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Aim To investigate the regulatory effects of 3-bromopyruvate (3-BrPA) on apoptosis and autophagy of fibroblast-like synoviocytes (FLS) in rats based on AMPK/mTOR signaling pathway and the underlying mechanism. Methods FLS of rats in vitro were cultured and induced by tumor necrosis factor-α (TNF-α) to construct a model of rheumatoid arthritis (R A). MTT assay was used to explore the optimal concentration of TNF-α and 3 -BrPA for induction and treatment of FLS. The effects of 3-BrPA on the migration and invasion of FLS were detected by Wound healing assay and Transwell assay. The apoptosis of FLS was tested by flow cytometry and mitochondrial membrane potential assay kit (JC-1). Moreover, FLS autophagic flux was detected by mCherry-EGFP-LC3B-overexpressed plasmids, and the expression of apoptosis/autophagy-related proteins as well as AMPK/mTOR pathway-related proteins were detected by Western blot. Results 3-BrPA (15 μmol • L) significantly inhibited the proliferation, migration, and invasion of FLS stimulated by TNF-a (25 μg • L
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Cervical cancer is the most common gynecological malignant carcinoma in Chinese women, which seriously threatens women's health. Lymph node metastasis is the most important factor affecting the prognosis and adjuvant treatment of patients with early cervical cancer. Conventional systematic lymphadenectomy has always been a reliable method to detect lymph node metastasis. However, this procedure may affect the quality of life of patients because of high risks of complications. Recently, sentinel lymph node biopsy (SLNB) has been researched aiming to assess the status of pelvic lymph node metastasis intraoperatively, avoid unnecessary lymphadenectomy and achieve the goal of individualized diagnosis and treatment. Exploring efficient tracing methods and pathological ultra-staging is the key to the clinical application of SLNB for cervical cancer, and understanding the latest relevant clinical research progress will help SLNB to be applied in the clinic as soon as possible to benefit patients with cervical cancer.
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Femelle , Humains , Lymphadénectomie/méthodes , Noeuds lymphatiques/chirurgie , Métastase lymphatique/anatomopathologie , Stadification tumorale , Qualité de vie , Noeud lymphatique sentinelle/chirurgie , Biopsie de noeud lymphatique sentinelle/méthodes , Tumeurs du col de l'utérus/chirurgieRÉSUMÉ
Objective: To investigate the expression of long non-coding RNA LOC101927476 (LncRNA LOC101927476) in ovarian cancer and its effect on the biological characteristics of ovarian cancer. Methods: Patients with ovarian cancer who underwent surgery in Cancer Hospital of Chinese Academy of Medical Sciences from 2018 to 2019 were selected. The expressions of LOC101927476 in ovarian cancer cells 3AO, OVCA429, TOV21G, A2780, SKOV3, as well as 22 primary tumor tissues and their matched metastatic tumor tissues were detected by real-time quantitative polymerase chain reaction (RT-PCR). Ovarian cancer transcriptome sequencing data from the TCGA database was used to verify the expressions of LOC101927476 and GATA4. 3AO and OVCA429 cells were infected with lentivirus plasmid containing OE-LOC101927476 and single guide RNA (sg-RNA) targeting LOC101927476, respectively. The effects of LOC101927476 on migration and invasion were detected by Transwell and wound healing assay. The effect of LOC101927476 on cell proliferation was detected by cell counting kit-8 (CCK-8) assay. Results: RT-PCR assay showed that 20 out of 22 patients had significantly lower expression of LOC101927476 in their metastatic tumors compared with primary tumors. Transwell assay showed that overexpression of LOC101927476 significantly inhibited the invasion and migration capacities of 3AO cells. The numbers of invading and migrating 3AO cells infected with OE-LOC101927476 lentivirus were (357±63) and (699±65), respectively, lower than (661±95) and (1 024±76) in OE-EV group (P<0.050). In contrast, the numbers of invading and migrating OVCA429 cells with LOC101927476 knockdown were (512±72) and (472±40), respectively, higher than (309±13) and (363±27) in sg-Control group (P<0.050). Wound healing assay results showed that after 48 hours, the percentage of scratch healing of 3AO cells in OE-LOC101927476 group was (10.86±0.63)%, significantly lower than (57.38±4.42)% of OE-EV group (P=0.009). After 24 hours, the percentage of scratch healing of OCVA429 cells in sg-LOC101927476 group was (59.98±1.34)%, significantly higher than (23.15±2.03)% of sg-Control group (P=0.004). CCK-8 assays showed that the OD value of 3AO cells in OE-LOC101927476 group was (2.07±0.08), significantly lower than (2.29±0.04) of OE-EV group (P=0.009). The OD value of OVCA429 cells in sg-LOC101927476 group was (2.13±0.03), significantly higher than (1.93±0.03) of sg-Control group (P=0.001). The relative expression of GATA4 in OE-LOC101927476 group was (1.86±0.25), significantly higher than 1.00 of OE-EV group (P=0.001). In patients with high expression of LncRNA LOC101927476, the expression level of GATA4 was (2.93±0.35), which was higher than (0.29±0.06) of LOC101927476 low expression group (P=0.001). Conclusion: LncRNA LOC101927476 can inhibit the invasion, migration and proliferation of ovarian cancer cells.
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Femelle , Humains , Lignée cellulaire tumorale , Mouvement cellulaire/génétique , Prolifération cellulaire/génétique , Invasion tumorale , Tumeurs de l'ovaire/génétique , ARN long non codant/génétiqueRÉSUMÉ
This study aims to develop multifunctional drug delivery system based on hollow mesoporous copper sulfide (HMCuS) nanoparticles. This type of nanoparticles is expected to achieve the synergistic treatment of tumor by targeted phototherapy and chemotherapy. The carrier was synthesized by a substitution method, and the anti-cancer drug doxorubicin (DOX) was loaded and then modified by hyaluronic acid (HA) to prepare the anti-cancer drug system DOX/HMCuS-HA. The results suggested that DOX/HMCuS-HA presented uniform spherical structure, with the drug loading efficiency of 33.6%, the particle size and zeta potential being 113.8 ± 6.9 nm and 18.4 ± 2.8 mV, respectively. When 100 μg·mL-1 HMCuS was irradiated under 808 nm laser (2 W·cm-2) for 8 min, the temperature can heat up 51 ℃, demonstrating high photothermal conversion efficacy. Electron spin resonance (ESR) tests and methylene blue degradation experiments showed that HMCuS nanoparticles could simultaneously produce hydroxyl radical (•OH) mediated photodynamic therapy. In addition, HA was responsible for minimizing premature drug release and increasing tumor targeting efficiency by acting as a smart gatekeeper with tumor specific targeting moiety. In vitro drug release experiments showed that the coated HA could be degraded by intracellular lysosomal enzyme hyaluronidase, which facilitated DOX release. The acidic micro-environment of tumor cell and external near infrared (NIR) stimulus could trigger further release of DOX from the nanoparticles. These results point to a new strategy for timely and effective anti-tumor treatment.
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<p><b>BACKGROUND</b>Erythromycin-resistant Streptococcus pneumoniae isolates that causing invasive pneumococcal diseases (IPD) in Chinese children remain uncharacterized. This study aims to identify the resistance genes associated with erythromycin resistance and to determine the genetic relationships of IPD isolates in Chinese children.</p><p><b>METHODS</b>A total of 171 S. pneumoniae strains were isolated from 11 medical centers in China from 2006 to 2008. All the isolates were characterized via serotyping and antibiotic susceptibility determination. The erythromycin-resistant isolates were further characterized via ermB and mefA gene detection, multi-locus sequence typing analysis, and pulsed-field gel electrophoresis.</p><p><b>RESULTS</b>A total of 164 (95.9%) isolates showed resistance to erythromycin, of which 162 strains with high high-level resistance (MIC ≥ 256 µg/ml). A total of 104 (63.4%) isolates carry the ermB gene alone, whereas 59 (36.0%) harbor both ermB and mefA genes. Of the 59 strains, 54 were of serotypes 19A and 19F and were identified as highly clonal and related to the Taiwan(19F)-14 clone.</p><p><b>CONCLUSIONS</b>The erythromycin resistance rate in IPD isolates is significantly high and is predominantly mediated by the ermB gene. Isolates that carry both ermB and mefA genes are predominantly of serotypes 19A and 19F.</p>
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Adolescent , Enfant , Enfant d'âge préscolaire , Humains , Nourrisson , Antibactériens , Pharmacologie , Résistance bactérienne aux médicaments , Électrophorèse en champ pulsé , Érythromycine , Pharmacologie , Typage par séquençage multilocus , Infections à pneumocoques , Microbiologie , Sérotypie , Streptococcus pneumoniae , Classification , GénétiqueRÉSUMÉ
<p><b>OBJECTIVE</b>To establish a bcr-abl(+) cell line resistance to nilotinib, and to investigate the possible mechanisms of resistance.</p><p><b>METHODS</b>K562 cells were treated with gradually increasing concentrations of nilotinib to generate resistance cell line K562-RN. The folder of drug-resistance was evaluated by MTT assay. Cells apoptosis rate was detected by flow cytometry, the mRNA level of bcr-abl fusion gene by FISH, and the expression of apoptosis relative gene mRNA and protein (such as bcr-abl, HO-1, mdr1, Bcl-2 and caspase-3) by RQ-PCR and western blot.</p><p><b>RESULTS</b>The resistant cell line K562-RN was successfully established, with 2.01 fold resistant to nilotinib compared with K562 cell line \[the IC(50) value of nilotinib to K562 and K562-RN were (12.320 ± 1.720) µmol/L and (24.742 ± 2.310) µmol/L, respectively\]. It also had the cross resistance to adriamycin, homoharringtonine, etoposide and imatinib. Treated with different concentrations of nilotinib, cell apoptosis rate of K562-RN was significantly lower than that of K562 cells. The rate of bcr-abl gene positive cells was 92% in K562-RN by FISH assay. The mRNA and protein levels of bcr-abl, HO-1 and mdr1 expression up-regulated in K562-RN cells, while those of caspase-3 expression down-regulated, being significantly statistical difference when compared with K562 cells (P < 0.05).</p><p><b>CONCLUSION</b>Human leukemic cell line resistance to nilotinib, K562-RN is established successfully by gradually increasing concentrations of drug. The mechanisms of resistance in K562-RN is probably associated with increasing expression of bcr-abl, HO-1, mdr1 and decreasing expression of caspase-3 mRNA and protein levels.</p>
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Humains , Sous-famille B de transporteurs à cassette liant l'ATP , Glycoprotéine P , Métabolisme , Caspase-3 , Métabolisme , Résistance aux médicaments antinéoplasiques , Protéines de fusion bcr-abl , Métabolisme , Régulation de l'expression des gènes dans la leucémie , Heme oxygenase-1 , Métabolisme , Cellules K562 , Pyrimidines , PharmacologieRÉSUMÉ
To determine the effect of compression-force on the expression of alkaline phosphatase [ALP], and ALP activity in cementoblasts. We performed this study in the State Key Laboratory of Oral Diseases, West China Stomatology Hospital, Sichuan University, Chengdu, Sichuan, China from October to December 2010. We exposed murine immortalized cementoblasts [OCCM-30] to 2000-microstrain compression-force at a frequency of 0.5 Hz for 1, 3, 6, 12, and 24 hours. We assayed the cellular ALP activity after the treatments. We used real-time polymerase chain reaction [RT-PCR] and western blot to examine the gene and protein expression of ALP in the OCCM-30 cells at each time point. Two-thousand micro strain compressive force significantly up-regulated the mRNA expression of ALP in OCCM-30 cells, which reached a peak at 12 hours loading, and the protein expression change of ALP in response to compression-force was consistent with the variation of gene level. We also noted marked enhancement of ALP activity in OCCM-30 cells during the application of mechanical stress. The compression-force increased the expression of ALP in OCCM-30 cells, suggesting that mechanical stimulation may affect the cellular function of cementoblasts by regulating ALP expression, which may participate in cementum metabolism during orthodontic tooth movement
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Animaux de laboratoire , Phosphatase alcaline , Réaction de polymérisation en chaine en temps réelRÉSUMÉ
<p><b>OBJECTIVE</b>To measure the utility value of different skeletal malocclusion for patients receiving orthodontic treatment.</p><p><b>METHODS</b>Utility value of different skeletal malocclusion for patients was measured by rating scale and time trade-off.</p><p><b>RESULTS</b>The youth group had higher utility values than adult group for skeletal malocclusion Class II (protruding facial type) with median mandibular angle. The utility value of skeletal malocclusion Class III (concave facial type) with low mandibular angle was the lowest, and the utility value of skeletal malocclusion Class II (protruding facial type) with median mandibular angle was the highest. There was no difference in the utility values by rating scale and by time trade-off.</p><p><b>CONCLUSION</b>For some skeletal malocclusion, the youth had different utility values with the adult.</p>
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Adolescent , Adulte , Femelle , Humains , Mâle , Céphalométrie , Face , Malocclusion dentaire , Malocclusion de classe III , MandibuleRÉSUMÉ
<p><b>OBJECTIVE</b>To determine the relative resistance to HIV-1 infection of CD4 + T lymphocytes in HIV-exposed seronegative individuals (ESNs) in China.</p><p><b>METHODS</b>HIV primary isolates were obtained from peripheral whole blood of HIV-infected persons. CD4 + T lymphocytes of Chinese ESNs were separated from peripheral blood mononuclear cells with magnetic cell sorting (MACS). The purified CD4 + T lymphocytes were cocultured with HIV primary isolates. The p24 level was detected and the culture medium was refreshed every 3 days within 2 weeks.</p><p><b>RESULTS</b>For M tropic HIV strains, p24 level was significantly lower in ESN group than in control group (P < 0.05); for some M tropic HIV strains, even no p24 replicated in ESN group. However, T tropic virus strains had no significant difference between these two groups (P > 0.05).</p><p><b>CONCLUSION</b>CD4 + T lymphocytes of Chinese ESNs may possess relative resistance to M tropic HIV strains, which may be one of the main influencing factors that result in ESN.</p>
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Adulte , Femelle , Humains , Mâle , Lymphocytes T CD4+ , Allergie et immunologie , Virologie , Chine , VIH (Virus de l'Immunodéficience Humaine) , Classification , Virulence , Infections à VIH , Virologie , Séronégativité VIH , Allergie et immunologie , Techniques in vitro , Partenaire sexuelRÉSUMÉ
Objective To observe the incidence of bronchial asthma thereafter by 3 to 10 years′ follow-up of 280 children with bronchio-litis and analyze the correlation factors affecting the development of asthma.Methods Two hundred and eighty children with bronchitis were chosen and treated from Jan.1995 to Dec.2002,220 males,and 60 females.The review research and clinical revisits were carried out to investigate and analyze the subjects in terms of whether receiving large dose of intravenous injection immunoglobulin(IVIG) treatment or not,whether getting vaccinated with bacille calmette guerin(BCG)as planed or not,whether having asthma family history and the allergic physique or not,the way of feeding,and so on.Results The rate of asthma was 29.29%(82/280),and the incidence of asthma was significantly lower in children with administering the large dose of IVIG,vaccinating BCG vaccine as planed,having no allergic physique,having no asthma family history or breast feeding than those without administering the large dose of IVIG,vaccinating BCG vaccine as planed,brest feeding,with allergic physique,asthma family history(Pa
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Objective To study pathogensand diagnostic value of beta2-microglobulin(?2-MG) in cerebrospinal fluid(CSF) in children with aseptic meningitis(AM).Methods Polymerase chain reaction (PCR) or reverse transcription-PCR (RT-PCR) assay,enzyme linked immunosorbent assay (ELISA) and passive particle agglutination were employed respectively to detect herpes simplex virus type-1(HSV-1) DNA,enterovirus(EV) RNA in CSF and the specific antibodies of HSV-1 and mycoplasma pneumonia(MP) in blood and CSF from 80 children with AM (AM group) and 20 cases of complete remission acute leukaemia(control group).Children with AM were divided into the EV meningitis group,HSV-1 meningitis group and MP meningitis group according to respective diagnostic criteria.The levels of ?2-MG in CSF from all groups were detected by radio immunoassay.SPSS 13.0 software was used to analyze the data.Results The infection rate of EV,HSV-1 and MP were 52.5%,11.25%and 5.0%,respectively in children with AM.The infection rate of EV was highest.The sensitivity of PCR was higher than ELISA.There could be false negative PCR results in bloody CSF specimens with HSV-1 DNA.The levels of ?2-MG in CSF was significantly higher in children with AM than that in control group(P
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Toll-like receptors are an important pattern recognition receptors that recognize specific components of microbes and certain host molecules.Through initiated cell signaling pathway,they can induce immunocyte.Their activation triggers the innate immune response,and is crucial to the successful induction of Th1/Th2-phenotyped adaptive immunity.This review will give a brief overview of our current knowledge of TLRs as key components in immune responses and their regulation in respiratory tract diseases such as asthma.
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Objective To study the efficiency of large dose intravenous immunoglobulin(IVIG) in decreasing incidence of asthma following bronchiolitis.Methods One hundred and sixty-eight children with bronchiolitis were chosen to use different treatment plans,81 children receiving large dose of IVIG[ 0.4 g/(kg?d)] daily,on the basis of routine treatment for 5 days,while other 87 children were reserved as control group receiving routine treatment only,which were followed up 5 to 10 years.Results After following up 5 to 10 years,only 17 cases in 81 patients with bronchiolitis receiving large dose IVIG developed into asthma.However,46 cases in control group developed into asthma.The incidence of asthma was 20.99% and(52.87%) in two groups,respectively,showing significant(differe-)nce between them(P
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<p><b>OBJECTIVE</b>Nuclear factor-kappa B (NF-kappaB) is a critical transcription factor governing the expression of many cytokines that are involved in the pathogenesis of inflammatory diseases, such as asthma and rheumatoid arthritis. Melatonin (MT), a relatively safe and potent antioxidant which has shown efficacy in several chronic inflammatory models, may inhibit the expression of NF-kappaB and therefore might have a therapeutic use in asthma. This study aimed at observing the effect of MT on the expression of NF-kappaB and airway inflammation in a rat model of bronchial asthma.</p><p><b>METHODS</b>Twenty-four male Sprague-Dawley (SD) rats weighing 120 g to 170 g were randomly divided into three experimental groups (8 in each): (1) Asthmatic group: Rats were immunized on day 1 by intraperitoneal injection of 100 mg ovalbumin (OVA) in 1 ml of saline with 100 mg of alu minum hydroxide. From day 15 the animals were challenged with aerosolized OVA (1% in saline) for 20 minutes per day for 7 consecutive days. (2) MT group: OVA-sensitized rats were injected intraperitoneally with 10 mg/kg MT 30 minutes before each OVA challenge. (3) CONTROL GROUP: OVA for inhalation and MT for intraperitoneal injection was replaced with normal saline (NS). Airway responsiveness to aerosolized acetylcholine of 24 rats was detected six hours after the last challenge. Then the rats were lavaged and total and differentiated leukocytes counts in bronchoalveolar lavage fluid (BALF) were performed after staining with Wright-Giemsa staining. At the same time, levels of nitric oxide (NO) in BALF, inducible nitric oxide synthesis (iNOS) and constitute nitric oxide synthesis (cNOS) in the lung tissues were assessed with the use of nitrate reductase and chemical colorimetry, respectively. The expression of NF-kappaB in the lung tissues was observed by means of immunohistochemical staining.</p><p><b>RESULTS</b>(1) After OVA challenge, there was a significant decrease in airway responsiveness, lymphocytes and eosinophils in BALF in MT group compared with asthmatic group (P < 0.01 respectively); (2) There was a significant decrease in amounts of NO(2)(-)/NO(3)(-) in the BALF and levels of iNOS in the lung tissues in MT group comparing with asthmatic group (P < 0.01 respectively); and the levels of iNOS in the lung tissues was correlated positively with NO(2)(-)/NO(3)(-) in the BALF (P < 0.01), but there were no significant differences in activity of cNOS in any of the groups analyzed. (3) There was a significant increase in expression of NF-kappaB in lung tissues in asthmatic group compared with the other groups (P < 0.01), and so was in MT group compared with control group (P < 0.05).</p><p><b>CONCLUSIONS</b>MT could partially inhibit the expression of NF-kappaB and down-regulate the activity of iNOS in lung tissue, decrease the production of NO in BALF. These data suggest that the inhibitory effect of MT probably play a role in decreasing airway hyperresponsiveness and airway inflammation of asthmatic rats model.</p>
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Animaux , Mâle , Rats , Antioxydants , Pharmacologie , Asthme , Traitement médicamenteux , Métabolisme , Liquide de lavage bronchoalvéolaire , Biologie cellulaire , Allergie et immunologie , Modèles animaux de maladie humaine , Poumon , Chimie , Anatomopathologie , Mélatonine , Pharmacologie , Facteur de transcription NF-kappa B , Monoxyde d'azote , Rat Sprague-DawleyRÉSUMÉ
Streptococcus pneumoniae is one of the most common cause of invasive disease,such as bacteremia,meningitis,and empyema,et al.But there has significant difference on the incidence of invasive pneumococcal disease among different regions and differernt groups of people.In addition the severity and mortality of invasive pneumococcal disease are closely related to the changes of serotypes,virulence of streptococcus pneumoniae and also the human immune response.The pneumococcal vaccination is an important measure to prevent streptococcus pneumoniae infection,providing good protection to vaccinees and createing herd immunity effect.This article briefly describes the pathogenesis,risk factors and preventive strategies of invasive pneumococcal disease.
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5-10 years old were 33 cases;febrile children were 55 cases,persistent cough in 65 cases,among of those cases,the wheezing were 8 cases,nonproductive cough 45 cases,nasal discharge 7 cases in early right lung pneumonia in 38 cases,left lung pneumonia in 23 cases,pulmones involved at same time were 8 cases.Among of those children who senerated plenritis were 9 cases,pleural effussion 1 case;complication besides lung appeared 32 children.Complication of digestive and urinary system had 8 cases(11.6%),respectively.Complication of cardiovascular system had 3 cases(4.3%).Complication of nervous system and skin had 4 cases(5.8%),respectively.Complication of hematological system had 5 cases(7.2%).Eight cases(11.6%) showed damage of no more one organ,Fifty-nine cases first treated by erythromycin in vein,then by azithromycin in oral,10 cases were treated by azithromycin oral medication.Thirty-three cases fully recovered,36 cases improved.Conclusions SMPP are more founded in school children,the complication of besides lung are more serious than that of non-segmental pneumonia,it is security and relatively satisfaction to treat with erythromycin and azithromycin in sequential therapy.
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Objective To investigate the proportion of streptococus pneumonia(SP), haemophilus influenzae (Hi),branhamella catarrhalis (BC), chlamydia pneumonia (CP) and mycoplasma pneumonia (MP) in children with acute pneumonia.Methods Fifty-three hospital-treated children with acute pneumofmonia were included in a prospective study. The enzyme immunoassay was used to detect antibodies in paired sera against non-capsulated Hi, Hi type-B capsular polysaccharide and whole cell, antibodies against pneumococcal pneumolysin, C-polysaccharide and pneumococcal pneumolysin, C-polysaccharide, surface protein A in circulating immune complexes and antibodies against BC and MP. Antibodies against CP by immunofluorescence in paired sera as well as blood cultures were detected in these cases.Results The evidence of bacteria infection was demonstrated in 32 cases (60.4 %) among children with acute pneumonia, of which 11 cases had 2 or 3 organisms coinfection. In these causative agents,SP and CP were the most common organisms (11/53 cases respectively,20.8 %), followed by MP (7/42,16.7 %),Hi(7/53,13.2 %).Conclusion SP,Hi,CP and MP are common causative agents of children with acute pneumonia and multi-agents coinfection isn′t ingnored in our empiric antibiotic therapy for the disease.