Cisplatin increases self-renewal ability of oral squamous cell carcinoma stem cells through Notch4 signaling pathway / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To study the effect of Notch4 signaling pathway on the self-renewal capacity of cancer stem cells (CSCs) of oral squamous cell carcinoma PJ15 and PJ41 cells and its mechanism. Methods: The expression of Notch4 gene in head and neck tumors was analyzed using TCGA database. 2 μmol/L cisplatin was used to treat oral squamous cell carcinoma PJ15 and PJ41 cells for 48 h, following with the treatment of Notch pathway inhibitor DAPT for 24 h. Flow cytometry was used to detect the ratio of CSCs, Western blotting and qPCR were used to detect the expressions of CSCs markers (Sox2, Bmi-1, Oct4, Nanog) and downstream genes in Notch4 signaling pathway (JAG1, HEY1, HEY2, HES1, HES2, DLL4, etc.). Notch4 was knocked down by siRNA technology. Notch4 was silenced with siRNA technology. Western blotting and qPCR were used to detect the effect of si-Notch4 and cisplatin on the expression level of CSCs markers. The stem cell pelletingtestwasusedtodetecttheself-renewalabilityof CSCs. Results: Notch4 gene was highly expressed in head and neck tumor tissues (P=0.046). After cisplatin treatment, compared with the control group, expressions of NICD4 and CSCs markers (Sox2, Bmi-1, Oct4, Nanog) as well as downstream genes of Notch4 signaling pathway (JAG1, HEY1, HEY2, HES1,HES2, DLL4, etc.) increased significantly (all P<0.05), and the proportion of ALDH1 positive cells increased significantly (P<0.05); with the further addition of DAPT, the expressions of the above genes decreased significantly (P<0.05 or P<0.01). After knocking down Notch4, compared with the control group, the size and number of spheres of PJ15 cells [1.33±0.47] vs [8.00±0.82], P<0.01) and PJ41 cells ([1.00±0.82] vs [7.67±1.25], P<0.01) were significantly lower than that of the control group; after the addition of 2 μmol/L DDP for 24 h, there was no significant difference in the expressions of Nanog and Bmi-1 gene between si-Notch4 group and control group. Conclusion:Activation of Notch4 signaling pathway can enhance the self-renewal ability of CSCs in oral squamous cell carcinoma PJ15 and PJ41 cells.

The effects of pH, surfactant, ion concentration, coformer, and molecular arrangement on the solubility behavior of myricetin cocrystals

Pharmaceutical cocrystals are a promising technology that can be used to improve the solubility of poor aqueous compounds. The objective of this study was to systematically investigate the solubility of myricetin (MYR) cocrystals, including their kinetic solubility, thermodynamic solubility, and intrinsic dissolution rate (IDR). The effects of pH, surfactant, ion concentration, and coformers on the cocrystal solubility were evaluated. Furthermore, single crystal structures of MYR, myricetin-isonicotinamide (MYR-INM) and myricetin-caffeine (MYR-CAF) cocrystals were analyzed to discuss the possible reasons for the enhancement of cocrystal solubility from the perspective of the spatial structure. The results indicated that the kinetic solubility of MYR cocrystals was modulated by pH and cocrystal coformer (CCF) ionization in buffer solution, while it primarily depended on the CCF solubility in pure water. In addition, the solubility of MYR cocrystals was increased in a concentration dependent fashion by the surfactant or ion concentration. The thermodynamic solubility of MYR-INM (1:3) cocrystals decreased with the increases of the pH value of the dissolution media. The IDR of MYR cocrystals was faster than that of MYR in the same medium and extremely fast in pH 4.5 buffer. The improved solubility of MYR cocrystals was probably related to the alternate arrangements of MYR and INM/CAF molecules and increased intermolecular distance. The present study provides some references to investigate the solubility behavior of pharmaceutical cocrystals.

Comparison of volatile organic compounds between cigarette smoke condensate (CSC) and extract (CSE) samples

Cigarette smoke is a major risk factor for several diseases, including chronic obstructive pulmonary and cardiovascular diseases. The toxicity of the cigarette smoke can be determined in vitro. The cytotoxicity test of the cigarette smoke is commonly conducted using the cigarette smoke condensate (CSC) and cigarette smoke extract (CSE). The CSC and CSE methods are well known for sampling of the particles and water-soluble compounds in the cigarette smoke, respectively. In this study, the CSC and CSE were analyzed by using a gas chromatography-mass spectrometry (GC-MS) system equipped with a wax column for separation of the volatile organic compounds. The cytotoxic effect of the CSC and CSE were evaluated thoroughly by comparing the analytical results of the CSC and CSE samples. The total concentration of the volatile organic compounds detected in the CSC sample was similar to that in the CSE sample based on the peak area. Except for the dimethyl sulfoxide solvent, nicotine had the highest concentration in the CSC sample, while acetonitrile had the highest concentration in the CSE sample. The compositions were as follows: (1) CSC sample: 55.8% nicotine, 18.0% nicotyrine, 3.20% 1,2,3-propanetriol, triacetate, 1.28% ethyl chloride, 1.22% phenol, etc. and (2) CSE sample: 18.7% acetonitrile, 18.0% acetone, 12.5% 2-hydroxy-2-methyl-propanenitrile, 8.98% nicotine, 5.86% nicotyrine, etc. In this manner, to accurately examine the cytotoxicity of the cigarette smoke using CSC or CSE, the components and their concentrations in the CSC and CSE samples should be considered.

Comparison of volatile organic compounds between cigarette smoke condensate (CSC) and extract (CSE) samples

Cigarette smoke is a major risk factor for several diseases, including chronic obstructive pulmonary and cardiovascular diseases. The toxicity of the cigarette smoke can be determined in vitro. The cytotoxicity test of the cigarette smoke is commonly conducted using the cigarette smoke condensate (CSC) and cigarette smoke extract (CSE). The CSC and CSE methods are well known for sampling of the particles and water-soluble compounds in the cigarette smoke, respectively. In this study, the CSC and CSE were analyzed by using a gas chromatography-mass spectrometry (GC-MS) system equipped with a wax column for separation of the volatile organic compounds. The cytotoxic effect of the CSC and CSE were evaluated thoroughly by comparing the analytical results of the CSC and CSE samples. The total concentration of the volatile organic compounds detected in the CSC sample was similar to that in the CSE sample based on the peak area. Except for the dimethyl sulfoxide solvent, nicotine had the highest concentration in the CSC sample, while acetonitrile had the highest concentration in the CSE sample. The compositions were as follows: (1) CSC sample: 55.8% nicotine, 18.0% nicotyrine, 3.20% 1,2,3-propanetriol, triacetate, 1.28% ethyl chloride, 1.22% phenol, etc. and (2) CSE sample: 18.7% acetonitrile, 18.0% acetone, 12.5% 2-hydroxy-2-methyl-propanenitrile, 8.98% nicotine, 5.86% nicotyrine, etc. In this manner, to accurately examine the cytotoxicity of the cigarette smoke using CSC or CSE, the components and their concentrations in the CSC and CSE samples should be considered.

Influence of RhoC on Apoptosis and Cancer-stem-cell-marker-ALDH1A1 and Morphology in Hep2 Laryngeal Squamous Carcinoma Cell / 医学研究杂志

Objective To investigate the effects of RhoC on biological behavior in the laryngeal squamous carcinoma.Methods By delivering exogenous gene into Hep2 laryngeal squamous carcinoma cell line,we alternatively repressed and strengthened the expression of RhoC.We tested the apoptosis of Hep2 tumor cell line with TUNEL,visualized tumor cells shape by staining cell skeleton with Alexa fluor phalloidin,measured the mRNA of CSC marker ALDH1A1 with QPCR.Results After repressing the expression of RhoC in Hep2 cell line,the apoptosis of cancer cells was elevated,the expression of CSC marker ALDH1A1 was significantly decreased.RhoC impacted the shapes of Hep2.Conclusion RhoC havd a positive role in LSCC metastasis.RhoC is a promising target of anti-metastases in LSC.

Coriorretinopatía serosa central en la Clínica Oftalmológica de Cartagena (Colombia) / Central serous chorioretinopathy at Clínica Oftalmológica de Cartagena (Colombia)

Objetivos: Determinar el comportamiento de la coriorretinopatía serosa central (CSC) en los pacientes que asistieron a consulta a la clínica Oftalmológica de Cartagena en el periodo comprendido de julio del 2013 a julio de 2014. Diseño del estudio: Estudio observacional-descriptivo de tipo transversal. Métodos: Se revisaron las historias clínicas y los reportes de las ayudas diagnosticas Tomografía Óptica Coherente (OCT) y Angiografía Fluoresceínica (AF) de los pacientes diagnosticados con CSC en el periodo de Julio de 2013 a Julio de 2014 en la Clínica Oftalmológica de Cartagena. Resultados: 42 pacientes que cumplieron con todos los criterios; 9 pacientes diagnosticados con AF, 10 pacientes con OCT y 23 con diagnóstico confirmado por ambas ayudas diagnósticas. La condición es más frecuente en el sexo masculino, en edades entre los 30 y 39 años, casados con estudios bachilleres completos y en la raza negra. Según el test de ansiedad aplicado, en la muestra de pacientes no se encontró relación entre la ansiedad y la aparición de CSC. Conclusiones: El comportamiento de la CSC en la Clínica Oftalmológica de Cartagena fue similar al reportado por la literatura médica. Se recomienda realizar más estudios en los que se evalúe con mayor enfoque la asociación de la CSC y el nivel de ansiedad.

Objective: Determine the behavior of the central serous chorioretinopathy in patients seen at Clínica Oftalmológica de Cartagena in the period of July 2013 to July 2014. Design: Descriptive observational crosssectional study. Methods: Revision of medical records and reports of diagnostic exams (OCT and FA) of patients diagnosed with CSC in the period from July 2013 to July 2014. Results: 42 patients who met all the criteria; 9 patients with FA, 10 patients with OCT and 23 with a diagnosis confi rmed by both methods. The condition was more common in males, aged between 30 and 39 years old, married with complete high school studies, and in blacks. According to the test anxiety applied in the sample there was no relationship between anxiety and the appearance of CSC. Conclusions: Th e trend of the CSC at Clínica Oftalmológica de Cartagena was similar to that reported in medical literature. We recommend further studies that evaluated the association with major focus of the CSC and the level of anxiety.

Reinforcing effect of calcium sulfate cement bovine bone morphogenetic protein on vertebral in the rabbit model of osteoporosis

OBJECTIVE@#To observe reinforcing effect of calcium sulfate cement (CSC) bovine bone morphogenetic protein (bBMP) on vertebral in the rabbit model of osteoporosis.@*METHODS@#A total of 48 New Zealand white rabbits were randomly divided into group I (blank control group), group II (CSC injection group), group III (CSC/bBMP injection group) and control group. White rabbit osteoporosis model was established rapidly by using castration method+methylprednisolone candidate. After modeling, groups II, III were given corresponding vertebral body injection material, and 4 animals were sacrificed respectively at 24 h, 6 weeks, 12 weeks after vertebral plasty. Tissue pathological status, vertebral mineral density and vertebral body bone mechanical strength were observed.@*RESULTS@#Vertebral body structure form was normal in the groups II and III. Trabecular bone coarsens, connection and repair were observed in micro fracture and bone defects, bone trabecular connectivity was superior to group I significantly; vertebral body compression strength in the group I was on the decline, vertebral compression strength in the groups II and III was on the rise, the largest vertebra. Postoperative BMC and BMD in groups II and III were increased, and significantly higher than group I after 6 weeks (P<0.05), BMC and BMD in group III after 12 weeks were higher than the other three groups.@*CONCLUSION@#Compound bBMP CSC has good bone induction. It can improve the three-dimensional construction effect for osteoporosis vertebral trabecula, and can significantly improve the vertebral strength, as a vertebral packing material with good application prospect.

Comparison of Therapeutic Effect Between Half-Energy Photodynamic Therapy and Intravitreal Bevacizumab Injection in Chronic Central Serous Chorioretinopathy for 12 Months

PURPOSE: In this study we compared the effectiveness between half energy photodynamic therapy (PDT) and intravitreal bevacizumab (IVB) injection for chronic central serous chorioretinopathy (CSC). METHODS: Forty-five eyes of 42 patients diagnosed as chronic CSC from March 2008 to April 2011 were treated with half energy PDT or IVB injection. The subjects were chosen for a retrospective study and analysis was performed on changes in best corrected visual acuity and existence of subretinal fluid, recurrence rate and changes in central macular thickness. RESULTS: Similar improvement of visual acuity was observed in both treatment groups 1 month after treatment and no meaningful difference was found in each stage for both groups (p = 0.001, p = 0.0012, respectively). However, 6 to 12 months after the treatment, the half energy PDT group showed more improvement in visual acuity compared to the IVB injection group (p = 0.019, p = 0.043, respectively). Nineteen out of 21 cases showed full recovery of subretinal fluid in the half energy PDT group with an average treatment period of 1.3 +/- 0.8 months and 7 out of 24 cases showed full recovery in the IVB injection group with an average treatment period of 3.2 +/- 2.8 months. There was a single case of recurrence in the half energy PDT group and 4 in the IVB injection group. The half energy PDT group showed a meaningful decline in central macular thickness at 1, 3, and 6 months after treatment (p = 0.001, p = 0.005, p = 0.007, respectively) compared to the IVB injection group and showed numerous cases with decline in central macular thickness below the 2 standard deviation from normal values (p = 0.002). CONCLUSIONS: Both half energy PDT and IVB injection were effective for the treatment of chronic central serous chorioretinopathy. However, the half energy PDT group comparatively showed better anatomical and functional outcomes. The thinning of central macular thickness below normal value was also observed, thus careful choice of treatment is necessary for patients with chronic central serous chorioretinopathy.

A Case of Central Serous Chorioretinopathy Associated With Retinal Detachment Improved by Intravitreal Bevacizumab Injection

PURPOSE: To report a case of chronic central serous chorioretinopathy treated with an intravitreal injection of 2.5 mg of bevacizumab. CASE SUMMARY: A 38-year-old male complained of visual disturbance in the right eye. He had a history of central serous chorioretinopathy which improved without treatment. Since the patient did not feel any ocular discomfort, he did not visit our clinic for approximately 11 months. At the time of presentation, fundus examination revealed exudative retinal detachment involving the macular area. Additionally, fluorescein angiography revealed multiple early phase hyperfluorescent lesions. Neurosensory detachment around the macula and increased central macular thickness were also observed using optical coherence tomography. Under the diagnosis of chronic central serous chorioretinoapthy, the patient was treated with 2.5 mg of intravitreal bevacizumab. Visual acuity improved two months after treatment, with improvement in both neurosensory retinal detachment and fluorescein leakage. No changes were observed at the six-month follow-up.

Photodynamic Therapy for Chronic Central Serous Chorioretinopathy: Multicenter Study of 65 Cases

PURPOSE: To determine the effect of photodynamic therapy (PDT) on patients with chronic central serous chorioretinopahty (CSC). METHODS: This was a retrospective multicenter study. We collected demographic and ophthalmic data for patients with chronic CSC who were treated with PDT from 16 institutions in Korea. The best corrected visual acuity (BCVA), ophthalmoscopic finding and optical coherence tomography at baseline and follow-up visits were compared longitudinally. RESULTS: Sixty-five eyes of 65 patients were recruited. Males (51 patients, 78.5%) outnumbered females (14 patients, 21.5%). The mean age was 46.4+/-10.0 years of age (28~69). By 1 month, mean BCVA improved from 0.47 to 0.37 (logarithm of the minimum angle of resolution) (P <0.01). This BCVA increased throughout 6 months. Subretinal fluid resolved partially or completely in 89.1% of the patients. Central macular thickness (CMT) decreased from 303.4 to 168.7 micrometer. The amount of change in CMT after PDT was correlated to the female sex and prePDT CMT. At 3 months after PDT, the amount of change in BCVA was significantly related to that of change of CMT (p <0.05). Adverse events were reported in 4 cases, however, most were mild. CONCLUSIONS: PDT was effective and safe for treating chronic CSC.

Comparation of magnetic activated cell sorting and chemotherapy agent DDP in enriching cancer stem cell of laryngeal carcinoma cell line Hep-2 / 肿瘤研究与临床

Objective To compare effect of chemotherapy agent DDP to MACS in sorting cancer stemcells (CSC) of laryngeal carcinoma cell line Hep-2. Methods CD133 magnetic beads were applied to sort Hep-2 cells. Different dosages of DDP were used to treat Hep-2 cells for 48 hours. Enrichment rate of CD133+ cells by MACS and after DDP treatment was detected by Flow Cytometer (FCM). Morphologic change was observed under inverse-phase microscope. Results FCM showed that the sorting rate of CD133+ cells through MACS was 64.33 %, while after DDP treatment for 48 hours, the rate of CD133+ cells was enriched significantly in each dosage of DDP, with the maximal rate was 50.7 %, in the dosage of 4 μg/ml. There was a significantly difference between MACS and each of DDP group (P <0.01). Cells treated with DDP were abnormal in morphology. Conclusion MACS and DDP sorting has respective advantages in enriching CSC in Hep-2 cell lines.

Isolation and detection of CK17~+,CD44v5~+ and Bcrp1~+ cells in HeLa cell line of cervical carinoma / 吉林大学学报(医学版)

Objective To identify and isolate the CK17+,CD44v5+ and Bcrp1+ cells in HeLa cells,and investigate the relationship between them and select cervical cancer stem cell markers.Methods Flow cytometry was used to identify and isolate the CK17+,CD44v5+ and Bcrp1+ cells in HeLa cells;after isolation,the expressions of CK17 and CD44v5 in Bcrp1+ and Bcrp1-phenotype HeLa cells were measured by immunocytochemistry.Results HeLa cells contained 11% Bcrp1+ cells,0.7% CK17+ cells and 0.1%CD44v5+ cells.Bcrp1+ HeLa cells contained CK17+ and CD44v5+ HeLa cells,but the Bcrp1-Hela cells did not(P

Choroidal Ne ovascularization in Patients with Chronic Central Serous Chorioretinopathy

Chronic central serous chorioretinopathy[CSC], which also has been termed diffuse retinal pigment epitheliopathy, is defined as a sensory retinal detachment associated with areas of RPE atrophy and pigment mottling. Fluorescein angiographic finding includes areas of granular hyperfluo-rescence and one or many subtle leaks. In older age group, choroidal neo-vascularization[CNV]may develop as a complication. We investigated the clinical features and performed fluorescein angiography[FAG], indocyanine green[ICG]angiography in 10 patients with CNV among 32 patients with chronic CSC. The location of CNV was extrafoveal[3 eyes]and juxtafoveal[7 eyes]. Associated fundus findings were RPE atrophy[8 eyes], PED[5 eyes], serous RD[4 eyes], subretinal hemorrhage[4eyes]. ICG findings were hypo-fluorescence[8 eyes], choroidal hyperpermeability[7 eyes], choroidal delayed filling[5 eyes]. Choroidal neovascularization was identified only with ICG angiography in 6 out of 10 cases. Laser photocoagulation was performed using ICG angiography in 6 cases. Visual acuity was improved in 4 out of 10 cases, and unchanged in 3 out of 10 cases. Therefore, we suggest that ICG angiography may be used as an important device for the diagnosis and management of CNV in patients with chronic CSC. Visual prognosis of CNV in chronic CSC seems to be relatively good.

Study on survivin mRNA and protein of BEP-2D cells during its malignant transformation by cigarette smoke condensate / 重庆医科大学学报

Objective:To study the change of the survivin mRNA and protein of BEP-2D cells during its malignant transformation.Methods:Normal BEP-2D cell and BEP-2D cells treated by cigarette smoke condensate(CSC)for 15 weeks(P-15),25 weeks(P-25)and 38 weeks(P-38)were respectively chosen to study the survivin gene and protein by RT-PCR(retro-translation PCR,RT-PCR)and immunohistochemical method.Results:The survivin mRNA was found in BEP-2D cells of P-15,P-25 and P-38,respectively with 0.56,0.80,and 0.81,but not in the normal BEP-2D cell.The survivin protein was found in the normal BEP-2D cell,but not in BEP-2D cells of P-15,P-25 and P-38.The levels of survivin protein expressian were different between BEP-2D cells of P-15,P-25 and P-38 and normal BEP-2D cell,with significant difference between P-15 BEP-2D cell and normal BEP-2D cell(P