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Sarcopenia, which is characterized by progressive and generalized loss of skeletal muscle mass and strength, has been well described to be associated with numerous poor postoperative outcomes, such as increased perioperative mortality, postoperative sepsis, prolonged length of stay, increased cost of care, decreased functional outcome, and poorer oncological outcomes in cancer surgery. Multimodal prehabilitation, as a concept that involves boosting and optimizing the preoperative condition of a patient prior to the upcoming stressors of a surgical procedure, has the purported benefits of reversing the effects of sarcopenia, shortening hospitalization, improving the rate of return to bowel activity, reducing the costs of hospitalization, and improving quality of life. This review aims to present the current literature surrounding the concept of sarcopenia, its implications pertaining to colorectal cancer and surgery, a summary of studied multimodal prehabilitation interventions, and potential future advances in the management of sarcopenia.
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OBJECTIVE@#To investigate the effect of PPP2R5C to the activity of Molt-4 cells in childhood acute T lymphocytic leukemia and its mechanism.@*METHODS@#The small interfering RNA (siRNA) technology targeting PPP2R5C gene was used to down-regulate the expression of PPP2R5C in Molt-4 cells. At the same time, a blank control group, a negative control group and a 17-DMAG group were set up. The cells in the negative control group were transfected with siRNA-NC, the cells in 17-DMAG group were treated with the HSP90 inhibitor 17-DMAG at a final concentration of 6.4 μmol/L for 48 h. Real-time fluorescent quantitative PCR (RT-qPCR) and Western blot were used to detect transfection efficiency; CCK-8 method was used to detect the proliferation activity of the cells in each group, EdU was used to detect the proliferation level of the cells in each group, flow cytometry was used to detect the cell cycle distribution ratio of the cells in each group, Annexin V-FITC/PI staining was used to detect the apoptosis of the cell, RT-qPCR and Western blot were used to detect the expression changes of heat shock protein 90 (HSP90) and glucocorticoid receptor (GR) of the cells in each group.@*RESULTS@#After Molt-4 cells were transfected with siRNA-PPP2R5C, the expression of PPP2R5C mRNA and protein in the cells were down-regulated significantly compared with those in the blank control group and the si-NC group (P<0.05); compared with cells in the blank control group and the si-NC group, the proliferation activity of the cells in the siRNA-PPP2R5C group and the 17-DMAG group significantly decreased (P<0.05), and the rate of EdU positive cells was significantly reduced (P<0.05); the proportion of the cells in G1 phase decreased while the proportion of the cells in G2 phase increased (P<0.05), the apoptosis rate of the cells also increased significantly (P<0.05); in addition, the expression of PPP2R5C mRNA and protein of the cells in siRNA-PPP2R5C group was significantly down-regulated compared with those in the blank control group and si-NC group (P<0.05). The expressions of PPP2R5C mRNA and protein in the 17-DMAG group were also significantly down-regulated compared with those in the blank control group and si-NC group (P<0.05).@*CONCLUSION@#Down-regulation of PPP2R5C gene expression can inhibit Molt-4 cell activity in childhood acute T lymphocytic leukemia, block the cells in G2 phase, and promote cell apoptosis, the mechanism may be related to the inhibition of HSP90-GR signaling pathway.
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Criança , Humanos , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Proteínas de Choque Térmico HSP90 , Leucemia-Linfoma Linfoblástico de Células T Precursoras , RNA Interferente Pequeno , Receptores de GlucocorticoidesRESUMO
OBJECTIVE@#To detect the expression level of suppressors of cytokine signaling 3 (SOCS3) in acute lymphoblastic leukemia (ALL), and to observe the effect of over-expresson of SOCS3 in Jurkat cells on the cytotoxicity of NK cells.@*METHODS@#The expression levels of SOCS3 mRNA in peripheral blood mononuclear cells of 20 children with ALL and 20 healthy children (normal control group) were detected by RT-PCR. The peripheral blood NK cells from healthy subjects were selected by immunomagnetic technique, and the purity was detected by flow cytometry. SOCS3 was overexpressed in Jurkat cells infected with lentivirus vector, and SOCS3 mRNA expression was detected by RT-PCR after lentivirus infection. The NK cells were co-cultured with the infected Jurkat, and LDH release method was used to detect the cytotoxicity of NK cells on the infected Jurkat cells. The concentrations of TNF-α and IFN-γ were determined by ELISA. The expression of NKG2D ligands MICA and MICB on the surface of Jurkat cells were detected by flow cytometry. Western blot was used to detect the effect of SOCS3 overexpression on STAT3 phosphorylation in Jurkat cells.@*RESULTS@#Compared with the control group, the mRNA expression of SOCS3 in the peripheral blood mononucleated cells of ALL children was significantly decreased. The purity of NK cells isolated by flow cytometry could reach more than 70%. The expression of SOCS3 mRNA in Jurkat cells increased significantly after lentivirus infection. Overexpression of SOCS3 in Jurkat cells significantly promoted the killing ability of NK cells and up-regulated the secretion of TNF-α and IFN-γ from NK cells. The results of flow cytometry showed that the expression of NKG2D ligands MICA and MICB on Jurkat cells increased significantly after SOCS3 overexpression. Western blot results showed that overexpression of SOCS3 significantly reduced the phosphorylation level of STAT3 protein in Jurkat cells.@*CONCLUSION@#SOCS3 mRNA expression was significantly decreased in ALL patients, and overexpression of SOCS3 may up-regulate the expression of MICA and MICB of NKG2D ligands on Jurkat cell surface through negative regulation of JAK/STAT signaling pathway, thereby promoting the cytotoxic function of NK cells.
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Criança , Humanos , Antígenos de Histocompatibilidade Classe I/metabolismo , Células Matadoras Naturais/citologia , Leucócitos Mononucleares/citologia , Ligantes , Subfamília K de Receptores Semelhantes a Lectina de Células NK/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , RNA Mensageiro/genética , Proteína 3 Supressora da Sinalização de Citocinas/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
OBJECTIVE@#To explore the prognostic factors of young and middle-aged patients with acute myeloid leukemia (AML) and the predictive value of minimal residual disease (MRD) before consolidation therapy.@*METHODS@#The clinical data of 262 middle-risk young and middle-aged patients with AML treated in our hospital from January 2010 to December 2018 were selected retrospectively. All the patients were reached morphological leukemia-free state (MLFS) after induction chemotherapy, the overall and subgroup clinical data of the selected patients were analyzed. Cox regression model was used to evaluate the independent prognostic factors of middle-risk newly diagnosed young and middle-aged patients.@*RESULTS@#Among the patients less than 40 years old treated by consolidation therapy with PR-CT and allo-HSCT regimens, the 5-year cumulative leukemia-free survival(LFS) rates were 40.92% and 63.51%(P=0.01)respectively, while those over 40 years old were 23.61% and 49.14%(P=0.00), respectively. The 5-year cumulative LFS rates of the patients treated by chemotherapy and achieved early remission and late remission were 63.51% and 41.33% (P=0.01), respectively. The 5-year cumulative overall survival(OS) rates of the patients treated by PR-CT and allo-HSCT regimens were 23.65% and 69.32% (P=0.00), respectively, and the 5-year cumulative LFS rates were 26.44% and 52.30% (P=0.01). Among the patients treated by PR-CT consolidation treatment, the MRD-negative and MRD-positive cases were 74 and 60 cases, respectively. The 5-year cumulative incidence of relapse rate in the MRD-negative subgroup was significantly lower than those in the MRD-positive subgroup (P<0.05), the 5-year LFS rate and OS rate of the patients in MRD-negative subgroup were significantly higher than those in MRD-positive subgroup (P<0.05). For the patients treated by allo-HSCT consolidation treatment, the MRD-negative and MRD-positive cases were 66 and 62 cases, respectively. The 5-year cumulative incidence of relapse rate of the patients in MRD-negative subgroup was significantly lower than those in MRD-positive subgroup(P<0.05), and the 5-year LFS and OS rates of the patients in MRD-negative subgroup were significantly higher than those in MRD-positive subgroup (P<0.05). The univariate analysis results showed that age, chromosome karyotype, MRD status after reaching MLFS, and consolidation treatment regime were all related to the prognosis of patients (P<0.05). The multivariate analysis results showed that age, MRD status after reaching MLFS, and consolidation therapy were the independent factors affecting the cumulative OS rate of the patients (P<0.05). Chromosome karyotype was an independent factor affecting the cumulative LFS rate of the patients (P<0.05). MRD status and consolidation treatment plan after reaching MLFS were the independent factors affecting the cumulative recurrence rate of the patients (P<0.05).@*CONCLUSION@#The OS rate of middle-risk young and middle-aged patients with newly diagnosed AML is independently related to age, MRD status after MLFS and consolidation therapy, while chromosome karyotype is independently related to cumulative LFS, and allo-HSCT consolidation therapy is recommended for middle-risk young and middle-aged AML patients after induction chemotherapy for MLFS, especially for those less than 40 years old and MRD positive before consolidation therapy.
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Adulto , Humanos , Pessoa de Meia-Idade , Quimioterapia de Consolidação , Transplante de Células-Tronco Hematopoéticas , Leucemia Mieloide Aguda , Neoplasia Residual , Prognóstico , Estudos RetrospectivosRESUMO
ObjectiveThe specific mechanism of microRNA-133a (miR-133a) involved in the pathological process of atherosclerosis (As) remains an open question. This study aims to explore the role of miR-133a in the regulation of endothelial cell apoptosis.MethodsCultured human coronary endothelial cells (HCAECs) were treated with oxidized low density lipoprotein (ox-LDL). The cell viability was detected by MTT assay. The mRNA levels of Bcl-xl and miRNA (miR-133a, etc) were detected by qRT-PCR method. The expression of anti-apoptotic protein Bcl-xl and cleaved-caspase3 was detected by Western blotting, and the apoptosis rate was detected by flow cytometry. The transient transfection technique was used to observe the effect of overexpression and silencing of miR-133a, on the expression of target gene Bcl-xl protein and endothelial cell apoptosis.Results Ox-LDL was observed to decrease the viability of HCAECs cells and induce HCAECs apoptosis; miR-133a increased abnormally in the apoptosis model; after silencing miR-133a, the decrease of Bcl-xl and the increase of apoptosis rate induced by ox-LDL were partially reversed; the overexpression of miR-133a, Bcl-xl decreased and the apoptosis rate increased, and the difference was statistically significant.Conclusion miR-133a might target and regulate the anti-apoptotic protein Bcl-xl, to induce endothelial cell apoptosis and promote the formation of AS.
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OBJECTIVE@#To explore the changes of autophagic activity after resistance of U266 cells to bortezomib (Bor) and its mechanisms.@*METHODS@#The proliferation inhibition rate, 50% inhibitory concentration (IC), drug-resistance coefficient, drug-resistance reversed multiple by 3-methyladenine (3-MA) in U266 and U266/Bor cells treated with Bor were detected and calculated by using MTT method, then the proliferation inhibition curve was drawed. The Western blot was used to detect the expression of LC3-I, IC3-II, p-mTOR, Beclin-1, ATG5 and ATG7 proteins.@*RESULTS@#The Bor showed the its proliferation inhibition effect on U266 cells and U266/Bor cells, IC of Bor on U266 and U266/Bor cells on 24 hours were 35.7 nmol/L and 526.5 nmol/L respectively; the drug-resistance coefficient was 14.7; the drug-resistance reversed multiple by 3-MA was 2.7. The expression of LC3-II, Beclin11, ATG5 and ATG7 in U266/Bor cells was higher than that in U266 cells; after the treatment with Bor for 24 h, the expression levels of LC3-II, Beclin-1, ATG5 and ATG7 in U266 cells all decreased, as compared with levels before treatment; while the expression levels of LC3-II, Beclin-1, ATG5 and ATG7 in U266/Bor cells were higher than those before treatment. There were no significant difference of p-mTOR expression among U266, U266+Bor, U266/Bor, U266/Bor+Bor cells.@*CONCLUSION@#The increase of autophagy closely relates with resistance of U266 cells to bortezomib, moreover with up-regulation of Beclin-1, ATG5 and ATG7 expression.
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Humanos , Apoptose , Autofagia , Proteína Beclina-1 , Bortezomib , Linhagem Celular Tumoral , Proliferação de Células , Resistencia a Medicamentos AntineoplásicosRESUMO
OBJECTIVE@#To explore the reversal effect of pioglitazone (PIO) on multidrug resistance in K562/ADR cells and its mechanism.@*METHODS@#The proliferation inhibition rate, half inhibition concentration (IC) and drug-resistance reversal multipe were detected and the curve of proliferation inhibition rate was drawn by MTT assay, the transcription of PPARγ, CYP2C8 and CYP2J2 genes was detected by RT-PCR; the expression of PPARγ, CYP2C8 and CYP2J2 proteins was detected by Western blot.@*RESULTS@#The IC of PIO on K562 and K562/ADR cells for 60 h was 326.7 μmol/L and 349.1 μmol/L respectively. The reversal multiple of 30 μmol/L PIO on ADR-resistance of K562/ADR cells was 6.4. After treatment of K562/ADR cells with PIO, the transcription of CYP2C8 and CYP2J2 and the protein expression of CYP2C8 and CYP2J2 significantly decreased, the transcription of PPARγ gene and the expression of PPARγ protein were not changed.@*CONCLUSIONS@#Pioglitazone can reverse the adriamycin-resistance in K562/ADR cells that is closely related to the decrease of protein expression of CYP2C8 and CYP2J2. Pioglitazone is an effective multidrug resistance reversal agent for tumors.
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Humanos , Doxorrubicina , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Células K562 , PioglitazonaRESUMO
Objective: To prepare a bacterial outer membrane vesicle (OMV) coated poly (lactic-co-glycolic acid) copolymer (PLGA) nanoparticle loaded with ovalbumin (OVA) and evaluate its intranasal immune effect in mice. Methods: OMV was prepared by ultrafiltration concentration method. OVA loaded PLGA nanoparticle (NP) was prepared by emulsion-solvent evaporation method. OMV coated PLGA nanoparticle (OMV-PLGA NP) loaded with OVA was prepared by extrusion method and characterized. BALB/c mice were intranasally immunized and specific sIgA levels in nasal wash, jejunum and fecal pellet were determined by ELISA. Results: Size of OVA loaded OMV-PLGA NP was (234.4±22.9) nm. The shell-core structure of OVA loaded OMV-PLGA NP was proved by transmission electron microscope. After 14 d of administration, sIgA antibody levels in nasal wash, jejunum and fecal pellet of OVA loaded OMV-PLGA NP treated group were the highest in all treated groups. Compared with the group treated with OMV and OVA, OVA-specific sIgA antibody level in nasal wash, jejunum and fecal pellet of OVA loaded OMV-PLGA NP treated group was increased 1.6, 2.1 and 1.7 times, respectively. Compared with the group treated with OMV and OVA, OMV-specific sIgA antibody level in nasal wash, jejunum and fecal pellet of OVA loaded OMV-PLGA NP treated group was all increased 1.5 times. Conclusion: This novel nanoparticle drug delivery system can simultaneously delivery OVA and OMV to antigen presenting cells, resulting in stronger mucosal immune response in mice.
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Objective·To prepare a bacterial outer membrane vesicle (OMV) coated poly (lactic-co-glycolic acid) copolymer (PLGA) nanoparticle loaded with ovalbumin (OVA) and evaluate its intranasal immune effect in mice.Methods· OMV was prepared by ultrafiltration concentration method.OVA loaded PLGA nanoparticle (NP) was prepared by emulsion-solvent evaporation method.OMV coated PLGA nanoparticle (OMV-PLGA NP) loaded with OVA was prepared by extrusion method and characterized.BALB/c mice were intranasally immunized and specific sIgA levels in nasal wash,jejunum and fecal pellet were determined by ELISA.Results· Size of OVA loaded OMV-PLGA NP was (234.4±22.9) nm.The shell-core structure of OVA loaded OMV-PLGA NP was proved by transmission electron microscope.After 14 d of administration,sIgA antibody levels in nasal wash,jejunum and fecal pellet of OVA loaded OMV-PLGA NP treated group were the highest in all treated groups.Compared with the group treated with OMV and OVA,OVA-specific sIgA antibody level in nasal wash,jejunum and fecal pellet of OVA loaded OMV-PLGA NP treated group was increased 1.6,2.1 and 1.7 times,respectively.Compared with the group treated with OMV and OVA,OMV-specific sIgA antibody level in nasal wash,jejunum and fecal pellet of OVA loaded OMV-PLGA NP treated group was all increased 1.5 times.Conclusion· This novel nanoparticle drug delivery system can simultaneously delivery OVA and OMV to antigen presenting cells,resulting in stronger mucosal immune response in mice.
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@#To investigate activities of three isomers of α-conotoxin TxID on human α3β4 and α6/α3β4 nicotinic acetylcholine receptors(nAChRs). The three isomers of α-conotoxin TxID were synthesized using solid phase Fmoc chemistry and fully folded by two-step oxidations. Human α3β4 and α6/α3β4 nAChRs were expressed in oocytes of Xenopus laevis, which were used for bioassay of the three isomers, including inhibition and washout reversibility. There were obvious differences between the inhibition potency of each isomers at human α3β4 and α6/α3β4 nAChRs. The blocking was reversible and washout rapidly. The most potent isomer is the globular form with an IC50 of 9. 3 nmol/L on human α3β4 and α6/α3β4 nAChRs respectively. The 2nd potent isomer was the ribbon form with much less potency, which had an IC50 of > 5 μmol/L. The bead isomer had little or no block on human α3β4 and α6/α3β4 nAChRs with an IC50 of > 10 μmol/L. The three isomers of α-conotoxin TxID were synthesized successfully with two pairs of desired disulfide bond. Inhibition activities of the 3 isomers on human α3β4 and α6/α3β4 nAChRs were obtained respectively, which would be basis for new marine drug development of α-conotoxin TxID.
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<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of the combination therapy of tamsulosin and solifenacin for mild and moderate benign prostatic hyperplasia (BPH) with overactive bladder (OAB).</p><p><b>METHODS</b>We randomly divided 166 patients with BPH and concomitant OAB into a mild obstruction symptom group (n = 88) and a moderate obstruction symptom group (n =78), 48 of the former group treated with 0. 2 mg tamsulosin + 5 mg solifenacin and the other 40 with 0. 2 mg tamsulosin; 36 of the latter group treated with 0. 2 mg tamsulosin + 5 mg solifenacin and the other 42 with 0. 2 mg tamsulosin, all administered once daily for 12 weeks. We obtained the International Prostate Symptom Score (IPSS), urine storage period symptom score (USPSS), voiding symptom score (VSS), Qmax, residual urine volume, OAB symptom score (OABSS) and adverse reactions, and compared them among different</p><p><b>RESULTS</b>Among the patients with mild obstruction symptoms, the combination of tamsulosin and solifenacin achieved remark-groups. able improvement in IPSS, USPSS, Qmax and OABSS as compared with the baseline (P < 0.05), but made no significant difference in the residual urine volume (P > 0. 05) , while tamsulosin improved IPSS only (P < 0.05). The combination therapy exhibited an obvious superiority over tamsulosin alone in improving IPSS (9.7 micro 3.0 vs 15.8 micro 3.3), USPSS (8. 1 micro 1.7 vs 12.3 micro 3.1), Qmax ([18.6 micro 2.3] ml/s vs [14.2 micro 2.3] ml/s ), and OABSS (5.3micro 1.3 vs 9.7 micro 2.7) (P < 0.05), but there were no obvious differences in residual urine, urine routine test results and adverse events between the two therapies ( P > 0. 05). In those with moderate obstruction symptoms, the combination therapy significantly improved IPSS, VSS, Qmax and OABSS (P < 0.05) but not the residual urine (P > 0. 05) in comparison with the baseline. The tamsulosin therapy achieved obvious improvement in IPSS, VSS, Qmax, OABSS and residual urine. The combination therapy showed a better effect than tamsulosin only in OABSS (4. 8 +/-1.5 vs 6.5 +/-2.5, P < 0.05), but no significant differences from the latter in IPSS, Qmax, VSS, routine urine test results, and adverse</p><p><b>CONCLUSION</b>Combination therapy of tamsulosin and solifenacin is obviously safe and efficacious in the treatment (P > 0.05). events of both mild and moderate BPH with concomitant OAB, and it is superior to tamsulosin alone.</p>
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Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Quimioterapia Combinada , Estudos Prospectivos , Hiperplasia Prostática , Tratamento Farmacológico , Quinuclidinas , Usos Terapêuticos , Succinato de Solifenacina , Sulfonamidas , Usos Terapêuticos , Tetra-Hidroisoquinolinas , Usos Terapêuticos , Bexiga Urinária Hiperativa , Tratamento FarmacológicoRESUMO
<p><b>OBJECTIVE</b>To investigate the epidemiological characteristics of tic disorders (TD) among pupils in the Shunde Longjiang area, and their relationship to trace elements.</p><p><b>METHODS</b>A cross-sectional study of 4062 children aged 6-12 years, who were selected from the Shunde Longjiang area by stratified cluster sampling to investigate the epidemiological characteristics of TD, was conducted, and blood concentrations of trace elements in children with TD were determined. Forty normal children were selected as controls.</p><p><b>RESULTS</b>The overall prevalence rate of TD was 2.98%; the prevalence rates of transient tic disorder, chronic motor or vocal tic disorder and Tourette's syndrome were 3.62%, 2.39% and 1.21% respectively. Boys had a significantly higher prevalence rate of TD than girls (3.92% vs 1.96%; P<0.05). There were no significant differences in blood copper, manganese and magnesium levels between children with TD and normal children (P>0.05), however, children with TD had a significantly increased blood lead level and significantly decreased blood zinc and iron levels compared with the normal children (P<0.05). No significant differences in trace elements were found between children with different subtypes of TD (P>0.05).</p><p><b>CONCLUSIONS</b>TD is common in children aged 6-12 years and more prevalent in boys than in girls. High blood lead level and zinc and iron deficiencies may be one of the causes of TD, and thus should be considered during therapy.</p>
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Criança , Feminino , Humanos , Masculino , China , Epidemiologia , Estudos Transversais , Ferro , Sangue , Chumbo , Sangue , Transtornos de Tique , Sangue , Epidemiologia , Oligoelementos , Sangue , Zinco , SangueRESUMO
<p><b>OBJECTIVE</b>To investigate the feasibility of bilateral same-day myringoplasty and the indications for myringoplasty for patients with bilateral tympanic membrane perforation, and to summarize relevant experience.</p><p><b>METHODS</b>Twenty-two patients underwent bilateral same-day underlay myringoplasty, and all cases were consistent with the indications for myringoplasty. The preoperative hearing and postoperative hearing at three months were compared, and the postoperative symptoms and complications were observed. Forty patients underwent monaural myringoplasty as the control group over the same period. All cases were followed up for 1 - 3 years.</p><p><b>RESULTS</b>The postoperative hearing was increased by an average of 18 dB, and the rate of closure of tympanic membrane perforation was 93.2% (41/44). There were seven patients with ear fullness after operation in the bilateral myringoplasty group and two patients in the control group (χ(2) = 4.5374, P = 0.0332). There were no differences in the postoperative hearing improvement, the rate of closure and the rates of other discomfort symptoms except for ear fullness between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>It was feasible and safe to perform bilateral same-day myringoplasty for bilateral tympanic membrane perforation, but the postoperative temporary discomfort of bilateral ear fullness should be informed the patients in advance.</p>
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Humanos , Audição , Testes Auditivos , Miringoplastia , Período Pós-Operatório , Resultado do Tratamento , Membrana Timpânica , Cirurgia Geral , Perfuração da Membrana Timpânica , Cirurgia GeralRESUMO
Objective To investigate the effect of IKKε knock-down on the biological characteristics of U251 glioblastoma cells. Methods IKKε small interfering RNA (IKKε siRNA) mediated by lipofectamine were transfected into U251 cells while cells transfected with scrambled siRNA and control cells were prepared.RT-PCR was employed to detect expressions of IKKε in the transfected cells.The cell proliferation was determined by MTT assay.Flow cytometry was used to monitor changes in cell cycle.Cell invasion was evaluated by Transwell assay.Moreover,the proliferating cell nuclear antigen (PCNA),cyclin D 1 and matrix metalloproteinase-9 (MMP9) that regulated proliferation,invasion and cycle progression of the transfected cells and the changes of NF-κB after transfection were examined by Western blotting. Results RT-PCR revealed that the proliferation of U251 cells was inhibited,the cell cycle was arrested in G0/G1 phase and the invasive activity was attenuated in cells transfected with IKKe siRNA,with significant differences as compared with the cells transfected with scrambled siRNA and control cells (P<0.05).The expressions ofPCNA,MMP9 and cyclin D1 were down-regulated in the IKKε knock-down cells, as compared with the other 2 groups. In addtion, transposition of NF-κB reduced from the cytoplasm to the nucleus after transfection. Conclusion As IKKε plays a vital role in proliferation and invasion of glioma cells,it may serve as a potential target ofgene therapy for glioma.
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<p><b>OBJECTIVE</b>To investigate the influence of the individual genotype differences of DC-SIGN and DC-SIGNR on the mother-to-neonate intrauterine infection of HBV.</p><p><b>METHODS</b>The genotypes of the gene DC-SIGN and DC-SIGNR in the pregnant women with HBV positive were detected by PCR and agarose gel electrophoresis. The significant difference of gene diversity of DC-SIGN and DC-SIGNR was analyzed by chi-square test.</p><p><b>RESULTS</b>(1) All of 29 cases in intrauterine infection group were 7/7 DC-SIGN genotype. In the non-intrauterine infection group, 7/5 genotype were observed in 2 of 54 cases, and the other 52 cases were 7/7 genotype. The two groups was no significant difference (P = 0.54). (2) 29 cases of intrauterine infection group was observed 4 genotypes of DC-SIGNR such as 7/7, 7/5, 9/7 and 6/5, the genotype frequencies were 0.3793, 0.3448, 0.2414 and 0.0345 respectively. 54 cases of non-intrauterine infection group was found 6 genotypes such as 7/7, 7/5, 9/5, 9/7, 7/6 and 6/5, genotype frequencies were 0.5186, 0.1481, 0.0926, 0.1852, 0.0370 and 0.0185 respectively. The distribution of 7/5 genotype in the intrauterine infection group (29 cases) and the non-intrauterine infection group (54 cases) was statistically significant (P = 0.038) , and no significant difference was found in other genotypes between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>The gene DC-SIGN showed relatively little variation in the pregnant women infected with HBV. On the countrary, there were multiple genotypes of the gene DC-SIGNR in these women, and the genotype "7/5" of DC-SIGNR might be one of the susceptibility genes associated with intrauterine infection.</p>
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Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Moléculas de Adesão Celular , Genética , Metabolismo , Predisposição Genética para Doença , Variação Genética , Hepatite B , Genética , Virologia , Vírus da Hepatite B , Genética , Transmissão Vertical de Doenças Infecciosas , Lectinas Tipo C , Genética , Metabolismo , Polimorfismo Genético , Complicações Infecciosas na Gravidez , Genética , Virologia , Receptores de Superfície Celular , Genética , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To construct an eukaryotic expression vector of human telomerase reverse transcriptase (hTERT) gene specific shRNA, and investigate the effect of pshRNA-hTERT combined with gamma-irradiation on cell survival and telomerase activity.</p><p><b>METHODS</b>According to the coding sequence of hTERT mRNA, the target of RNAi was designed, and recombinant expression plasmid pshRNA-hTERT was constructed. The vector was transfected into Hep-2 cells. The radiosensitivity of Hep-2 cells was determined by clonogenic assay. Telomeric repeat amplification protocol (TRAP-PCR-ELISA) was used to observe the telomerase activity in each group. Results Recombinant expression vector pshRNA-hTERT was successfully transfected into Hep-2 cells. The hTERT expression inhibition rate reached 60. 8%. pshRNA-hTERT not only inhibited telomerase activity of Hep-2, but also inhibited the raise of telomerase activity induced by gamma-irradiation. Exposure of Hep-2 cells to pshRNA-hTERT for 24 hrs before irradiation resulted in a decrease in mean surviving fraction of Hep-2 cells compared with cells of group with irradiation alone (67. 7% vs 85. 7%, P <0. 05) .</p><p><b>CONCLUSION</b>RNAi showed a significant inhibitory effect to the expression of hTERT. The results indicate that pshRNA-hTERT can effectively inhibit telomerase activity of Hep-2 cells treated or untreated with 2 Gy gamma-irradiation and significantly enhance the radiosensitivity of Hep-2 cells in vitro. The role of radiosensitization of pshRNA-hTERT may be related with the inhibition of telomerase activity.</p>
Assuntos
Humanos , Carcinoma de Células Escamosas , Genética , Patologia , Linhagem Celular Tumoral , Sobrevivência Celular , Genética , Efeitos da Radiação , Radioisótopos de Cobalto , Ensaio de Imunoadsorção Enzimática , Raios gama , Neoplasias Laríngeas , Genética , Patologia , Plasmídeos , Interferência de RNA , RNA Mensageiro , Genética , RNA Interferente Pequeno , Genética , Proteínas Recombinantes , Telomerase , Genética , Metabolismo , TransfecçãoRESUMO
<p><b>OBJECTIVE</b>To examine the changes of expressions of orexin A, orexin receptor-1 (OX1R), prepro-orexin (Prepro-OX) mRNA, OX1R mRNA and ob-R of hypothalamus in rats with chronic renal failure (CRF).</p><p><b>METHODS</b>Sixty-two male Wister rats weighing 200-250 g were divided into three groups, including group 1 (normal, n = 5), group 2 (sham-operated, n = 25) and group 3 (CRF, n = 32). Hypothalamus orexin A was assayed by radioimmunoassay. Serum leptin was assayed by enzyme linked immunosorbent assay. The expression of Prepro-OX mRNA and OX1R mRNA of hypothalamus were measured by reverse transcription polymerase chain reaction, and expression of orexin A, OX1R and ob-R by immunohistochemistry. Automatic biochemical analyzer was used to measure the serum creatinine.</p><p><b>RESULTS</b>Hypothalamus orexin A levels were negatively correlated (r = -0.63, P < 0.001) with serum leptin levels in the rats. The expression of hypothalamus Prepro-OX mRNA in CRF rats was significantly lower than that of sham-operation at week 12 (P < 0.01). Hypothalamus Prepro-OX mRNA levels were negatively correlated (r = -0.81, P < 0.001) with the levels of serum leptin and serum creatinine (r = -0.68, P < 0.05) in the rats at week 12. The expression of hypothalamus OX1R mRNA in CRF rats was lower than that of sham-operation at week 12 (P > 0.05). Specific immunoreactivity for orexin A was present in perikeryon of the hypothalamus neuron. Specific OX1R-like immunoreactivity was observed in some nerve fibres. Specific immunoreactivity for ob-R was present in membranes of the hypothalamus neuron. Hypothalamus neurons of orexin A-like specific immunoreactivity in CRF rats were significantly fewer than those in shamoperated rats at week 8. Hypothalamus neurons of OX1R-like specific immunoreactivity in CRF rats were similar to those in sham-operated rat at week 8. Hypothalamus neurons of ob-R-like specific immunoreactivity in CRF rats were significantly more than those in sham-operated rats at week 8.</p><p><b>CONCLUSIONS</b>The lower hypothalamus orexin A levels may be induced by high serum leptin level in CRF rats. The lower expression of hypothalamus Prepro-OX mRNA in CRF rats may be one of the main causes inducing lower hypothalamus orexin A. The expression of OX1R in hypothalamus neurons is somewhat reduced and the expression of ob-R in hypothalamus neurons is somewhat raised in CRF rats. These remain to be studied further.</p>
Assuntos
Animais , Masculino , Ratos , Proteínas de Transporte , Genética , Metabolismo , Hipotálamo , Metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Falência Renal Crônica , Metabolismo , Leptina , Genética , Metabolismo , Neuropeptídeos , Genética , Metabolismo , Neurotransmissores , Genética , Metabolismo , Receptores de Orexina , Orexinas , Precursores de Proteínas , Genética , Metabolismo , RNA Mensageiro , Genética , Metabolismo , Distribuição Aleatória , Ratos Wistar , Receptores de Superfície Celular , Genética , Metabolismo , Receptores Acoplados a Proteínas G , Receptores para Leptina , Receptores de Neuropeptídeos , Genética , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To culture dendritic cells (DC) from peripheral blood of patients with laryngeal carcinoma for therapeutic aid.</p><p><b>METHODS</b>Adherent peripheral blood mononuclear cells from peripheral blood were cultured with 15 ng/ml rhGM-CSF and 7 ng/ml rhIL-4 for one or two weeks. The purity of DC was detected by immunocytochemistry method. The mixed leukocyte reactions stimulated by DC loaded with laryngeal carcinoma antigen were tested by measuring 3H-TdR uptake.</p><p><b>RESULTS</b>A considerable number of suspended cells with spicular or dendritic appearance were observed after 1 week of culture, and their mitochondria were rich in cytoplasm. The positivity of DC was about 30%-60%. DC loaded with laryngeal antigen could induce proliferation of syngeneic T lymphocytes.</p><p><b>CONCLUSION</b>A large number of DC with high purity can be cultured from peripheral blood of patients with laryngeal carcinoma in vitro. It may be used in further experimental studies for clinical applications.</p>