ABSTRACT
Iron overload and oxidative stress have been reported to contribute to ferroptosis in endometriotic lesions. However, the possible roles of iron overload on macrophages in endometriosis (EMs) remain unknown. Based on recent reports by single-cell sequencing data of endometriosis, here we found significant upregulations of ferroptosis-associated genes in the macrophage of the endometriotic lesion. Additionally, there was an elevated expression of HMOX1, FTH1, and FTL in macrophages of peritoneal fluid in EMs, as well as iron accumulation in the endometriotic lesions. Notably, cyst fluid significantly up-regulated levels of intracellular iron and ferroptosis in Phorbol-12-myristate-13-acetate (PMA)-stimulated THP-1 cells. Additionally, high iron-induced ferroptosis obviously reduced PMA-stimulated THP-1 cells' phagocytosis and increased the expression of angiogenic cytokines, such as vascular endothelial growth factor A (VEGFA) and interleukin 8 (IL8). Baicalein, a potential anti-ferroptosis compound, increased GPX4 expression, significantly inhibited ferroptosis, and restored phagocytosis of THP-1 cells in vitro. Collectively, our study reveals that ferroptosis triggered by high iron in cyst fluid promotes the development of EMs by impairing macrophage phagocytosis and producing more angiogenic cytokines (e.g., IL8 and VEGFA). Baicalein displays the potential for the treatment of EMs, especially in patients with high ferroptosis and low phagocytosis of macrophages.
ABSTRACT
Evidence is mounting that abnormal vascular remodeling (VR) is a vital pathological event that precedes many cardiovascular diseases (CVD). This provides us with a new research perspective that VR can be a pivotal target for CVD treatment and prevention. However, the current drugs for treating CVD do not fundamentally reverse VR and repair vascular function. The reason may be that a complicated regulatory network is formed between the various signaling pathways involved in VR. Recently, ginsenoside, the main active substance of ginseng, has become increasingly the focus of many researchers for its multiple targets, multiple pathways, and few side effects. Several data have revealed that ginsenosides can improve VR caused by vasodilation dysfunction, abnormal vascular structure and blood pressure. This review is intended to discuss the therapeutic effects and mechanisms of ginsenosides in some diseases involved in VR. Besides, we herein also give a new and contradictory insight into intracellular and molecular signaling of ginsenosides in all kinds of vascular cells. Most importantly, we also discuss the feasibility of ginsenosides Rb1/Rg1/Rg3 in drug development by combining the pharmacodynamics and pharmacokinetics of ginsenosides, and provide a pharmacological basis for the development of ginsenosides in clinical applications.
Subject(s)
Ginsenosides/pharmacology , Vascular Remodeling/drug effects , Animals , Cardiovascular Diseases/drug therapy , Ginsenosides/chemistry , Ginsenosides/therapeutic use , Humans , Stereoisomerism , Structure-Activity RelationshipABSTRACT
Ultrasonic guided waves are sensitive to many different types of defects and have been studied for defect recognition in rail. However, most fault recognition algorithms need to extract features from the time domain, frequency domain, or time-frequency domain based on experience or professional knowledge. This paper proposes a new method for identifying many different types of rail defects. The segment principal components analysis (S-PCA) is developed to extract characteristics from signals collected by sensors located at different positions. Then, the Support Vector Machine (SVM) model is used to identify different defects depending on the features extracted. Combining simulations and experiments of the rails with different kinds of defects are established to verify the effectiveness of the proposed defect identification techniques, such as crack, corrosion, and transverse crack under the shelling. There are nine channels of the excitation-reception to acquire guided wave detection signals. The results show that the defect classification accuracy rates are 96.29% and 96.15% for combining multiple signals, such as the method of single-point excitation and multi-point reception, or the method of multi-point excitation and reception at a single point.
Subject(s)
Algorithms , Support Vector Machine , Intelligence , Principal Component Analysis , Ultrasonic WavesABSTRACT
In order to evaluate effects of extraction techniques on the physicochemical characteristics and antioxidant activities of kiwifruit polysaccharides (KPS), and further explore KPS as functional food ingredients, both microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) were optimized for the extraction of KPS. Furthermore, the physicochemical structures and antioxidant activities of KPS extracted by different techniques were investigated. The optimal extraction conditions of UAE and MAE for the extraction of KPS were obtained by response surface methodology. Different extraction techniques significantly affected the contents of uronic acids, molecular weights, molar ratios of constituent monosaccharides, and the degree of esterification of KPS. Results showed that KPS exhibited remarkable DPPH and ABTS radical scavenging activities, and reducing power. The high antioxidant activities observed in KPS extracted by the MAE method (KPS-M) might be partially attributed to its low molecular weight and high content of unmethylated galacturonic acid. Results suggested that the MAE method could be a good potential technique for the extraction of KPS with high antioxidant activity, and KPS could be further explored as functional food ingredients.
Subject(s)
Actinidia/chemistry , Antioxidants/chemistry , Antioxidants/pharmacology , Fruit/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Polysaccharides/chemistry , Polysaccharides/pharmacology , Chemical Fractionation/methods , Free Radical Scavengers/chemistry , Free Radical Scavengers/pharmacology , Microwaves , Molecular Weight , Monosaccharides/chemistry , Phytochemicals/chemistry , Spectroscopy, Fourier Transform Infrared , Ultrasonic WavesABSTRACT
Madecassoside is one of increasingly used constituent of Centella asiatica, a frequently prescribed crude drug in South eastern Asia and China for wound healing. In the present experiment, it exposes the neuroprotective nature of Madecassoside in GT1-7 cell lines, further, which the antioxidant activities are performed. The cellular toxicity was assessed using 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay with increased cell viability with IC50 2.5µg/ml. the regulation of antioxidant levels showed changes in madecassoside treated cell lysate viz., SOD assay. Also, the antioxidative assays confirmed the negligible cellular damage caused to the GT1-7 cell lines. Hence, the results advocate that the current antioxidant and antitumor activity be justified by the high concentration of phenolic constituents, primarily the triterpene present in the C. asiatica.
Subject(s)
Antioxidants/pharmacology , Centella/chemistry , Hypoxia-Ischemia, Brain/prevention & control , Neurons/drug effects , Neuroprotective Agents/pharmacology , Triterpenes/pharmacology , Animals , Antioxidants/chemistry , Antioxidants/isolation & purification , Antioxidants/toxicity , Biphenyl Compounds/chemistry , Cell Line , Cell Survival/drug effects , Dose-Response Relationship, Drug , Hypoxia-Ischemia, Brain/metabolism , Hypoxia-Ischemia, Brain/pathology , L-Lactate Dehydrogenase/metabolism , Mice , Neurons/metabolism , Neurons/pathology , Neuroprotective Agents/chemistry , Neuroprotective Agents/isolation & purification , Neuroprotective Agents/toxicity , Phytotherapy , Picrates/chemistry , Plant Extracts , Plants, Medicinal , Superoxide Dismutase/metabolism , Triterpenes/chemistry , Triterpenes/isolation & purification , Triterpenes/toxicityABSTRACT
OBJECTIVE: To compare the diagnostic accuracy of five internationally used indolent prostate cancer screen protocols in Chinese prostate cancer patients. METHODS: Retrospective analysis was made of the consecutive cohort of 314 patients, from Jan. 2006 to Apr. 2014, who had both prostate biopsy and radical prostatectomy in Peking University First Hospital. The Gleason score≤6, pT2, tumor volume≤0.5 mL, margin negative and lymph nodes negative were defined as indolent prostate cancer. The predictive value of five indolent screen criteria including Epstein, Memorial Sloan-Kettering Cancer Center (MSKCC), Prostate Cancer Research International: Active Surveillance (PRIAS), University of California, San Francisco (UCSF), and University of Miami (UM) were evaluated in Chinese prostate cancer patients. Measures of diagnostic accuracy and areas under the receiver-operating curve (AUC) were calculated for each protocol and compared. RESULTS: A total of 16% (49 cases) of the patients met the inclusion criteria of at least one protocol, including 24 cases in Epstein, 33 cases in MSKCC, 28 cases in PRIAS, 34 cases in UCSF, and 22 cases in UM. Three percent were eligible for all the studied criteria. UCSF and MSKCC protocols had the highest sensitivity and specificity than the others. The Epstein and PRIAS protocols demonstrated acceptable positive predictive value, but the specificity and sensitivity were inefficient. The UM protocol was performed unsatisfiedly on sensitivity, positive predictive value and AUC. A strict limited protocol which contained all the five protocols could not improve the predictive accuracy. CONCLUSION: The UCSF protocol had better diagnostic accuracy than the others, but the results were not satisfied. A further investigation on indolent prostate cancer screening in Chinese patients is needed.
Subject(s)
Early Detection of Cancer/methods , Prostatic Neoplasms/diagnosis , Asian People , Biopsy , Humans , Male , Neoplasm Grading , Prostatectomy , Retrospective StudiesABSTRACT
OBJECTIVE: To develop a duplex PCR method for identifying Metagonimus yokogawai and Haplorchis taichui. METHODS: ITS1 sequences of M. yokogawai and H. taichui, as well as those of their homologous species were obtained from GenBank, and two sets of specific primer pairs for M. yokogawai and H. taichui were designed accordingly using Primer Premier 5.0 software. PCR reaction system and conditions were optimized. The established duplex PCR method was applied in a pool of M. yokogawai, H. taichui, and 17 related species to examine its specificity. Sensitivity was evaluated through serial dilutions of plasmids containing their specific sequences. Finally, the duplex PCR was applied to identify M. yokogawai and H. taichui among trematodes collected from the viscera of 47 cats and 40 dogs to test its practicality. RESULTS: The duplex PCR method amplified target sequences of M. yokogawai and H. taichui, generating 648 bp and 279 bp products, respectively. No cross reaction was found with the following 17 related species: Haplorchis pumilio, Clonorchis sinensis, Pharyngostomum cordatum, the metacercaria of Metorchis sp. and Exorchis sp., Echinochasmus liliputanus, Echinochasmus perfoliatus, Echinostoma friedi, Hypoderaeum conoideum, Holostephanus sp., Diplodiscus sp., Anisakis sp., Metorchis orientais, Paragonimus westermani, Watsonius watsoni, Notocotylus sp. and Hysterothylacium sp, indicating a high specificity of this method. The detection limits for DNAs of M. yokogawai and H. taichui were 1.49 x 10(-1) pg and 1.14 x 10(-1) pg, suggesting a good sensitivity for this method. Further, the duplex PCR successfully identified M. yokogawai and H. taichui from cat and dog viscera, with no cross amplification of other trematodes. CONCLUSION: The duplex PCR is effective in identifying Metagonimus yokogawai and Haplorchis taichui.
Subject(s)
Heterophyidae , Metacercariae , Trematode Infections , Animals , Anisakis , Cats , Clonorchis sinensis , DNA Primers , Dogs , Paragonimus westermani , Paramphistomatidae , Polymerase Chain ReactionABSTRACT
BACKGROUND/AIMS: To investigate the protective effect of SGK1 (serum- and glucocorticoid-inducible protein kinase 1) in rat hippocampal neurons in vitro and in vivo following ischemia reperfusion (I/R). METHODS: Isolated rat hippocampal neurons were subjected to 2 h of oxygen and glucose deprivation (OGD) then returned to normoxic conditions for 10, 30 or 60 min. Cell apoptosis and protein expression of SGK1 were analyzed. To examine SGK1 function, we overexpressed SGK1 in rat hippocampal neurons. Finally we examined the involvement of PI3K/Akt/GSK3ß signaling by treating the cells (untransfected or transfected with expression vector encoding SGK1) with the PI3K inhibitor LY294002. Findings were confirmed in vivo in a rat model of middle cerebral artery occlusion. RESULTS: I/R caused a time-dependent increase in apoptosis, both in vitro and in vivo. SGK1 protein levels decreased significantly under the same conditions. Overexpression of SGK1 reduced apoptosis following OGD or I/R compared to cells transfected with empty vector and subjected to the same treatment, or sham-operated animals. Addition of LY294002 revealed that the action of SGK1 in suppressing apoptosis was mediated by the PI3K/Akt/GSK3ß pathway. CONCLUSION: SGK1 plays a protective role in ischemia reperfusion in rat hippocampal neurons, exerting its effects via the PI3K/Akt/GSK3ß pathway.
Subject(s)
Hippocampus/pathology , Immediate-Early Proteins/metabolism , Neurons/pathology , Protein Serine-Threonine Kinases/metabolism , Reperfusion Injury/prevention & control , Animals , Base Sequence , Cells, Cultured , DNA Primers , Hippocampus/enzymology , In Situ Nick-End Labeling , Male , Neurons/enzymology , Rats , Rats, Sprague-Dawley , Real-Time Polymerase Chain ReactionABSTRACT
OBJECTIVE: To investigate the relationship of prostate-specific antigen (PSA) with age in men older than 50 years in Beijing. METHODS: A cross-sectional study was performed in men aged over 50 years at communities in Beijing, and serum total PSA (T-PSA) and free PSA (F-PSA) levels were assayed. The relationship between age and PSA was analyzed with simple linear regression. RESULTS: A total of 1 027 subjects were enrolled. Serum T-PSA and F-PSA levels were positively correlated with age (r=0.208,P<0.001; r=0.230, P<0.001), whereas F/T ratio wasn't correlated with age (r=0.055, P>0.079). The upper limit (95% CI) for serum PSA: 1.55 µg/L for 50-59 years, 2.08 µg/L for 60-69 years, 2.40 µg/L for 70-79 years, 3.52 µg/L for ≥80 years. CONCLUSION: Of the men aged over 50 years in Beijing, there is a positive correlation between serum T-PSA, F-PSA and age; whereas F/T ratio isn't correlated with age.
Subject(s)
Mass Screening/statistics & numerical data , Prostate-Specific Antigen/blood , Age Factors , Aged , Aged, 80 and over , Asian People , Cross-Sectional Studies , Humans , Male , Middle Aged , Prostatic Neoplasms/prevention & control , Sampling StudiesABSTRACT
OBJECTIVE: To analyze the clinicopathologic features of 10 Chinese prostate adenocarcinoma patients under 50 years. METHODS: Between January 2007 and April 2012, 10 cases of prostate adenocarcinoma patients under 50 years (age: 36-49) were diagnosed at our institution through prostate biopsy. The indications of biopsy were abnormal digital rectal examination (7 cases) and/or a total prostate specific antigen (PSA) over 4.0 µg/L (4 cases). The clinicopathological data of these patients were reviewed. The results of biopsy were 1 case with Gleason score (GS) 6, 4 cases with GS 7 and 5 cases with GS 9. In the study, 7 patients were diagnosed with cT4. Lymph node metastasis and bone metastasis were found in 5 and 4 cases, respectively. The treatments were hormonal therapy in 5 cases, radical prostatectomy in 2 cases and combination therapy in 3 cases. RESULTS: During the follow-up, 6 patients were in progression within 3 to 7 months, 1 patient died after 3 years, and 4 patients were lost of the follow-up. CONCLUSION: These findings suggest that the Chinese prostate adenocarcinoma patients with age under 50 years are likely to have more aggressive tumor and combination therapy might be appropriate.
Subject(s)
Adenocarcinoma/surgery , Antineoplastic Agents, Hormonal/therapeutic use , Prostatectomy/methods , Prostatic Neoplasms/surgery , Adenocarcinoma/pathology , Adult , Age Factors , Biopsy, Needle , Combined Modality Therapy , Humans , Male , Middle Aged , Neoplasm Metastasis , Prognosis , Prostatic Neoplasms/pathologyABSTRACT
OBJECTIVE: To determine the distribution characteristics of cancerous foci in the prostate by retrospective analysis on the radical prostatectomy (RP) samples from patients with prostate cancer diagnosed by single positive core biopsy and treated by RP. METHODS: Thirty-seven patients with prostate cancer diagnosed by ultrasound-guided biopsy and single positive core biopsy underwent RP. We reviewed the pre- and post-operative data of the patients, compared the results of biopsies and pathological examination of the RP samples, and analyzed the factors that led to the underestimation of the overall prostate cancer risks. RESULTS: Post-operative pathological results showed multifocal distribution of the tumors in 70% of the patients (26/37) and obviously increased Gleason score (7-9) in 56% (21/37). The clinical stages of the tumors had been significantly underestimated preoperatively. The underestimation of their clinical stages might be due to a larger proportion of cancer tissues in a single positive core biopsy, and that of the overall cancer risks attributed to PSAD > 0.2 microg/L. Larger prostate volume (> or = 40 ml) increased the possibility of multifocal distribution. CONCLUSION: The risk of prostate cancer diagnosed by single positive core biopsy might be underestimated, and the cancerous foci were characterized by multifocal distribution in the prostate.
Subject(s)
Biopsy, Needle/methods , Prostate/pathology , Prostatic Neoplasms/pathology , Aged , Humans , Male , Middle Aged , Neoplasm Staging , Retrospective StudiesABSTRACT
OBJECTIVE: To evaluate the effects of acupuncture on hypoglycaemic outcomes in type 2 diabetes mellitus (T2DM). METHODS: PubMed, Embase, Cochrane Library, and ClinicalTrials.gov were searched from inception up to July 2020, to identify randomised controlled trials (RCTs) that enrolled patients with T2DM and compared acupuncture combined with antidiabetic drugs to antidiabetic drugs alone. The primary outcomes were haemoglobin A1c (HbA1c) and fasting blood glucose (FBG). The secondary outcomes included 2-h blood glucose (2hBG), fasting insulin (FINS), homeostatic model assessment for insulin resistance (HOMA-IR), and acupuncture-related adverse events. Mean difference (MD) and 95% confidence interval (CI) were used as the effect measure in the meta-analysis. The quality of evidence was assessed using the Grading of Recommendations Assessment, Development and Evaluation tool. RESULTS: Twenty-one RCTs (n=1,188) were included. The meta-analytic results showed that the acupuncture group had greater reductions in FBG (MD -6.46 mg/dL, 95% CI -11.95 to -0.98; moderate-quality evidence) and HOMA-IR (MD -1.23, 95% CI -2.16 to -0.31; low-quality evidence), but comparable changes in HbA1c (MD -0.39%, 95% CI -0.84 to 1.61; very-low-quality evidence), 2hBG (MD -4.99 mg/dL, 95% CI -20.74 to 10.76; low-quality evidence), and FINS (MD -1.32 µIU/mL, 95% CI -3.76 to 1.12; low-quality evidence). No data on the incidence of diabetic complications were found. All acupuncture-related adverse events reported were mild. CONCLUSIONS: The current evidence suggests that acupuncture, as a complementary therapy to antidiabetic drugs, has a small but statistically significant effect on decreasing FBG and improving insulin resistance. The effects of acupuncture on HbA1c, 2hBG, and FINS remain uncertain. Acupuncture is generally safe in patients with mild diabetes. More evidence for the long-term effects of acupuncture on T2DM is needed. (Trial registration No. CRD42018115639).
Subject(s)
Acupuncture Therapy , Diabetes Mellitus, Type 2 , Insulin Resistance , Acupuncture Therapy/adverse effects , Acupuncture Therapy/methods , Blood Glucose , Diabetes Mellitus, Type 2/drug therapy , Humans , Hypoglycemic Agents/adverse effects , Randomized Controlled Trials as TopicABSTRACT
Heat-sensitive moxibustion is the appropriate technique of the external treatment in traditional Chinese medicine and it is widely used in community because of its "easy learning, simple operation and clear curative effect". Pragmatic randomized controlled trial is a main intervention design in the real world study, which provides a high-level evidence for the effectiveness assessment of heat-sensitive moxibustion in community management. Focusing on the key links of randomization, e.g. block randomization, stratified randomization, cluster randomization, sample size allocation, allocation concealment and blinding, the paper elaborates the advantages, disadvantages and technical details of two-stage randomization with consideration of patient preference in pragmatic randomized controlled trials of heat-sensitive moxibustion in community. It facilitates improving the quality of evidence, reproducibility and methodological homogeneity among different trials.
Subject(s)
Moxibustion , Hot Temperature , Humans , Patient Preference , Random Allocation , Reproducibility of ResultsABSTRACT
Porous spherical silica-based chiral stationary phases (CSPs) have been commercially used in the field of chiral separation, however, the scope of their application is, to some extent, limited by the instability of silica towards mobile phase containing strong base or acid. As such, developing new matrix-based CSPs is one of the effective strategies to overcome this bottleneck in studies of chiral separation materials. In this work, we have demonstrated that stable spherical covalent organic frameworks (SCOFs) can be utilized as matrixes for the fabrication of new CSPs for the first time. Specifically, a porous imine-linked SCOF with good crystallinity, large surface area, and high chemical stability is synthesized at room temperature. Then, cellulose-tris (3,5-dimethylphenylcarbamate) (CDMPC), a typical cellulose derivative, is selected as a potential chiral selector and coated onto the robust SCOFs, giving rise to the fabrication of new CDMPC@SCOF CSPs. The as-synthesized stable SCOF-based CSPs are exploited for high-performance liquid chromatographic (HPLC) enantioseparation, showing high resolution abilities for the separation of racemic compounds such as metalaxyl, 1-(1-naphthalenyl)ethanol, epoxiconazol, trans-stilbene oxide, and so on. Moreover, the prepared SCOF-based CSPs exhibit more superior acid and base stability than those of the silica-based CSPs. Our work not only uncovers the great potential of SCOFs as matrixes for constructing novel CSPs, but also expands the application of COFs in the field of enantiomeric separation under harsh base and acid conditions.
Subject(s)
Metal-Organic Frameworks , Cellulose/chemistry , Chromatography, High Pressure Liquid/methods , Silicon Dioxide/chemistry , StereoisomerismABSTRACT
Based on the community research experience of heat-sensitive moxibustion, this study explained technical recommendations for pragmatic randomized controlled trials (pRCTs) of heat-sensitive moxibustion in community from 7 aspects: selection of community research sites, ethical approval and registration, patient recruitment, training of standard operating procedures, ensuring patient compliance, quality control of follow-up visits and patient safety, which aimed to reduce the difficulty of research execution and improve the quality of pRCTs implementation and follow-up visits of heat-sensitive moxibustion.
Subject(s)
Moxibustion , Follow-Up Studies , Hot Temperature , Humans , Moxibustion/methods , Randomized Controlled Trials as TopicABSTRACT
BACKGROUND: Activation of amoeboid microglial cells (AMC) and its related inflammatory response have been linked to the periventricular white matter damage after hypoxia in neonatal brain. Hypoxia increases free ATP in the brain and then induces various effects through ATP receptors. The present study explored the possible mechanism in ATP induced AMC activation in hypoxia. RESULTS: We first examined the immunoexpression of P2X4, P2X7 and P2Y12 in the corpus callosum (CC) and subependyma associated with the lateral ventricles where both areas are rich in AMC. Among the three purinergic receptors, P2X4 was most intensely expressed. By double immunofluorescence, P2X4 was specifically localized in AMC (from P0 to P7) but the immunofluorescence in AMC was progressively diminished with advancing age (P14). It was further shown that P2X4 expression was noticeably enhanced in P0 day rats subjected to hypoxia and killed at 4, 24, 72 h and 7 d versus their matching controls by double labeling and western blotting analysis. P2X4 expression was most intense at 7 d whence the inflammatory response was drastic after hypoxia. We then studied the association of P2X4 with cytokine release in AMC after hypoxic exposure. In primary microglial cells exposed to hypoxia, IL-1ß and TNF-α protein levels were up-regulated. Blockade of P2X4 receptor with 2', 3'-0-(2, 4, 6-Trinitrophenyl) adenosine 5'-triphosphate, a selective P2X1-7 blocker resulted in partial suppression of IL-1ß (24% vs hypoxic group) and TNF-α expression (40% vs hypoxic group). However, pyridoxal phosphate-6-azo (benzene-2, 4-disulfonic acid) tetrasodium salt hydrate, a selective P2X1-3, 5-7 blocker did not exert any significant effect on the cytokine expression. CONCLUSIONS: It is concluded that P2X4 which is constitutively expressed by AMC in postnatal rats was enhanced in hypoxia. Hypoxia induced increase in IL-1ß and TNF-α expression was reversed by 2', 3'-0-(2, 4, 6-Trinitrophenyl) adenosine 5'-triphosphate suggesting that P2X4 mediates ATP induced AMC activation and its production of proinflammatory cytokines.
Subject(s)
Adenosine Triphosphate/metabolism , Cell Movement/physiology , Hypoxia-Ischemia, Brain/metabolism , Microglia/metabolism , Receptors, Purinergic P2X4/physiology , Amoeba , Animals , Animals, Newborn , Cell Movement/drug effects , Disease Models, Animal , Humans , Hypoxia-Ischemia, Brain/pathology , Infant, Newborn , Inflammation/metabolism , Inflammation/pathology , Interleukin-1beta/biosynthesis , Microglia/cytology , Microglia/pathology , Primary Cell Culture , Rats , Rats, Sprague-Dawley , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
The aim of this study is to establish a method to culture and purify cerebral astrocyte of tree shrew (Tupaia belangeri), a kind of new laboratorial animal which is a relative of primates. Newborn tree shrews were used in this experiment. The cortex of cerebrum was isolated and placed in 4°C for 20 min to injure neurons. The cortical tissue was disaggregated by trypsin digestion. Differential attachment method was used to remove fibroblasts. The mixed culture was rinsed by trypsin (0.005%) solution to remove neurons. Upon reaching 70% confluence, the culture was subjected to static trypsin digestion until a white slice film exfoliated from the bottom of culture bottle. This film, i.e. astrocyte layer, was taken out and cultured, and the third passage was identified by immunocytochemical staining and immunofluorescence with anti-glial fibrillary acidic protein (GFAP) antibody. The result showed the purity of tree shrew astrocytes was more than 98%. Thus the method to culture highly purified astrocyte of tree shrew was successfully established, which would contribute to further study in central nervous system physiology and diseases in this new laboratorial animal.
Subject(s)
Astrocytes/cytology , Brain/cytology , Primary Cell Culture/methods , Tupaiidae , Animals , Cell Separation/methodsABSTRACT
OBJECTIVE: To inspect the third stage larvae of Gnathostoma in imported Monopterus albus, and identify its species. METHODS: Ten batches of M. albus imported to Shanghai were detected for nematode Gnathostoma from January 2010 to March 2011. Fifty-two M. albus imported from the Philippines (25), Indonesia (24) and Bangladesh (3) were sampled (3-10/batch), which were dissected, minced, and digested. The suspension was filtered with 10 mesh screen to take the deposit. The complete parasites were picked out under stereoscope followed by morphological identification. The rate and intensity of infection were calculated. Genomic DNA of Gnathostoma was extracted to amplify internal transcribed spacer region 2 (ITS-2) and cytochrome C oxidase subunit 1 (cox1) by PCR, the product of which was analyzed by electrophoresis and sequencing. The sequences were aligned with corresponding sequences in GenBank. RESULTS: The third stage larvae of Gnathostoma were detected in M. albus from Indonesia and Philippines with infection rate of 36.0% (9/25) and 50.0% (12/24) and average infectiosity of 7.8 (70/9) and 2.8 (34/12), respectively. No Gnathostoma was found in M. albus imported from Bangladesh. Under microscope, the larvae showed one cephalic bulb with 4 rings of hooklets on it, cross striations and small spines on the body surface. The front body spines were bigger and denser, while the rear spines were smaller and sparser. It had 1 cervical papilla and 4 cervical capsules. Morphological characteristics were similar to the third stage larvae of G. spinigerum. PCR results showed that the length of the ITS-2 and cox1 PCR products was 647 bp and 441 bp, respectively. Sequence alignment analysis showed that the two PCR products had 99%-100% consistency with G. spinigerum ITS-2 (GenBank Accession No. AB181155 and Z97175) and cox1 (GenBank Accession No. AY501388, AB180099, and AB551552). CONCLUSION: All the larvae detected in M. albus imported from the Philippines and Indonesia have been identified as G. spinigerum.
Subject(s)
Fishes/parasitology , Gnathostoma/classification , Gnathostoma/isolation & purification , Quarantine/methods , Animals , China , LarvaABSTRACT
OBJECTIVE: The aim of this study was to quantify the expression of melanoma-antigen family A proteins (MAGE-A) and New York esophageal squamous cell carcinoma-1 (NY-ESO-1) in breast cancer and establish the prognosis of breast cancer patients with MAGE-A and NY-ESO-1 co-expression. METHODS: A total of 122 patients with breast cancer were recruited for this study. Their clinicopathological data were collected retrospectively, and the MAGE-A and NY-ESO-1 expressions in paraffin-embedded specimens from the 122 patients were evaluated using immunohistochemical analysis. In addition, the survival states of the patients were recorded. RESULTS: Fifty-four patients (44.26%) were MAGE-A positive and 46 (37.70%) were NY-ESO-1 positive. Interestingly, 22 of the 46 NY-ESO-1-positive cases co-expressed MAGE-A. The expression of MAGE-A was positively associated with estrogen-receptor status (χ2 = 4.026, p = 0.045) and human epidermal growth factor receptor 2 status (χ2 = 5.482, p = 0.019), while the expression of NY-ESO-1 was positively associated with p53 expression (χ2 = 4.541, p = 0.033). Of the 122 patients, the lowest survival rate was observed in patients with NY-ESO-1 (+)/MAGE-A (+), with a 5-year survival rate of 59.09% and a median survival of 97 months. CONCLUSION: The results showed that MAGE-A and NY-ESO-1 were frequently expressed in breast cancer patients. The co-expression of MAGE-A and NY-ESO-1 occurred in about 18% of these patients, and it may indicate a poor prognosis.
ABSTRACT
Amine-functionalized covalent organic frameworks (COFs) hold great potential in diversified applications. However, the synthesis is dominated by postsynthetic modification, while the de novo synthesis allowing for direct installation of amine groups remains a formidable challenge. Herein, we develop a site-selective synthetic strategy for the facile preparation of amine-functionalized hydrazone-linked COF for the first time. A new monomer 2-aminoterephthalohydrazide (NH2-Th) bearing both amine and hydrazide functionalities is designed to react with benzene-1,3,5-tricarbaldehyde (Bta). Remarkably, the different activity of amine and hydrazide groups toward aldehyde underpin the highly site-selective synthesis of an unprecedented NH2-Th-Bta COF with abundant free amine groups anchored in the well-defined pore channels. Interestingly, NH2-Th-Bta COF exhibits dramatically enhanced iodine uptake capacity (3.58 g g-1) in comparison to that of the nonfunctionalized Th-Bta COF counterpart (0.68 g g-1), and many reported porous adsorbents, despite its low specific surface area. Moreover, NH2-Th-Bta COF possesses exceptional cycling capability and retained high iodine uptake, even after six cycles. This work not only provides a simple and straightforward route for the de novo synthesis of amine-functionalized COFs but also uncovers the great potential of amine-functionalized COFs as adsorbents in the efficient removal of radioiodine and beyond.