab resistance in gastric cancer

Background Recent studies have shown that the activation of PI3K/AKT signaling pathway is an essential molecular mechanism participating in trastuzumab resistance in HER2 + GC (gastric cancer). However, how can we effectively inhibit AKT activity associated with drug resistance during trastuzumab treatment? Screening inhibitors against the upstream receptors of PI3K/AKT signaling pathway or interacting proteins of members has become an important way. Methods In this study, western blot, qRT-PCR, CCK8, Co-IP and other techniques were used to explore possible mechanisms participating in trastuzumab resistance in vitro. Besides, the xenograft mouse model and GC tissue samples from patients were used to further validate the in-vitro results. Results The expression of XB130 adaptor protein was remarkably increased in GC cell lines resistant to trastuzumab, and knockdown of XB130 could reverse the resistance via downregulating p-AKT. In addition, p-SRC (Tyr416) was increased in resistant cells, which could facilitate the binding of XB130 to PI3K p85α. It was also discovered that XB130 could negatively regulate PTEN gene transcription, and thus a positive feedback loop was formed between SRC-XB130-PTEN. Conclusions In HER2 + GC, XB130 contributes to trastuzumab resistance by stimulating the PI3K/AKT signaling pathway through binding to PI3K p85α under the mediation of SRC kinase and regulating PTEN gene transcription, and in turn forming a positive feedback loop between SRC-XB130-PTEN (AU)

The N6-methyladenosine METTL3 regulates tumorigenesis and glycolysis by mediating m6A methylation of the tumor suppressor LATS1 in breast cancer.

BACKGROUND: Posttranscriptional modification of tumor-associated factors plays a pivotal role in breast cancer progression. However, the underlying mechanism remains unknown. M6A modifications in cancer cells are dynamic and reversible and have been found to impact tumor initiation and progression through various mechanisms. In this study, we explored the regulatory mechanism of breast cancer cell proliferation and metabolism through m6A methylation in the Hippo pathway.  METHODS: A combination of MeRIP-seq, RNA-seq and metabolomics-seq was utilized to reveal a map of m6A modifications in breast cancer tissues and cells. We conducted RNA pull-down assays, RIP-qPCR, MeRIP-qPCR, and RNA stability analysis to identify the relationship between m6A proteins and LATS1 in m6A regulation in breast cancer cells. The expression and biological functions of m6A proteins were confirmed in breast cancer cells in vitro and in vivo. Furthermore, we investigated the phosphorylation levels and localization of YAP/TAZ to reveal that the activity of the Hippo pathway was affected by m6A regulation of LATS1 in breast cancer cells.  RESULTS: We demonstrated that m6A regulation plays an important role in proliferation and glycolytic metabolism in breast cancer through the Hippo pathway factor, LATS1. METTL3 was identified as the m6A writer, with YTHDF2 as the reader protein of LATS1 mRNA, which plays a positive role in promoting both tumorigenesis and glycolysis in breast cancer. High levels of m6A modification were induced by METTL3 in LATS1 mRNA. YTHDF2 identified m6A sites in LATS1 mRNA and reduced its stability. Knockout of the protein expression of METTL3 or YTHDF2 increased the expression of LATS1 mRNA and suppressed breast cancer tumorigenesis by activating YAP/TAZ in the Hippo pathway. CONCLUSIONS: In summary, we discovered that the METTL3-LATS1-YTHDF2 pathway plays an important role in the progression of breast cancer by activating YAP/TAZ in the Hippo pathway.

Molecular mechanism of XB130 adaptor protein mediates trastuzumab resistance in gastric cancer.

BACKGROUND: Recent studies have shown that the activation of PI3K/AKT signaling pathway is an essential molecular mechanism participating in trastuzumab resistance in HER2 + GC (gastric cancer). However, how can we effectively inhibit AKT activity associated with drug resistance during trastuzumab treatment? Screening inhibitors against the upstream receptors of PI3K/AKT signaling pathway or interacting proteins of members has become an important way. METHODS: In this study, western blot, qRT-PCR, CCK8, Co-IP and other techniques were used to explore possible mechanisms participating in trastuzumab resistance in vitro. Besides, the xenograft mouse model and GC tissue samples from patients were used to further validate the in-vitro results. RESULTS: The expression of XB130 adaptor protein was remarkably increased in GC cell lines resistant to trastuzumab, and knockdown of XB130 could reverse the resistance via downregulating p-AKT. In addition, p-SRC (Tyr416) was increased in resistant cells, which could facilitate the binding of XB130 to PI3K p85α. It was also discovered that XB130 could negatively regulate PTEN gene transcription, and thus a positive feedback loop was formed between SRC-XB130-PTEN. CONCLUSIONS: In HER2 + GC, XB130 contributes to trastuzumab resistance by stimulating the PI3K/AKT signaling pathway through binding to PI3K p85α under the mediation of SRC kinase and regulating PTEN gene transcription, and in turn forming a positive feedback loop between SRC-XB130-PTEN.

[Community structure and species composition of typical Quercus variabilis natural secondary forest at the northern foothills of the Qinling Mountains, China]. / ç§¦å²­åŒ—éº“å ¸åž‹æ “çš®æ Žå¤©ç„¶æ¬¡ç”Ÿæž—ç¾¤è½ç»“æž„ä¸Žç‰©ç§ç»„æˆ.

We investigated species composition and community structure of a typical Quercus variabilis natural secondary forest in the northern foothills of the Qinling Mountains, within the dynamic monitoring plot of deciduous broad-leaved forest at the Louguantai experimental forest farm in Zhouzhi County, Shaanxi Province. The results showed that there were 3162 individual woody plants with diameter at breast height ≥1 cm in the plot, which were belonged to 42 species, 36 genera, and 25 families. The community genus's areal type was dominated by the temperate component, which accounted for 44.4%, and followed by the tropical component. The community was dominated by several tree species. The top three species with respect to importance value were Q. variabilis, Pinus tabuliformis, and Quercus aliena, with the sum of their importance value being 64.7%. The average DBH of all woody plants was 7.58 cm. The distribution of all individuals and dominant species in the tree layer was approximately normal, with more medium-size individuals. The community structure was stable. The community was poorly renewed, with a trend of population decline. Biodiversity indices varied considerably among different plots, being lower than those of subtropical broad-leaved evergreen forests. There was a significant correlation between community species distribution and environmental factors. Soil and topography explained 42.4% of the variation in community distribution. Altitude and soil alkali hydrolysable nitrogen had a significant effect on community distribution. Altitude, soil total phosphorus, and organic matter content significantly affected the species diversity of Q. variabilis communities. The stronger adaptability of Q. variabilis populations allowed them to become dominant in low-nutrient environments, which limited species diversity in the community.

[Short-term effectiveness of staged management in treatment of complex tibial plateau fracture with severe soft tissue injury].

Objective: To evaluate short-term effectiveness of staged management for complex tibial plateau fracture with severe soft tissue injury. Methods: A clinical data of 12 patients with complex tibial plateau fractures and severe soft tissue injuries between July 2017 and March 2021 and met the selection criteria was retrospectively analyzed. There were 7 males and 5 females with an average age of 43.1 years (range, 33-58 years). All patients were traffic accident injuries and admitted to hospital within 24 hours after injury. The tibial plateau fractures were closed fractures. According to the Schatzker classification standard, the fractures were rated as type Ⅳ in 3 cases, type Ⅴ in 4 cases, and type Ⅵ in 5 cases. According to the Tscherne classification standard, the soft tissue injuries were rated as grade Ⅱ in 4 cases and grade Ⅲ in 8 cases. The treatment of all patients was divided into 3 stages. In the first stage, emergency trans-articular fracture fixation with external fixator was performed; in the second stage, the fracture reduction and internal fixation were performed and bone cement was implanted to fill the bone defect; in the third stage, the bone cement was removed and the bone graft was performed to repair defect. All patients performed joint function exercise after operation as early as possible. Results: There was no neurological symptom after all staged managements, the incisions healed by first intention, and no complications such as incision infection or necrosis occurred. All patients were followed up 6-32 months (mean, 16.9 months). The fractures were all anatomical reduction confirmed by the X-ray films after operation. During follow-up, there was no obvious loss of reduction, loosening and rupture of internal fixator, or collapse of the articular surface. All fractures healed after 14-20 weeks (mean, 17.6 weeks). The posterior slope angle of the tibial plateau was (9.7±2.3)° and the varus angle was (3.9±1.9)° immediately after bone grafting, and were (8.5±2.9)° and (4.3±1.9)° respectively at 6 months after operation. There was no significant difference between the two time points ( t=0.658, P=0.514; t=-1.167, P=0.103). At last follow-up, the Hospital for Special Surgery (HSS) score was 85-96 (mean, 91.2), and the range of motion of knee was 110°-135° (mean, 120.9°). Conclusion: The staged management for complex tibial plateau fracture with severe soft tissue injury can obtain good short-term effectiveness, but the long-term effectiveness needs to be further followed up.

[Community structure of phyllosphere fungi associated with dominant tree species in a broad-leaved Korean pine forest of Changbai Mountain, Northeast China]. / é•¿ç™½å±±é˜”å¶çº¢æ¾æž—ä¼˜åŠ¿æ ‘ç§å¶é™ çœŸèŒç¾¤è½ç»“æž„.

Forest is the main component of terrestrial ecosystems that harbors about 40% of the existing species on the earth. As a vital component of biodiversity, phyllosphere microbes in the canopy play a critical and unique role in maintaining plant health, improving host resistance, and influencing global biogeochemical cycle. However, the studies on the community structure of phyllosphere fungi in natural forests are scarce as compared to that on rhizosphere microbes. Consequently, we know litter about how phyllosphere fungi associates with leaf traits. In this study, we analyzed fungal community composition of canopy leaves of six dominant tree species (i.e., Pinus koraiensis, Tilia amurensis, Quercus mongolica, Acer mono, Fraxinus mandshurica, and Ulmus japonica), in a broad-leaved Korean pine forest of Changbai Mountain Nature Reserve in Jilin Province, using high-throughput sequencing. We compared the differences of phyllosphere fungal community structure and functional groups of different dominant tree species. Moreover, 14 key leaf functional traits of their host trees were measured to investigate the relationships between fungal community composition and leaf functional traits. We found that the dominant phyla and class of phyllosphere fungi were Ascomycota and Basidiomycota, and Dothideomycetes and Taphrinomycetes, respectively. Results of LEfSe analysis indicated that all the tree species except Ulmus japonica had significant biomarkers, such as the Eurotiomycetes of Pinus koraiensis and the Ascomycetes of Quercus mongolica. The main functional groups of phyllosphere fungi were pathotroph. The results of redundancy and envfit analysis showed that functional traits related to plant nutrient acquisition as well as resistance to diseases and pests were the main factors influencing the community structure of phyllosphere fungi.

Soil bacterial community and metabolism showed a more sensitive response to PBAT biodegradable mulch residues than that of LDPE mulch residues.

Biodegradable mulch film (BDM) is considered as an environmentally sustainable alternative to low density polyethylene (LDPE) mulch film. However, the low degradation rate of BDM resulted in residues in soil after service period which were similar to LDPE mulch film. Distinguishing the differential responses of crop growth, soil bacteria and metabolism to residues of BDM and LDPE mulch films is favourable for comparing the environmental toxicities of the two materials. The results indicated that emergence rate and yield of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino) were significantly inhibited by two types mulch residues. BDM residues significantly decreased bacterial diversity by 1.2-2.3% through the enrichment of dominant phyla and inhibition of inferior phyla, while LDPE mulch residues not. The effects of BDM residues on soil metabolite spectrum were stronger than LDPE mulch residues with significant increase (3.9% 5.8%) in the abundance of total metabolites. Besides the pathways of metabolism, organismal systems, environmental information processing influenced by LDPE mulch resides, differential pathways including human diseases and cellular processes were also determined in soil with BDM residues. According to all the results of the present study, prior to the promotion of BDM, its influences on soil safety must be carefully investigated through critical and systematic research.

Baseline serum uric acid level is associated with progression-free survival, disease control rate, and safety in postoperative patients with colorectal cancer treated by FOLFOX, FOLFIRI, or XELOX.

Background: High serum uric acid (SUA) levels increase the risk of overall cancer morbidity and mortality, particularly for digestive malignancies. Nevertheless, the correlation between SUA level and clinical outcomes of the postoperative patients with colorectal cancer (CRC) treated by chemotherapy is unclear. This study aimed at exploring the relationship between baseline SUA level and progression-free survival (PFS), disease control rate (DCR), and safety in postoperative CRC patients receiving chemotherapy. Patients and Methods: We conducted a retrospective study to evaluate the relationship between baseline SUA level and PFS, DCR, and incidence of serious adverse events of 736 postoperative CRC patients treated with FOLFOX, FOLFIRI or XELOX at our center. Results: Data from our center suggested that high baseline SUA level is linked to poor PFS in non-metastatic CRC patients using FOLFOX (HR=2.59, 95%CI: 1.29-11.31, p=0.018) and in male patients using FOLFIRI (HR=3.77, 95%CI: 1.57-39.49, p=0.012). In patients treated by FOLFIRI, a high SUA is also linked to a low DCR (p=0.035). In patients using FOLFOX, high baseline SUA level is also linked to a high incidence of neutropenia (p=0.0037). For patients using XELOX, there is no significant correlation between SUA level and PFS, effectiveness, or safety. Conclusions: These findings imply that a high SUA level is a promising biomarker associated with poor PFS, DCR and safety of postoperative CRC patients when treated with FOLFOX or FOLFIRI.

YAP knockdown in combination with ferroptosis induction increases the sensitivity of HOS human osteosarcoma cells to pyropheophorbide-α methyl ester-mediated photodynamic therapy.

BACKGROUND AND AIMS: This study was designed to explore the effects of Yes-associated protein (YAP) knockdown on human osteosarcoma (HOS) cell sensitivity to Pyropheophorbide-α methyl ester-mediated photodynamic therapy (MPPa-PDT), and to assess how YAP silencing in combination with treatment with the ferroptosis inducer Erastin improves HOS cell sensitivity to MPPa-PDT in an effort to better clarify the molecular mechanisms underlying these phenotypes. METHODS: At 12 h post-MPPa-PDT, Hoechst staining and flow cytometry were conducted to evaluate the apoptotic death of HOS cells. The expression of YAP in these cells at 12 h post-MPPa-PDT treatment was assessed via Western blotting and immunofluorescent staining. BODIPY581/591-C11 was used to evaluate lipid peroxidation. Following shYAP lentiviral transduction, Western blotting was conducted to assess the expression of proteins associated with proliferation, apoptosis, and ferroptosis. EdU assays and clonogenic assays were performed to analyze cellular proliferation. Erastin-treated HOS cells were used to establish a ferroptosis model. Western blotting was used to measure ferroptosis-associated protein levels following shYAP and erastin treatment, while changes in proliferation and MDA levels in each group were examined using an MDA kit. RESULTS: At 12 h post-MPPa-PDT, HOS cells exhibited apoptotic characteristics including nuclear fragmentation and pyknosis, with concomitant increases in apoptosis-associated proteins as detected via Western blotting and apoptotic induction as measured via flow cytometry. Phosphorylated YAP levels fell and non-phosphorylated YAP levels rose following such treatment. Transfection with shYAP was successful as a means of generating stable HOS cell lines, and Western blotting analyses of these cells revealed reductions in proteins associated with cellular proliferation together with the upregulation of apoptosis-related proteins.  MDA assays indicated that erastin combined with YAP knockdown enhanced the sensitivity of HOS cells to MPPa-PDT treatment. CONCLUSIONS: These data indicate that ferroptosis and YAP knockdown can enhance osteosarcoma cell sensitivity to MPPa-PDT therapy.

Overexpression of miR-328-5p influences cell growth and migration to promote NSCLC progression by targeting LOXL4.

Background: Lung cancer is the leading cause of cancer-associated mortality worldwide, and most lung cancers are classified as non-small cell lung cancer (NSCLC). MiR-328 influence the progression of multiple tumors, but the role of miR-328-5p in NSCLC has not been elucidated. The aim of this study was to illuminate the oncogenic role and potential molecular mechanisms of the miR-328-5p and lysyl oxidase like 4 (LOXL4) in NSCLC. Methods: Expression of miR-328-5p was detected by real-time quantitative polymerase chain reaction (qRT-PCR) in tumor and non-tumor adjacent tissues. After Lentivirus-miR-328-5p was employed to intervene this miRNA in NSCLC cell lines, RT-qPCR was used to detect the expression levels of miR-328-5p. Cell Counting Kit-8 (CCK-8), cell colony formation, flow cytometry, wound healing, Transwell assays were used to determine the malignant phenotypes of NSCLC cells. Nude mice models of subcutaneous tumors were established to observe the effect of miR-328-5p on tumorigenesis. Targeting the 3'UTR of LOXL4 by miR-328-5p was verified by integrated analysis including transcriptome sequencing, dual-luciferase and western-blot assays. Results: High miR-328-5p level was observed in NSCLC cells from The Cancer Genome Atlas (TCGA) database and tumor tissues collected from NSCLC patients. Overexpressed miR-328-5p promoted NSCLC cell proliferation, survival, and migration, and promoted tumor growth in vivo. Knockdown of miR-328-5p suppressed tumorigenic activities. Transcriptome sequencing analysis revealed that LOXL4 was downregulated by miR-328-5p, which was confirmed by dual-luciferase reporter and western-blot assays. Conclusions: miR-328-5p showed targeted regulation of LOXL4 to promote cell proliferation and migration in NSCLC.

Kindlin-2 Promotes Chondrogenesis and Ameliorates IL-1beta-Induced Inflammation in Chondrocytes Cocultured with BMSCs in the Direct Contact Coculture System.

The osteoarthritis caused by trauma or inflammation is associated with severe patient morbidity and economic burden. Accumulating studies are focusing on the repair of articular cartilage defects by constructing tissue-engineered cartilage. Recent evidence suggests that optimizing the source and quality of seed cells is one of the key points of cartilage tissue engineering. In this study, we demonstrated that Kindlin-2 and its activated PI3K/AKT signaling played an essential role in promoting extracellular matrix (ECM) secretion and ameliorating IL-1beta-induced inflammation in chondrocytes cocultured with bone marrow stem cells (BMSCs). In vivo experiments revealed that coculture significantly promoted hyaline cartilage regeneration. In vitro studies further uncovered that chondrocytes cocultured with BMSCs in the direct contact coculture system upregulated Kindlin-2 expression and subsequently activated the PI3K/AKT signaling pathway, which not only increases Sox9 and Col2 expression but also restores mitochondrial membrane potential and reduces ROS levels and apoptosis under inflammatory conditions. Overall, our findings indicated that direct contact BMSC-chondrocyte coculture system could promote chondrogenesis, and identified Kindlin-2 represents a key regulator in this process.

The Mechanisms Responsible for N Deficiency in Well-Watered Wheat Under Elevated CO2.

Elevated CO2 concentration [e(CO2)] often promotes plant growth with a decrease in tissue N concentration. In this study, three experiments, two under hydroponic and one in well-watered soil, including various levels or patterns of CO2, humidity, and N supply were conducted on wheat (Triticum aestivum L.) to explore the mechanisms of e[CO2]-induced N deficiency (ECIND). Under hydroponic conditions, N uptake remained constant even as transpiration was limited 40% by raising air relative humidity and only was reduced about 20% by supplying N during nighttime rather than daytime with a reduction of 85% in transpiration. Compared to ambient CO2 concentration, whether under hydroponic or well-watered soil conditions, and whether transpiration was kept stable or decreased to 12%, e[CO2] consistently led to more N uptake and higher biomass, while lower N concentration was observed in aboveground organs, especially leaves, as long as N supply was insufficient. These results show that, due to compensation caused by active uptake, N uptake can be uncoupled from water uptake under well-watered conditions, and changes in transpiration therefore do not account for ECIND. Similar or lower tissue NO 3 - -N concentration under e[CO2] indicated that NO 3 - assimilation was not limited and could therefore also be eliminated as a major cause of ECIND under our conditions. Active uptake has the potential to bridge the gap between N taken up passively and plant demand, but is limited by the energy required to drive it. Compared to ambient CO2 concentration, the increase in N uptake under e[CO2] failed to match the increase of carbohydrates, leading to N dilution in plant tissues, the apparent dominant mechanism explaining ECIND. Lower N concentration in leaves rather than roots under e[CO2] validated that ECIND was at least partially also related to changes in resource allocation, apparently to maintain root uptake activity and prevent more serious N deficiency.

External penetrating laryngeal trauma caused by a metal fragment: A Case Report.

BACKGROUND: Although external penetrating laryngeal trauma is rare in the clinic, such cases often result in a high mortality rate. The early recognition of injury, protection of the airway, one-stage laryngeal reconstruction with miniplates and interdisciplinary cooperation are important in the treatment of such patients. CASE SUMMARY: A 58-year-old male worker sustained a penetrating injury in the left neck. After computed tomography scanning at a local hospital, he was transferred to our hospital, where he underwent tracheotomy, neck exploration, extraction of the foreign object, debridement and repair of the thyroid cartilage using titanium miniplates. An endo laryngeal stent was inserted, which was removed 12 days later. The patient recovered well and his voice rapidly improved after surgery. CONCLUSION: Penetrating laryngeal trauma is uncommon. We successfully treated a patient with early laryngeal reconstruction and management by interdisciplinary cooperation.

Exosomes derived from hypoxia preconditioned mesenchymal stem cells laden in a silk hydrogel promote cartilage regeneration via the miR-205-5p/PTEN/AKT pathway.

Tissue engineering has promising prospects for cartilage regeneration. However, there remains an urgent need to harvest high quality seed cells. Bone marrow mesenchymal cells (BMSCs), and in particular their exosomes, might promote the function of articular chondrocytes (ACs) via paracrine mechanisms. Furthermore, preconditioned BMSCs could provide an enhanced therapeutic effect. BMSCs naturally exist in a relatively hypoxic environment (1%-5% O2); however, they are usually cultured under higher oxygen concentrations (21% O2). Herein, we hypothesized that hypoxia preconditioned exosomes (H-Exos) could improve the quality of ACs and be more conducive to cartilage repair. In our study, we compared the effects of exosomes derived from BMSCs preconditioned with hypoxia and normoxia (N-Exos) on ACs, demonstrating that H-Exos significantly promoted the proliferation, migration, anabolism and anti-inflammation effects of ACs. Furthermore, we confirmed that hypoxia preconditioning upregulated the expression of miR-205-5p in H-Exos, suggesting that ACs were promoted via the miR-205-5p/PTEN/AKT pathway. Finally, an injectable silk fibroin (SF) hydrogel containing ACs and H-Exos (SF/ACs/H-Exos) was utilized to repair cartilage defects and effectively promote cartilage regeneration in vivo. The application of SF/ACs/H-Exos hydrogel in cartilage regeneration therefore has promising prospects. STATEMENT OF SIGNIFICANCE: Cartilage tissue engineering (CTE) has presented a promising prospect. However, the quality of seed cells is an important factor affecting the repair efficiency. Our study demonstrates for the first time that the exosomes derived from hypoxia preconditioned BMSCs (H-Exos) effectively promote the proliferation, migration and anabolism of chondrocytes and inhibit inflammation through miR-205-5p/PTEN/AKT pathway. Furthermore, we fabricated an injectable silk fibrion (SF) hydrogel to preserve and sustained release H-Exos. A complex composed of SF hydrogel, H-Exos and chondrocytes can effectively promote the regeneration of cartilage defects. Therefore, this study demonstrates that hypoxia pretreatment could optimize the therapeutic effects of BMSCs-derived exosomes, and the combination of exosomes and SF hydrogel could be a promising therapeutic method for cartilage regeneration.

Virtual surgical planning is a useful tool in the surgical management of mandibular condylar fractures.

PURPOSE: The aim of this study is to evaluate the application value of virtual surgical planning in the management of mandibular condylar fractures and to provide a reliable reference. METHODS: This was a prospective randomized controlled study and recruited 50 patients requiring surgical treatment for their mandibular condylar fractures. The inclusion criteria were patients (1) diagnosed with a condylar fracture by two clinically experienced doctors and required surgical treatment; (2) have given consent for the surgical treatment; and (3) had no contraindications to the surgery. Patients were excluded from this study if: (1) they were diagnosed with a non-dislocated or only slightly dislocated condylar fracture; (2) the comminuted condylar fracture was too severe to be treated with internal reduction and fixation; or (3) patients could not complete follow-up for 3 months. There were 33 male and 17 female patients with 33 unilateral condylar fractures and 17 bilateral condylar fractures included. The 50 patients were randomly (random number) divided into control group (25 patients with 35 sides of condylar fractures) and experimental group (25 patients with 32 sides of condylar fractures). Virtual surgical planning was used in the experimental group, but only clinical experience was used in the control group. The patients were followed up for 1, 3, 6 and 12 months after operation. Variables including the rate of perfect reduction by radiological analysis, the average distance of deviation between preoperative and postoperative CT measurements using Geomagic software and postoperative clinical examinations (e.g., mouth opening, occlusion) were investigated for outcome measurement. SPSS 19 was adopted for data analysis. RESULTS: The average operation time was 180.60 min in the experimental group and 223.2 min in the control group. One week postoperatively, CT images showed that the anatomic reduction rate was 90.63% (29/32) in the experimental group and 68.57% (24/35) in the control group, revealing significant difference (X2 = 4.919, p = 0.027). Geomagic comparative analysis revealed that the average distance of deviation was also much smaller in the experimental group than that in the control group (0.639 mm vs. 0.995 mm; t = 3.824, p < 0.001). CONCLUSION: These findings suggest that virtual surgical planning can assist surgeons in surgical procedures, reduce operative time, and improve the anatomic reduction rate & accuracy, and thus of value in the diagnosis and treatment of condylar fractures.