New perspectives on macular hole surgery at three years of follow-up / Novas perspectivas em cirurgia de buraco macular com de 3 anos de seguimento

ABSTRACT Purpose: The aim of this study was to determine the functional and anatomical success rates as well as the safety of sutureless combined surgery involving vitreous base removal and internal limiting membrane peeling after Brilliant Blue G (0.5 mg/mL) staining for the management of idiopathic macular holes after three years. Methods: Forty-six eyes of 46 patients with an idiopathic macular hole were enrolled in this retrospective study. The inclusion criteria were macular holes with a minimum linear diameter below 1,500 mm, 0.05 or better decimal best-corrected visual acuity and duration of symptoms less than two years. The exclusion criteria included pregnancy, optic nerve atrophy, advanced glaucoma, and other chronic ocular diseases. The surgical procedure included internal limiting membrane peeling after Brilliant Blue G (0.5 mg/mL) staining, along with C3F8 tamponade and face-down positioning for three days postoperatively. Ophthalmologic examinations and optical coherence tomography were performed at 1 and 7 days and 1, 6, 12, 24, and 36 months postoperatively. If no anatomic closure of the macular holes occurred within the first month, the area of the internal limiting membrane peeling was enlarged in a second procedure. Multiple logistic regression and chi-squared tests were used for data analyses, and p-values of <0.05 were considered significant. Results: Out of 46 eyes with a preoperative idiopathic macular hole, anatomic closure was achieved in 42 (91.3%) after one procedure and in 45 (97.8%) after an additional surgery. The median postoperative best-corrected visual acuity improvement was 0.378 (range: 0.050-0.900) decimal. None of the patients experienced macular hole reopening, surgery-related complications, or ocular complications related to the dye. Conclusion: Combined surgery including vitreous base removal and internal limiting membrane peeling after staining with Brilliant Blue G (0.5 mg/mL) for the management of idiopathic macular holes resulted in adequate staining, best-corrected visual acuity improvement, and macular hole closure with no signs of ocular toxicity at the three-year follow-up examination.

RESUMO Objetivo: Determinar, após 3 anos de seguimento, as taxas de sucesso funcional e anatômico e a segurança da cirurgia combinada sem sutura, incluindo remoção da base vítrea e da membrana limitante interna após coloração com azul brilhante (0,5 mg/ml) para o manejo de buracos maculares idiopáticos. Métodos: Quarenta e seis olhos de 46 pacientes com buraco macular idiopático foram incluídos neste estudo retrospectivo. Os critérios de inclusão foram: buraco macular com diâmetro linear mínimo menor que 1500 micrômetros, acuidade visual com melhor correção de 0,05 decimal ou melhor e tempo de sintomas menor que 2 anos. Os critérios de exclusão foram gravidez, atrofia do nervo óptico, glaucoma avançado ou outra doença ocular crônica. A técnica cirúrgica incluiu a remoção da membrana limitante interna após coloração com Azul Brilhante 0,5 mg/ml, tamponamento com C3F8 posicionamento em prona ção durante 3 dias de pós-operatório. O seguimento foi realizado por exame oftalmológico e Tomografia de Coerência Óptica no 1 e 7 dias, 1, 6, 12, 24 e 36 meses de pós-operatório. Se o fechamento anatômico do buraco macular não fosse atingido na visita de um mês, realizava-se um segundo procedimento no qual a área do peeling da membrana limitante interna era ampliada. Para análise estatística, foram utilizados testes de regressão logística múltipla e Qui-quadrado. Valores de p menores que 0.05 foram considerados estatisticamente significativos. Resultados: Dos 46 olhos com buraco macular idiopático, 42 (91,3%) obtiveram fechamento do buraco macular após um procedimento cirúrgico e 45 (97,8%) após uma cirurgia adicional. A média de melhora da acuidade visual com melhor correção no pós-operatório foi de 0.378 (0.050-0.900) decimal. Não foram observados: reabertura do buraco macular, complicações relacionadas ao procedimento cirúrgico ou complicações relacionadas ao corante. Conclusão: A cirurgia combinada sem sutura que incluiu remoção da base vítrea e remoção membrana limitante interna após coloração com Azul Brilhante (0,5 mg/ml) para o tratamento de buracos maculares idiopáticos foi realizada com adequada capacidade de coloração, melhora da acuidade visual e fechamento do buraco macular sem sinais de toxicidade ocular no seguimento de 3 anos.

Determination of common dyes in dyed safflower by near infrared spectroscopy / 中国中药杂志

Because the red and bright color of corolla is the main indicator for the quality assessment of good safflower,the dyed safflower is sometimes found at the herbal market,what is influence on this herb quality and efficacy. A total of 127 safflower samples was therefore collected from different cultivating areas and herbal markets in China to develop a rapid method to identify the dyed safflower. Near-infrared spectroscopy(NIRS) combined with characteristic identification,high performance liquid chromatography(HPLC),principal component analysis(PCA) and partial least squares regression analysis(PLS) were employed to differentiate safflower from dyed safflower samples,and further quantify the levels of the 6 dyes,i.e. tartrazine,carmine,sunset yellow,azorubine,acid red 73 and orange Ⅱ in the dyed safflower. The results indicated that the 50 safflower samples and 77 dyed safflower samples were located at different regions in PCA cluster diagram by NIR spectra. Tartrazine,carmineand and sunset yellow were found in the 77 dyed safflower samples with the amounts of 0. 60-3. 66,0. 11-1. 37,0. 10-0. 71 mg·g-1,respectively. It indicated that the three dyes were the common and main dyes in the dyed safflower. However,azorubine,acid red 73 and orange Ⅱ were not detected in all herb samples. A total of 62 dyed safflower samples were chosen as calibration samples to develop the model for estimating the amount of dyes in dyed safflower. The estimating accuracy was verified by another 15 dyed safflower samples. The values of tartrazine,carmine and sunset yellow in dyed safflower samples were compared between the NIRS and HPLC methods. Each value of mean absolute difference(MAD) was less than 5%. The correlation coefficients of tartrazine,carmineand and sunset yellow were 0. 970,0. 975,0. 971,respectively. It indicated the data quantified by NIRS and HPLC were consistence. It is concluded that NIRS can not only differentiate safflower from dyed safflower,but also quantify the amount of the dyes. NIRS is suitable for rapidly identify the quality of safflower.

Comparisons of retinal nerve fiber layer thickness changes after macular hole surgery / Comparação nas alterações da espessura da camada de fibras nervosas retinianas após cirurgia de buraco macular

ABSTRACT Purpose: To compare postoperative changes in retinal nerve fiber layer thickness in patients with macular holes treated with vitrectomy with Brilliant Blue-assisted internal limiting membrane peeling. Methods: Twenty-two eyes of 20 patients with macular holes were studied. Each eye was selected to undergo Brilliant Blue-assisted internal limiting membrane peeling. The circumferential retinal nerve fiber layer thickness was determined using spectral domain optical coherence tomography preoperatively and 2 months postoperatively. Mean overall and sectoral retinal nerve fiber layer thicknesses were obtained for each patient. Results: There was no statistically significant difference (p≥0.05) between the pre- and post-treatment measurements in relation to each CFN variable, i.e., on average, pre-treatment measures were the same as post-treatment measures. Furthermore, despite the differences between the pre- and post-treatment measures always being positive (pre-post >0), they are not statistically significant. Conclusions: This study showed no significant decrease in retinal nerve fiber layer thickness measurements after macular holes surgery, regardless of age or sex.

RESUMO Objetivo: Comparar as alterações pós-operatórias na espessura da camada de fibras nervosas da retina em pacientes com buracos maculares submetidos à vitrectomia via pars-plana associada à remoção de membrana limitante interna. Métodos: Foram estudados 22 olhos de 20 pacientes consecutivos diagnosticados com buraco macular. Todos os pacientes foram submetidos à vitrectomia via pars-plana e remoção de membrana limitante interna corada com azul brilhante. A espessura da camada de fibras nervosas da retina em região peripapilar foi determinada por tomografia de coerência óptica de domínio espectral antes e 2 meses após a cirurgia. As espessuras totais e espessuras setoriais da camada de fibras nervosas da retina foram obtidas para cada paciente. Resultados: Os resultados mostram que não existe diferença estatisticamente significativa (p≥0,05) entre as medidas pré e pós-operatórias em relação a cada uma das variáveis. Conclusão: Este estudo não demonstrou diminuição significativa nas medidas da espessura da camada de fibras nervosas retinianas após a cirurgia de buraco macular, independente da faixa etária ou sexo.

Ehretia microphylla (Tsaang gubat) versus loratadine as treatment for allergic rhinitis: A randomized controlled trial

OBJECTIVE: To determine if Ehretia microphylla (Tsaang Gubat) decoction tea and placebo can improve the symptoms of mild intermittent allergic rhinitis in comparison to loratadine and control tea.METHODS:Design: Double-Blind, Randomized ControlledTrial Setting: Tertiary-Government Training HospitalParticipants: Twenty-four patients diagnosed with mild intermittent allergic rhinitis from October 2015 to July 2016 were randomly divided into a treatment group given Ehretia microphylla (Tsaang Gubat) decoction tea and placebo, and a control group given control tea and loratadine, both taken for 7 days. Patients underwent pre- and post-intervention evaluation by anterior rhinoscopy, Sino-nasal Outcome Test 22 (SNOT 22) Questionnaire and 10-point Visual Analog Scale (VAS). Data were encoded and subjected to statistical analysis using Mann Whitney U test and Wilcoxon Signed Rank test.RESULTS: Age and gender of the treatment and control group participants were comparable. Prior to intervention, no differences in symptoms were noted between both groups on SNOT 22 and VAS scores. After intervention, no differences in symptoms were noted between the 2 groups on SNOT 22 and VAS scores either. Comparison of pre- (30.4 ± 17.3) and post- (7.2 ± 6.5) intervention mean SNOT 22 scores of the loratadine control group with pre- (32.5 ± 23.7) and post- (7.8 ± 10.4) intervention mean SNOT 22 scores of the Ehretia Microphylla treatment group showed significant improvement of symptoms in both groups. Likewise, comparison of pre- and post-intervention mean VAS scores of the loratadine control group and pre- and post-intervention mean VAS scores of the Ehretia Microphylla treatment group based on symptoms of sneezing, rhinorrhea, nasal congestion and pruritus showed significant improvement of symptoms in both groups (p-values of CONCLUSION: Ehretia microphylla (Tsaang Gubat) decoction tea may improve symptoms of allergic rhinitis (sneezing, rhinorrhea, pruritus and nasal congestion) and be taken as an alternative to loratadine in patients with mild intermittent allergic rhinitis. Further clinical trials with more participants may provide stronger evidence for this conclusion.

Ehretia microphylla (Tsaang gubat) versus loratadine as treatment for allergic rhinitis: A randomized controlled trial

@#<p style="text-align: justify;"><strong>OBJECTIVE:</strong> To determine if Ehretia microphylla (Tsaang Gubat) decoction tea and placebo can improve the symptoms of mild intermittent allergic rhinitis in comparison to loratadine and control tea.<br /><strong>METHODS:</strong><br /><strong>Design:</strong> Double-Blind, Randomized Controlled<br /><strong>Trial Setting:</strong> Tertiary-Government Training Hospital<br /><strong>Participants:</strong> Twenty-four patients diagnosed with mild intermittent allergic rhinitis from October 2015 to July 2016 were randomly divided into a treatment group given Ehretia microphylla (Tsaang Gubat) decoction tea and placebo, and a control group given control tea and loratadine, both taken for 7 days. Patients underwent pre- and post-intervention evaluation by anterior rhinoscopy, Sino-nasal Outcome Test 22 (SNOT 22) Questionnaire and 10-point Visual Analog Scale (VAS). Data were encoded and subjected to statistical analysis using Mann Whitney U test and Wilcoxon Signed Rank test.<br /><strong>RESULTS:</strong> Age and gender of the treatment and control group participants were comparable. Prior to intervention, no differences in symptoms were noted between both groups on SNOT 22 and VAS scores. After intervention, no differences in symptoms were noted between the 2 groups on SNOT 22 and VAS scores either. Comparison of pre- (30.4 ± 17.3) and post- (7.2 ± 6.5) intervention mean SNOT 22 scores of the loratadine control group with pre- (32.5 ± 23.7) and post- (7.8 ± 10.4) intervention mean SNOT 22 scores of the Ehretia Microphylla treatment group showed significant improvement of symptoms in both groups. Likewise, comparison of pre- and post-intervention mean VAS scores of the loratadine control group and pre- and post-intervention mean VAS scores of the Ehretia Microphylla treatment group based on symptoms of sneezing, rhinorrhea, nasal congestion and pruritus showed significant improvement of symptoms in both groups (p-values of < .001).<br /><strong>CONCLUSION:</strong> Ehretia microphylla (Tsaang Gubat) decoction tea may improve symptoms of allergic rhinitis (sneezing, rhinorrhea, pruritus and nasal congestion) and be taken as an alternative to loratadine in patients with mild intermittent allergic rhinitis. Further clinical trials with more participants may provide stronger evidence for this conclusion.</p>

STAT signaling pathway mediates high glucose induced cardiac fibroblasts proliferation and collagen deposition in vitro / 中华心血管病杂志

<p><b>OBJECTIVE</b>To observe the signal transducers and activator of transcriptions (STATs) protein expression changes and investigate the functional role of STATs pathway in case of high glucose-induced cardiac fibroblasts (CFs) proliferation and collagen deposition in vitro.</p><p><b>METHODS</b>Rat cardiac fibroblasts were isolated from 1- to 3-day-old SD rats, cells from the second to fourth passages were used for the experiment. CFs were cultured in Dulbecco's modified Eagle's medium, supplemented with 5.5 mmol/L glucose (NG), 5.5 mmol/L glucose plus 19.4 mmol/L mannose (OC) or 25 mmol/L glucose (HG) in the presence of absence of STAT1 inhibitor (fludarabine, FLU) and STAT3 inhibitor (S3I-201). After 24 h and 48 h culture in vitro, the proliferation of CFs was measured by 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. After 12 h and 24 h culture in vitro, the production of type I and III collagen was evaluated using real-time quantitative PCR and ELISA. After 0, 30, 60 and 120 min culture in vitro, the phosphorylated expression of STAT1 and STAT3 was analyzed by Western blot.</p><p><b>RESULTS</b>CFs proliferation was significantly enhanced post 24 h and 48 h HG stimulation, and procollagen I and III mRNA expression was significantly upregulated post 12 h and 24 h HG stimulation. Deposition of collagen I and III was also significantly increased post 24 h and 72 h HG stimulation. STAT1 phosphorylation in CFs was increased after 120 min HG stimulation and STAT3 phosphorylation in CFs was increased post 60 min and 120 min HG stimulation. FLU and S3I-201 could inhibit HG-induced CFs proliferation and suppress of which was stimulated by FLU and S3I-201 could both suppress upregulated procollagen I and III mRNA expression and the deposition of collagen types I and III post HG stimulation. STAT1 phosphorylation inhibition resulted in less mRNA downregulation of procollagen type III than that of procollagen type I post 12 h HG stimulation. The STAT3 phosphorylation inhibition resulted in more significantly upregulated procollagen type III mRNA expression than procollagen type I mRNA expression at 12 h post HG stimulation.</p><p><b>CONCLUSION</b>HG could enhance the protein expression of phosphorylated STAT1 and STAT3 in CFs, which are responsible for HG-induced increased CFs proliferation and collagen deposition in vitro.</p>

Genotoxic and cytotoxic effects of sunset yellow and brilliant blue, colorant food additives, on human blood lymphocytes

The synthetic dyes over fifty are used in many areas including the food industry around the world. Sunset Yellow FCF and Brilliant Blue FCF are used as colorant food additives in many food products. The present study investigated the genotoxic and cytotoxic effects of Sunset Yellow and Brilliant Blue. Genotoxic and cytotoxic activities of the food additives were evaluated in lymphocyte cell cultures using mitotic index, replication index and micronucleus assay. Mitotic index frequencies and replication index values were decreased and micronucleus frequency was increased with increasing concentrations of Sunset Yellow and Brilliant Blue. The changes in mitotic index and micronucleus are statistically significant [p<0.05]. The results show that the Sunset Yellow and Brilliant Blue can have cytotoxic and genotoxic potential. It care must be taken when using these materials as a food additive

Cytotoxic effects of sodium dodecyl benzene sulfonate on human keratinocytes are not associated with proinflammatory cytokines expression / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Keratinocytes play a crucial role in the biological function of skin barrier. The relationship between sodium lauryl sulfate (SLS) and keratinocytes has been studied. However, the cytotoxicity and effects of sodium dodecyl benzene sulfonate (SDBS), a common detergent similar to SLS, on keratinocytes are still not known. This study aimed to investigate the effects of SDBS on cytotoxicity and expression of proinflammatory cytokines in cultured human keratinocytes.</p><p><b>METHODS</b>This study was carried out using the keratinocytes cell line, HaCaT cells. The cytotoxicity of SDBS on HaCaT cells was evaluated with cell counting kit-8 (CCK-8) and phase-contrast microscopy. After exposure to different concentrations of SDBS, the total RNA of the HaCaT cells was extracted for evaluating the relative mRNA expression of IL-1α, IL-6, IL-8, and TNF-α by qPCR. The supernatants of cells were collected for measuring the levels of IL-6 and IL-8 by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>SDBS at concentrations of 20 µg/ml and over showed direct cytotoxicity and induced morphological changes of the HaCaT cells. The mRNA expressions of IL-1α, IL-6, IL-8, and TNF-a in different concentrations of SDBS at different time were comparable with that of controls. SDBS at concentrations of 5, 10, and 15 µg/ml had no significant effects on IL-6 and IL-8 excretion from HaCaT cells after 24-hour exposure. Moreover, no significant effects on the IL-6 and IL-8 excretion were found after 10 and 15 µg/ml SDBS stimulations for 6, 12, and 24 hours, respectively.</p><p><b>CONCLUSION</b>SDBS at higher concentrations had cytotoxicity on HaCaT cells but had no effects on the mRNA expression of IL-1α, IL-6, IL-8, and TNF-a, that was different from SLS.</p>

Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba

Acanthamoeba cysts are resistant to unfavorable physiological conditions and various disinfectants. Acanthamoeba cysts have 2 walls containing various sugar moieties, and in particular, one third of the inner wall is composed of cellulose. In this study, it has been shown that down-regulation of cellulose synthase by small interfering RNA (siRNA) significantly inhibits the formation of mature Acanthamoeba castellanii cysts. Calcofluor white staining and transmission electron microscopy revealed that siRNA transfected amoeba failed to form an inner wall during encystation and thus are likely to be more vulnerable. In addition, the expression of xylose isomerase, which is involved in cyst wall formation, was not altered in cellulose synthase down-regulated amoeba, indicating that cellulose synthase is a crucial factor for inner wall formation by Acanthamoeba during encystation.

Comparison of three staining methods for the detection of intestinal microspora Spp.

This study aimed to compare three staining methods including: Calcofluor white, Chromotrope and Quick Hot Gram chromotrope used in diagnosis of intestinal microsporidial spores. One hundred and seventy five stool specimens were collected from patients referred to Laboratory of Intestinal Protozoology at the School of Public Health, Tehran University of Medical Sciences during 2012-2013. All of specimens were evaluated by nested PCR. The formalin-fixed stool samples were prepared from each specimen and dried at room temperature for 10 min, followed by 10 min methanol fixation. All the collected stool samples were evaluated blindly by calcofluor white, Chromotrope and Quick Hot Gram chromotrope staining methods separately. Microsporidial spores were recognized using Chromotrope, Quick Hot Gram chromotrope and Calcofluor white, in16 of 18 [88.8%], 17 of 18 [94.4%] and 18 of 18[100%] samples that were positive by nested PCR respectively. Regarding 14 stool samples that were negative by nested PCR, 14 cases were negative by chromotrope and Quick hot Gram chromotrope and 13 samples were negative by Calcofluor white. One discordant sample interpreted as false positive. Calcofluor white staining had the best performance for the detection of intestinal Microsprora spores and can be used as initial screen test for the detection of intestinal Microspora spp

simple technique for staining of platyhelminths with the lactopenol cotton blue stain

This paper describes a simple technique for staining of flatworms using lactophenol cotton blue [LPCB]. The staining was tested on 2 trematode species: Heterophyes heterophyes and Mesostephanus appendiculatus, and one cestode: Diplopylidium acanthotetra, which were collected from the intestine of stray cats in Kuwait. The specimens were mounted in a small amount of the LPCB stain on a clean slide for 2-3 minutes before covering with a cover slip. The technique rapidly and clearly differentiated the internal structures of the helminthes. Its speed and simplicity are advantages over other staining methods. It is easily used in wide-scale surveys where a large number of platyhelminths have to be identified and it is suitable for field studies

Sorafenib and octreotide combination therapy can inhibit proliferation of and induce apoptosis in human hepatoma cells / 中华肝脏病杂志

To investigate the effects of sorafenib and octreotide combination treatment on cellular proliferation and explore the underlying molecular mechanisms by using an in vitro cell culture system with the human hepatoma cell line, HepG2. HepG2 cells were treated with different concentrations of sorafenib and octreotide alone or in combination. Untreated HepG2 cells were used as controls. Treatment-induced cytotoxicity was determined with the cell counting kit-8 by Sigma-Aldrich, and rate of apoptosis was detected by flow cytometry. Fluorescent microscopy was used to observe rates of cell growth under the various treatments. Treatment-induced changes in protein expressions were detected by enzyme-linked immunosorbent assay (for vascular endothelial growth factor (VEGF)) and Western blotting (for the Mcl-1 apoptosis mediator and the ERK1/2 and PERK1/2 kinases). Sorafenib and octreotide, used alone or in combination, inhibited proliferation and induced apoptosis in HepG2 cells. Combination treatment was more effective than either mono-treatment (F = 200.398, P less than 0.05). Fluorescent microscopy showed that combination treatment stimulated phosphatidylserine, the marker of early apoptosis, better than either mono-treatment. VEGF expression in cultures exposed to combination treatment was also significantly lower than in mono-treatment or untreated control cultures (F = 1019.725, P less than 0.05). Western blotting showed that octreotide mono-treatment had no effect on Mcl-1 expression (vs. control group; P more than 0.05) and that combination treatment significantly lowered Mcl-1 expression (vs. mono-treatment and control groups; P less than 0.05). None of the treatments affected ERK1/2 expression (all, P more than 0.05), while all treatments significantly lowered PERK1/2 expression (vs. control group; F = 2.401, P less than 0.05) and the combination treatment lowered PERK1/2 significantly more than either mono-treatment (P less than 0.05). Sorafenib and octreotide can inhibit proliferation and induce apoptosis in the human hepatoma cell line, HepG2. Combination treatment is significantly more efficacious (P less than 0.05) and produced synergistic effects. The mechanism underlying this phenomenon may depend on synergistic inhibition of VEGF, the anti-apoptotic protein Mcl-1, and the proliferation-inducing PERK1/2.

Role of cetuximab and sorafenib in treatment of metastatic colorectal cancer.

Background: The relationship of epidermal growth factor receptors (EGFR) pathway, such as PI3K, K-ras, and B-raf, with response to EGFR-targeted antibodies is less well studied. Aim: To assess sorafenib with cetuximab in treating metastatic colorectal cancer. Settings and Design: Thirty-five patients with metastatic colorectal cancer were randomized to receive cetuximab with or without oral sorafenib. Patients and Methods: Patients received cetuximab IV weekly for four weeks and oral sorafenib twice daily on days 1 - 28, with recycling every four weeks. The primary end point was the response rate (partial and complete), while the secondary end points were the adverse effects, time to progression and overall survival. Statistical Analysis was made using the Statistical Product and Service Solutions, using SPSS 10.0, with estimation of both time to progression and overall survival time by the Kaplan-Meier method and comparing the two groups with the use of a log-rank test. Results: Partial response was higher in cetuximab-sorafenib (EN), which constituted 33.3% compared to 17.6% in the cetuximab group (P = 0.44). Progression-free survival had a statistically higher significant difference in wild K-ras compared to mutant K-ras cases (P = .0001). Median overall survival was seven and five months in the (EN) and (E) groups respectively (P = 0.49). Conclusion: K-ras and B-raf was a predictor of response, so genotyping of tumors was needed for defining the patient population that was likely to benefit from the targeted therapy. A combination of therapy that simultaneously targets K-ras and B-raf could be a useful approach to increase the number of patients who may benefit from anti-EGFR therapy.

Novel EGFR-TK Inhibitor EKB-569 Inhibits Hepatocellular Carcinoma Cell Proliferation by AKT and MAPK Pathways

Epidermal growth factor receptor (EGFR)-targeted therapies have been effective in some cancers, but not in hepatocellular carcinoma (HCC). The aim of this study was to investigate the drug potential to overcome multi-drug resistance in HCC cells. Thirteen drug-sensitive HCC cells were assessed using the CCK-8 assay. G0-G1 arrest was measured by FACS. Western blot analysis was used to detect the key enzymes in both the Ras/Raf and PI3K pathways. When establishing the IC50 of HCC to several drugs, including EKB-569, sorafenib, erlotinib, gefitinib, pazopanib, and brivanib, SK-Hep1 cells treated with EKB-569 have shown the highest (72.8%-86.4%) G0-G1 arrest and decreased the phosphorylation of AKT and ERK at the protein level. We found that EKB-569 had higher efficacy in HCC, compared to first generation, reversible EGFR-TK inhibitors. Furthermore, the combination of sorafenib and EKB-569 showed a synergistic effect to inhibit proliferation of SNU-475, previously the most resistant cell to EGFR-TKIs. Therefore, novel EKB-569 in combination with sorafenib may be able to overcome HCC resistance to EGFR-TK inhibitors.

Sorafenib induces apoptosis of U937 cells via inhibiting WNT signal pathway / 中国实验血液学杂志

This study was aimed to investigate the effect of multikinase inhibitor sorafenib on the proliferation and apoptosis of U937 cells and its possible mechanism. U937 cells were treated with different concentrations of sorafenib for 48 hours. Cell viability was determined by Cell Counting Kit-8; cell apoptosis and cell ratio in cell cycle were detected by flow cytometry with Annexin V/PI staining and PI staining respectively; expressions of GSK-3β, β-catenin and cyclin-D1 were assayed by Western blot. The results showed that the proliferation of U937 cells was inhibited by sorafenib in a dose-dependent manner (p < 0.05). Sorafenib induced cell apoptosis and cell cycle G(1)/G(0) arrest. Compared with results of Western blot before treatment, expression of inactivated GSK-3β, β-catenin and Cyclin-D1 down-regulated in a dose-dependent manner after treatment with sorafenib, this same changes were observed after up-regulation of inactivated GSK-3β by LiCl (p < 0.05). It is concluded that sorafenib inhibits the proliferation of U937 cells and induces cell apoptosis through reducing negative regulation of WNT signal pathway on inactivated GSK-3β and down-regulating β-catenin and cyclin-D1 level, which result in U937 cell cycle G(1)/G(0) arrest.

Sorafenib in combination with chemotherapy in the induction therapy for FLT3-ITD positive acute monocytic leukemia: a case report and literature review / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the safety and efficacy of sorafenib in combination with chemotherapy for the treatment of FLT3 positive acute myeloid leukemia (AML), to highlight the impact of FLT3 mutations and targeting therapy on response of AML.</p><p><b>METHODS</b>The clinical and laboratory features and the treatment response, especially the safety profile of sorafenib in an acute monocytic leukemia patient with FLT-ITD were reported.</p><p><b>RESULTS</b>The patient achieved clinical and molecular CR after sorafenib was added to the second course of combination chemotherapy. The side effects of sorafenib were mild and tolerable.</p><p><b>CONCLUSION</b>The patient responded well to the combination of sorafenib and standard chemotherapy of AML without significant adverse effects.</p>

Optimal Media Conditions for the Detection of Extracellular Cellulase Activity in Ganoderma neo-japonicum

To determine the optimal media conditions for the detection of the extracellular cellulase activity in Ganoderma neo-japonicum, we varied three media conditions: dye reagent, pH, and temperature. We evaluated the use of four dyes, Congo red, phenol red, remazol brilliant blue, and trypan blue. To observe the effect of pH on the chromogenic reaction, we tested media ranging from 4.5 to 8.0. To research the effect of temperature on the clear zone and the fungus growing zone, we tested temperatures ranging from 15 to 35degrees C. On the whole, the best protocol called for Ganoderma neo-japonicum transfer onto media containing Congo red with a pH of 7.0, followed by incubation at 25degrees C for 5 days. Our results will be useful to researchers who study extracellular enzyme activity in Ganoderma neo-japonicum.