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AIM To explore the radioprotective function of Xihuang Capsules (Bovis Calculus artijactus,Moschus,Olibanum,Myrrha) in patients with acute radiation-induced oral mucositis and its mechanisms.METHODS Eighty patients with nasopharyngeal carcinoma undergone radiotherapy were randomized into two groups:treatment group (radiation plus Xihuang Capsules) and control group (radiation alone).The comparison of two groups in the onset and the tolerated dose of mucositis was made according to the acute radiation injury classification standard.The levels of TNF-α and IL-6 in saliva were detected by ELISA.RESULTS After the treatment with Xihuang Capsules,the onset of oral mucositis delayed significantly and the tolerated dose elevated markedly comparing with the control group (t =2.180,12.930,P ＜ 0.05).The morbidity rate of Ⅲ-N-degree of oral mucositis in the treatment group was lower than that of the control group at the dosage of 40 Gy and 70 Gy,respectively (Z =3.661,4.270,P ＜0.01).Furthermore,there was no difference in the levels of TNF-α and IL-6 in the pre-treatment in the two groups (t =1.010,1.469,P ＞ 0.05).With the increase in radiant dose,the levels of TNF-α and IL-6 in the two groups both elevated dramatically,and peaked at the dose of 40 Gy,but both levels in the treatment group were lower than those of the control group (t =8.305,6.069,P ＜ 0.05).When DT =70 Gy,the levels of TNF-α and IL-6 were lower than pre-radiation,and the levels of TNF-α and IL-6 in the treatment group were lower than those in the radiation alone group (t =3.835,2.488,P ＜ 0.05).CONCLUSION Xihuang Capsules can delay and reduce acute radiation-induced oral mucositis and improve radiation tolerated dose,so it may involve the release of TNF-oα and IL-6 in saliva.
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Objective To analysis the difference of lymph node cleaning and operative complication rate between thoracoscope surgery and routine thoracotomy on patients with thoracic segment esophageal cancer.Methods A summary of 62 patients with thoracic segment esophageal cancer in Xiaogan Hospital Affiliated to Wuhan University of Science and Technology from August 2012 to August 2014,who were carried with thoracoscope surgery,were randomly chosed and designed as the thoracosc0Pe group,and 62 patients with thoracic segment esophageal cancer over the same period,carried out with routine thoracotomy,were designed as the control group.All the clinical data of the two groups were collected.The total of thoracic lymph node cleaned and the group of thoracic lymph node cleaned were compared between the two groups.The operation time,intraoperative blood loss,chest tube placement time and postoperative hospital duration were collected and compared.All the patients were followed up at least for one year.The incidence of postoperative complications such as pulmonary infection,pneumothorax,atelectasis,recurrent laryngeal nerve injury and anastomotic leakage in the follow-up period were compared.The follow-up time,mortality and recurrence rate were compared.Results The total of thoracic lymph node cleaned(13.36±3.28) and the group of thoracic lymph node cleaned(3.35±0.84) in the thoracoscope group were lower then these of the control group ((14.22± 2.78) and (3.58±0.75)),but with no statistical difference (t =1.57,1.61,P＞ 0.05).The operation time of the thoracoscope group((314.63±38.72) min) were higher then that of the control((217.46±41.54) min),and the intraoperative blood loss ((205.73 ± 114.38) ml),chest tube placement time ((6.83 ± 1.92) d) and postoperative hospital duration((18.47±5.36) d) of the thoracoscope group were remarkably lower then these of the control ((345.72 ±175.62) m1,(10.04±2.41) d,(22.65±6.84) d,t=13.47,5.26,8.20,3.79,P＜0.05).The incidence of pulmonary infection (4.8％ (3/62)) and atelectasis (1.6％ (1/62)) of the thoracoscope group,were evidently lower then these of the control (17.7％ (11/62),1.3％ (7/62),x2 =5.15,4.81,P＜0.05).There was no significant difference in mortality and recurrence ratebetween the two groups during the follow-up period (3.2％ (2/62) vs.8.1％ (5/62),11.3％ (7/62) vs.14.5％ (9/62),x2 =1.36,0.29,P＞0.05).Conclusion There are no significant difference inlymph node cleaning between thoracoscope surgery and routine thoracotomy on patients with thoracic segment esophageal cancer,but thoracoscope surgery can shorten the length of hospital duration,reduce the intraoperative blood loss,chest tube placement time and postoperative complications.So the thoracoscope surgery is a safe and feasible operation for patients with thoracic segment esophageal cancer.
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AIM: To investigate the role of chloride channels in the apoptosis of human poorly differentiated nasopharyngeal carcinoma CNE-2Z cells induced by arsenic trioxide (As2O3).METHODS: The apoptotic rates of CNE-2Z cells induced by As2O3 for 24 h or 48 h were monitored by flow cytometry.The technique of whole-cell patch clamp was used to record the currents activated by As2O3 in the CNE-2Z cells.The inhibition of As2O3-induced apoptosis by chloride channel blocker DIDS in the CNE-2Z cells was analyzed by flow cytometry.RESULTS: As2O3 at 5 μmol/L induced apoptosis of CNE-2Z cells in time-dependent manner.The currents with outward rectification were activated when the cells were exposed to 5 μmol/L As2O3.No obvious time-and voltage-dependent inactivation of the currents was observed.The reverse potential of the currents was close to the equilibrium potential for chloride.The activated currents were inhibited by the chloride channel blockers NPPB and DIDS.The 47% hypertonic solution inhibited the activated currents completely.Chloride channel blocker DIDS inhibited the apoptosis of CNE-2Z cells induced by As2O3.CONCLUSION: As2O3 activates volume-sensitive chloride channels, and chloride channels may play an important role in the apoptosis of CNE-2Z cells induced by As2O3.
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[ ABSTRACT] AIM:To investigate the effect of the overexpression of voltage-gated chloride channel family protein 3 ( ClC-3) gene on bones of mice .METHODS: The tail gene detection assay was used to confirm the overexpression of ClC-3.The male FVB mice of three months old were divided into two groups , the wild type ( WT) group and the ClC-3 overexpressed (ClC-3 transgene) group.The body weight, length and weight of the right tibias were measured .The upper and middle parts of the tibias were dissected , decalcified, paraffin-imbed, sectioned and stained with HE staining .The bone morphology metrology was used to analyze the changes of bone structures .The percent trabecular area (%Tb.Ar), trabecular number ( Tb.N) , trabecular width ( Tb.Wi) and trabecular separation ( Tb.Sp) of cancellous bone in the upper part of the tibia were measured.The total tissue area (T.Ar), cortical area (Ct.Ar), percent cortical area (%Ct.Ar), marrow area ( Ma.Ar) and percent marrow area (%Ma.Ar) of the cortical bone in the middle part of the tibia were detec-ted .RESULTS:The wild type mice and the ClC-3-overexpressed mice were verified by the tail gene detection assay . Compared with WT group , the body weight and the length and weight of the tibia were decreased in ClC -3 transgene mice (P<0.05).In the cancellous bones of ClC-3 transgene mice, the%Tb.Ar and Tb.Wi were decreased (P<0.05), the Tb.Sp was increased (P<0.05) and the Tb.N was not significantly changed .In the cortical bones of ClC-3 transgene mice, the T.Ar, Ct.Ar and%Ct.Ar were decreased (P<0.05), the%Ma.Ar was increased (P<0.05), and the Ma. Ar was not significantly changed .CONCLUSION:ClC-3 overexpression may lead to the reduction of the bone mass and the destructure of the cancellous and cortical bones .The results suggest that ClC-3 may be involved in the regulation of bone resorption and/or formation.
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Aim To study the mechanism of DSF-Cu induced apoptosis of human nasopharyngeal carcinoma CNE-2 Z cells by affecting the function of mitochondria and cytoskeleton. Methods The cell cycle,the rate of apotosis,the levels of intracellular ROS and MMP in CNE-2 Z cells were tested by flow cytometry after trea-ted with different concentration of DSF-Cu. The chan-ges of the cell surface morphology, ultrastructure, cell height, width and roughness were detected by AFM. The distribution and reorganization of cytoskeleton F-actin were observed by Laser scanning confocal micro-scope. Results Cells were incubated with different concentration of DSF-Cu ( 0 ~200 nmol · L-1 ) for 24 h, the apoptotic ratio increased significantly and the treatment of DSF-Cu resulted in a concentration-de-pendent accumulation of CNE-2Z cells in G2/M phase. Furthermore,the treatment of DSF-Cu was able to in-crease the production of intracellular ROS and decrease the MMP in CNE-2Z cells. In addition,AFM imaging showed that compared to the control group,with the in-crease of DSF-Cu concentration,the CNE-2Z cells be-came smaller, cytoplasm condensed, the height in-creased,and the surface roughness reduced. Moreover, the filopodia became shorter, shrinked and even com-pletely destroyed after treated with different concentra-tion of DSF-Cu. At last,the LSCM image showed that the fluorescence intensity of F-actin networks was de-creased, then the structure was rearranged and de-stroyed obviously by treated with DSF-Cu. Conclusion DSF-Cu can induce apoptosis and arrest cell cycle at G2/M phase in CNE-2Z cell through a mitochondria-dependent pathway. Above findings highlight the appli-cations of AFM at the single cell level for the investiga-tion of antineoplastic drug in nasopharyngeal carcinoma therapy.
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AIM: To study the effects of disulfiram/copper complex (DSF/Cu) on ultrastructures and mechan-ical properties of human breast cancer and normal breast epithelial cells by atomic force microscopy (AFM) based on the nanoscale resolution and piconewton force measurement level.METHODS: The change of cell cycle and apoptotic rate of MCF-7 cells and MCF-10A cells induced by DSF/Cu were compared by flow cytometry.The cell surface morphology, ultra-structure, height, width and roughness were detected by AFM.The effects of DSF/Cu on the hardness (Young’s modu-lus) of the 2 kinds of cells were determined by AFM with indentation technique.RESULTS: DSF/Cu significantly in-duced apoptosis of the MCF-7 cells in a dose-dependent manner, whereas had little effect on the MCF-10A cells.The cell cycle analysis showed that DSF/Cu induced G2 /M arrest in the MCF-7 cells, but led to G0 /G1 arrest in the MCF-10A cells.The AFM images showed that the MCF-7 cells shrank and showed smaller and smoother morphology, and the filopo-dia were retracted obviously, even some became into lamellipodia, or disappeared completely after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.The quantitative analysis indicated that the MCF-7 cells showed smaller width and larger height, and the root mean square roughness and average roughness were decreased significantly in a dose-dependent manner after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.However, little effect in the MCF-10A cells was observed.The biomechanics test at a single cell level demonstrated that the Young’s modulus of the MCF-7 cells and MCF-10A cells were both increased, yet the proportion increased in the MCF-7 cells was much higher than that in the MCF-10A cells after treated with DSF/Cu at concentrations of 400 and 800 nmol/L.CONCLUSION: DSF/Cu has strong antitumor effect on breast cancer with high efficiency and low toxicity by changing the properties of the biomechanics specifically.
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Aim To investigate the roles of chloride channels in the apoptosis and apoptotic volume de-crease (AVD)induced by adriamycin in nasopharyn-geal carcinoma CNE-2Z cells.Methods Apoptotic rates were detected by flow cytometry,and the volume changes were measured by the time-lapse live cell ima-ging technique.The patch clamp technique was used to record whole-cell chloride currents.Results Adria-mycin induced apoptosis of CNE-2Z cells.An early ap-optotic volume decrease was observed in the cell trea-ted with adriamycin.The cell volume was decreased by about 10% in 2 h.Adriamycin activated a chloride current which showed outward rectification.The chlo-ride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB ) could inhibit the adriamycin-in-duced chloride currents,apoptosis and prevent cell shrinkage.Conclusions Our findings suggest that ad-riamycin causes cell apoptosis by activation of chloride channels.Chloride channels may be involved in the apoptosis and apoptotic volume decrease induced by adriamycin in CNE-2Z cells.
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Aim To investigate the effect of gelsemium alkaloids on chloride channels and cell volume in he-patic carcinoma cells. Methods The time-lapse live cell imaging and whole-cell patch clamp techniques were used respectively to detect the volume changes and currents induced by gelsemium alkaloids in HepG2 cells. Results It was found that the cell volume was decreased by (12. 48 ± 2. 2) % (P<0. 01) when ex-posed to gelsemium alkaloids for 50 min and this phe-nomenon could be inhibited by the chloride channel blocker tamoxifen. It was shown by whole-cell patch clamping that a chloride current could be evoked by extracellular application of gelsemium alkaloids ( 2μmol·L-1 ) . The current was outward-rectified with-out obvious voltage- and time-dependent inactivation. The reversal potential of the current was ( -3. 21 ± 0. 67) mV ,which was close to the equilibrium poten-tial of chloride. The extracellular application of the chloride blockers, tamoxifen and 5-notro-2-(3-phenyl-propylamino)benzoic acid (NPPB), and 47% hyper-tonic solution inhibited the current significantly ( P <0. 01 ) . Conclusion Gelsemium alkaloids could acti-vate chloride channels and induce a volume decrease ( named apoptotic volume decrease, AVD) , and these effect could be inhibited by chloride channel blockers. The results suggest that the chloride channel can be one of the targets of gelsemium alkaloids in their anti-cancer action.
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Aim To clarify the effect of Borneol on the chloride channels and cell volume in poorly differentia-ted nasopharyngeal carcinoma CNE-2Z cells.Methods The technique of whole-cell patch clamp was used to detect the chloride currents and analyze the character-istics of the currents in CNE-2Z cells.The volume changes caused by Borneol were measured by the meth-od of time-lapse live cell imaging.Results The chlo-ride currents were induced by extracellular application of Borneol (20 μmol·L -1 )isotonic condition.The currents showed a characteristic of outward rectification and did not show voltage-dependent or time-dependent inactivation.The reversal potential of the currents was close to the CI-equilibrium potential. The currents were inhibited by the chloride channel blocker tamox-ifen.The currents were also inhibited by 47% hyper-tonic solution.Borneol decreased the cell volume by 9.4% in 30 min.Tamoxifen completely inhibited the Borneol-induced cell volume decrease.Conclusion Borneol can activate volume-sensitive chloride channels and induce volume decrease in CNE-2Z cells.Chloride channels play a pivotal role in the process of volume decrease caused by Borneol.
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AIM:To investigate the type of chloride channel activated by cisplatin in poorly differentiated na -sopharyngeal carcinoma cells (CNE-2Z cells).METHODS:The technique of whole-cell patch-clamp was used to investi-gate the role of Ca 2+in the activation of cisplatin-activated chloride currents and to analyze the effect of hypertonic stress on these currents in CNE-2Z cells.RESULTS:Chloride currents were induced when the cells were exposed to the calcium -free cisplatin solution , showing the similar density to the currents induced by cisplatin with the presence of extracellular cal -cium.However , the latency and the peak time of cisplatin-activated currents in the absence of extracellular calcium were prolonged.The activation of cisplatin-activated chloride currents was insensitive to the depletion of intra-and extracellular calcium.Calcium channel antagonist nifedipine had no effect on the cisplatin -activated chloride currents , while hypertonic solution completely inhibited those currents .CONCLUSION:The cisplatin-activated chloride currents are independent on intra/extracellular calcium .The chloride channels activated by cisplatin are not calcium-activated chloride channels , but are probably volume-sensitive chloride channels .
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Aim To explore the role of heat shock protein 90(Hsp90)in hydrogen sulfide-induced protection against PC12 cells injuries elicited by cobalt chloride(CoCl_2).Methods Hydrogen sulfide preconditioning against CoCl_2-induced injury model was set up in PC12 cells.Cell viability was tested by using cell counter kit-8.Morphological changes in apoptotic PC12 cells were detected by Hoechst 33258 staining and photofluorography.Apoptotic rate was evaluated by propidium iodide(PI)staining and flow cytometry(FCM).The expression of Hsp90 was evaluated by Western blot.Results Hsp90 expression was upregulated after treatment with 400 μmol·L~(-1) sodium hydrosulfide(a H_2S donor),peaking at 3 h,returning to the basal level at 24 h.Furthermore,H_2S preconditioning obviously enhanced the upregulation of Hsp90 expression induced by CoCl_2.H_2S preconditioning markedly protected PC12 cells against injuries induced by CoCl_2,increasing the viability of cells and decreasing the percentage of apoptotic cells.17-allylamino-17-demethoxygeldanamycin(17-AAG),an inhibitor of Hsp90,antagonized H_2S preconditioning-induced Hsp90 activation and the adaptive cytoprotection.Conclusion H_2S can protect PC12 cells against CoCl_2-induced injuries,and upregulating the expression of Hsp90 may be one of the mechanisms underlying cytoprotection induced by H_2S preconditioning.
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AIM: The roles of Cl-channels in regulatory volume decrease (RVD), cell proliferation and cell cycle progression in nasopharyngeal carcinoma cells (CNE-2Z) were investigated. METHODS: Image analysis of living cells was used to detect the volume changes following exposure to hypotonic solutions. Cell viability was determined by the trypan blue assay. MTT method was applied to detected cell proliferation. The effect of the blocker on the cell cycle distribution was monitored by the flow cytometry. RESULTS: 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) inhibited RVD and cell proliferation in a dose-dependent manner. NPPB at the concentration of 100 ?mol/L arrested cells in G 1 phase (G 1 population increased from 54% to 71% at 48 h after treatments), but did not significantly alter cell viability. CONCLUSION: Block of chloride channels suppressed cell proliferation by arresting cells in G 1 phase. The results suggest that activation of Cl-channels and RVD is necessary for facilitating cells to proceed to the S phase from G 1 phase and maintaining cell proliferation.
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AIM: To clarify the role of Cl - in regulatory volume decrease (RVD) of nasopharyngeal carcinoma cells (CNE-2Z).METHODS: Analysis of living cell images was used to detect the volume changes following exposure to hypotonic solution. Iron replacement and block of iron channels were also applied in the present study. RESULTS: Extracelluar hypotonic treatment made the cells swell and induced RVD. The RVD was correlated positively to the swelling in the range of 160-230 mOsmol/L. Substitution of gluconate for Cl - in perfusing solutions markedly increased RVD. Depletion of cellular Cl - abolished, and chloride channel blockers inhibited RVD. CONCLUSION: Cl - is the key iron to establish the RVD in CNE-2Z cells. Activation of Cl - channels and Cl - efflux are the major mechanisms of RVD.