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Objective To investigate the effect of curcumin on proliferation and apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-2.Methods The CNE-2 cells were treated by by different concentrations (0,10,20,40,60 μmol/L) of curcumin.The proliferation activity of CNE-2 cells was detected by MTT assay,the cell cycle and apoptosis rate of CNE-2 were detected by using the flow cytometry (FCM),and the apoptosis was observed by Hoechest33258 fluorescence staining.Results Curcumin could significantly inhibit the proliferation of CNE-2 cells,moreover which was increased with curcumin concentration increase,the inhibitory rate of CNE-2 cells showed an increasing trend (P＜0.05),the half inhibitory concentrations (IC50) of curcumin acting on CNE-2 cells at 24,48,72 h were (23.54 ± 0.36),(18.31 ± 0.42) and (8.56 ± 0.37) μmol/L respectively.Curcumin could significantly inhibit the proliferation effect of CNE-2 cells,showing the apparent concentration and time dependence.The FCM detection results showed that in treating CNE-2 cells by 0,10,20,40,60 μmol/L of curcumin,the apoptosis rate was increased with the curcumin concentration increase;the fluorescence staining results showed that CNE-2 cells without curcumin treatment were round or oval,the cell nucleis were uniform in size,chromatin distribution showed the homogeneous light blue fluorescence;after 24 h of 10 μmol/L curcumin treatment,the CNE-2 cell body was shrunk and cell nuclear chromatin was condensed,showing granular bright blue fluorescence;after 24 h of 20 μmol/L curcumin treatment,the cell body was shrunk,nuclear was condensed,chromatin was uneven,apoptotic bodies appeared,and even the nuclear fragmentation appeared;after 24 h of 40 μmol/L and 60 μmol/L curcumin treatment,the number of apoptotic cells was increased,a large number of nuclear fragmentation appeared.Conclusion Curcumin has a significant inhibitory effect on the proliferation of NPC cell line CNE-2,moreover promotes the apoptosis of CNE-2 cells.
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OBJECTIVE@#To explore the changes of microRNAs in nasal mucosa after the specific immunotherapy (SIT) for allergic rhinitis (AR) in mice.@*METHOD@#Female BALB/c mice, 6-8 weeks of age, were randomly divided into control group, model group and treatment group. AR model were established by intraperitoneal injection and intranasal challenge of ovalbumin and SIT was performed by inguinal subcutaneous injections. AR symptom scores were documented. The eosinophils (EOS) in the nasal mucosa were measured. Ovalbumin-specific IgE (OVA-sIgE) in the serum and expression of interferon-γ and interleukin-4 in the nasal lavage were measured by enzyme-linked immunosorbent assay meanwhile the ratio of interferon-γ and interleukin-4 was calculated. The microRNAs in the nasal mucosa were preliminary screened by microRNA gene microarray. Comparing with model group, the Fold changes of microRNA of the treatment group were ≥ 2.0 and the P < 0.05. MicroRNA target genes were predicted with GeneSpring 12.5 software. We took the intersection between genes in the signal pathway which associated with immune response,inflammation and target genes. The MEV-4-6-0 and Cytoscape_v2. 8. 2. software was applied to perform the cluster analysis and target gene regulatory networks maps.@*RESULT@#The model of AR in mice and its SIT were successful. Comparing with the model group, the Fold changes of 15 microRNAs, of which 9 microRNAs were up-regulated and 6 microRNAs were down-regulated, were ≥ 2.0 in treatment group (P < 0.05). Cluste analysis showed clearly that microRNAs in the treatment group and model group respectively aggregated in two branches. The 15 microRNAs had 5302 target genes, of which, 451 genes were related more with SIT by the intersection. One microRNA can regulate many target genes, and one gene can also be affected by many microRNAs. Their synergistic effects may be involved in the mechanism of SIT.@*CONCLUSION@#The expressions of microRNAs are changed in nasal mucosa after SIT for AR in mice and we can speculate that microRNAs are involved in the process of SIT for AR. Bioinformatics methods can diminish the scope of target genes of microRNAs, which will help us studying the effect of changed microRNA on its relative target genes after SIT, and make us better understanding the mechanism of the disease and its SIT.
Subject(s)
Animals , Female , Mice , Administration, Intranasal , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Eosinophils , Allergy and Immunology , Immunoglobulin E , Blood , Immunotherapy , Interferon-gamma , Allergy and Immunology , Interleukin-4 , Allergy and Immunology , Mice, Inbred BALB C , MicroRNAs , Metabolism , Nasal Mucosa , Metabolism , Ovalbumin , Rhinitis, Allergic , TherapeuticsABSTRACT
OBJECTIVE@#To explore the effects of simultaneous multi-level surgery intervention on the changes of the oxidative stress and the cognitive function in young and middle-aged patients with moderate-severe OSAHS.@*METHOD@#One hundred and sixteen young and middle-aged patients with moderate-severe OSAHS received simultaneous multi-level surgery. PSG,malondialdehyde (MDA), advanced oxidation protein products (AOPP) and 8-isoprostaneF2α (8-iso-PGF2α) were tested, together with the evaluation of the questionnaires of Montreal cognitive assessment (MoCA), mini-mental state examination (MMSE) and Epworth sleepiness scale (ESS) before and 6 months after surgery.@*RESULT@#(1) The AHI, TS90% and LSaO₂ were all improved significantly 6 months after operation. No severe complications were observed in any patient. (2) The preoperative MoCA and MMSE scores were (23.52 ± 2.64) and (26.54 ± 1.07) respectively. The cognitive impairment were detected in OSAHS patients. MoCA was negatively correlated with AHI, TS90%, MDA, AOPP and 8-iso-PGF2a, while positively correlated with LSaO₂ (P < 0.05, r = -0.233, -0.190, -0.256, -0.247, -0.221 and 0.215, respectively). Cognitive impairment was mainly manifested as disorder in the visual space and execu- tive function, memory and delayed recall (mainly short-term memory). (3) MoCA and MMSE scores were both increased (P < 0.05) after the operation. MoCA showed negative correlations with MDA, AOPP, 8-iso-PGF2α, AHI and TS90%, and positive correlation with LSaO₂ (P < 0.05, r = -0.495, -0.463, -0.397, -0.332, -0.292 and 0.336, respectively).@*CONCLUSION@#Oxidative stress and cognitive dysfunction are closely related in young and middle-aged patients with Moderate-severe OSAHS. Oxidative stress may play an important role in cognitive function of young and middle-aged patients with Moderate-severe OSAHS. Multi-level surgery can improve oxidative stress so that to improve cognitive impairment.
Subject(s)
Adult , Humans , Middle Aged , Cognition , Cognition Disorders , Oxidative Stress , Sleep Apnea, Obstructive , General SurgeryABSTRACT
Objective:To explore the feasibility of inguinal subcutaneous immunotherapy for allergic rhinitis ( AR ) in mice. Methods:36 female BALB/c mice were divided randomly into six groups( n=6 per group) including the control A,the model A, the treatment A groups,and the control B,the model B,the treatment B groups(inguinal subcutaneous immunotherapy for group A, cervical back subcutaneous immunotherapy for for group B). AR model was established with ovalbumin. At 25 to 55 days,ovalbumin im-munotherapy were performed in treatment groups,once two days,15 times totally. After intranasal rechallenge was performed at 56 to 62 days the AR symptom scores were documented. The eosinophils(EOS)in the nasal mucosa were measured by chromotropic acid 2R staining. Ovalbumin-specific IgE( OVA-sIgE) in the serum and expression of interferon-γ and interleukin-4 in the nasal lavage were measured by enzyme-linked immunosorbent assay meanwhile the ratio of interferon-γ and interleukin-4 was calc μlated. SPSS17. 0 software was used to analyze the data. Results:Before treatment ,the AR symptom scores of the model and treatment groups were more than 5. After treatment,the treatment A group were less than 5. The EOS count of the control A,model A,treatment A groups and the control B,model B, treatment B groups was 0. 78 ± 0. 31, 21. 60 ± 2. 90, 10. 43 ± 2. 56, 0. 83 ± 0. 46, 22. 44 ± 3. 39, 23. 40 ± 4. 24, respectively. The EOS count of the treatment A group was significantly lower than those in model A group ( P0. 05 ) . OVA-sIgE expressed was negative in control groups and positive in other groups. The ratio of interferon-γ and interleukin-4 was 10. 75 ± 3. 38,10. 38 ± 3. 08,3. 02 ± 0. 69,2. 71 ± 0. 89,2. 52 ± 0. 30,5. 45±1. 41,respectively. The ratio in treatment A group was significantly higher than those in model A group(P0. 05 ) . Conclusion: Inguinal subcutaneous immunotherapy has a good effect on this disease. It spends short time ,has simple operation and good feasibility,which is a novel treatment method for AR in mice.
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Objective To investigate the effects of simultaneous multi-level surgery for moderate to severe obstructive sleep apnea hypopnea syndrome (OSAHS). Methods A retrospective analysis was made on surgical cases of one hundred and thirty seven patients with moderate to severe OSAHS diagnosed by polysomnography (PSG). They were divided into multi-level group (n = 95) and UPPP group (n = 42). The two groups were compared in terms of postoperative complications as well as the related indicators of PSG , calgary sleep apnea quality of life index (SAQLI), epworth sleepiness scale (ESS), snore scales (SS) before operation and after operation. Results Just one patient in the multi-level group had difficulties in respiration and was rescued by timely tracheotomy. The AHI, LSaO2, TS90%, the total score and the scores on the four dimensions of SAQLI, ESS score, SS score in the multi-level group were significantly improved as compared both to the results after operation (P < 0.01) and to the UPPP group (P < 0.05). But only the AHI, LSaO2 and TS90% in the UPPP group were improved (P < 0.05). Conclusions The multi-level surgery is a safe and feasible therapy or moderate to severe OSAHS. The evaluation in subjective and objective ways can be more accurate in comprehensive reflecting the surgical efficacy and effects of OSAHS on patients′ of life quality.
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<p><b>OBJECTIVE</b>To investigate the ultrastructure of ciliated epithelia and inflammatory changes upon repeated exposure to staphylococcal enterotoxin A (SEA) of different concentrations in the maxillary sinus mucosa of rabbits.</p><p><b>METHODS</b>The rabbits were randomly divided into 2 groups (24 rabbits per group): low-dose SEA group and high-dose SEA group. The low-dose SEA group and high-dose SEA group received daily injections of 0.6 ng of SEA (2 ml) and 60 ng of SEA (2 ml) into the left maxillary sinus of rabbits for 28 days, respectively. Concurrent treatment of the right maxillary sinus with normal saline was used as control. Six rabbits chosen randomly in two groups were examined by computed tomography (CT) scans and then sacrificed to obtain the sinus mucosa from the two-side of maxillary sinuses for histological assessment on days 3, 7, 14 and 28. To characterize the inflammatory changes of the sinus mucosa examined using light microscope, hematoxylin and eosin (HE) and toluidine blue staining was performed. Scanning and transmission electron microscopy were performed to observe ultrastructure of ciliated epithelia in the maxillary sinus mucosa. SPSS 13.0 software was used to analyze the data.</p><p><b>RESULTS</b>On days 14 and 28, CT images showed opacification of the left maxillary sinus in the high-dose SEA group. The percentage of epithelial disruption was (22.73 ± 5.72) % and (30.79 ± 4.30)% in the high-dose SEA group respectively, and were significantly greater than those in the low-dose SEA group (5.12% ± 1.98% and 5.38% ± 1.64%, q value was 10.079 and 19.132) and control group (4.08% ± 1.29% and 4.81% ± 1.62%, q value was 11.016 and 19.592, respectively, all P < 0.01). The subepithelial thickness in the high-dose SEA group was (113.34 ± 14.81)µm and (120.86 ± 12.35) µm respectively, and were significantly different from those of the low-dose SEA group [(71.08 ± 10.39)µm and (81.63 ± 9.32)µm, q value was 8.090 and 8.782] and control group [(37.45 ± 7.67)µm and (38.79 ± 7.68)µm, q value was 15.759 and 19.541, all P < 0.01]. Viewed under the electron microscope, loss of cilia was observed, a few compound cilia and cytoplasmic protrusion were found, an obvious stretching of the endoplasmic reticulum and an obvious turgescence of the mitochondria was also observed. However, in the low-dose SEA group on days 14 and 28, CT scan of the left maxillary sinus showed transparency; light microscopy observations of the maxillary sinus mucosa showed the number of eosinophils was markedly increased as compared with the high-dose SEA and control groups, the differences were significant (q value was 5.871 and 6.766 on day 14, and q value was 7.572 and 8.970 on day 28, respectively, all P < 0.05). But no significant differences were observed in epithelial disruption between the low-dose SEA and the control groups on days 14 and 28 (q value was 1.512 and 0.859 respectively, all P > 0.05); inordinate array and adhesion of cilia was observed, but cilia loss, compound cilia, cytoplasmic protrusions, mitochondrial swelling and endoplasmic reticulum stretching were not found.</p><p><b>CONCLUSIONS</b>SEA may induce allergic inflammation of the sinus mucosa without damaging the structure of ciliated epithelia at low concentration. Whereas SEA impairs the structure of mitochondria and endoplasmic reticulum in ciliated epithelial cells at high concentration, and results in cilia loss and epithelial disruption, which may be one of the main reasons to induce acute sinusitis.</p>
Subject(s)
Animals , Rabbits , Cilia , Physiology , Cost of Illness , Enterotoxins , Toxicity , Eosinophils , Epithelial Cells , Physiology , Leukocyte Count , Maxillary Sinus , Metabolism , Mucous Membrane , Physiology , SinusitisABSTRACT
OBJECTIVE@#The aim of this study was to determine the expression of GATA-3 and the level of Th1 and Th2 cytokines upon repeated exposure to staphylococcal enterotoxin B(SEB) of different concentrations in the maxillary sinus mucosa of rabbits.@*METHOD@#The rabbits were randomly divided into 2 groups (24 rabbits per group): low-dose SEB group and high-dose SEB group. The low-dose SEB group and high-dose SEB group received daily injections of 0.6 ng of SEB (2 ml) and 60 ng of SEB (2 ml) into the left maxillary sinus of rabbits for 28 days, respectively. Concurrent treatment of the right maxillary sinus with normal saline was used as a control. Six rabbits chosen randomly in two groups were killed on days 3, 7, 14, and 28, and to obtain the sinus mucosa from the two-side maxillary sinuses for measurement. Mucosal levels of IL-2, IL-4, IL-5, and IFN-γ were measured using ABC-ELISA. Tissue expression of GATA-3 were examined using Real-time PCR and immunohistochemistry.@*RESULT@#IFN-γ and IL-2 levels were significantly elevated in the high-dose SEB group compared with the low-dose SEB and control groups on days 7, 14, and 28 (P 0.05).@*CONCLUSION@#SEB promoted Th1 cytokines production at high concentrations, and enhanced Th2 cytokines expression and Th2 immune response at low concentrations.
Subject(s)
Animals , Male , Rabbits , Cytokines , Metabolism , Enterotoxins , Pharmacology , Enzyme-Linked Immunosorbent Assay , Interferon-gamma , Metabolism , Interleukin-2 , Metabolism , Interleukin-4 , Metabolism , Interleukin-5 , Metabolism , Maxillary Sinus , Metabolism , Nasal Mucosa , Metabolism , RNA, Messenger , Metabolism , Th1 Cells , Th2 Cells , Transcription Factors , MetabolismABSTRACT
OBJECTIVE@#To investigate the inhibitory effect of elemene on the tumor growth of human laryngeal squamous carcinoma (LSCC) and to explore the possible mechanism of its inhibitory effect on LSCC.@*METHOD@#Nude mice model engrafted with laryngocarcinoma was established by using human laryngeal squamous carcinoma cell line (Hep-2). When the transplantable tumor size is more than 10 mm, the mice were given elemene (120 mg/kg) in the treatment group or PBS in the control group, at the day of 1, 4, 7, 10, 13, 16, 19 and 22 respectively. The animals were killed three days after the last treatment. The tumor volume, tumor weight,mice net weight and tumor inhibition rate were measured. The expression of vascular endothelial growth factor-C (VEGF-C) and vascular endothelial growth factor receptor-3 (VEGFR-3) mRNA were quantitated by using reverse transcription and polymerase chain reaction technique.@*RESULT@#Elemene could inhibit the tumor growth in vivo. The differences were statistically significant for the mice net weight, tumor weight, and tumor volume between the treatment group and the control group. The tumor inhibition percentage was 52.24%. The gene expression of VEGF-C and VEGFR-3 of the treatment group is lower than that of the control group and the differences were statistically significant (P < 0.01).@*CONCLUSION@#Elemene could inhibit the subcutaneous plantation of human laryngeal carcinoma in nude mice and its mechanism may be associated with inhibited expression of VEGF-C and VEGFR-3.