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ObjectiveBy observing the effect of Xiaoluowan on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin complex 1 (mTORC1) pathway in experimental goiter rats, this study aims to explore its therapeutic effect on experimental goiter rats. MethodSixty 5-month-old SD rats of SPF grade were purchased, half males and half females, of which 10 were used as a normal group, and the remaining rats were administrated with propylthiouracil (PTU) solution to induce nodular goiter. After successful modeling, rats were randomly divided into a model group, levothyroxine sodium tablets group, Xiaoluowan low-dose group, medium-dose group, and high-dose group, ten rats each. The levothyroxine sodium tablets group was given 15 μg·kg-1 levothyroxine sodium tablets by gavage. The Xiaoluowan low-, medium-, and high-dose groups were given (ig) Xiaoluowan low-dose (10 g·kg-1), medium-dose (20 g·kg-1), and high-dose (30 g·kg-1) Xiaoluowan, and the normal group and model group were administered (ig) with the same volume of 0.9% sodium chloride solution. Four weeks after the intervention, rats were sacrificed by routine intraperitoneal anesthesia using 5% phenobarbital. Subsequently, the histopathology was observed under a microscope, and serum thyroid hormone levels were measured using a Roche electrochemiluminescence immunoassay analyzer. Serum cytokines were detected by enzyme-linked immunosorbent assay (ELISA), and neurotransmitters were measured using a high-performance liquid chromatograph. The protein level of PI3K/Akt/mTORC1 pathway was determined by Western blot. ResultAs compared with the normal group, the levels of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), insulin-like growth factor (IGF-1), 5-hydroxytryptamine (5-HT), and thyroid stimulating hormone (TSH) were increased, and PI3K, Akt, and mTORC1 protein levels were up-regulated in the model group, while the levels of norepinephrine (NE), triiodothyronine (T3), tetraiodothyronine (T4), free triiodothyronine (FT3), and free thyroid hormone (FT4) were decreased (P<0.05). Compared with the model group, the levothyroxine sodium tablets group, and Xiaoluowan low-, medium-, and high-dose groups exhibited reduced levels of bFGF, VEGF, IGF-1, 5-HT, and TSH, and down-regulated PI3K, Akt, and mTORC1 protein levels, and increased NE, T3, T4, FT3, and FT4 levels (P<0.05). ConclusionXiaoluowan may act on the PI3K/Akt/mTORC1 signaling pathway to play its role in the treatment of nodular goiter, and it is dose-dependent.
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ObjectiveThere are few studies on the correlation between the concentration of oncoembryonic antigen associated cell adhesion molecule 1(CEACAM1) and osteonecrosis of the femoral head (ONFH). The purpose of this study was to investigate the value of CEACAM1 in the early diagnosis of ONFH and the monitoring of the disease by detecting the CEACAM1 concentration in the serum of patients with ONFH and healthy subjects respectively.Methods95 patients, who were hospitalized and diagnosed as ONFH in the Department of No.3 Orthopaedic Ward, the First Affiliated Hospital of Guangzhou University of Chinese Medicine from May 2016 to November 2016, were selected as the experimental group. In addition, 56 genders and age-matched healthy subjects in our hospital were selected as the control group. The peripheral venous blood was taken and separated by a centrifuge. Their CEACAM1 concentrations were measured by enzyme linked immunosorbent assay (ELISA). The differences in CEACAM1 concentrations were analyzed between the two groups, and between patients with ONFH before (ARCO stage I or II) and after (ARCO stage III or IV) collapse as well.Results①The concentration of CEACAM1 in the experimental group was significantly lower than that in the control group [(6.11±2.07)ng/mL vs (7.21±3.76)ng/mL, P=0.022]. ②The concentration of CEACAM1 in Arco stage II[(7.33±1.90) ng/mL] was significantly higher than that in stage III [(6.08±2.26) ng/mL], P=0.037.③The difference of CEACAM1 concentration between before(stage II) and after collapse (stage III or stage IV) was statistically different [(7.33±1.90)ng/mL vs (5.86±2.02)ng/mL, P=0.007].④ROC curve analysis showed that the area under the curve was 0.710 (0.608-0.798), the sensitivity was 71.79%, the specificity was 58.82%, and the cut off value was ≤ 6.757ng/mL in the diagnosis of collapse of ONFH.ConclusionThe concentration of serum CEACAM1 can be used as a biochemical marker to assist in the diagnosis and monitoring of ONFH, which can provide reference for early diagnosis and monitoring of ONFH.
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Objective@#To explore the relationship between psychological resilience and cognitive bias towards school violence in grade 3-5 primary school students in Luzhou city, so as to provide scientific basis for prevention and control of school violence in primary school students.@*Methods@#Students from grade 3-5 in primary schools in Luzhou were selected through stratified cluster random sampling method and were investigated with questionnaire survey.@*Results@#A total of 5 976 valid questionnaires were included, with an average score of psychological resilience (40.08±8.05) and an average score of school violence cognition (62.55±6.38). Multivariate results showed that psychological resilience was an independently associated with school violence perception (OR=1.04, P<0.01). The awareness of campus violence increased with resilience score. In addition, public school (OR=0.45) was associated with low awareness of school violence; senior grades (OR=1.77), girls (OR=1.20), and a greater number of friends(OR=1.37), student cadre(OR=1.37), middle/upper score in class(OR=2.13), no game playing(OR=1.33), no off-campus wandering(OR=1.78), timely parenting (OR=1.45) was associated with high awareness of school violence(P<0.05).@*Conclusion@#Psychological resilience positively correlates with cognition bias towards school violence. The higher the psychological resilience, the more positive perception of campus violence. Family, school and community-based interventions to enhance the resilience of students, increasing awareness towards school violence and ultimately reducing potential adverse impacts of school violence.
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AIM:To study the effect of ClC-3 gene over-expression on thyroid structure and function in mice. METHODS:Three-months-old FVB mice were used to study the difference of thyroid structure and function between wild-type(WT)mouse and ClC-3 transgene mice.The expression and distribution of ClC-3 in the thyroid of mice were deter-mined by the methods of qPCR,Western blot and immunofluorescence.Behavioral monitoring was performed on the daily activities of mice.Serum concentrations of total triiodothyronine(TT3), total thyroxine(TT4)and thyrotropin(TSH) were measured by ELISA.RESULTS:Compared with the WT group,the expression of ClC-3 in the thyroid of ClC-3 trans-gene group was significantly increased(P<0.05).The thyroid gland showed obvious hyperplasia and the folliculi glandu-lae thyreoideae was significantly bigger in ClC-3 transgene mice(P<0.05).The weight loss was increased in ClC-3 trans-gene mice(P<0.05).The expression of TT3 and TT4 were significantly higher than that of WT group(P<0.05),but the change of TSH was not obvious.CONCLUSION:ClC-3 over-expression results in thyroid hyperplasia and thyroid hor-mone secretion.This study suggests that ClC-3 is likely to be involved in the synthesis of thyroid hormones.
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Aim To study the effects of L-borneol on the chloride channel and cell volume of human umbili-cal vein endothelial cells (HUVECs). Methods Whole-cell patch-clamp technique was used to record chloride currents. The expression of ClC-3 protein was down-regulated by siRNA interference technique. The cell volume was measured by dynamic image analysis. Results 20 nmol·L-1L-borneol significantly activa-ted chloride current in HUVEC (79.59 ± 4.90) pA/pF, which could be inhibited by chloride channel blockers,NPPB and DIDS. The outward current inhib-itory rate of NPPB was (95.57 ± 2.57)%, while that of DIDS was (97.28 ± 6.36)%. The chloride current activated by L-borneol significantly decreased after the silence of ClC-3 (27.03 ± 3.89) pA/pF. Cell volume was markedly reduced by L-borneol (14.38 ± 1.58)%,which was inhibited after NPPB appliance. Conclusion L-borneol can activate ClC-3 chloride channel in HUVECs, which induces Cl- outflow then cell volume decrease.
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AIM:To compare the influence of anterior chamber maintainer and viscoelastic agent on corneal astigmatism and endothelial cells after phacoemulsification.METHODS:Totally 70 patients(70 eyes) of cataract from April 2013 to April 2015 were randomly divided into the study group and the control group, with 35 cases in each group.The study group were treated with anterior chamber maintainer during phacoemulsification with support system approach, and the control group were treated with phacoemulsification under viscoelastic agent.RESULTS:The age (t=0.215, P=0.831), the density of corneal endothelial cells (t=-0.352, P=0.726) and corneal luminosity (t=-0.162, P=0.872) of two groups had no significant difference before surgery;there were no significant difference in preoperative visual acuity (t=0.463, P=0.599) and visual acuity (t=1.616, P=0.124) at 1mo after operation.And patients in the study group (t=-21.129, P<0.01) and the control group (t=-12.780, P<0.01) before surgery and 1mo postoperative when compared with the naked eye eyesight showed significant difference.The visual acuity after operations improved significantly.There were significant differences of corneal endothelial cells density (t=8.489, P< 0.01) and corneal astigmatism (t=-2.032, P=0.046) in the study group before surgery and 1mo after surgery;corneal endothelial cell density (t=8.999, P<0.01) and corneal astigmatism (t=-2.167, P=0.034) in the control group before surgery and 1mo after surgery also had significant differences.There was no significant difference in the rate of corneal endothelial cell loss between the two groups (t=0.410, P=0.683).CONCLUSION:Compared with viscoelastic agent, anterior chamber maintainer during phacoemulsification in patients with cataract won't increase the damage of postoperative surgically induced astigmatism and corneal endothelial cells, which mean the method of anterior chamber maintainer during phacoemulsification in the treatment of cataract is safe and effective.
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<p><b>OBJECTIVE</b>To investigate the value of serum fructosamine (FMN) measurement in clinical typing and staging of multiple myeloma(MM) as well as evaluation of curative efficacy and prognosis of IgA type MM.</p><p><b>METHODS</b>The levels of FMN were detected by nitro-blue-tetrazolium colorimetric method in 62 IgA type MM patients, 65 IgG type MM patients, 24 IgM type MM patients. The difference between the serum FMN levels and different MM types and its relationship with IgA type MM, patients' stages, efficacy and as well as, the effect of FMN level on overall survival (OS) time of IgA type MM patients were analyzed.</p><p><b>RESULTS</b>FMN level of IgA type patients was obviously higher than that of IgG type and IgM type patients. Patients of DS stage III had higher FMN level than that of stage II, and patients of DS stage II had higher FMN level than that of stage I, the differences were statistical significant(P<0.05). Patients of ISS stage III also had higher FMN level than that of stage II, and patients of ISS stage II also had higher FMN level than that of stage I, the differences were statistical significant(P<0.05). After treatment, FMN level of patients obtained complete remission (CR) or very good partial remission(VGPR) obviously decreased, FMN level of patients obtained partial remission(PR) decreased, FMN level of patients obtained less than PR increased. Patients with normal FMN level had longer OS time, while patients with high FMN level had obviously short OS time.</p><p><b>CONCLUSIONS</b>Serum FMN is obviously elevated in the IgA type MM, there is a certain association between staging and serum FMN level, the IgA type MM patients with FMN normalization after treatment have better prognosis.</p>
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The aim of this study was to investigate the relationship between the acetylcholine concentration in the blood and gelsenicine-induced death in mice. Kunming mice were given intraperitoneal injections of normal saline, gelsenicine or different doses of acetylcholine chloride. Atropine was given to the mice which received gelsenicine or medium dose acetylcholine chloride injection. The blood was sampled immediately when the mice died or survived for 20 min after injection. The acetylcholine concentration and acetylcholinesterase activity in the blood were measured by the testing kits, and the mortality was calculated and analyzed. The results showed that half lethal dose of gelsenicine (0.15 mg/kg) reduced the acetylcholinesterase activity and increased the blood acetylcholine concentration. The blood acetylcholine concentration of the dead mice in the gelsenicine group was increased to 43.0 μg/mL (from 31.1 μg/mL in the control), which was lower than that (53.9 μg/mL) of the dead mice in the medium dose acetylcholine chloride group, but almost equal to that (42.7 μg/mL) of the survival mice in the medium dose acetylcholine chloride group. Atropine could successfully rescue the mice from acetylcholine poisoning, but its efficiency of rescuing the mice from gelsenicine intoxication was weak. These results suggest that gelsenicine can inhibit acetylcholinesterase activity and increase blood acetylcholine concentration, but the accumulation of acetylcholine may not be the only or main cause of the death induced by gelsenicine in mice.
الموضوعات
Animals , Mice , Acetylcholine , Death , Indole Alkaloidsالملخص
AIM:To disscus the clinical effect of the silicone tube implantation under the guidance of memory wire in the treatment of lacrimal canaliculus. METHODS:One hundred and fifteen cases (115 eyes ) of traumatic canalicular laceration were treated by canaliculoplasty from September 2012 to June 2014. Finding the end of lacrimal canaliculus under microscope, guided by memory wire which was probed in lacrimal passage to the nasal cavity, intubating double-passage silicone tube as a support and end-to-end anastomosis. The condition of epiphora and irrigation of lacrimal passage were observed after extubation. RESULTS: All 115 cases were experienced successful operation. All patients were followed up for 6mo ~ 1a (mean 9. 3mo) after extubation. Lacrimal passage was unobstructed in 96 cases 96 eyes(83. 5%), stricture in 13 cases 13 eyes (11. 3%), and blocked in 6 cases 6 eyes (5. 2%). CONCLUSION:Double-passage silicone tube guided by memory wire may be an optional technique in the treatment of traumatic lacrimal duct laceration, which is a feasible, minimally - invasive, safe and effective method.
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<p><b>OBJECTIVE</b>To investigate the relationship between degree of abnormal deeper lateral femoral notch between anterior cruciate ligament tear.</p><p><b>METHODS</b>The radiograph and MRI image material of 16 patients with anterior cruciate ligament injury from January 2013 to November 2013 were reviewed including 14 males and 2 females with an average age of 28.3 years old ranging from 18 to 52 years. Eleven cases was on right side and 5 on left. Survey tool of PASC imaging system was used to measure the depth of lateral femoral notch in patients with abnormal indicated by lateral X-ray view or sagittal view of MRI in knee joint,while clinical data,physical examination,image material in arthroscopy of these patients were retrospective researched.</p><p><b>RESULTS</b>Four patients had an abnormal lateral femoral notch with the depth of 2 mm on lateral X-ray and sagittal MRI, while positive anterior drawer sign and Lachman test as well as anterior cruciate ligament tears on MRI, and completed tears were comfirmed on the operation of arthroscopy. Two patients without abnomal lateral femoral notch on lateral view of X-ray while with the depth of 1 mm on sagittal view of MRI were also coupled with positive anterior drawer sign and Lachman test as well as anterior cruciate ligament tears on MRI, and one of them were comfirmed completed anterior cruciate ligament tears on the arthroscopy operation and completed tear could not comfirmed on another one because of disagreed with arthroscopy operation.</p><p><b>CONCLUSION</b>There appears to be an association between abnormal lateral femoral notch on lateral view of knee with anterior cruciate ligament tears. An abnormal deeper lateral femoral notch is an indirect evidence for anterior cruciate ligament tears.</p>
الموضوعات
Adult , Female , Humans , Male , Middle Aged , Anterior Cruciate Ligament , General Surgery , Anterior Cruciate Ligament Injuries , Femur , Pathology , Magnetic Resonance Imagingالملخص
The present study aimed to investigate the effects of ursolic acid on the chloride channels and cell volume in nasopharyngeal carcinoma cells (CNE-2Z). The whole-cell patch clamp technique was used to detect the current, and cell imaging technique was applied to measure cell volume. The properties of the currents induced by ursolic acid were investigated by changing the extracellular osmotic pressure, replacing the extracellular anions and applying chloride channel blockers. The results showed that, under isotonic conditions, the background current was weak and stable. When perfusing the cells with ursolic acid (100 nmol/L), a large current (-59.86 pA/pF ± 4.86 pA/pF at -80 mV, 78.92 pA/pF ± 6.39 pA/pF at +80 mV) was induced. The chloride current showed outward rectification and negligible time- and voltage-dependent inactivation. The reversal potential (-4.83 mV ± 0.30 mV) of the current was close to the calculated equilibrium potential for Cl⁻ (-0.9 mV). The permeabilities of the channel to different anions were ranked in order as follows: Cl⁻ = I⁻ > Br⁻ > gluconate. Hypertonic solutions inhibited the current induced by ursolic acid. The chloride channel blockers, tamoxifen (20 μmol/L) and 5-nitro-2-(3-phenylpro-pylamino) benzoic acid (NPPB, 100 μmol/L), suppressed the current. Furthermore, ursolic acid decreased the cell volume by (11.78 ± 1.20)% in 1 h, and the effect was inhibited by NPPB. These results suggest that ursolic acid can activate chloride channels, resulting in outflow of Cl⁻ and decrease of cell volume in nasopharyngeal carcinoma cells.
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Humans , Carcinoma , Cell Differentiation , Cell Line, Tumor , Cell Size , Chloride Channels , Metabolism , Nasopharyngeal Neoplasms , Metabolism , Patch-Clamp Techniques , Tamoxifen , Pharmacology , Triterpenes , Pharmacologyالملخص
<p><b>OBJECTIVE</b>To investigate the role of chloride channels in the apoptosis of poorly differentiated nasopharyngeal carcinoma CNE-2Z cells induced by gambogic acid (GA).</p><p><b>METHODS</b>MTT assay was applied to detect the proliferation of CNE-2Z cells after GA treatment, and the cell apoptosis was detected by Hoechst 33342 staining. Whole-cell patch clamp technique was employed to record GA-activated Cl(-) currents in the cells.</p><p><b>RESULTS</b>GA inhibited the cell proliferation in a time- and concentration-dependent manner with an IC(50) of 3.1 µmol/L for a 48-h treatment. The apoptosis-inducing effect of 8 µmol/L GA was attenuated by the chloride channel blocker NPPB (100 µmol/L) and tamoxifen (20 µmol/L). GA induced an outward-rectified Cl(-) current in the cells, which was significantly inhibited by NPPB.</p><p><b>CONCLUSION</b>GA suppresses cell proliferation and induces apoptosis by activating Cl(-) channels in CNE-2Z cells, suggesting the important role of Cl(-) channels in GA-induced apoptosis.</p>
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Humans , Apoptosis , Cell Line, Tumor , Chloride Channels , Physiology , Nasopharyngeal Neoplasms , Pathology , Patch-Clamp Techniques , Xanthones , Pharmacologyالملخص
<p><b>BACKGROUND</b>A growing number of studies worldwide have advocated the replacement of whole-breast irradiation with accelerated partial breast irradiation using three-dimensional conformal external-beam radiation (APBI-3DCRT) for early-stage breast cancer. But APBI can be only used in selected population of patients with early-staged breast cancer. It is not replacing the whole breast radiotherapy. This study aimed to examine the feasibility and acute normal tissue toxicity of the APBI-3DCRT technique in Chinese female patients who generally have smaller breasts compared to their Western counterparts.</p><p><b>METHODS</b>From May 2006 to December 2009, a total of 48 Chinese female patients (with early-stage breast cancer who met the inclusion criteria) received APBI-3DCRT after breast-conserving surgery at Sun Yat-sen University Cancer Center. The total dosage from APBI-3DCRT was 34 Gy, delivered in 3.4 Gy per fractions, twice per day at intervals of at least six hours. The radiation dose, volume of the target area and volume of irradiated normal tissues were calculated. Acute toxicity was evaluated according to the Common Toxicity Criteria (CTC) 3.0.</p><p><b>RESULTS</b>Among the 48 patients, the planning target volume for evaluation (PTVE) was (90.42 ± 9.26) cm³, the ipsilateral breast volume (IBV) was (421.74 ± 28.53) cm³, and the ratio between the two was (20.74 ± 5.86)%. Evaluation of the dosimetric characteristics of the PTVE revealed excellent dosimetric results in 14 patients and acceptable results in 34 patients. The dose delivered to the PTVE ranged from 93% to 110% of the prescribed dose. The average ratio of the volume of PTVE receiving 95% of the prescription dose (V95) was (99.26 ± 0.37)%. The habituation index (HI) and the conformity index (CI) were 1.08 ± 0.01 and 0.72 ± 0.02, respectively, suggesting good homogeneity and conformity of the dose delivered to the target field. The radiation dose to normal tissues and organs was within the dose limitation. Subjects experienced mild acute toxicity. The main manifestations were breast edema in 22 patients, breast pain in 7, skin erythema in 25, general malaise in 22 and cytopenia in 8. No acute radiological cardiac or pulmonary injury was found.</p><p><b>CONCLUSIONS</b>The results of our short-term follow-up showed that it is feasible to perform APBI-3DCRT for early-stage breast cancer after breast-conserving surgery in Chinese female patients with smaller breasts. However, further studies are required to elucidate its efficacy and long-term side effects.</p>
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Adult , Female , Humans , Middle Aged , Asian People , Breast Neoplasms , Radiotherapy , General Surgery , Radiotherapy, Conformalالملخص
The present study aimed to clarify the effect of berberine on the chloride channels in human colorectal carcinoma cells (SW480). The whole-cell patch clamp technique was used to detect the Cl(-) current activated by berberine. The physiological and pharmacological characteristics of the current were clarified by changing the osmotic pressure of extracellular perfusate and applying chloride channel blockers. The results showed that, under isotonic conditions, the background current of SW480 cells was weak and stable. A large current was induced by perfusing the cells with the isotonic solution containing berberine (10 nmol/L), current density being (85.8 ± 4.6) pA/pF at +80 mV, (-71.9 ± 3.5) pA/pF at -80 mV, with a latency of (115.6 ± 21.7) s. The chloride current showed weak outward rectification and negligible time- and voltage-dependent inactivation. The reversal potential (-5.5 mV ± 1.2 mV) of the current was close to the calculated equilibrium potential for Cl(-) (ECl = -0.9 mV). Experiments under different osmotic pressures showed that the properties of hypotonicity-activated current recorded in SW480 cells were similar to those of the current induced by berberine, and hypertonic solutions suppressed the berberine-induced current by (98.6 ± 2.3)%. On the other hand, berberine-induced Cl(-) current was significantly inhibited by the chloride channel blockers NPPB (100 µmol/L) and tamoxifen (20 μmol/L), with the inhibition ratios of (83.1 ± 3.6)% and (95.6 ± 1.2)% respectively. These results suggest that berberine can activate the chloride channels that are sensitive to NPPB and tamoxifen, as well as the changes of cell volume in human colorectal carcinoma cells.
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Humans , Berberine , Pharmacology , Cell Line, Tumor , Chloride Channels , Colorectal Neoplasms , Metabolism , Pathology , Nitrobenzoates , Pharmacology , Osmotic Pressure , Patch-Clamp Techniques , Tamoxifen , Pharmacologyالملخص
<p><b>OBJECTIVE</b>To investigate the role of ClC-3 chloride channels in regulatory volume decrease (RVD) of nasopharyngeal carcinoma (NPC) CNE-2Z cells.?</p><p><b>METHODS</b>ClC-3 siRNA was transfected into CNE-2Z cells in the presence of the transfection reagent HiPerFect Reagent(TM). The transfection efficiency of ClC-3 siRNA was detected by flow cytometry. The expression of ClC-3 protein was detected by Western blotting, and the changes of cell volume in 160 mOsmol/L hypotonic solution were determined by image analysis.</p><p><b>RESULTS</b>The transfection efficiency of ClC-3 siRNA was (63.8∓3.8)% (n=3, P<0.01), and compared with the control group, ClC-3 siRNA transfection resulted in a reduction of ClC-3 expression by (60.9∓4.0)% (n=3, P<0.01). The hypotonic challege (160 mOsmol/L) caused cell swelling and induced RVD. In the control group, hypotonic solution bath for 35 min resulted in a RVD of (42.6∓2.8)% (n=20), which was significantly decreased to (10.5∓4.8)% (n=16) in ClC-3 siRNA-transfected cells, demonstrating a reduction of RVD capacity by 75.4% (P<0.01).?</p><p><b>CONCLUSION</b>The capacity of RVD is significantly reduced in CNE-2Z cells by ClC-3 chloride channel protein knock-down via ClC-3 siRNA transfection, indicating an important role of ClC-3 chloride channels in the RVD of CNE-2Z cells.</p>
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Humans , Cell Line, Tumor , Cell Size , Chloride Channels , Genetics , Nasopharyngeal Neoplasms , Genetics , Pathology , RNA Interference , RNA, Small Interfering , Genetics , Transfectionالملخص
<p><b>OBJECTIVE</b>To construct the eukaryotic expression vectors of human cyclin D1 gene and express them in poorly differentiated nasopharyngeal carcinoma cells (CNE-2Z cells).</p><p><b>METHODS</b>The full-length cyclin D1 was cloned from CNE-2Z cells by RT-PCR. The cDNA fragments were inserted into pIRES2-EGFP plasmids and pEGFP-C2 plasmids and confirmed by restriction enzyme digestion, PCR and sequencing. The recombinant vectors were transfected into CNE-2Z cells via Lipofectamine 2000, and the expression of cyclin D1 in the cells was examined by immunofluorescence and Western blotting.</p><p><b>RESULTS</b>Agarose gel electrophoresis showed a 918 bp band of the RT-PCR products, which matched the expected size. Restriction enzyme digestion, PCR and sequencing demonstrated successful construction of the recombinant vectors. CNE-2Z cells transfected with the recombinant vectors expressed cyclin D1 protein or cyclin D1-GFP protein as were verified by immunofluorescence and Western blotting.</p><p><b>CONCLUSION</b>We have cloned cyclin D1 gene and constructed its eukaryotic expression vectors that can be expressed in nasopharyngeal carcinoma cells, which may facilitate the study of the role of cyclin D1 in the development of nasopharyngeal carcinoma.</p>
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Humans , Cell Line, Tumor , Cloning, Molecular , Cyclin D1 , Genetics , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Nasopharyngeal Neoplasms , Metabolism , Pathology , Plasmids , Genetics , Recombinant Proteins , Genetics , Transfectionالملخص
<p><b>BACKGROUND AND OBJECTIVE</b>The planning dose distribution of intensity-modulated radiation therapy (IMRT) has to be verified before clinical implementation. The commonly used verification method is to measure the beam fluency at 0 degree gantry angle with a 2-dimensional (2D) detector array, but not the composite dose distribution of the real delivery in the planned gantry angles. This study was to investigate the angular dependence of a 2D diode array (2D array) and the feasibility of using it to verify the composite dose distribution of IMRT.</p><p><b>METHODS</b>Angular response of the central detector in the 2D array was measured for 6 MV X-ray, 10 cmx10 cm field and 100 cm source axis distance (SAD) in different depths. With the beam incidence angle of 0-60 degrees, at intervals of 10 degrees, and inherent buildup of the 2D array (2 g/cm2), the array was irradiated and the readings of the central diode were compared with the measurement of thimble ionization chamber. Using a combined 30 cmx30 cmx30 cm phantom which consisted of solid water slabs on top and underlying the 2D array, with the diode detectors placed at 8 g/cm2 depth, measurements were taken for beam angles of 0 degrees-180 degrees at intervals of 10 degrees and compared with the calculation of treatment planning system (TPS) that pre-verified with ion chamber measuring.</p><p><b>RESULTS</b>Differences between the array detector and thimble chamber measurements were greater than 1% and 3.5% when the beam angle was larger than 30 degrees and 60 degrees, respectively. The measurements in the combined phantom were different from the calculation as high as 20% for 90 degrees beam angle, 2% at 90 degrees+/-5 degrees and less than 1% for all the other beam angles.</p><p><b>CONCLUSIONS</b>The 2D diode array is capable of being used in composite dose verification of IMRT when the beam angles of 90 degrees+/-5 degrees and 270 degrees+/-5 degrees are avoided.</p>
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Calibration , Feasibility Studies , Particle Accelerators , Phantoms, Imaging , Radiometry , Radiotherapy Dosage , Radiotherapy Planning, Computer-Assisted , Radiotherapy, Intensity-Modulated , Methodsالملخص
<p><b>BACKGROUND AND OBJECTIVE</b>To ensure the accuracy of dose calculation for radiation treatment plans is an important part of quality assurance (QA) procedures for radiotherapy. This study evaluated the Monitor Units (MU) calculation accuracy of a third-party QA software and a 3-dimensional treatment planning system (3D TPS), to investigate the feasibility and reliability of independent verification for radiation treatment planning.</p><p><b>METHODS</b>Test plans in a homogenous phantom were designed with 3-D TPS, according to the International Atomic Energy Agency (IAEA) Technical Report No. 430, including open, blocked, wedge, and multileaf collimator (MLC) fields. Test plans were delivered and measured in the phantom. The delivered doses were input to the QA software and the independent calculated MUs were compared with delivery. All test plans were verified with independent calculation and phantom measurements separately, and the differences of the two kinds of verification were then compared.</p><p><b>RESULTS</b>The deviation of the independent calculation to the measurements was (0.1 +/- 0.9)%, the biggest difference fell onto the plans that used block and wedge fields (2.0%). The mean MU difference between the TPS and the QA software was (0.6 +/- 1.0)%, ranging from -0.8% to 2.8%. The deviation in dose of the TPS calculation compared to the measurements was (-0.2 +/- 1.7)%, ranging from -3.9% to 2.9%.</p><p><b>CONCLUSIONS</b>MU accuracy of the third-party QA software is clinically acceptable. Similar results were achieved with the independent calculations and the phantom measurements for all test plans. The tested independent calculation software can be used as an efficient tool for TPS plan verification.</p>
الموضوعات
Algorithms , Phantoms, Imaging , Quality Control , Radiometry , Methods , Radiotherapy Dosage , Radiotherapy Planning, Computer-Assisted , Methods , Softwareالملخص
<p><b>BACKGROUND AND OBJECTIVE</b>X-ray stereotactic radiotherapy (SRT) is one of the effective treatments for brain metastases (BM). This study was to evaluate the efficacy of SRT on BM, and investigate prognostic factors.</p><p><b>METHODS</b>Between July 1999 and December 2004, a total of 122 intracranial lesions in 78 patients with BM were treated using SRT in our Center. Forty-nine patients had a solitary lesion and 29 had multiple (2-6) lesions. The median SRT dose was 15 Gy (11-24 Gy) in single fraction for 38 lesions, and 24 Gy (11-40 Gy) in 2-6 fractions for 84 lesions. SRT was combined with whole brain radiotherapy (WBRT) of 30-40 Gy for 39 patients. Progression-free survival (PFS) and overall survival (OS) were estimated by Kaplan-Meier method. Univariate and multivariate analyses were performed by the log-rank test and Cox model, respectively.</p><p><b>RESULTS</b>The median survival time was 12.9 months (1.7-77.4 months). The 1-year intracranial PFS rate was 87.4%. The 1-and 2-year OS rates were 53.9% and 25.8%, respectively. Univariate analysis showed that the 1-year OS rates were higher in the patients with pretreatment KPS of >/= 70, extracranial lesions controlled, or SRT combined with WBRT than in those with KPS of < 70 (60.7% vs. 29.4%, P = 0.002), extracranial lesions uncontrolled (69% vs. 44.9%, P = 0.005), or SRT alone (64.1% vs. 43.4%, P = 0.03). The benefit of treating with WBRT in combination was mainly achieved in the patients with extracranial lesions controlled or with more than one intracranial lesion. Multivariate analysis showed that KPS score and status of extracranial lesions were independent prognostic factors for OS.</p><p><b>CONCLUSIONS</b>SRT is an effective and safe modality for BM. SRT combined with WBRT may prolong the survival time of the patients with extracranial lesions controlled or multiple intracranial lesions. Independent prognostic factors for OS are KPS score and status of extracranial lesions.</p>
الموضوعات
Adult , Aged , Female , Humans , Male , Middle Aged , Brain Neoplasms , Radiotherapy , General Surgery , Breast Neoplasms , Pathology , Cranial Irradiation , Methods , Disease-Free Survival , Follow-Up Studies , Karnofsky Performance Status , Lung Neoplasms , Pathology , Neoplasms, Glandular and Epithelial , Radiotherapy , General Surgery , Proportional Hazards Models , Radiosurgery , Methods , Radiotherapy Dosage , Survival Rateالملخص
It has been shown that cell volume regulation mechanisms play important roles in various cell functions. We demonstrated previously that volume-activated chloride channels were involved in cell volume regulation. The present study aimed to clarify the roles of various types of potassium channels in regulatory volume decrease (RVD) induced by hypotonic challenges in human nasopharyngeal carcinoma cells (CNE-2Z cells). The whole-cell patch clamp technique was used to record hypotonic challenge-induced potassium currents. During current recordings, cells were held at 0 mV and stepped to +/-46 and +/-92 mV, repeatedly. The cell volume was computed from cell diameters. The changes of cell volume were monitored and analyzed by the time-lapse imaging technique. The results showed that the exposure to 160 mOsm/L hypotonic solution caused the cells to swell by (144.5+/-4.2)%, activated a potassium current (59.2 pA/pF+/-13.3 pA/pF at 92 mV), and induced RVD. Cell volume was recovered from hypotonic challenge-induced swelling by (48.9+/-4.6)% after 20 min. The potassium current (at 92 mV) and RVD were inhibited by the calcium-dependent potassium channel blocker, clotrimazole (100 mumol/L), by (98.5+/-2.8)% and (89.3+/-4.9)%, respectively. Depletion of extracellular calcium prevented the activation of the hypotonic challenge-induced potassium current and inhibited the process of RVD. The voltage-gated potassium channel blocker, 4-AP (5 mmol/L), partially inhibited the hypotonic challenge-activated potassium currents by (66.6+/-5.3)% (at 92 mV). These results suggest that the Ca(2+)-dependent potassium channel is the main component of volume-activated potassium channels and plays an important role in volume regulation of CNE-2Z cells. The voltage-gated potassium channels may also contribute in part to the formation of the volume-activated potassium current.