Multiomics integration-based immunological characterizations of adamantinomatous craniopharyngioma in relation to keratinization.

Although adamantinomatous craniopharyngioma (ACP) is a tumour with low histological malignancy, there are very few therapeutic options other than surgery. ACP has high histological complexity, and the unique features of the immunological microenvironment within ACP remain elusive. Further elucidation of the tumour microenvironment is particularly important to expand our knowledge of potential therapeutic targets. Here, we performed integrative analysis of 58,081 nuclei through single-nucleus RNA sequencing and spatial transcriptomics on ACP specimens to characterize the features and intercellular network within the microenvironment. The ACP environment is highly immunosuppressive with low levels of T-cell infiltration/cytotoxicity. Moreover, tumour-associated macrophages (TAMs), which originate from distinct sources, highly infiltrate the microenvironment. Using spatial transcriptomic data, we observed one kind of non-microglial derived TAM that highly expressed GPNMB close to the terminally differentiated epithelial cell characterized by RHCG, and this colocalization was verified by asmFISH. We also found the positive correlation of infiltration between these two cell types in datasets with larger cohort. According to intercellular communication analysis, we report a regulatory network that could facilitate the keratinization of RHCG+ epithelial cells, eventually causing tumour progression. Our findings provide a comprehensive analysis of the ACP immune microenvironment and reveal a potential therapeutic strategy base on interfering with these two types of cells.

Identification of core genes of craniopharyngioma angiogenesis based on single-cell nuclear transcriptome sequencing.

This study aimed to explore the core genes of craniopharyngioma angiogenesis for targeted vascular therapy based on single-cell nuclear transcriptome sequencing. For single-cell nuclear transcriptome sequencing, we collected six samples from the tumor center and adjacent hypothalamic tumor tissues from three patients with craniopharyngioma, as well as four normal brain tissues based on Gene Expression Omnibus. We screened genes with differential up-regulation between vascular endothelial cells of craniopharyngioma and those of normal brain tissues, performed GO and KEGG analysis, constructed the protein-protein interaction network, and selected key genes verified using immunofluorescence. After data cleaning and quality control, 623 craniopharyngioma endothelial cells and 439 healthy brain endothelial cells were obtained. Compared with normal brain endothelial cells, craniopharyngioma endothelial cells were screened for 394 differentially up-expressed genes (DEGs). GO and KEGG results showed that DEGs probably modulated endothelial cells, adherens junction, focal adhesion, migration, actin cytoskeleton, and invasion via the PI3K-AKT, Rap1, Ras, Wnt, and Hippo pathways. The core genes screened were CTNNB1, PTK2, ITGB1, STAT3, FYN, HIF1A, VCL, SMAD3, PECAM1, FOS, and CDH5. This study obtained possible anti-angiogenic genes in craniopharyngioma. Our results shed novel insights into molecular mechanisms and craniopharyngioma treatment.

Cumulative summation analysis of learning curve for endoscopic endonasal transsphenoidal resection of craniopharyngiomas.

Background: To evaluate the cumulative summation (CUSUM) analysis of the learning curve for Endoscopic Endonasal Transsphenoidal resection of craniopharyngioma (EETC). Methods: Retrospectively analyzed the clinical data of 113 patients who underwent EETC by the same neurosurgery team of the first affiliated Hospital of Nanchang University from June 2012 to November 2020. The learning curve was created by the CUSUM method and analyzed, which was divided into two groups: the learning stage and stable stage based on the learning curve trend. The median operation time and minimum surgical case number was calculated and the operation time and postoperative complications were compared between the two groups. Results: The median operation time was 318 min. The best fitting curve equation was y = 227.72 + 49.06x + 0.14x2 - 0.05x3, R2 = 0.949, (p < 0.001). The minimum number of surgical cases was 65. Between the two groups, the operation time decreased from 360.8 ± 106.4 min in the learning group to 281.6 ± 69.9 min in the stable group (p < 0.05). The incidence of postoperative complications (intracranial infection, cerebrospinal fluid rhinorrhea, and diabetes insipidus) was significantly reduced (p < 0.05). Conclusion: The CUSUM learning curve of craniopharyngioma resection via endoscope endonasal transsphenoidal approach could better describe the learning process for a neurosurgeon. The frequency of surgery could be a good factor for strengthening the learning effect and help to shorten the learning time. After 65 cases of EETC, the surgical skills can reach a stable stage, the operation time is obviously shortened, and the postoperative complications are significantly reduced.

Machine learning-enabled performance prediction and optimization for iron-chromium redox flow batteries.

Iron-chromium flow batteries (ICRFBs) are regarded as one of the most promising large-scale energy storage devices with broad application prospects in recent years. However, transitioning from laboratory-scale development to industrial-scale deployment can be a time-consuming process due to the multitude of complex factors that impact ICRFB stack performance. Herein, a data-driven optimization methodology applying active learning, informed by an extensive survey of the literature encompassing diverse experimental conditions, is proposed to enable exceptional precision in predicting ICRFB system performance considering both operation conditions and key materials selection. Specifically, multitask ML models are trained on experimental data with a high prediction accuracy (R2 > 0.92) to link ICRFB properties to energy efficiency, coulombic efficiency, and capacity. We also interpret the ML models based on Shapley additive explanations and extract valuable insights into the importance of descriptors. It is noted that the operation conditions (current density and cycle number) and the electrode type are the most critical descriptors affecting the voltage efficiency and coulombic efficiency while the electrode size strongly affects the capacity. Moreover, active learning is used to explore the most optimized cases considering the highest energy efficiency and capacity. The versatility and robustness of the approach are demonstrated by the successful validation between ML prediction and our experiments of energy efficiency (±0.15%) and capacity (±0.8%). This work not only affords fruitful data-driven insight into the property-performance relationship, but also unveils the explainability of critical properties on the performance of ICRFBs, which accelerates the rational design of next-generation ICRFBs.

Self-Assembly Dual Active Site Nanocomposite Anode Ce0.6Mn0.3Fe0.1O2-δ/NiFe/MnOx for Electrooxidative Dehydrogenation of Ethane to Ethylene.

As the demand for ethylene grows continuously in industry, conversion of ethane to ethylene has become more and more important; however, it still faces fundamental challenges of low ethane conversion, low ethylene selectivity, overoxidation, and instability of catalysts. Electrooxidative dehydrogenation of ethane (EODHE) in a solid oxide electrolysis cell (SOEC) is an alternative process. Here, a multiphase oxide Ce0.6Mn0.3Fe0.1O2-δ-NiFe-MnOx has been fabricated by a self-assembly process and utilized as the SOEC anode material for EODHE. The highest ethane conversions reached 52.23% with 94.11% ethylene selectivity at the anode side and CO with 10.9 mL min-1 cm-2 at the cathode side, at 1.8 V at 700 °C. The remarkable electrooxidative performance of CMF-NiFe-MnOx is ascribed to the NiFe alloy and MnOx nanoparticles and improvement of the concentration of oxygen vacancies within the fluorite substrate, generating dual active sites for C2H6 adsorption, dehydrogenation, and selective transformation of hydrogen without overoxidizing the ethylene generated. Such a tailored strategy achieves no significant degradation observed after 120 h of operation and constitutes a promising basis for EODHE.

Harpagide inhibits the TNF-α-induced inflammatory response in rat articular chondrocytes by the glycolytic pathways for alleviating osteoarthritis.

Osteoarthritis (OA) causes severe and functional dysfunction due to abnormal inflammation. The objective of this study was to evaluate the effect of Harpagide (HPG) on TNF-α-induced inflammation in vitro and in vivo. The effect of HPG on the proliferation of rat chondrocytes was studied. The anti-inflammatory effect of HPG and its molecular mechanisms were elucidated by qPCR, Western blotting, flow cytometry, metabolome analysis in vitro. In addition, the OA rat model was established, and the effect of HPG on OA was verified in vivo. We revealed 10 µM HPG demonstrated biocompatibility. The results demonstrated that HPG restored the upregulation of MMP-13, COX2, IL-1ß and IL-6 induced by TNF-α. Moreover, HPG reversed TNF-α induced degradation of the extracellular matrix of chondrocytes. TNF-α treatment induced down-regulation of the mRNA/protein levels of proliferative markers Bcl2, CDK1 and Cyclin D1 were also recovered. HPG can inhibit TNF-α-induced inflammatory response through glycolytic metabolic pathways. HPG can restore TNF-α-induced upregulation of GRP78/IRE1α, and downregulation of AMPK proteins. In vivo experiments demonstrated that after HPG treatment, the appearance and physiological structure of articular cartilage were more integrated with highly organized chondrocytes and rich cartilage matrix compared with OA group. Finally, the molecular docking of HPG and selected key factors in glycolysis results showed that HPG had good binding potential with PFKM, PFKP, PFKFB3, PKM, HK2, and PFKL. In conclusion, the results shown HPG protects and activates chondrocytes, inhibits TNF-α-induced inflammatory response by glycolysis pathway in rat articular chondrocytes, and plays a role in the treatment of OA.

Vitamin B ameliorates PM2.5-induced kidney damage by reducing endoplasmic reticulum stress and oxidative stress in pregnant mice and HK-2.

As an air pollutant, particulate matters 2.5 (PM2.5) poses a severe risk to kidney and the mechanism involves oxidative stress and endoplasmic reticulum (ER) stress. As an essential nutrient for human health, Vitamin B performs anti-inflammatory and antioxidant functions. In order to study the effect of Vitamin B on PM2.5-induced kidney damage during pregnancy, the pregnant mice were divided into the four experimental groups randomly: control group, model group, treatment group and VB group. PM2.5 was sprayed on the trachea of pregnant mice once each three days for six times from pregnancy until delivery. The model group was given 30 µL PM2.5 suspension of 3.456 µg/µL and 10 mL/(kg·d) PBS. The treatment group was given 30 µL PM2.5 suspension of 3.456 µg/µL and 10 mL/(kg·d) Vitamin B. The VB group was given 10 mL/(kg·d) Vitamin B and the control group was given the same dose of PBS. Vitamin B was composed of Vitamin B6, Vitamin B12 and folic acid, with final concentrations are 1.14, 0.02 and 0.06 mg/mL, respectively. The results showed Vitamin B ameliorated PM2.5-induced kidney damage such as improving histopathological change, decreasing expressions of Bip and Chop, increasing expressions of Nrf2, HO-1 and Nqo1. In addition, HK-2 cells were used for cell experiments and were divided into the four groups, in which the dosage of PM2.5 was 75 µg/mL for 24 h and Vitamin B was 5 µL/100 µL. The results showed Vitamin B ameliorated PM2.5-induced HK-2 damage, such as decreasing expressions of Bip, Chop, P47phox and ROS, increasing expressions of Nrf2, HO-1, Nqo1 and NO. Our findings showed Vitamin B ameliorated PM2.5-induced kidney damage by reducing ER stress and oxidative stress in pregnant mice and in HK-2.

Patterns of Chromosomal Variation, Homoeologous Exchange, and Their Relationship with Genomic Features in Early Generations of a Synthetic Rice Segmental Allotetraploid.

Polyploidization is a driving force in plant evolution. Chromosomal variation often occurs at early generations following polyploid formation due to meiotic pairing irregularity that may compromise segregation fidelity and cause homoeologous exchange (HE). The trends of chromosomal variation and especially factors affecting HE remain to be fully deciphered. Here, by whole-genome resequencing, we performed nuanced analyses of patterns of chromosomal number variation and explored genomic features that affect HE in two early generations of a synthetic rice segmental allotetraploid. We found a wide occurrence of whole-chromosome aneuploidy and, to a lesser extent, also large segment gains/losses in both generations (S2 and S4) of the tetraploids. However, while the number of chromosome gains was similar between S2 and S4, that of losses in S4 was lower than in S2. HEs were abundant across all chromosomes in both generations and showed variable correlations with different genomic features at chromosomal and/or local scales. Contents of genes and transposable elements (TEs) were positively and negatively correlated with HE frequencies, respectively. By dissecting TEs into different classes, retrotransposons were found to be negatively correlated with HE frequency to a stronger extent than DNA transposons, whereas miniature terminal inverted elements (MITEs) showed a strong positive correlation. Local HE frequencies in the tetraploids and homologous recombination (HR) rates in diploids within 1 Mb sliding windows were significantly correlated with each other and showed similar overall distribution profiles. Nonetheless, non-concordant trends between HE and HR rates were found at distal regions in some chromosomes. At local scale, both shared and polymorphic retrotransposons between parents were negatively correlated with HE frequency; in contrast, both shared and polymorphic MITEs showed positive correlations with HE frequency. Our results shed new light on the patterns of chromosomal number variation and reveal genomic features influencing HE frequency in early generations following plant polyploidization.

The impact of PM2.5 on kidney.

PM2.5 poses a severe risk to kidneys, inducing kidney function decline, increasing the risk of suffering from chronic kidney diseases and promoting the occurrence and development of various renal tumors. The mechanism of PM2.5-induced renal injury may involve oxidative stress, inflammatory response, and cytotoxicity. This paper elaborated PM2.5-induced kidney damage and the corresponding possible mechanism so as to raise awareness of air pollution and reduce the damage to human body.

Molecular selectivity in the water flooding heavy oil process from porous rocks.

Water flooding increases the recovery factor of crude oil and has been proven to be an economical and viable technique for enhancing the oil recovery of oil fields. The process has been systematically studied previously, in which the oil was considered a substance of constant composition. However, the molecular selectivity during the water flooding process has rarely been addressed, especially for heavy oil. Herein, the properties and compositional changes of heavy oil have been investigated in a simulated water flooding process at 60 °C and 85 °C. The crude oil and produced oils from different water flooding stages were characterized by gas chromatography, gas chromatography-mass spectrometry, and Fourier transform ion cyclotron resonance mass spectrometry. The results show that with the increase in temperature, the content of resins in the produced oils from different water flooding stages decreases, and the content of asphaltenes increases slightly. The viscosity of the produced oils increases at low temperatures and decreases at high temperatures as the water cut increases. The composition of the produced oils from different water flooding stages is different. Compared with the no water cut and high water cut stages, the changes in the produced oils of the low water cut stages are significant at different temperatures. The molecular selectivity of heteroatoms is higher than that of hydrocarbons in the water flooding process. Water flooding preferentially extracts small-molecule low-carbon hydrocarbons and small-molecule heteroatoms with low condensation degrees. The compositional differences between the produced oils were characterized by the double bond equivalent versus carbon number distribution of the S, N, and O-containing compounds. This study can not only provide some explanations on the viscosity-forming mechanism of heavy oil but also explains the watered-out phenomena in the development of oilfields.

Mechanism of kidney injury induced by cisplatin.

Cisplatin is a common metal platinum complex. The platinum atom in the molecule is of great significance to its antitumor effect. Clinically, it can show curative effect on a variety of solid tumors. However, cisplatin has certain adverse effects in treatment, one among which is acute renal injury (AKI). Except for the nuclear DNA damage caused by cisplatin, damage of organelles, and cytoplasm also contribute to side effects. Endoplasmic reticulum stress, mitochondrial apoptosis pathway or cascade reaction caused by complement and caspase protein also play important roles in cisplatin induced renal injury. Therefore, the damage studies of organelles and cytoplasm are also necessary for exploring adverse effects of cisplatin. This paper reviews the damage of endoplasmic reticulum, mitochondria, and indirect DNA apoptosis pathways induced by cisplatin. It also explains in detail why cisplatin is easy to cause kidney damage. Deep understanding of such interactions could be helpful to exploit better drugs which would minimize kidney injury and maximize anti-tumor effects of cisplatin.

Efficacy of Apatinib plus S-1 Therapy in the Treatment of Advanced Gastric Cancer Patients and the Effect on the Levels of Tumor Markers and Th1 and Th2-Like Cytokines.

Objective: To assess the efficacy of apatinib plus S-1 therapy in the treatment of advanced gastric cancer patients and the effect on the levels of tumor markers and Th1 and Th2-like cytokines. Methods: From October 2019 to December 2020, 100 patients with advanced gastric cancer assessed for eligibility were recruited and assigned at a ratio of 1 : 1 to receive either S-1 regimen (tegafur, gimeracil, and oteracil potassium capsules) (observation group) or apatinib plus S-1 therapy (experimental group). Outcome measures included clinical efficacy serum tumor marker levels, Th1 and Th2-like cytokine levels, time to progression (TTP), overall survival (OS), and adverse events. Results: The S-1 therapy plus apatinib was associated with a significantly higher efficacy versus S-1 therapy alone (P < 0.05). The eligible patients given S-1 therapy plus apatinib showed significantly lower levels of serum carcinoembryonic antigen (CEA), glycoantigen 199 (CA199), and glycoantigen 125 (CA125) versus those receiving S-1 therapy (P < 0.05). S-1 therapy plus apatinib outperformed the single therapy of S-1 therapy in mitigating the levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), and interleukin-10 (IL-10) (P < 0.05). There was no statistically significant difference in the incidence of adverse reactions between the two groups (P > 0.05). S-1 therapy plus apatinib was associated with a significantly shorter TTP (5.2 ± 0.7 months) and a longer OS (9.3 ± 2.5 months) versus S-1 therapy alone (7.1 ± 1.3, 5.1 ± 1.3 months) (P < 0.05). Conclusion: The efficacy of apatinib plus S-1 therapy showed better improvement in lowering the serum tumor marker levels and ameliorating the Th1 and Th2-like cytokine levels versus S-1 therapy alone, so it is worthy of clinical application.

[Intradermal needling combined with heat-sensitive moxibustion for moderate to severe cancer pain].

OBJECTIVE: To observe the effect of intradermal needling combined with heat-sensitive moxibustion for moderate to severe cancer pain. METHODS: A total of 60 patients with moderate to severe cancer pain were randomly divided into an observation group and a control group，30 cases in each one. In the control group，opioids were taken to relief pain according to the three-step analgesic method of World Health Organization. On the base of the treatment as the control group, intradermal needling combined with heat-sensitive moxibustion were applied at Neiguan (PC 6), Hegu (LI 4), Zusanli (ST 36), Taichong (LR 3), etc. in the observation group, 14 days of treatment were required. The equivalent morphine consumption at the first day and whole course, the scores of cancer quality of life questionnaire-C30 (QLQ-C30) and Hamilton anxiety scale before and after treatment, and the adverse reaction rate were compared in the two groups. The total analgesic effective rate was evaluated. RESULTS: The total analgesic effective rate was 93.3% (28/30) in the observation group, higher than 73.3% (22/30) in the control group (P<0.05). The total equivalent morphine consumption in the observation group was less than the control group (P<0.05). After treatment, the QLQ-C30 scores were increased (P<0.001) and the HAMA scores were decreased (P<0.001) in the both groups, and those in the observation group were superior to the control group (P<0.001). The adverse reaction rates of fatigue, dizziness, nausea and vomiting, constipation in the observation group were lower than the control group (P<0.05). CONCLUSION: Intradermal needling combined with heat-sensitive moxibustion can reduce the dose of opioids, improve the quality of life, relief the anxiety in patients with moderate to severe cancer pain, and reduce the incidence of common adverse reaction of opioids.

Evolution of Homeologous Gene Expression in Polyploid Wheat.

Polyploidization has played a prominent role in the evolutionary history of plants. Two recent and sequential allopolyploidization events have resulted in the formation of wheat species with different ploidies, and which provide a model to study the effects of polyploidization on the evolution of gene expression. In this study, we identified differentially expressed genes (DEGs) between four BBAA tetraploid wheats of three different ploidy backgrounds. DEGs were found to be unevenly distributed among functional categories and duplication modes. We observed more DEGs in the extracted tetraploid wheat (ETW) than in natural tetraploid wheats (TD and TTR13) as compared to a synthetic tetraploid (AT2). Furthermore, DEGs showed higher Ka/Ks ratios than those that did not show expression changes (non-DEGs) between genotypes, indicating DEGs and non-DEGs experienced different selection pressures. For A-B homeolog pairs with DEGs, most of them had only one differentially expressed copy, however, when both copies of a homeolog pair were DEGs, the A and B copies were more likely to be regulated to the same direction. Our results suggest that both cis- and inter-subgenome trans-regulatory changes are important drivers in the evolution of homeologous gene expression in polyploid wheat, with ploidy playing a significant role in the process.

Molecular transformation of dissolved organic matter in process water from oil and gas operation during UV/H2O2, UV/chlorine, and UV/persulfate processes.

This paper reported the impact of UV/H2O2, UV/chlorine, and UV/persulfate advanced oxidation processes on the molecular transformations of dissolved organic matter (DOM), removal of naphthenic acids (NAs) and acute toxicity in oil sands process water (OSPW). The UV/persulfate process exhibited the highest removal (81.2% with 2 mM dose) towards classical NAs and highest reduction in acute toxicity to Vibrio fischeri among the three processes. The fraction of DOM such as CHOS class species decreased along with the increase of the oxidant doses in all processes. The increase in O/C ratio and lack of change in the H/C and double bond equivalence indicated that H-abstraction followed by the OH-addition was the main reaction pathway for all processes. This observation aligned with previous studies using model compounds and proved that OSPW DOM reacted similarly to model compounds. Sulfur containing organic matters were the most liable compounds in OSPW NOM, while UV/chlorine was the most effective process to oxidize nitrogen containing organic matters. Overall results revealed that the UV/persulfate process could be used as a promising technique for the removal of OSPW NA and reduction of acute toxicity towards Vibrio fischeri. In addition, this DOM characterization approach could be utilized to investigate the transformation of complicated OSPW DOM and to identify the byproducts generated during different water treatment processes.

Correlations of coagulation indexes and inflammatory changes with the prognosis of lung cancer complicated with thromboembolic disease.

PURPOSE: To investigate the correlations of coagulation indexes and inflammatory changes with the prognosis of lung cancer (LC) patients complicated with thromboembolic (TE) disease. METHODS: A total of 84 LC patients complicated with TE disease admitted to hospital from January 2010 to January 2016 were enrolled in this study and their clinical data were retrospectively analyzed. A 2-year post-treatment follow-up was carried out. According to the prognosis, patients were divided into 2 groups as dead group (n=25) and alive group (n=59). The coagulation indexes and inflammatory factor levels before low-molecular weight heparin (LMWH) treatment and on the 1st, 3rd, and 7th day after treatment were compared between the two groups. Their relations with the prognosis of patients were analyzed using Pearson method. RESULTS: No statistically significant difference was found in the prothrombin time (PT), levels of Fibrinogen (FIB), D-Dimer (D-D), Interleukin-6 (IL-6) and Procalcitonin (PCT), and activated partial thromboplastin time (APTT) before treatment between the two groups (p>0.05). The PT and levels of FIB, D-D, IL-6, and PCT on the 1st, 3rd, and 7th day after treatment were significantly increased in the dead group compared to those in the alive group, while the APTT was remarkably shortened. Moreover, the PT was gradually prolonged and FIB, D-D, IL-6 and PCT levels were increased in the dead group , but the APTT was gradually shortened over time (p<0.05). The poor prognosis of LC patients complicated with TE disease was positively correlated with PT, FIB, D-D, IL-6 and PCT, but negatively correlated with APTT (p<0.05). CONCLUSION: The poor prognosis of LC complicated with TE disease has positive correlations with PT, FIB, D-D, IL-6 and PCT, and a negative association with APTT, providing a certain reference as a prognostic value in the diagnosis and treatment.

DNA methylation repatterning accompanying hybridization, whole genome doubling and homoeolog exchange in nascent segmental rice allotetraploids.

Allopolyploidization, which entails interspecific hybridization and whole genome duplication (WGD), is associated with emergent genetic and epigenetic instabilities that are thought to contribute to adaptation and evolution. One frequent genomic consequence of nascent allopolyploidization is homoeologous exchange (HE), which arises from compromised meiotic fidelity and generates genetically and phenotypically variable progenies. Here, we used a genetically tractable synthetic rice segmental allotetraploid system to interrogate genome-wide DNA methylation and gene expression responses and outcomes to the separate and combined effects of hybridization, WGD and HEs. Progenies of the tetraploid rice were genomically diverse due to genome-wide HEs that affected all chromosomes, yet they exhibited overall methylome stability. Nonetheless, regional variation of cytosine methylation states was widespread in the tetraploids. Transcriptome profiling revealed genome-wide alteration of gene expression, which at least in part associates with changes in DNA methylation. Intriguingly, changes of DNA methylation and gene expression could be decoupled from hybridity and sustained and amplified by HEs. Our results suggest that HEs, a prominent genetic consequence of nascent allopolyploidy, can exacerbate, diversify and perpetuate the effects of allopolyploidization on epigenetic and gene expression variation, and hence may contribute to allopolyploid evolution.

Expression of LOXs and MMP-1, 2, 3 by ACL Fibroblasts and Synoviocytes Impact of Coculture and TNF-α.

This study aims to confirm the effects of synoviocytes (SCs) on regulating lysyl oxidases (LOXs) and matrix metalloproteinase (MMP)-1, 2, 3 in the normal and injured anterior cruciate ligament (ACL) fibroblasts response to tumor necrosis factor-α(TNF-α). The gene and protein expression levels of LOXs and MMP-1, 2, 3 in SCs cocultured ACL fibroblasts (ACLfs) induced by TNF-α and mechanical injury were analyzed by real-time polymerase chain reaction (PCR) and western bolting; the MMP-2 activity were analyzed by zymography. The results exhibited that TNF-α alone slightly downregulated the expressions of LOXs and upregulated the expression of MMP-1, 2, 3 in both normal and injured ACL fibroblasts. The decrease of LOXs and increase of MMP-1, 2, 3 in ACLfs response to TNF-α were further promoted by coculture. Taken together, these results show for the first time that the crosstalk between ACLfs and SCs could modulate the LOXs and MMP-1, 2, 3 synthesis in ACLfs in the presence of TNF-α. Accumulation of MMPs in the isolated fluid-containing space not only disrupts the balance of ACL healing, but also increases cartilage degradation and accelerates osteoarthritis (OA) in injured joint. Based on this mechanism, targeting inhibition of MMPs could provide a promising therapeutic strategy for acute ligament injury.

Bay11-7082 facilitates wound healing by antagonizing mechanical injury- and TNF-α-induced expression of MMPs in posterior cruciate ligament.

Purposes: To investigate the ability of synoviocytes (SCs) in regulating MMPs expression in the posterior cruciate ligament fibroblasts (PCLfs) after TNF-α treatment, to test whether a specific inflammation inhibitor Bay11-7082 can antagonize the expression of MMPs in PCLfs after injury. Methods: The microenvironment of knee joint cavity after PCL injury was mimicked in an in vitro co-culture system. The effects of TNF-α treatment on the expression of MMPs in PCL fibroblasts (PCLfs) were studied. The expression of MMPs mRNA and protein was detected by qRT-PCR and western blot. For the in vivo study, the Bay11-7082 inhibitor was injected into the knee joint cavity after injury, and then were performed on histological analysis. Results: In the mono-culture conditions, 6% mechanical injury upregulated the expression of MMP-2, whereas downregulated MMP-1 and -3, additionally 12% mechanical injury were upregulated all. However, in co-culture conditions, 6% and 12% both significantly increased MMPs expressions. Stretch injury and TNF-α treatment significantly upregulated expression of MMPs mRNA and protein levels in mono-cultured PCLfs. This effect was more significant in PCLfs Plus SCs co-culture system, in which the cells were treated by combination of stretch injury and TNF-α. In addition, Bay11-7082, a specific inflammation inhibitor, could significantly decrease the expression of MMPs induced by stretch injury and/or TNF-α treatment. Less infiltrated inflammatory cells and more integrated tissues were detected in injury PCL 2 weeks after Bay11-7082 treatment, compared to injury group. Immunofluorescent staining showed very low expression levels of MMPs in PCL of Bay11-7082-treated group, compared to the injury groups. Conclusions: SCs sever as the supporting cells that aggravate the TNF-α-induced MMPs accumulation in PCLfs. Inhibition of the expression of MMPs by Bay11-7082 is a promising way to facilitate the self-healing of PCL.

Molecular Characterization of Organics Removed by a Covalently Bound Inorganic-Organic Hybrid Coagulant for Advanced Treatment of Municipal Sewage.

Coagulation is an important process to remove organics from water. The molecular composition and structure of organic matter influence water quality in many ways, and the lack of information regarding the organics removed by different coagulants makes it challenging to optimize coagulation processes and ensure reclaimed water safety. In this paper, we investigated coagulation of secondary biological effluent from a municipal sewage treatment plant with different coagulants. We emphasized investigation of organics removal characteristics at the molecular level using Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS) coupled with electrospray ionization (ESI). We found that conventional coagulants can only partially remove condensed polycyclic aromatics and polyphenols with low H/C (H/C < 0.7) and highly unsaturated and phenolic compounds and aliphatic compounds with high O/C (O/C > 0.6). A new coagulant, CBHyC, had better removal efficiencies for all organics with different element compositions and molecular structures, especially organics that are resistant to conventional coagulants such as highly unsaturated and phenolic compounds and aliphatic compounds located in 0.3 < O/C < 0.8 and 1.0 < H/C < 2.0 regions and sulfur-containing compounds with higher O/C (e.g., anionic surfactants and their metabolites or coproducts). This study provides molecular insights into the organics removed by different coagulants and provides data supporting the possible optimization of advanced wastewater treatment processes.