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Objective:To investigate the effect of LRRK2G2019S mutation on activation of microglia after iron deprivation and its mechanism.Methods:(1) Microglia were differentiated from human induced pluripotent stem cells (IPSC) with the help of hematopoietic progenitor cells (HPC) and identified by immunofluorescent staining, and α-synuclein (α-syn) A53T mutant protein was obtained by protein purification technology. (2) Microglia were divided into control group, α-syn group, α-syn+ deferoxamine (DFO) group; phosphate buffer solution (PBS), 1 μmol/L purified α-syn A53T mutant protein, 1 μmol/L purified α-syn A53T mutant protein+30 mmol/L DFO were given respectively for 24 h. Fe 2+ concentration was detected by colorimetry, Rab35 protein expression was detected by Western blotting, intracellular reactive oxygen species (ROS) level was detected by flow cytometry, and interleukin-6 ( IL-6), tumor necrosis factor-α ( TNF-α) and transforming growth factor-β ( TGF-β) mRNA expressions were detected by real time-PCR (RT-PCR); microglia culture supernatant (MCS) in the 3 groups were transfered to SH-SY5Y cells, and SH-SY5Y cell apoptosis was detected by flow cytometry. (3) Bidirectional DNA sequencing was used to detect leucine rich repeat kinase 2 ( LRRK2) gene mutations in microglia treated with 1 μmol/L purified α-syn A53T mutant protein. Microglia were divided into control group, α-syn group and α-syn+GSK3357679A group, and treated with corresponding drugs for 24 h, respectively (LRRK2 inhibitor GSK3357679A concentration: 10 nmol/L), and LRRK2 protein expression was detected by Western blotting; microglia were divided into control group, α-syn group, α-syn+GSK3357679A, and α-syn+GSK3357679A+DFO group, and treated with corresponding drugs for 24 h, Rab35 protein expression was detected by Western blotting, intracellular ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. (4) Microglia were divided into control group, α-syn group, α-syn+rapamycin (RAPA) group, and treated with corresponding drugs for 24 h (concentration of autophagy inducer RAPA: 50 nmol/L); protein expressions of Rab35, P62 and microtubule-associated protein light chain 3 II (LC3II) were detected by Western blotting; intracellular ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. (5) Microglia were divided into control group, α-syn group, and α-syn+Rab35 group, and treated with corresponding drugs for 24 h (concentration of Rab35 overexpressed plasmids: 1 μg/mL); Rab35, P62, and LC3II protein expressions were detected by Western blotting; ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. Results:(1) Immunofluorescent staining showed negative neuronal nuclei (NeuN) expression and positive ionized calcium-binding adapter molecule 1 (Iba1) expression in microglia, and high LRRK2 expression; PcDNA3.1-SNCA-A53T expression plasmid was constructed and α-syn A53T mutant protein was purified. (2) The Fe 2+ concentration in α-syn group was significantly higher than that in control group, and the Fe 2+ concentration in α-syn+DFO group was significantly lower than that in α-syn group ( P<0.05); the Rab35 protein and TGF-β mRNA expressions in control group, α-syn group and α-syn+DFO group were decreased successively, while the IL-6 and TNF-α mRNA expressions were increased successively, with significant differences ( P<0.05); ROS level and SH-SY5Y cell apoptosis rate in control group, α-syn group, α-syn+DFO group were increased successively. (3) Bidirectional DNA sequencing showed that the LRRK2G2019S mutation in microglia was the most obvious after α-syn A53T mutant protein stimulation; compared with the control group, the α-syn group had significantly increased LRRK2 protein expression, while the α-syn+GSK3357679A group had significantly decreased LRRK2 protein expression compared with α-syn group ( P<0.05); compared with the control group, the α-syn group had significantly decreased Rab35 protein and TGF-β mRNA expressions, and statistically increased IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn group, the α-syn+GSK3357679A group had significantly increased Rab35 protein and TGF-β mRNA expressions, and statistically decreased IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn+GSK3357679A group, α-syn+GSK3357679A+DFO group had significantly increased IL-6 and TNF-α mRNA expressions, and significantly decreased Rab35 protein and TGF-β mRNA expressions ( P<0.05). The α-syn group had higher ROS level than the control group, the α-syn+GSK3357679A group had lower ROS level than the α-syn group, and the α-syn+GSK3357679A+DFO group had higher ROS level than the α-syn+GSK3357679A group. (4) Compared with the control group, the α-syn group had significantly decreased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly increased P62 protein, IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn group, the α-syn+RAPA group had significantly increased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly decreased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05); the α-syn group had higher ROS level than the control group and α-syn+RAPA group. (5) Compared with the control group, the α-syn group had significantly decreased Rab35 and LC3II protein, and TGF-β mRNA expressions, and statistically increased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05); compared with the α-syn group, the α-syn+Rab35 group had significantly increased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly decreased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05). The α-syn group had higher ROS level than the control group and α-syn+Rab35 group. Conclusion:LRRK2G2019S can induce neuroinflammation by inhibiting Rab35-related autophagy under iron deprivation, and Rab35 is expected to be a key factor in intervening neuroinflammation.
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BACKGROUND@#The presence of fibrosis is a criterion for subtype classification in the newly updated hypersensitivity pneumonitis (HP) guidelines. The present study aimed to summarize differences in clinical characteristics and prognosis of non-fibrotic hypersensitivity pneumonitis (NFHP) and fibrotic hypersensitivity pneumonitis (FHP) and explore factors associated with the presence of fibrosis.@*METHODS@#In this prospective cohort study, patients diagnosed with HP through a multidisciplinary discussion were enrolled. Collected data included demographic and clinical characteristics, laboratory findings, and radiologic and histopathological features. Logistic regression analyses were performed to explore factors related to the presence of fibrosis.@*RESULTS@#A total of 202 patients with HP were enrolled, including 87 (43.1%) NFHP patients and 115 (56.9%) FHP patients. Patients with FHP were older and more frequently presented with dyspnea, crackles, and digital clubbing than patients with NFHP. Serum levels of carcinoembryonic antigen, carbohydrate antigen 125, carbohydrate antigen 153, gastrin-releasing peptide precursor, squamous cell carcinoma antigen, and antigen cytokeratin 21-1, and count of bronchoalveolar lavage (BAL) eosinophils were higher in the FHP group than in the NFHP group. BAL lymphocytosis was present in both groups, but less pronounced in the FHP group. Multivariable regression analyses revealed that older age, <20% of lymphocyte in BAL, and ≥1.75% of eosinophil in BAL were risk factors for the development of FHP. Twelve patients developed adverse outcomes, with a median survival time of 12.5 months, all of whom had FHP.@*CONCLUSIONS@#Older age, <20% of lymphocyte in BAL, and ≥1.75% of eosinophil in BAL were risk factors associated with the development of FHP. Prognosis of patients with NFHP was better than that of patients with FHP. These results may provide insights into the mechanisms of fibrosis in HP.
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Humains , Liquide de lavage bronchoalvéolaire , Études prospectives , Alvéolite allergique extrinsèque/diagnostic , Fibrose , GlucidesRésumé
Objective:To study the application value of flexible endoscopic examination of swallowing(FEES) for the aspiration screening, the diagnosis of dysphagia and evaluation of the therapeutic effectin acute stroke patients with dysphagia.Methods:Three hundred and seventy-three patients with acute stroke who hospitalized from October 2015 to January 2020 in Dalian Municipal Central Hospital Affiliated of Dalian Medical University and underwent FEES for analyzing the characteristic performance were enrolled, and 11 cases of them were examined by video fluoroscopic swallowing study (VFSS). The results of the reliability of diagnosis dysphagia of the two methods were compared. The results of FEES for assessing the recovery effect after treatment were evaluated.Results:In 373 patients, the FEES revealed 268 cases(71.85%) of aspiration (99 cases) were recessive aspiration, which was better than that in water swallow test (50.94%, 190/373). Patients with potential cricopharyngeus achalasia got the same results through both of VFSS and FEES. FEES could provide more positive indicators and guide clinical rehabilitation treatment and objective assessment of rehabilitation effectiveness.Conclusions:Acute stroke patients with dysphagia have characteristic laryngeal performance. FEES is simple to operate and has high application value.
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Objective@#To understand the current situation of intimate partner violence (IPV) among young students in Chengdu and its relationship with emotion regulation self-efficacy,and to provide a reference for conducting the education on close relationship.@*Methods@#Totally 1 041 young students with love experience in Chengdu were selected by by stratified cluster random sampling to explore potentional factors related to IPV.@*Results@#The incidence of IPV perpetration among young students with love experience was as high as 69.6% and the incidence of IPV victimization was 62.2%. Young students had committed(65.4%) or been subjected(64.0%) to more than three intimate partner violence. 59.92% young students were both perpetrators and victims of IPV. Multiple Logistic regression analysis showed that compared with young female students, young male students were not prone to commit violence in intimate relationships(OR=0.59), but may become victims of IPV(OR=1.91). More than half a year in love(OR=1.70), cohabitation(OR=2.47), bullying by peers (OR=1.54) and interference by parents (OR=1.63) were risk factors for IPV perpetration. Among them, more than half a year in love (OR=1.51) and cohabitants (OR=2.52) were positively associated with IPV victimization. The efficacy of managing negative emotions was a negatively associated with IPV perpetration (OR=0.96) and victimization(OR=0.97)(P<0.05).@*Conclusion@#The phenomenon of intimate partner violence among young students is more common, which is closely related to the rearing style of young students, peer relationship, love relationship and the ability to manage negative emotions, which should be paid attention to.
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The presence of D-amino acid in proteins is thought to be a result of racemization of L-amino acid,which may influence the space structure and the functions of proteins and the connection to the occurence of some diseases has been demonstrated.Recently,tremendous advancement in instrumental techniques has brought the rapid development to analytical methods and detecting techniques in the research on racemization of protein amino acid.The advances in the relationship between racemizadon of protein amino acid and diseases,the influence factors on racemization of protein amino acid and the analytical techniques are reviewed in this article,and the research directions of racemization of protein amino acid are also given.
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Objective To construct a recombinant lentivirus containing human beta defensins -3 ( hBD3 ) , connective tissue growth factor gene (CTGF) and enhanced green fluorescent protein (EGFP), and to detect its translation in rabbit bone marrow mesenchymal stem cells (BMSC).Methods The lentivirus containing hBD3, CTGF and EGFP genes was constructed in vitro.The titer of lentivirus was tested with end-paint dilution assay .Rabbit BMSCs were transfected with recombinant virus.The best value of multiplicity of infection (MOI) was tested.The expression condition, transfection efficacy and genetic stability of the target genes were evaluated by using fluorescence microscopy and flow cytometry . Western blotting was used to detect the expression of the target protein .Results Recombinant lentivirus vectors: Lenti-CTGF-hBD3-EGFP, Lenti-hBD3-EGFP, and Lenti-EGFP, were successfully obtained . The titer of the recombinant lentiviruses was 3.21 ×108, 5.80 ×108, and 1.16 ×109, respectively.The best MOI value to transfect BMSCs was 150. The transfection efficacy of these lentivirus vectors was high , reaching 79.72%as assessed by flow cytometry , and it could be stably inherited .Western blotting displayed that target protein expression was successful .Conclusion The construction of recombinant lentiviruses carrying hBD3 and CTGF genes is successful and can be effectively transfected into BMSCs .
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Objective To investigate the effects of the eucalyptol,limonene and pinene enteric soft capsules (mucoregulatory agents) in preventing from tympanum fluid in children.Methods A total of 341 children (497 ears) with acute otitis media were randomly divided into two groups:the experimental group and the control group.The experimental group was given symptomatic treatment and oral in-take of the eucalyptol,limonene and pinene enteric soft capsules,while the control group with acute otitis media received conventional symptomatic treatment.All the patients were followed-up 7 and 14 days after treatment.Clinical data were collected,including symptoms,and tympanogram changes.Results Seven days after the initial treatment,tympanum fluid occurred in 110 cases of 170 ears(65.09%,110/170) in the control group,and in 51 cases of 70 ears (29.65%,51/172) in the experimental group.The difference was statistically significant (P<0.01).After 14 days,41 cases (66 ears)of tympanum fluid occurred in the control group (24.26%,41/169),32 cases(36 ears) of otitis media (18.60%,32/172) occurred in the experimental group.The difference was statistically significant (P<0.01).In the control group,the tympanum fluid disappeared in 54 cases(60 ears) after regular treatment 72.0%,54/75),and reliefed in 15 cases(22 ears) with irregular treatment (42.86%,15/35).The difference was statistically significant (P<0.01).Conclusion Eucalyptol,limonene and pinene enteric soft capsules can effectively reduce the incidence of tympanum fluid in children with acute otitis media.Mucoregulatory agents may promote the eduction of effusion.
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Objective To probe the periodontal ligament regeneration following the implantation of bone marrow mesenchymal cells ( BMSCs ) sheet-collagen membrane-BMSCs sheet sandwich complex.Methods BMSCs cell sheet-collagen membrane-BMSCs sheet complexes were compounded on the root surface of teeth of Beagle dogs.All the dogs were killed on 4 and 12 weeks after implantation.Periodontal ligament regeneration was observed by radiological means, HE staining and Sirus-red staining.Results Compared with collagen membrane group and blank control group, there was a clearly periodontal ligament like tissue and Sharpey′s like fibres formation in test group only.Conclustion Cell sheet-collagen membrane-cell sheet sandwich complex can effectively improve the periodontal ligament regeneration.
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Objective To study on whitening and anti -aging effect of ginseng saponin and its safety.Methods Whitening effect:using vitamin C as the control drug, the inhibition rate of tyrosinase was determined.Anti senescence effect: the aging model of D-was established, and the DPPH was applied to the skin of the rat's neck, and the drug was prepared by 2 times a day.Skin safety evaluation: the skin changes of the skin of the ginseng saponins were observed after the skin was given ginseng saponin and control drugs.Results When the concentration of ginsenoside was 7 mg/mL, the inhibition rate order was, the water solution of vitamin C >ginsenoside nano >ginsenoside aqueous solution ; In the aging model, the surface of the elastic fibers and the skin surface was parallel to the skin, but the elastic fibers were arranged in a more tortuous and non parallel to the skin;DPPH free radical scavenging effect order was, Vitamin C aqueous solution>ginsenoside nano>ginsenoside aqueous solution, and with the dosage of ginsenoside increasing, the efficiency of ginsenoside DPPH scavenging free radical increased significantly;Skin safety evaluation results showed that ginseng saponin nanometer milk smeared skin, no redness, irritation and other phenomena occured.Conclusion Ginseng saponin nanometer milk has obvious whitening effect, and can not cause damage to the skin, is safe and reliable.
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OBJECTIVE@#To investigate the therapeutic efficacy of coblation-assisting uvulopalatopharyngoplasty (CAUPPP) combining with coblation-channeling of the tongue (CCT) for patients with severe obstructive sleep apnea-hypopnea syndrome (OSAHS).@*METHOD@#A retrospective analysis was made on patients with severe OSAHS treated by CAUPPP combining with CCT. The adult with severe OSAHS were involved in the study and the apnea-hypopnea index (AHI) were beyond 40/h and lowest arterial oxygen saturation (LSaO2) were under or equal to 80%. Surgical approach: The UPPP was performed to decrease the size of soft palate with coblation and coblation-channeling in the soft palate and decrease the size of tongue by CCT. All patients were followed up for 6 to 12 months and underwent polysomnography (PSG).@*RESULT@#Subjective symptoms of patients improved more significantly than that of preoperation. The function of soft palate is normal without significant nasopharyngeal regurgitation. Compared with the preoperative data, AHI values were significantly decreased (P<0.01) and the lowest oxygen saturations increased significantly (P<0.01) and the effective rate was 91.7%.@*CONCLUSION@#There are usually multiple obstruction levels in patients with severe OSAHS and the traditional surgical treatment is not ideal. This study demonstrated that with the assistance of coblation, combining CAUPPP with CCT for patients with severe OSAHS is an effective surgical method. It has less blood loss, minimally invasive, retained the normal function of the soft palate, etc., should be widely applied.
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Adulte , Sujet âgé , Femelle , Humains , Mâle , Adulte d'âge moyen , Ablation par cathéter , Palais , Chirurgie générale , Palais mou , Chirurgie générale , Pharynx , Chirurgie générale , Études rétrospectives , Syndrome d'apnées obstructives du sommeil , Chirurgie générale , Langue , Chirurgie générale , Luette , Chirurgie généraleRésumé
OBJECTIVE@#To investigate the relationship between the protein expression of MDM2 and P16 in laryngeal squamous cell carcinoma (LSCC), and their relationship with clinic pathological feature.@*METHOD@#The expression of MDM2 and P16 were detected by S-P immunohistochemical method with mouse monoclonal anti MDM2 antibody and anti P16 antibody in 35 LSCC specimens, 21 specimens of precancerous larynx, 20 normal laryngeal specimens.@*RESULT@#Expression of MDM2 positive in normal laryngeal specimens, precancerous larynx and LSCC specimens were 10.0%, 38.1%, 54.3% respectively (P < 0.05), and in I - III pathological grade laryngeal carcinoma were 38.1%, 66.7%, 100.0% respectively (P < 0.05); Expression of P16 positive in normal laryngeal specimens, precancerous larynx and LSCC specimens were 85.1%, 52.4%, 34.3% respectively (P < 0.05), and in I - III pathological grade laryngeal carcinoma were 52.4%, 11.1%, 0% respectively (P < 0.05).@*CONCLUSION@#Expression of MDM2 positive were higher with lower differentiation, so it's maybe helpful to diagnose LSCC. Expression of P16 positive were lower with lower differentiation, and there's no expression in III pathological grade LSCC, so it has positive effect on malignancy determination. Abnormal expression of MDM2 and P16 have different effects on LSCC development. Expression of MDM2 and P16 have negative correlation in LSCC.
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Adulte , Sujet âgé , Humains , Adulte d'âge moyen , Carcinome épidermoïde , Métabolisme , Anatomopathologie , Inhibiteur p16 de kinase cycline-dépendante , Métabolisme , Tumeurs du larynx , Métabolisme , Anatomopathologie , Pronostic , Protéines proto-oncogènes c-mdm2 , MétabolismeRésumé
OBJECTIVE@#To investigate the efficacy and feasibility of coblation treatment of partial tonsillectomy in children with obstructive sleep apnea-hypopnea syndrome (OSAHS).@*METHOD@#The clinical datum of 91 children with OSAHS were retrospective analysis during the period from January 2009 to November 2009. All subjects, the main obstructive lesions were hypertrophy of tonsils and adenoids ,and which without recurrent pharyngodynia and tonsillitis, were applied with coblation treatment of partial tonsillectomy and adenoid ablation.@*RESULT@#Intraoperative blood loss was about 1-2 ml in all subjects,no primary and delayed hemorrhage happened and postoperative pain happened. All subjects were followed-up for 12 months to 22 months, symptoms of snoring and mouth breathing disappeared. No tonsil regeneration and repeated inflammatory episode was found.@*CONCLUSION@#Coblation treatment of partial tonsillectomy in children with OSAHS is micro-invasive, safe and effective, which is suitable for children of different age groups with hypertrophy of tonsils.
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Enfant , Enfant d'âge préscolaire , Femelle , Humains , Mâle , Ablation par cathéter , Méthodes , Études rétrospectives , Syndrome d'apnées obstructives du sommeil , Chirurgie générale , Amygdalectomie , MéthodesRésumé
<p><b>OBJECTIVE</b>To evaluate the effect of endoscopic surgery using the low-temperature plasma radiofrequency for nasal inverted papilloma. Methods In a retrospective study, clinical data of 8 patients underwent endoscopic surgery were analyzed since July 2008. Tumor stage according to Krouse was T1:1, T2:6, T3:1, T4:0, respectively. T1 and T2 lesions were completed excised using the low-temperature plasma radiofrequency, T3 lesions were excised using microdebrider combined with plasma radiofrequency.</p><p><b>RESULTS</b>The average blood loss was about 20 ml, with the exception of one patients with recurrent nasal inverted papilloma about 200 ml. Shorter operative time, minimal postoperative pain, epithelization of nasal cavity was seen 2 months postoperatively. There were no complications. All patients were followed-up for 2-6 months, with no recurrence.</p><p><b>CONCLUSIONS</b>Surgical treatment of nasal inverted papilloma using low-temperature plasma radiofrequency had following advantages: less blood loss, complete en bloc tumor resection, mild injury, short surgery time, mild postoperative pain. In summary, low-temperature plasma radiofrequency is a minimally invasive treatment for nasal inverted papilloma.</p>
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Sujet âgé , Sujet âgé de 80 ans ou plus , Femelle , Humains , Mâle , Adulte d'âge moyen , Ablation par cathéter , Méthodes , Basse température , Débridement , Endoscopie , Tumeurs du nez , Chirurgie générale , Papillome inversé , Chirurgie générale , Études rétrospectives , Résultat thérapeutiqueRésumé
Objective To investigate the use of the endotracheal silicone stent with silver wire in repair of stricture of laryngotrachea. Methods A retrospective analysis was performed on nine cases with stricture of laryngotracheal stenosis that were treated with endotracheal silicone stent with silver wire that was then decannulated three months later. Results Endotracheal silicone stent with silver wire were successfully decannulated in all cases. The follow up ranging from 11 months to three years showed no stricture happened again. Conclusion Endotracheal silicone stent with silver wire can effectively and minimally invasively treat the stricture of laryngotrachea.