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Aim To investigate the mechanism by which formononetin (FN) inhibits mitochondrial dynamic-related protein 1 (DRP1) -NLRP3 axis via intervening the generation of ROS to reduce allergic airway inflammation. Methods In order to establish allergic asthma mouse model, 50 BALB/c mice aged 8 weeks were divided into the control group, model group, FN treatment group and dexamethasone group after ovalbumin (OVA) induction. Airway inflammation and collagen deposition were detected by HampE and Masson staining. Th2 cytokines and superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and IgE levels in bronchoalveolar lavage fluid (BALF) were measured by ELISA, ROS in BEAS-2B cells was assessed by DCFH-DA staining, DRP1 expression in lung tissue and BEAS-2B cells was detected by immunohistochemistry and immunofluorescence, and the DRP1-NLRP3 pathway was analyzed by immunoblotting. Results FN treatment could effectively ameliorate the symptoms of asthmatic mouse model, including reducing eosinophil accumulation, airway collagen deposition, decreasing Th2 cytokine and IgE levels, reducing ROS and MDA production, increasing SOD and CAT activities, and regulating DRP1-NLRP3 pathway-related protein expression, thereby relieving inflammation. Conclusion FN ameliorates airway inflammation in asthma by regulating DRP1-NLRP3 pathway.
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Aim To investigate the protective effect and mechanism of JTE-013 on allergic rhinitis (AR) by regulating mitochondrial injury and apoptosis through RhoA/ROCKl/Drpl pathway. Methods AR model was established by ovalbumin (OVA) in mice. Nasal tissue sections were then stained with HE, TUNEL and DHE. Western blot assay. In vitro, human nasal epithelial cells (HNEpCs) were stimulated with human recombinant interleukin-13 (IL-13), and the effects of JTE-013 and Y27632-related protein expression were detected by Western blot. Immunofluorescence was used to observe the effects of JTE-013 and Y 27632 on total ROS, mitochondrial membrane potential and mitochondrial ROS generation, Drpl translocation and Cyt-c expression in cells. Results JTE-013 reduced the frequency of nose rubbing and sneezing, reduced nasal mucosal thickening and decreased eosinophil infiltration in AR mice. TUNEL and DHE staining results suggested that JTE-013 could inhibit apoptosis and reduce ROS expression in mouse nasal epithelial cells. Western blot showed that both JTE-013 and Y 27632 could significantly reduce RhoA, ROCK1, Drpl and p-Drpl(616), inhibit the expression of apoptotic proteins Bax, cleaved-caspase-3, Cyt-c, cleavedcaspase-9 and up-regulate the expression of p-Drpl (637) and Bcl-2. Immunofluorescence showed that inhibitors of JTE-013 or ROCK1 almost blocked IL-13mediated increase in ROS and mtROS production, inhibited decrease in mitochondrial membrane potential, and blocked Cyt-c expression and Drpl translocation in nasal mucosal epithelial cells. Conclusion JTE-013 can regulate the morphology and function of mitochondria by inhibiting RhoA/ROCKl/Drpl signaling axis, thereby alleviating nasal epithelial cell inflammation in mice with allergic rhinitis.
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Aim To investigate whether notoginsenoside Rl (PNS-R1) alleviates allergic rhinitis (AR) through AMP-activated protein kinase (AMPK)/mitochondrial fission critical protein (DRP1) -mediated mitochondrial fission. Methods Different doses of PNSRl were used to treat ovalbumin (OVA) -induced AR model mice,and the inhibitory effect of PNS-R1 on AR was investigated by observing allergic symptoms such as nasal rubbing and sneezing, as well as HE staining of nasal tissues. Serum IgE levels and nasal lavage fluid (NLF) inflammatory cytokine levels were detected by enzyme-linked immunosorbent assay (ELISA) and apoptosis-related proteins were detected by Western blot. In vitro human nasal epithelial cells (HNEpC) were stimulated with IL-13 to observe apoptosis, mitochondrial membrane potential, cellular ROS and mitochondrial ROS production, as well as the expression levels of AMPK/DRP1, expression levels of the TXNIP/NLRP3 inflammasomes and the translocation of DRP1. Results PNS-R1 attenuated allergic symptoms in AR mice, HE staining reduced inflammatory cells and reduced the levels of OVA-specific IgE in serum, and the levels of IL-4, IL-6, and IL-8 in NLF. PNS-R1 attenuated the apoptosis and ROS production of nasal epithelial cells in AR. In vitro PNS-R1 could up-regulate mitochondrial membrane potential after IL-13 stimulation, reduce ROS and mtROS production, the proportion of apoptotic positive cells, and reduce cleaved caspase-3, Bax, and up-regulate Bcl-2 expression, down-regulate DRP1 phosphorylation (Ser 616) and DRP1 translocation at the mitochondrial membrane in an AMPK-dependent manner, reducing TXNIP/NLRP3 expression. Conclusions PNS-R1 can protect mitochondrial integrity by inhibiting the AMPK/DRP1 signaling axis and its subsequent TXNIP/NLRP3 signaling axis,thereby alleviating rhinitis in AR mice.
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Aim To discuss the mechanism of Lurong Dabu Decoction on cough variant asthma. Methods Guinea pigs were divided into normal group(CON), model group(OVA), Lurong Dabu Decoction high-dose group(HIGH),low-dose group(LOW), and dexamethasone group(DEX)at random. The CVA model was established by smoking plus injection of OVA, aluminum hydroxide solution and nebulized inhalation to stimulate cough. Gguinea pigs were dissected 24 hours after the last challenge to obtain alveolar lavage fluid(BALF)and lung tissues. Immunoadsorption(ELISA)method was applied to detect the types of inflammatory cells and the content of inflammatory cytokines in BALF; HE and Masson staining of the middle lobe of the left lung were used to observe the pathological changes in lung tissues; immunohistochemical staining was used to observe TLR4 and WNT-5A protein expression and distribution of lung tissues; the protein extracted from the upper lobe of the left lung was used to measure the level of TLR4 and WNT-5A protein in lung tissues by Western blot; immunofluorescence was employed to measure the fluorescence intensity of TLR4 and WNT-5A in lung tissues; flow cytometry was used to detect IL-4 and IFN-γ in guinea pig lung tissues. Results Lurong Dabu Decoction could improve guinea pig airway inflammation, inhibit collagen fiber deposition, reduce the content of IL-4, IL-5, and IL-13 in BALF, and inhibit the protein expression of TLR4 and WNT-5A in lung tissues and increase IFN-γ levels in lung tissues while decreasing IL-4 levels. Conclusion Lurong Dabu Decoction may inhibit the occurrence of CVA through TLR4/WNT-5A signaling pathway.
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Aim To explore the important role of HDAC5 in P-gp expression in rats in high-altitude low oxygen environment and its effect on phenytoin sodium pharmacokinetics. Methods Wistar rats were transported to Batang, Yushu, Qinghai, at an altitude of 4010 m, with 6 rats in each group, divided into 1 d and 3 d groups. Different groups were given phenytoin, phenytoin combined with hypericin, and phenytoin combined with verapamil. Plasma and liver tissues were collected at different time after taking the drug in the plateau area. The concentration of phenytoin sodium in plasma was determined by UFLC-MS method. Changes in protein expression were detected by Western blot. Results The results of UFLC-MS showed that the AUC
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Objective To investigate the protective effect and mechanism of polydatin ( PD) on allergic rhinitis (AH) by regulating mitophagy through PINK1- Parkin signaling pathway, and to provide a new target for clinical treatment of AH.Methods Thirty-two BALB/c murine were randomly divided into 4 groups: control group, OVA group, PD low-dose (30 mg • kg 1 ) and high-dose ( 45 mg • kg 1 ) treatment groups.At the end of modeling, the total number of sneezing and nasal nibbing of murine was recorded.HE staining was used to observe the morphology of na- sal mucosal epithelium and eosinophil infiltration.Western blot was used to detect the expression of P1NK1 , Parkin, TOM20 and mitochondrial apoptosis- related proteins Bax, Bcl-2, caspase-3, cleaved- caspase-3 and Cytochrome C.Hie expression of P1NK1 and cleaved-caspase-3 in nasal epithelial cells (HNEpC) was observed by immunofluorescence.Re¬sults The frequency of sneezing and nasal rubbing movements was significantly increased, the nasal mu¬cosa epithelium was thickened, and eosinophils were accumulated in AH murine , these results were reversed after PD treatment.Western blot results shower] that signaling proteins PINK1/Parkin anrl pro-apoptotie pro¬teins, including Bax, caspase-3, cleaved-caspase-3 and Cytochrome C were significantly overexpressed, the expression of TOM20 and Bcl-2 was decreased in OVA group, and PD up-regulated the levels of P1NK1 , Parkin, as well as Bcl-2 and inhibited the expression of TDM20 and pro-apoptotic proteins, while after pre- treatment with mitochondrial division inhibitor 1 ( Mdi- vi-1 ) , the expression of P1NK1 and Parkin was re¬duced , the expression of TOM20 was increased, while PD treatment did not significantly affect this effect.From the immunofluorescence results, it can be seen that the level of P1NK1 was increased after IL-13 stim¬ulation of HNEpCs compared with the control group, and PD further up-regulated the expression of P1NK1 , which was suppressed after pretreatment with Mdivi-1 , while PD did not change this phenomenon.Western blot results for P1NK1 and cleaved-caspase-3 were con¬firmed by immunofluorescence.Conclusion PD may activate mitophagy through the P1NK1 -Parkin signaling pathway, thereby protecting against AR.
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Objective: To systematically evaluate and integrate the real experience of burn patients during rehabilitation, and to provide theoretical guidance for the continual nursing care for burn patients. Methods: The systematic evaluation method was adopted. Databases including the China National Knowledge Internet, Wanfang Database, China Biology Medicine disc were retrieved with the search terms in Chinese version of "/, //, ////", and PubMed, Embase, CINAHL, PsycINFO, Cochrane Library were retrieved with the search terms of "burn/burns, rehabilitation/recovery/survivor/survive, experience/views/perceptions/, qualitative/phenomenon/interview/grounded theory". The qualitative studies on the real experience of burn patients during rehabilitation published from the establishment of each database to June 2020 were searched. The quality of the included studies was evaluated according to the quality evaluation criteria for qualitative research of the Joanna Briggs Institute Evidence-Based Health Care Center. The research country, research method, research object, research content, and main research result were summarized, and meta-synthesis of the research results was conducted with the aggregative integration method. Results: A total of 12 studies were included, and the quality of all the studies was grade B. The studies were conducted in 8 countries including Australia, Canada, Norway, etc., the research method mainly was phenomenological research method, and all the studies were focused on adult burn patients. A total of 46 specific themes were extracted with totally 10 new categories formed after summarization, and 3 integrated results were obtained as follows: burn patients suffered from both physical and psychological burdens, and their normal life was broken; burn patients gained post-traumatic growth and could actively adjust to cope with life difficulties; burn patients had multiple needs. Conclusions: Burn patients experience both physical and psychological pains during rehabilitation, so they long for multiple support from family and society. Medical staff, social groups, and family members should pay attention to the psychological experience and needs of burn patients with different characteristics during rehabilitation, and build a multi-directional social support system to help patients return to the society and rebuild their lives.
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Adult , Humans , Adaptation, Psychological , Australia , Burns , China , Qualitative ResearchABSTRACT
Objective: To explore the application effects of feedforward control theory in the rollover bed treatment of mass patients with burn-explosion combined injury. Methods: A retrospective observational research was conducted. From June 13 to 14, 2020, 15 patients with severe burn-explosion combined injury caused by liquefied natural gas tank car explosion and conforming to the inclusion criteria were admitted to the Second Affiliated Hospital of Zhejiang University School of Medicine. There were 13 males and 2 females, aged 33-92 (66±17) years. All the patients were treated with rollover bed from 48 h post admission, and the feedforward control theory was introduced, including establishing a special feedforward control management team for rollover bed, clarifying the duties of the medical staff in the rollover bed treatment of patients, implementing the cooperation strategy of multidisciplinary physician, training and examining for 80 nurses in the temporarily organized nurse team in the form of "rollover bed workshop", and formulating the checklist and valuation list of rollover bed treatment for continuous quality control. The frequency and the total number of turning over, and successful rate of one-time posture change with the rollover bed of patients within 30 days of admission were recorded, the occurrences of adverse events caused by improper operation for the rollover bed during the treatment were observed, including respiratory and cardiac arrests, treatment interruption, unplanned extubation, bed falling, and skin graft displacement. The lowest levels of arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2), the number of patients with oxygenation index>300 mmHg (1 mmHg=0.133 kPa), and the occurrence of acute respiratory distress syndrome (ARDS) of patients within 2 days of admission and on the 30th day of admission were recorded. Results: Within 30 days of admission, the patients were turned over with the rollover bed for 2 to 6 times each day, with a total of 1 320 turning over operations, the successful rate of one-time posture change reached 99.9% (1 319/1 320), and no adverse event occurred. Within 2 days of admission, the lowest levels of PaO2 and PaCO2 of the patients were (100±19) and (42±4) mmHg, respectively, and the number of patients with mild, moderate, and severe ARDS were 10, 2, and 3, respectively, and none of the patients had oxygenation index>300 mmHg. On the 30th day of admission, the lowest levels of PaO2 and PaCO2 of the patients were (135±28) and (37±8) mmHg, respectively, 3 patients developed moderate ARDS, 1 patient developed severe ARDS, and 11 patients had oxygenation index>300 mmHg. Conclusions: The introduction of feedforward control theory in the treatment of rollover bed of mass patients with burn-explosion combined injury can ensure safe and successful completion of turning over with the rollover bed, promote the repair of burn wound, and improve respiratory function, and therefore improve the treatment quality of patients.
Subject(s)
Female , Humans , Male , Blood Gas Analysis , Burns/therapy , Explosions , Respiratory Distress Syndrome, Newborn , Retrospective StudiesABSTRACT
Acute lung injury is a common critical respiratory disease, which can further develop into acute respiratory distress syndrome (ARDS) with a high fatality rate, but there is no effective drug at present. Phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway is involved in cell proliferation, metabolism, survival, and motility through phosphorylation of numerous downstream effector molecules. It plays an important role in the development of acute lung injury by regulating mitochondrial function, apoptosis, autophagy, oxidative stress, and inflammatory response. The active ingredients in Chinese medicinals alleviate acute lung injury by targeting the PI3K/Akt signaling pathway. There has been an explosion of research on the treatment of acute lung injury by active ingredients in Chinese medicinals via PI3K/Akt signaling pathway, which is of great clinical research value. The article presented the first summary of studies exploring the correlation between PI3K/Akt signaling pathway and acute lung injury in recent years and summed up the protective effect of the active ingredients in Chinese medicinals against acute lung injury via PI3K/Akt signaling pathway, providing innovative mindsets and strategies for clinical application of active ingredients in Chinese medicinals in the treatment of acute lung injury.
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The study is to investigate the effect of glaucocalyxin A (GLA) on mast cell-mediated anaphylaxis. The animal welfare and experimental process of this experiment followed the regulations of the Animal Ethics Committee of Yanbian University. BALB/c mice were used in the animal experiment and randomly divided into five groups, control group, model group, and GLA low, medium, and high dose groups (10, 20, and 40 mg·kg-1). Mice were sensitized by intradermal injection of anti-dinitrophenyl-immunoglobulin E (DNP-IgE) into the ears and challenged with a mixture of DNP-human serum albumin (HSA) and 4% evans blue into the tail veins to prepare an animal skin passive cutaneous anaphylaxis (PCA) model, which was collected from both ears for measurement of dye staining and histology. Rat peritoneal mast cells (RPMCs) were used in the cell experiment and divided into control, IgE + antigen (Ag), and IgE + Ag + GLA groups to determine histamine release as well as calcium influx levels. High-affinity IgE receptor (FcεRI)-mediated signaling pathway proteins and HMGB1/TLR4/NF-κB (high mobility group box 1/toll like receptor 4/nuclear transcription factor kappa B) signaling proteins were detected by Western blot. The results of animal experiments suggest that GLA inhibits PCA, reduces evans blue dye exudation, and reduces ear inflammation and ear thickness in mice. The results of cellular experiments suggested that GLA could reduce histamine release and calcium influx, and inhibit tumor necrosis factor-α (TNF-α), interleukin (IL)-4, IL-13, and IL-1β production; Western blot results showed that GLA inhibited FcεRI-mediated phosphorylation levels of spleen tyrosine kinase (Syk), Lck/Yes novel tyrosine kinase (Lyn), tyrosine kinase Fyn (Fyn), growth-factor receptor-bound protein 2 (Gab2), and phospholipase C (PLC) γ1, while GLA inhibited HMGB1/TLR4 signaling pathway to limit NF-κB p65 nuclear metastasis. The results indicate that GLA inhibits mast cell degranulation and attenuates allergic inflammation through the HMGB1/TLR4/NF-κB signaling pathway.
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Aim To investigate the efficacy of arctigenin on airway inflammation in a mouse model of asthma and the mechanism related to the SIRT1/NLRP3 signaling pathway. Methods Forty female BALB/c mice of clean grade were selected and divided into control group, OVA model group and ATG group (5, 10 and 20 mg · kg
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Aim To investigate the effect of astragalin (AG) on airway inflammation in asthmatic mice and its mechanism. Methods Fifty SPF male mice were randomly divided into normal group, asthma model group, and astragalin low (AG25), medium (AG50), and high (AG100) dose groups. A mouse model of asthma was prepared by egg albumin inhalation, the number of cells was counted by Diff-Quik staining after collecting bronchoalveolar lavage fluid (BALF), and IL4, 5, and 13 levels in BALF were measured by ELISA. The inflammatory changes in mouse lung tissues were observed using HE staining. Reactive oxygen species (ROS) levels were measured using a DCFH-DA probe, and superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured by colorimetry. IL-4, IL-5, IL-13, NOX2, p47phox, p-NF-κBp65, NF-κBp65, IκBα, p-IκBα and β-actin expressions in lung tissues were detected by Western blot. Results AG significantly reduced the number of inflammatory cells and total cells in BALF, decreased IL-4, IL-5, and IL-13 contents in BALF and lung tissues, and reduced inflammatory cell infiltration in lung tissues. AG inhibited nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) and p47phox expression, decreased ROS levels and MDA levels, increased SOD activity, and inhibited IκBα and NF-κBp65 phosphorylation. Conclusion AG attenuates airway inflammation in asthma by modulating the oxidative stress response through the NOX2/ROS/NF-κB signaling pathway.
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Objective:To observe the effect of Jianpi Xiaoai prescription on long non-coding RNA Hox transcript antisense intergenic RNA (lncRNA HOTAIR)/Janus kinase 2 (JAK2) /signal transducer and activator of transcription 3 (STAT3) signaling pathway and to explore the potential mechanism of Jianpi Xiaoai prescription in suppressing the metastasis of colon cancer. Method:The expression of lncRNA HOTAIR in different cells was analyzed. Following the treatment of HCT116 cells with 10%,15%,and 20% Jianpi Xiaoai prescription -containing serum, the invasive ability of Jianpi Xiaoai prescription on HCT116 cells was assessed by transwell assay. The mRNA expression levels of lncRNA HOTAIR,JAK2,and STAT3 were measured by Real-time polymerase chain reaction (Real-time PCR), and the protein expression levels of JAK2, phosphorylated STAT3 (p-STAT3) and STAT3 by Western blot. Result:The highest expression of lncRNA HOTAIR was detected in HCT116 cells. Compared with the blank group, each Jianpi Xiaoai prescription group exhibited a decreased number of invasive cells (<italic>P</italic><0.05, <italic>P</italic><0.01). The relative JAK2 mRNA expression in the middle-dose Jianpi Xiaoai prescription group was down-regulated (<italic>P</italic><0.05), and the relative lncRNA HOTAIR mRNA expression in the middle- and high-dose Jianpi Xiaoai prescription groups and the relative JAK2 mRNA expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated (<italic>P</italic><0.01). Compared with the blank group,the relative p-STAT3 protein expression was down-regulated in the middle-dose Jianpi Xiaoai prescription group (<italic>P</italic><0.05), and the relative JAK2 protein expression in the middle- and high-dose Jianpi Xiaoai prescription groups and the relative p-STAT3 protein expression in the high-dose Jianpi Xiaoai prescription group were remarkably down-regulated (<italic>P</italic><0.01). Conclusion:Jianpi Xiaoai prescription effectively inhibits the metastasis of colon cancer cells, which may be related to the inhibition of lncRNA HOTAIR/JAK2/STAT3 signaling pathway.
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Objective:To observe and evaluate the efficacy and safety of Ganduqing Granules in the treatment of common cold due to qi deficiency and pathogen invasion. Method:A multi-center, randomized, double-blind, and placebo-controlled clinical trial was conducted. One hundred and twenty patients were randomly divided into an experimental group (<italic>n</italic>=60) and a control group (<italic>n</italic>=60). Patients in the experimental group were treated with oral Ganduqing Granules, 6 g per time, 3 times per day, while those in the control group received placebo, 6 g per time, 3 times per day. After treatment for 5 successive days in both groups, the overall remission rate, total score of primary symptoms, total score of secondary symptoms, total score of all symptoms, time required for overall symptom remission, and therapeutic efficacy against traditional Chinese medicine (TCM) syndrome in the two groups were analyzed in both full analysis set (FAS) and per-protocol set (PPS), followed by the evaluation of safety in the safety set (SS). Result:No drop-out was found in the experimental group, whereas 3 cases in the control group dropped out. After 5 days of treatment, the overall remission rate of the experimental group was significantly higher than that of the control group (<italic>P</italic><0.01). The analysis in FAS and PDS revealed identical results. The experimental group was obviously better than the control group in improving the total scores of primary symptoms, secondary symptoms, and all symptoms (<italic>P</italic><0.01). The analysis results in FAS and PDS were consistent. There was no significant difference in overall remission time between the two groups. The experimental group was remarkably superior to the control group in alleviating such symptoms as aversion to wind and cold, nasal congestion, runny nose, fatigue, shortness of breath, laziness to speak, dry throat, sore throat, cough, and expectoration (<italic>P</italic><0.05,<italic>P</italic><0.01). The analysis results in FAS were the same as those in PDS. All the patients did not present with significant abnormalities in vital signs, blood routine test, or liver and kidney function tests after medication. There was no significant difference in the incidence of adverse events between the experimental group and the control group. Conclusion:Ganduqing Granules effectively alleviate the symptoms of patients with common cold and shorten the course of disease, without inducing obvious side effects.
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Objective::To investigate the effect of drug-containing serum of Jianpi Xiaoai prescription on protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathways in colorectal cells HCT116. Method::The HTC116 cells were treated by 15%concentration of drug-contained serum, and then the cell migration and invasion were detected by Transwell assay, the protein expression levels of Akt, phosphorylated protein kinase B (p-Akt), mTOR, phosphorylated mammalian target of rapamycin (p-mTOR), ribosomal protein S6 kinase, polypeptide1(S6K1), phosphorylated ribosomal protein S6 kinase, polypeptide1 (p-S6K1), 4E-binding protein1(4EBP1), and phosphorylated 4E-binding protein1(p-4EBP1) in HCT116 cells were detected by Western blot. The control group was treated by untreated serum (15%), and 10%fetal bovine serum(FBS). Result::As compared with the control group, the number of migration and invasion cells was significantly reduced in drug-contained serum group (P<0.01), the expression of Akt had no obvious decrease, p-Akt protein expression was significantly lowered in the drug-contained serum group (P<0.01), the expression of mTOR had no obvious decrease, but p-mTOR protein expression was significantly lowered in drug-contained serum group (P<0.01), the expression of S6K1 had no obvious decrease, but p-S6K1 protein expression was significantly lowered in the drug-contained serum group (P<0.01), the protein expression of 4EBP1 had no obvious decrease, but p-4EBP1 protein expression was significantly lowered in the drug-contained serum group (P<0.01). Conclusion::The anti-tumor mechanism and transfer of Jianpi Xiaoai prescription may be related to inhibiting the activation of Akt/mTOR signaling pathways in colorectal cancer.
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Objective:To observe the effect of Jianpi Xiaoai prescription on the activation of normal human embryonic lung fibroblasts (HFL1) into tumor-associated fibroblasts (CAFs) induced by human colon cancer cells (HCT116) derived exosomes. Method:SD rats were gavaged with 13.1 g·kg-1 of Jianpi Xiaoai prescription to prepare drug-containing serum, and HCT116 cell exosomes-containing 10% exosomes-free serum and 20% Jianpi Xiaoai prescription drug serum were isolated by ultra-high speed centrifugation. The particle size distribution of exosomes were detected by Nanoparticle tracking analyzer (Zetaview), and the exosomes' marker proteins apoptotic transfer gene 2 interaction protein X (Alix), heat shock protein 70 (HSP70), and tumor-susceptibility gene 101 (TSG101) were identified by Western blot, and the uptake of exosomes labeled with cell membrane staining kit (PKH67) by HFL1 was observed by fluorescence microscope. HFL1 cells were divided into six groups: the blank group, the transforming growth factor-β1 (TGF-β1) group, the TGF-β1 combined with HCT116 exosomes of 2 mg·L-1 group, the TGF-β1 combined with HCT116 exosomes of 4 mg·L-1 group, the TGF-β1 combined with Jianpi Xiaoai prescription exosomes of 2 mg·L-1 group, and the TGF-β1 combined with Jianpi Xiaoai prescription exosomes of 4 mg·L-1 group, and all groups were cultivated for 48 h. Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to determine the protein and mRNA expressions of α-smooth muscle actin (α-SMA). Result:The particle size distribution detected by Zetaview was mainly between 50-100 nm, and the exosomes were verified based on the expressions of marker proteins Alix, HSP70 and TSG101. After co-incubation of HFL1 cells with exosomes, a large number of exosomes were absorbed by HFL1 cells under fluorescence microscope. Compared with the blank control group, the protein and mRNA expressions of α-SMA in the TGF-β1 group and TGF-β1 combined with HCT116 exosome groups were increased (P<0.01). Compared with the TGF-β1 combined with HCT116 exosome groups, the protein and mRNA expressions of α-SMA were decreased in the TGF-β1 combined with Jianpi Xiaoai prescription exosome groups (P<0.01). Conclusion:Human colon cancer cell exosomes combined with TGF-β1 can induce the activation of HFL1 into CAFs, and Jianpi Xiaoai prescription can reduce the activation of HFL1 by affecting the expressions of α-SMA, thus antagonizing the lung metastasis of colon cancer.
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Objective:To observe the effect of polyphyllin Ⅰ on the expressions of forkhead box Q1(FOXQ1)and epithelial-mesenchymal transition (EMT)-related factors, in order to explore the possible mechanism of polyphyllin Ⅰ in inhibiting the metastasis of colon cancer. Method:After the treatment with 1.25,2.50 μmol·L-1 polyphyllin Ⅰ on HCT116 cells, Western blot and Real-time PCR were used to detect the expressions of FOXQ1,E-cadherin,Vimentin protein and mRNA. Result:Compared with the blank group, relative expressions of FOXQ1 protein and mRNA in low-concentration polyphyllin Ⅰ group were decreased, while relative expression of E-cadherin mRNA was increased, the differences were not statistically significant, and relative expressions of Vimentin protein and mRNA in low-concentration polyphyllin Ⅰ group were decreased (P<0.05), and relative expression of E-cadherin protein in low-concentration polyphyllin Ⅰ group was increased (P<0.01). Compared with the blank group, relative expressions of FOXQ1, Vimentin protein and mRNA in high-concentration polyphyllin Ⅰ group were decreased,while relative expressions of E-cadherin protein and mRNA were increased (P<0.05, P<0.01). Compared with low-concentration polyphyllin Ⅰ group, relative expressions of Vimentin protein and mRNA in high-concentration polyphyllin Ⅰ group were decreased, but the difference was not statistically significant, relative expressions of E-cadherin protein and mRNA in high-concentration polyphyllin Ⅰ group were increased, whereas relative expressions of FOXQ1 protein and mRNA were decreased (P<0.05, P<0.01). Conclusion:Mechanism of polyphyllin Ⅰ inhibiting the metastasis of colon cancer may be related to the decrease of FOXQ1 and Vimentin expressions, and the up-regulation of E-cadherin.
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OBJECTIVE@#To investigate the influencing factors on prognosis of adult patients with chronic primary immune thrombocytopeuia (ITP) after rituximab treatment and predictive value of platelet (Plt) count.@*METHODS@#Clinical data of 52 adult patients with chronic primary ITP treated with rituximab from January 2012 to December 2016 were retrospectively analyzed, including 32 patients for failed in treatment as group A and 20 patients for succeeded in treatment as group B. The independent risk factors influencing the clinical efficacy of rituximab were analyzed. The influence of CD41 megakaryocyte count in bone marrow diagnosed for first time on the response rate of patients with 1-year followed-up were observed, and the Plt count were calculated to predict the clinical efficacy index and the best cut-off point.@*RESULTS@#The CD41 megakaryocyte count in bone marrow for first time treatment in group B were significantly higher than that in group A (P<0.05). Multivariate Logistic regression analysis showed that the number of CD41 megakaryocytes in bone marrow<150 at first diagnosis was the independent risk factor influencing the clinical efficacy of rituximab (OR=5.40,95%CI:1.82-15.66,P=0.00). The response rate of 1-year followed-up in patients with CD41 megakaryocyte count ≥150 at first diagnosis was significantly higher than that of CD41 megakaryocyte count <150 (P<0.05). The Plt count level in group B was significantly lower than that in group A at the 3rd, 14th, 21th, 30th, 60th, 90th, 180th, 270th and 360th days after first treatment with rituximab (P<0.05). ROC curve analysis showed that the best cut-off point for Plt count was 50×10/L and AUC was 0.68 at the 14th day after first treatment with rituximab (95%CI: 0.57-0.78, P=0.00). The predictive sensitivity and specificity of clinical efficacy in adult patients with chronic primary ITP treated with rituximab were separately 48.73% and 87.58%, and the AUC in 30th and 60th day after rituximab treatment were separately 0.74 (95%CI: 0.64-0.87, P=0.00), 0.93 (95%CI:0.82-0.98,P=0.00).@*CONCLUSION@#Adult patients with chronic primary ITP may possess long-term remission after rituximab treatment, but the prognosis is poor for patients with bone marrow megakaryocyte count <150. The Plt counts in 14th, 30th and 60th days after rituximab treatment can effectively predict the long-term clinical efficacy and guide the formulation of treatment plans.
Subject(s)
Adult , Humans , Megakaryocytes , Platelet Count , Prognosis , Purpura, Thrombocytopenic, Idiopathic , Retrospective Studies , RituximabABSTRACT
Periodontal tissue, especially the alveolar bone, are closely associated with the progress and efficacy of orthodontic treatment. Prior to and during orthodontic treatment, dentists should fully evaluate the status of periodontal hard tissues to prevent clinical problems. This article aims to discuss bone issues associated with orthodontic treatment, including gingival papilla absence, alveolar bone insufficiency, excessive cortical resistance, and altered passive eruption, etc. The mechanism and prevention methods of these problems are also described.
Subject(s)
Gingiva , PeriodontiumABSTRACT
OBJECTIVE@#To evaluate the association between Chinese medicine (CM) therapy and disease-free survival (DFS) outcomes in postoperative patients with non-small cell lung cancer (NSCLC).@*METHODS@#This multiple-center prospective cohort study was conducted in 13 medical centers in China. Patients with stage I, II, or IIIA NSCLC who had undergone radical resection and received conventional postoperative treatment according to the National Comprehensive Cancer Network (NCCN) guidelines were recruited. The recruited patients were divided into a CM treatment group and a control group according to their wishes. Patients in the CM treatment group received continuous CM therapy for more than 6 months or until disease progression. Patients in the control group received CM therapy for less than 1 month. Follow-up was conducted over 3 years. The primary outcome was DFS, with recurrence/metastasis rates as a secondary outcome.@*RESULTS@#Between May 2013 and August 2016, 503 patients were enrolled into the cohort; 266 were classified in the CM treatment group and 237 in the control group. Adjusting for covariates, high exposure to CM was associated with better DFS [hazard ratio (HR) = 0.417, 95% confidential interval (CI): 0.307-0.567)]. A longer duration of CM therapy (6-12 months, 12-18 months, >24 months) was associated with lower recurrence and metastasis rates (HR = 0.225, 0.119 and 0.083, respectively). In a subgroup exploratory analysis, CM therapy was also a protective factor of cancer recurrence and metastasis in both stage I-IIIA (HR=0.50, 95% CI: 0.37-0.67) and stage IIIA NSCLC postoperative patients (HR = 0.48, 95% CI: 0.33-0.71), DFS was even longer among CM treatment group patients.@*CONCLUSIONS@#Longer duration of CM therapy could be considered a protective factor of cancer recurrence and metastasis. CM treatment is associated with improving survival outcomes of postoperative NSCLC patients in China. (Registration No. ChiCTR-OOC-14005398).