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Objective:To investigate the effect of different collection procedures of COBE Spectra blood cell separator on collection of autologous peripheral blood hematopoietic stem cells in children.Methods:The clinical data of 10 children who were collected autologous peripheral blood hematopoietic stem cells with the monocytes (MNC) or fully automatic peripheral blood stem cells (AutoPBSC) programs of COBE Spectra blood cell separator in the Children's Hospital of Soochow University from July 2018 to January 2020 were retrospectively analyzed, and the children were 3-10 years old with the body weight of 15-31kg. There were 5 cases in MNC group and 5 cases in AutoPBSC group.Results:Autologous peripheral blood hematopoietic stem cells were collected for 25 times, with an average of 2.5 times (1-4 times), 10 times in MNC group and 15 times in AutoPBSC group. The number of stem cells [the median (the range)] before collection was 19.3/μl (3.5-129.0/μl) in MNC group and 9.4/μl (2.2-38.7/μl) in AutoPBSC group. The number of CD34 + cells of single collection was 1.22×10 6/kg (0.18×10 6/kg-6.30×10 6/kg) in MCN group and 0.85×10 6/kg (0.13×10 6/kg-2.64×10 6/kg) in AutoPBSC group. Correlation analysis showed that there was a positive correlation between the number of collected CD34 + cells and the number of stem cells before collection (AutoPBSC group: r=0.921, P < 0.01; MNC group: r=0.833, P=0.003). The collection efficiency was 5.4% (3.4%-11.2%) in MNC group and 10.4% (4.7%-13.9%) in AutoPBSC group, and the difference was statistically significant ( Z=2.163, P = 0.031). Conclusion:The collection effect of children's autologous peripheral blood hematopoietic stem cells with COBE Spectra blood cell separator AutoPBSC program is better than that with MNC program.
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Objective:To develop a geriatric nursing core literacy evaluation scale and to test its validity and reliability.Methods:The scale item pool was constructed by referring to relevant literatures and expert consultation method was used to form initial scale. A preliminary investigation was conducted among 40 nurses to form the formal scale, and then 252 nurses were selected to test the validity and reliability of the formal scale.Results:The formal scale consisted of four dimensions and 37 items. The Cronbach α coefficient of the formal scale was 0.980, the content validity index was 0.912, exploratory factor analysis identified 4 principal factors (knowledge geriatric nursing core literacy, skill geriatric nursing core literacy, cognitive geriatric nursing core literacy, belief geriatric nursing core literacy) and explained 73.135% of the total variance.Conclusion:The geriatric nursing core literacy evaluation scale has good reliability and validity, which can be used to evaluate the geriatric nursing core literacy of nursing staffs.
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Objective To investigate the effects of compound Haishe capsule combined with rivastigmine capsule on serum thyroid hormone ,inflammatory factors and coagulation function in patients with Alzheimer＇s disease (AD).Methods From January 2016 to January 2018,74 patients with AD in the Seventh People＇s Hospital of Hangzhou were randomly divided into control group and observation group according to the digital table ,with 37 cases in each group.The control group was treated with rivastigmine capsule ,while the observation group was treated with compound Haishe capsule on the basis of the control group.The two groups were treated for 6 months.The therapeutic effects of the two groups were compared.The changes of BEHAVE-AD and ADAS-Cog scores,thyroid hormones,inflammatory factors and coagulation function were compared before and after treatment.Results The total effective rate of the observation group was 94.59%(35／37),which was higher than 72.97%(27／37) of the control group(u＝6.366,P＜0.05).After treatment, the scores of BEHAVE -AD and ADAS -Cog in the observation group were (4.87 ± 0.64)points,(14.28 ±1.82 ) points, respectively, which were lower than those in the control group [( 8.91 ± 1.07)points,(18.93 ±1.25)points] ( t＝19.710,12.811,all P＜0.05).After treatment,the level of FT4 in the observation group was (13.09 ±0.71) pmol／L,which was higher than (12.30 ±0.52) pmol／L in the control group (t＝5.460,P＜0.05).After treatment,the levels of CRP and IL-6 in the observation group were (2.13 ±0.38)mg／L, (31.32 ±5.64)ng／L,respectively,which were lower than those in the control group [(3.47 ±0.61) mg／L,(58.93 ± 12.15)ng／L](t＝11.342,12.538,all P＜0.05).After treatment,the levels of D-dimer and Fib in the observation group were (519.82 ±10.29) μg／L,(3.21 ±0.32) ng／L,respectively,which were lower than those in the control group[(552.31 ±8.31) μg／L,(4.12 ±0.49) ng／L] ( t＝14.942,9.458,all P＜0.05).Conclusion Compound Haishe capsule combined with rivastigmine capsule is effective in the treatment of AD , and can improve thyroid hormone and coagulation function ,and reduce the level of inflammatory factors.
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Objective@#To investigate the effects of compound Haishe capsule combined with rivastigmine capsule on serum thyroid hormone, inflammatory factors and coagulation function in patients with Alzheimer's disease(AD).@*Methods@#From January 2016 to January 2018, 74 patients with AD in the Seventh People's Hospital of Hangzhou were randomly divided into control group and observation group according to the digital table, with 37 cases in each group.The control group was treated with rivastigmine capsule, while the observation group was treated with compound Haishe capsule on the basis of the control group.The two groups were treated for 6 months.The therapeutic effects of the two groups were compared.The changes of BEHAVE-AD and ADAS-Cog scores, thyroid hormones, inflammatory factors and coagulation function were compared before and after treatment.@*Results@#The total effective rate of the observation group was 94.59%(35/37), which was higher than 72.97%(27/37) of the control group(u=6.366, P<0.05). After treatment, the scores of BEHAVE-AD and ADAS-Cog in the observation group were (4.87±0.64)points, (14.28±1.82)points, respectively, which were lower than those in the control group[(8.91±1.07)points, (18.93±1.25)points](t=19.710, 12.811, all P<0.05). After treatment, the level of FT4 in the observation group was (13.09±0.71)pmol/L, which was higher than (12.30±0.52)pmol/L in the control group (t=5.460, P<0.05). After treatment, the levels of CRP and IL-6 in the observation group were (2.13±0.38)mg/L, (31.32±5.64)ng/L, respectively, which were lower than those in the control group[(3.47±0.61)mg/L, (58.93±12.15)ng/L](t=11.342, 12.538, all P<0.05). After treatment, the levels of D-dimer and Fib in the observation group were (519.82±10.29)μg/L, (3.21±0.32)ng/L, respectively, which were lower than those in the control group[(552.31±8.31)μg/L, (4.12±0.49)ng/L](t=14.942, 9.458, all P<0.05).@*Conclusion@#Compound Haishe capsule combined with rivastigmine capsule is effective in the treatment of AD, and can improve thyroid hormone and coagulation function, and reduce the level of inflammatory factors.
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OBJECTIVE@#To study the regulatory effect of IL-35 on the balance of Treg/Th17 cells in AR patients.@*METHOD@#In this study, 30 cases were randomly selected from outpatients of otolaryngological department in the second hospital of Hebei Medical university who were diagnosed as AR. Another 20 healthy cases enrolled from physical examination branch of our hospital were control group. The expression level of IL-35 and IL-17 in peripheral blood were detected by using ELISA and defeced CD4+CD25+Foxp3+ T cell and CD4+IL-17+T cell expression level were identified via flow cytometry.@*RESULT@#The expression level of IL-35 in AR group was obviously lower than that in control group, and the difference was a statistically significance (t = -8.145, P < 0.01). The expression level of IL-17 in AR group was obviously higher than that in control group, and the difference was a statistically significance (t = 14.969, P < 0.01). There was a remarkable negative correlation between the IL-35 and IL-17 expression in the serum of AR group (r = -0.773, P < 0.01). The percentage of CD4+CD25+Foxp3+ T cell in CD4+ T cell was significant lower in AR group than that in control group (t = -13.678, P < 0.01). The percentage of CD4+IL-17+ T cell in CD4+ T cell was much higher in AR group than that in control group (t = 5.632, P < 0.01). There was a remarkable negative correlation between the Treg and Th17 expression in the peripheral blood of AR group (r = -0.613, P < 0.01). There was a positive correlation between the expression of CD4+ CD25+Foxp3+ T cell and IL-35. There was a negative correlation between the IL-35 and Th17 in AR group (r = 0. -594, P < 0.01).@*CONCLUSION@#The lower expression of IL-35 was related to the incidence of AR, and it was an important cytokines for that. The lower expression of IL-35 may inhibit the proliferation of Treg cells, lead to hyper function of Th17 cells, increase secretion of s IL-17 and result in unbalance of Treg/Th17 cells; these may be the important mechanism of the occurrence of AR, thus regulation of IL-35 may become a new target for the immunological therapy of AR.
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Humans , Case-Control Studies , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Interleukin-17 , Blood , Interleukins , Rhinitis, Allergic , Allergy and Immunology , T-Lymphocytes, Regulatory , Allergy and Immunology , Th17 Cells , Allergy and ImmunologyABSTRACT
BACKGROUND:Damage to nasal ciliated epithelial cels can lead to a severe injury in nasal biological function. Compared with other adult stem cels, human umbilical cord blood stem cels have better differentiation potential. OBJECTIVE: To explore the feasibility of human umbilical cord blood stem cels differentiating into nasal ciliated epithelial cels through in vitro culture and induction techniques. METHODS:Normal and healthy umbilical cord blood samples were colected to isolate human umbilical cord blood stem cels, folowed by identification and subculturein vitro. Umbilical cord blood stem cels at passage 3 were infected with recombinant adeno-associated virus carrying enhanced green fluorescent protein and cultured using air liquid interface culture method. Thereafter, PCR assay was employed for detecting MUCS expression in cultured stem cels at 1 and 2 weeks after induction, and immunofluorescent staining for FOXJ1 was performed at 3 weeks. RESULTS AND CONCLUSION:After subculture, passage 3 umbilical cord blood stem cels that could express stem cel surface markers were visible in a uniform shape and had good refraction. After 3 hours of gene transfection, green fluorescence issued from the passage 3 cels were visible, and the cel positive rate was up to 96.2% until 48 hours, indicating good transfection efficiency. RT-PCR findings showed that MUC8 mRNA had no expression in the umbilical cord blood stem cels, but expressed strongly in the nasal ciliated epithelial cels, whose expression was weak at 1 week of culture and increased at 2 weeks. Additionaly, the positive expression of FOXJ1 red fluorescence was observed under the transfection of green fluorescent protein. These results suggest that human umbilical cord blood stem cels could differentiate into nasal epithelial cels under suitable conditions.
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Objective To investigate the expression of GRP78 and caspase-12 on the auditory cortex and to study the effects of endoplasmic reticulum stress in the auditory cortex neuron apoptosis after the brain ischemia reperfusion injury in guinea pig.Methods Fifty healthy male guinea pigs were selected and randomly divided into 5 groups which were hormal group group A(reperfusion for 6 hours),group B(reperfusion for 12 hours),group C (reperfusion for 24 hours),group D(reperfusion for 72 hours).The cerebral ischemia reperfusion injury model was produced by the occlusion of bilateral common carotid arteries.The guinea pigs were sacrificed at reperfusion of 6, 12,24,and 72 hours respectively after they received ABR tests.The pathological changes were observed by HE and the levels of GRP78 and caspase-12 protein were detected by immunohistochemistry and Western blot.Results The hearing thresholds increased gradually from the normal group to group B,and decreased gradually from group C to group D,but the thresholds of group D were still higher than that of the normal group.HE staining showed that the neurons in the normal control group were arranged in order,the cytoplasm was abundant,large and round.The cells were stained clearly.After reperfusion,the number of neurons in each time point was decreased,the nucleus presented atrophic,fragmented,the disappeared.The expression of GRP78 and caspase-12 protein in normal con-trol group was only a trace or a small amount by immunohistochemistry and Western blot.The expression of GRP78 and caspase-12 began to inerease.The expression of GRP78 reached the peak at reperfusion of 12 hours and de-creased gradually.There were significant statistic differences between each group comparison.The expression of caspase-12 reached the peak at reperfusion of 24 hours,then decreased gradually.There was no statistic difference between group A and group B.There were significant statistic differences anong other groups.Conclusion Endo-plasmic reticulum stress(ERS)may be induced by brain ischemia reperfusion injury,and can increase the expression of GRP78 and caspase-12.GRP78 and caspase-12 participate in the process of neuron apoptosis on auditory cor-tex caused by ERS.
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<p><b>OBJECTIVE</b>To compare the effect of three cannulated screws and dynamic hip screw (DHS) combined with antirotation screw in treating patients with comminuted fractures of femoral neck.</p><p><b>METHODS</b>From January 2008 to December 2012,67 patients with comminuted fractures of femoral neck were treated with three cannulated screws or DHS combined with antirotation screw. Among them, 36 patients were treated with DHS combined with antirotation screw including 24 males and 12 females with an average of 43.7 years old ranging from 22 to 58 years old; 31 patients were treated with three cannulated screws including 22 males and 9 females with an average of 43.0 years old ranging from 24 to 56 years old. The incision size, operation time, blood loss, blood transfusion rate, postoperative hospital stay, walking time, weight-bearing time, union time, incidence of complication were recorded and compared between two groups, and functional outcomes were evaluated by Harris scoring.</p><p><b>RESULTS</b>All incision healed at stage I. The differences in operation time, blood loss, blood transfusion rate and postoperative hospital stay between two groups were not significant (P > 0.05). Incision size of DHS group (5.00 ± 0.86) cm was larger than that of cannulated screw group (3.30 ± 0.57) cm (P < 0.01). All patients were followed up more than one year, and the follow-up time showed no significant difference between two groups (P > 0.05). Walking time of DHS group (19.0 ± 3.5) d was less than that of cannulated screw group (37.0 ± 6.3) d (P < 0.01. Partial and full weight-bearing time of DHS group (23.0 ± 7.0) d and (138.0 ± 13.0) d was less than that of cannulated screw group (38.0 ± 5.7) d and (164.0 ± 12.0) d (P < 0.01). Union time of DHS group (151 ± 11) d was less than that of cannulated screw group (162 ± 11) d (P < 0.01). Harris hip score of DHS group (91.0 ± 5.7) was higher than that of cannulated screw group (85.0 ± 12.0) (P < 0.01). No significant differences were found in the incidence of postoperative complications between two groups (P > 0.05).</p><p><b>CONCLUSION</b>The DHS combined with antirotation screw is superior to three cannulated screws for treatment of comminuted fractures of femoral neck due to earlier weight-bearing, faster fracture healing and better hip function</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Bone Screws , Femoral Neck Fractures , General Surgery , Follow-Up Studies , Fracture Fixation, Internal , Methods , Fractures, Comminuted , General SurgeryABSTRACT
OBJECTIVE@#To evaluate the effect of hypoxia improvement in Hep-2 cell on cisplatin-induced apoptosis.@*METHOD@#Human laryngeal squamous cell carcinoma Hep-2 cells and HIF-1α-RNAi-Hep-2 cells were cultured in normoxic, hypoxic and reoxygenation condition. The inhibition of cisplatin on cell proliferation was evaluated by MTT assay. The influence of cisplatin on cell cycle and apoptosis were detected by flow cytometry.@*RESULT@#The inhibition of cisplatin on cell proliferation was reduced by hypoxia. After HIF-1α gene was silenced, the inhibition of cisplatin on Hep-2 cell proliferation was increased apparently, but was still interfered partly by hypoxia. Hypoxia could induce cell apoptosis decreased and enhance chemotherapeutic resistance. After reoxygenation, cell apoptosis induced by cisplatin was increased significantly (P < 0.05). HIF-1α-RNAi-Hep-2 cells under hypoxia also showed certain resistance to apoptosis but the sensitivity to cisplatin was higher than that of Hep-2 cells. When cells were returned from hypoxic condition to normoxic condition for some time, the apoptosis induced by cisplatin was increased significantly (P < 0.05).@*CONCLUSION@#The improvement of hypoxic microenvironment with HIF-1α gene knockout could increase the sensitivity of Hep-2 cells to chemotherapy.
Subject(s)
Humans , Apoptosis , Carcinoma, Squamous Cell , Pathology , Cell Cycle , Cell Hypoxia , Cell Line, Tumor , Cell Proliferation , Cisplatin , Pharmacology , Flow Cytometry , Gene Knockout Techniques , Head and Neck Neoplasms , Pathology , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Laryngeal Neoplasms , Pathology , RNA Interference , Squamous Cell Carcinoma of Head and NeckABSTRACT
OBJECTIVE@#The purpose of this study was to investigate the expression of PKM2 and HIF-1α in laryngeal squamous cell carcinoma and to analyze their correlation in laryngeal squamous cell carcinoma.@*METHOD@#Total 37 laryngeal carcinoma samples and para-carcinoma tissues were taken from the patients who accepted operation treatment in the Second Hospital of HeBei Medical University from 06/2013 to 06/2014. The protein expression levels of PKM2 and HIF-1α were detected with SP immunohistochemical methods. The data were analyzed by the SPSS 13.0 statistical software.@*RESULT@#The positive expression of PKM2 in laryngeal carcinoma tissues and adjacent tissues were 62.16% and 13.15%. The difference was statistically significant (P 0.05). The expression of PKM2 was positively related with HIF-1α in laryngeal squamous cell carcinoma (P < 0.01).@*CONCLUSION@#The expression of PKM2 and HIF-1α are related with the proliferation, invasion and metastasis of laryngeal squamous cell carcinoma. It provides a certain theoretical basis for laryngeal cancer diagnosis and screening to measure the expression of PKM2 and HIF-1α as biological indicators.
Subject(s)
Humans , Biomarkers, Tumor , Metabolism , Carcinoma, Squamous Cell , Metabolism , Carrier Proteins , Metabolism , Head and Neck Neoplasms , Metabolism , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Laryngeal Neoplasms , Metabolism , Lymphatic Metastasis , Membrane Proteins , Metabolism , Squamous Cell Carcinoma of Head and Neck , Thyroid Hormones , MetabolismABSTRACT
OBJECTIVE@#To investigate the functional mechanism of metadherin (MTDH), hypoxia-inducible factor-1 alpha (HIF-1α) and transketolase-like gene 1 (TKTL1) and their interaction with each other in laryngeal carcinoma development.@*METHOD@#Thirty laryngeal carcinoma samples and 20 samples of para-carcinoma tissue were taken from the patients received operation treatment. The expression levels of MTDH, HIF-1α and TKTL1 were detected in all the samples by SP immunohistochemical methods. The data were analyzed by the SPSS13.0 statistical software.@*RESULT@#The positive expression rate of MTDH, HIF-1α and TKTL1 in the 30 cases of laryngeal carcinoma was 56.67%, 60.00% and 63.33%, respectively, which was 20.00%,10.00% and 15.00% respectively in the para-carcinoma tissue, the difference of the positive expression rate of the tested objects between these two groups was statistically significant (P 0.05). The Spearman's rank correlation analysis showed that the positive expression rates of MTDH and HIF-1α in laryngeal carcinoma had positive correlation (r = 0.384, P 0.05).@*CONCLUSION@#It suggested that these three proteins may have close relationship with the occurrence, invasion and metastasis of the laryngeal carcinoma. MTDH and TKTL1 may be expected to be new clinical targets for laryngeal carcinoma treatment and it could offer theoretical basis for the prognosis of the laryngeal carcinoma.
Subject(s)
Humans , Carcinoma, Squamous Cell , Diagnosis , Metabolism , Cell Adhesion Molecules , Metabolism , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Laryngeal Neoplasms , Diagnosis , Metabolism , Lymphatic Metastasis , Neoplasm Staging , Prognosis , Transketolase , MetabolismABSTRACT
OBJECTIVE@#To observe the change of auditory brainstem response (ABR) in hearing deprivation rat model induced by bilateral cochlears ablation at different time points.@*METHOD@#Forty SD rats were randomly divided into four experiment groups including 2-week group, 4-week group, 6-week group, 8-week group and four control groups with 5 rats (n = 10) in each group. Then bilateral cochlears ablation was applied to experiment groups. The threshold value of ABR was measured at different time and latent period of each wave was compared.@*RESULT@#The threshold of ABR in experiment group was elevated significantly. The latent period of each wave was pro longed significantly (P < 0.01). In experiment group, the threshold value of ABR in 2-week and 4-week group was significantly greater than that in 6-week group and 8-week group (P < 0.01).@*CONCLUSION@#Bilateral cochlears ablation surgery could elevate the threshold of ABR and latent period of each wave prolonged. The effect of hearing deprivation became apparent after surgery for 4 weeks.
Subject(s)
Animals , Male , Rats , Auditory Threshold , Cochlea , General Surgery , Disease Models, Animal , Evoked Potentials, Auditory, Brain Stem , Physiology , Hearing Loss , Rats, Sprague-DawleyABSTRACT
Erbin, a member of Leucine-rich repeat and PDZ-containing protein family, was found to inhibit TGF-β-induced epithelial-mesenchymal transition (EMT) in our previous study. However, the mechanism of Erbin in regulating EMT is unclear. Semaphorin protein Sema4C, with PDZ binding site at C-terminal has been recognized as a positive regulator of EMT. Here, we aimed to examine the interaction between Erbin and Sema4C. HK2 cells were treated with TGF-β1, or transfected with Erbin and (or) Sema4C. Interaction of Erbin and Sema4C was identified by immunoprecipitation. RT-PCR was used to detect the expression of Erbin and Sema4C at mRNA level after transfection. The expression levels of Erbin, Sema4C, and markers of EMT were measured by using Western blotting or ELISA. After HK2 cells were stimulated with 10 ng/mL TGF-β1 for 72 h, the protein expression levels of Erbin and Sema4C were both up-regulated, and immunoprecipitation results showed Erbin interacted with Sema4C in HK2 cells both at endogenous and exogenous levels. Furthermore, overexpression of Sema4C suppressed E-cadherin, induced vimentin and promoted fibronectin secretion, indicating Sema4C promotes the process of EMT. However, HK2 cells overexpressing Erbin were resistant to Sema4C-induced EMT. In contrast, Erbin specific siRNA promoted EMT induced by Sema4C. Taken together, these results suggest that Erbin can interact with Sema4C, and co-expression of Erbin blocks the process of Sema4C-induced EMT.
Subject(s)
Humans , Adaptor Proteins, Signal Transducing , Genetics , Metabolism , Blotting, Western , Cadherins , Metabolism , Cell Line , Epithelial-Mesenchymal Transition , Immunoprecipitation , Kidney Tubules, Proximal , Cell Biology , Metabolism , Protein Binding , RNA Interference , Reverse Transcriptase Polymerase Chain Reaction , Semaphorins , Genetics , Metabolism , Transfection , Transforming Growth Factor beta1 , Pharmacology , Vimentin , MetabolismABSTRACT
Objective To investigate the morphological change and SHP -2(src -homology domain contai-ning protein tygosine phosphatase type 2) gene expression in auditory cortical neuron after auditory deprivation in rat .Methods A total of 48 SD rats were randomly divided into 2-week group ,4-week group ,6-week group ,8-week group and 4 corresponding control groups with 6 rats in each group .Bilateral cochlear ablation was done to traumatized groups to ensure their postoperative ABR threshold was above 90 dB SPL .Then paraffin sectioning and HE and Nissl staining were used to detect morphological change of auditory cortical neuron ,simultaneously the RT-PCR was used to measure the expression of SHP -2 gene in auditory cortical neuron of each group ,and relative quantitative analysis was used .Results The HE and Nissl staining revealed that apoptotic shape of auditory cortical neuron became serious by time ,with the diversified cellular morphology .The relative quantity of SHP -2 gene showed statistical differences between any two groups by the method of one -way ANOVA ,showing a rising trend by time (P<0 .05) .Conclusion Along with time ,auditory cortex ,auditory deprivation showed increasing neuronic apoptosis and neuronic proliferation .Apoptosis was the final result from the mutual antagonism by the 2 factors .
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OBJECTIVE@#To explore the expression of phospholipase Cgamma-2 (PLCgamma-2), lipoxygenase-12 (12-LOX) and arachidonic acid (AA) in laryngeal squamous cell carcinoma and to study the the relationship between lipid metabolism and laryngeal squamous cell carcinoma.@*METHOD@#In 30 cases of carcinoma tissue and peritumoral laryngeal mucosa tissues (confirmed to be normal laryngeal tissues by pathology), immunohistochemical method (Streptavidin-peroxidase method, SP method) was used for the detection of expression of PLCgamma-2 and 12-LOX, and gas chromatography/mass spectrometry (GC/MS) for the content of the arachidonic acid in carcinoma tissue and peritumoral normal laryngeal mucosa tissues.@*RESULT@#The positive rates of PLCgamma-2 and 12-LOX in carcinoma tissue were higher than in peritumoral normal laryngeal mucosa tissues with statistically significance differences (P 0.05). Both the expression of PLCgamma-2 and 12-LOX and the content of arachidonic acid had no statistically significant correlation with age (P > 0.05).@*CONCLUSION@#PLCgamma-2, AA and 12-LOX play important roles in laryngeal squamous cell carcinoma. It may be meaningful to the treatment of laryngeal carcinoma by suppressing this passway.
Subject(s)
Aged , Humans , Male , Middle Aged , Arachidonate 12-Lipoxygenase , Metabolism , Arachidonic Acid , Metabolism , Carcinoma, Squamous Cell , Metabolism , Pathology , Head and Neck Neoplasms , Metabolism , Pathology , Laryngeal Neoplasms , Metabolism , Pathology , Neoplasm Staging , Phospholipase C gamma , Metabolism , Squamous Cell Carcinoma of Head and NeckABSTRACT
Erbin, a member of Leucine-rich repeat and PDZ-containing protein family, was found to inhibit TGF-β-induced epithelial-mesenchymal transition (EMT) in our previous study. However, the mechanism of Erbin in regulating EMT is unclear. Semaphorin protein Sema4C, with PDZ binding site at C-terminal has been recognized as a positive regulator of EMT. Here, we aimed to examine the interaction between Erbin and Sema4C. HK2 cells were treated with TGF-β1, or transfected with Erbin and (or) Sema4C. Interaction of Erbin and Sema4C was identified by immunoprecipitation. RT-PCR was used to detect the expression of Erbin and Sema4C at mRNA level after transfection. The expression levels of Erbin, Sema4C, and markers of EMT were measured by using Western blotting or ELISA. After HK2 cells were stimulated with 10 ng/mL TGF-β1 for 72 h, the protein expression levels of Erbin and Sema4C were both up-regulated, and immunoprecipitation results showed Erbin interacted with Sema4C in HK2 cells both at endogenous and exogenous levels. Furthermore, overexpression of Sema4C suppressed E-cadherin, induced vimentin and promoted fibronectin secretion, indicating Sema4C promotes the process of EMT. However, HK2 cells overexpressing Erbin were resistant to Sema4C-induced EMT. In contrast, Erbin specific siRNA promoted EMT induced by Sema4C. Taken together, these results suggest that Erbin can interact with Sema4C, and co-expression of Erbin blocks the process of Sema4C-induced EMT.
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<p><b>OBJECTIVE</b>To investigate the effects of hypoxia on the features and chemoresistance of cancer stem cells in Hep-2 cells and underlying mechanism.</p><p><b>METHODS</b>The shRNA interference recombinant plasmid targeting HIF-1α was synthesized and transfected into Hep-2 cells. The HIF-1α knockdown Hep-2 cells were established after clonal selection and the expression of HIF-1α was measured. The cellular features including proliferation, clonal formation, cell cycle, apoptosis and CD133 phenotype were measured in Hep-2 cells cultured under hypoxic condition in vitro. CD133+ cells were sorted from Hep-2 cells with flow cytometry. Clonal formation test and cisplatin treatment were carried out, and the expressions of related genes (Oct-4, suvivin and p53) in CD133+ cells were measured.</p><p><b>RESULTS</b>HIF-1α knockdown Hep-2 cells was successfully established, as evidenced by the reduced mRNA and protein expressions of HIF-1α. The Hep-2 cells cultured under hypoxic microenvironment showed higher proliferation and clonal formation activity, cell cycle arrest in G0/G1, lower apoptosis, up-regulated CD133, however the effects of hypoxia reduced in HIF-1α knockdown Hep-2 cells. CD133+ cells were successfully sorted from Hep-2 cells, and the CD133+ cells showed increased clonal formation activity and cisplatin treatment resistance in hypoxia. Also the effects of hypoxia on CD133+ cells decreased with HIF-1α knockdown, showing down-regulated Oct-4 and survivin and up-regulated p53.</p><p><b>CONCLUSIONS</b>Hypoixa can induce the features of cancer stem cells in Hep-2 cells and increase proliferation, differentiation and chemoresistant ability of CD133+ cells, which might be correlated with the changes in expressions of HIF-1α and related genes regulated by HIF-1α.</p>
Subject(s)
Humans , Apoptosis , Carcinoma, Squamous Cell , Pathology , Cell Cycle , Cell Differentiation , Cell Hypoxia , Cell Line, Tumor , Cell Proliferation , Drug Resistance, Neoplasm , Gene Knockdown Techniques , Hypoxia-Inducible Factor 1, alpha Subunit , Genetics , Laryngeal Neoplasms , Pathology , Neoplastic Stem Cells , Cell Biology , RNA, Small Interfering , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of hypoxia inducible factor 1 alpha (HIF-1α), glucose transporter protein-1 (GLUT-1) and vascular endothelial growth factor (VEGF) in human laryngeal carcinoma tissue, and to study the relationship between hypoxia and HIF-1α, GLUT-1, VEGF in human laryngeal carcinoma Hep-2 cells and to explore the effect of HIF-1α, GLUT-1 and VEGF as endogenous hypoxic markers on laryngeal carcinoma.</p><p><b>METHODS</b>The expression levels of HIF-1α, GLUT-1 and VEGF were detected in 35 cases of laryngeal carcinoma by SP immunohistochemical methods and in Hep-2 cells by SP immunocytochemical methods. The relationship between HIF-1α and GLUT-1, VEGF protein expression was analyzed.</p><p><b>RESULTS</b>Of the 35 cases, 16 cases expressed HIF-1α, 16 cases expressed GLUT-1, 19 cases expressed VEGF. The expression of HIF-1α and VEGF were closely correlated with pathologic grading and lymphnode metastasis. GLUT-1 was correlated with lymphnode metastasis. The expression levels of HIF-1α, GLUT-1 and VEGF in Hep-2 cells under hypoxic condition were higher than those under normoxic condition.</p><p><b>CONCLUSION</b>HIF-1α may promote the expression of GLUT-1 and VEGF in laryngeal carcinoma, furthermore promote tumor angiogenesis, invasion, and metastasis of the laryngeal carcinoma.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , Metabolism , Pathology , Cell Line, Tumor , Glucose Transporter Type 1 , Metabolism , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Laryngeal Neoplasms , Metabolism , Pathology , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
Hypoxia could not only induce tumor chemoradiotherapeutic resistance, but also cause strong invasiveness which make tumor inclined to recurrence and metastasis. With regard to the detection of oxygenation condition in tumor tissue, there are many methods, but each has its advantages and disadvantages. Accurate monitoring of tumor oxygenation state plays an important role in therapeutic schedule formulation and implementation which would improve curative effect of tumor.
Subject(s)
Animals , Humans , Cell Hypoxia , Hypoxia , Neoplasms , Blood , Oximetry , MethodsABSTRACT
<p><b>OBJECTIVE</b>To study whether laryngeal cancer stem cells in hypoxia have the characteristic of resistance to irradiation and underlying mechanism.</p><p><b>METHODS</b>CD133(+) cells were separated from Hep-2 cells with flow cytometry (FCM) and the purity was 92.8%. The separated CD133(+) cells were cultured in serum-free medium in hypoxia in normoxia environment respectively, and hypoxia-inducible factor 1 alpha (HIF-1α) expression was detected by FCM. The cells were exposed respectively to X-rays emitted by linear accelerator with a dose of 0, 5, 10, 15 or 20 Gy for 24 hours, with additional time points of 12, 36, and 48 hours for the cells exposed to 10 Gy. Then the growth inhibition ratios of cells in hypoxia and normoxia groups were detected with MTT assay at different time points. Soft agar colony formation assay was used to detect colony formation ratios of cells in hypoxia and normoxia groups. DNA dependent protein kinase catalytic subunit (DNA-PKcs), ataxia telangiectasia mutate (ATM), Survivin and P53 were detected by FCM.</p><p><b>RESULTS</b>Growth inhibition ratio of CD133(+) cells in hypoxia group was lower than that in normoxia group (P < 0.05). Colony formation ratio of CD133(+) cells was higher than that of CD133(-) cells (P < 0.01) and the ratio of CD133(+) cells in hypoxia group was higher than that in normoxia group (P < 0.05). The ratio of hypoxia group was not affected by irradiation, while the ratio of normoxia group decreased significantly after irradiation (P < 0.05). The expressions of DNA-PKcs, ATM, Survivin and P53 in CD133(+) cells were higher than those in CD133(-) cells respectively (P < 0.01). In CD133(+) cells with radiation, the expressions of DNA-PKcs and Survivin of hypoxia group were higher those of normoxia group (P < 0.05), but no difference in the expression of ATM or P53 between the two groups.</p><p><b>CONCLUSIONS</b>Laryngeal cancer stem cells play an important role in radioresistance mediated by hypoxia.</p>