To investigate the feasibility of using Real-Time PCR to evaluate the effectiveness of Sindbis virus inactivation by Methylene Blue with visible light. Sindbis virus was treated by Methylene Blue with different intensity of visible light and the transcribed cDNA was quantified by Real-Time PCR. Residual infectivity of treated virus was tested by cell infection method as parallel control at the same time. The residual infectivity of virus decreased from 6.50 lgTCID50/mL to under the limit of detection as light intensity increased. Meanwhile, the quantity of virus cDNA decreased significantly (P < 0.05), which correlated to the decline of virus infectivity (R2 > 0.98). Methylene Blue with visible light could cause lesion to nucleic acid of Sindbis virus, the extent of which was light intensity-dependent and correlated to the decrease of virus infectivity. The results demonstrated that Real-Time PCR can be a useful tool for evaluating effect of virus inactivation after Methylene Blue treatment with light.
Light ; Methylene Blue ; pharmacology ; Polymerase Chain Reaction ; methods ; Sindbis Virus ; drug effects ; genetics ; physiology ; radiation effects ; Virus Inactivation ; drug effects ; radiation effects

OBJECTIVETo evaluate the therapeutic efficacy of three-dimensional conformal radiotherapy (3DCRT) combined with stereotactic radiotherapy (SRT) and the radiation-induced complications in patients with locally advanced non-small cell lung cancer (NSCLC).

METHODSSixty-eight patients with locally advanced NSCLC were randomly divided into two groups. The 33 patients in groups A received 3DCRT at the total dose of 66 to 70 Gy in 33 to 35 fractions, and the 35 patents in group B received stereotactic radiotherapy (SRT) at the dose of 20 Gy in 4 fractions (500 cGy per fraction every other day) after 60 Gy 3DCRT.

RESULTSThe total response rates in groups A and B were 56.3% and 80.0%, with the complete remission rates of 9.4% and 28.6%, respectively, both showing significant differences between the two groups (P<0.05). The 1- and 2-year survival rates of the patients in group A were 65.6% and 46.8%, respectively, which were comparable to those in group B (74.3% and 51.4%, respectively, P>0.05). The median survival time in groups A and B were 12.6 and 18.3 months, respectively. The major radiation-induced complications in the two groups included grade I to II acute radiation esophagitis and hematopoietic toxicity. The complications in later stages following the radiation were grade I to II radiation lung fibrosis, occurring at a similar incidence between the two groups.

CONCLUSIONThe combination of 3DCRT and SRT produces better therapeutic effects than 3DCRT alone in patients with locally advanced NSCLC.

Adolescent ; Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; radiotherapy ; Female ; Humans ; Lung Neoplasms ; radiotherapy ; Male ; Middle Aged ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Conformal ; methods ; Stereotaxic Techniques ; Young Adult

Rosmarinic acid (RA) is a natural polyphenolic compound that exists in many medicinal species of Boraginaceae and Lamiaceae. The previous studies have revealed that RA had therapeutic effects on hepatocellular carcinoma (HCC) in the H22-xenograft models by inhibiting the inflammatory cytokines and NF-κB p65 pathway in the tumor microenvironment. However, its molecular mechanisms of immunoregulation and pro-apoptotic effect in HCC have not been fully explored. In the present study, RA at 75, 150, and 300 mg/kg was given to H22 tumor-bearing mice via gavage once a day for 10 days. The results showed that RA can effectively inhibit the tumor growth through regulating the ratio of CD4⁺/CD8⁺ and the secretion of interleukin (IL)-2 and interferon-γ, inhibiting the expressions of IL-6, IL-10 and signal transducer and activator of transcription 3, thereby up-regulating Bax and Caspase-3 and down-regulating Bcl-2. The underlying mechanisms involved regulation of immune response and induction of HCC cell apoptosis. These results may provide a more comprehensive perspective to clarify the anti-tumor mechanism of RA in HCC.
Animals ; Apoptosis ; Boraginaceae ; Carcinoma, Hepatocellular ; Caspase 3 ; Cytokines ; Interleukin-10 ; Interleukin-6 ; Interleukins ; Lamiaceae ; Mice ; STAT3 Transcription Factor ; Therapeutic Uses ; Tumor Microenvironment

OBJECTIVETo investigate the mechanism by which heat shock protein 90 (HSP90) regulates 26S proteasome in hyperthermia.

METHODSHyperthermic HepG2 cell models established by exposure of the cells to 42 degrees celsius; for 3, 6, 12, and 24 h were examined for production of reactive oxygen species (ROS) and cell proliferation, and the changes in Hsp90α and 26S proteasome were analyzed.

RESULTSROS production in the cells increased significantly after hyperthermia (F=28.958, P<0.001), and the cell proliferation was suppressed progressively as the heat exposure time extended (F=621.704, P<0.001). Hyperthermia up-regulated Hsp90α but decreased the expression level (F=164.174, P<0.001) and activity (F=133.043, P<0.001) of 26S proteasome. The cells transfected with a small interfering RNA targeting Hsp90α also showed significantly decreased expression of 26S proteasome (F=180.231, P<0.001).

CONCLUSIONThe intracellular ROS production increases as the hyperthermia time extends. Heat stress and ROS together cause protein denature, leading to increased HSP90 consumption and further to HSP90 deficiency for maintaining 26S proteasome assembly and stability. The accumulation of denatured protein causes unfolded protein reaction in the cells to eventually result in cell death.

HSP90 Heat-Shock Proteins ; metabolism ; Hep G2 Cells ; Hot Temperature ; Humans ; Proteasome Endopeptidase Complex ; metabolism ; RNA, Small Interfering ; genetics ; Reactive Oxygen Species ; metabolism ; Up-Regulation

OBJECTIVETo evaluate the efficacy and toxicity of the combined chemotherapy with docetaxel, capecitabine and cisplatin (TXP) in the treatment of metastatic nasopharyngeal carcinoma (NPC).

METHODSThis retrospective analysis involved 22 patients with metastatic NPC receiving treatment with the TXP regimen. The patients were given docetaxel at 60 mg/m² on day 1, cisplatin at 20 mg/m² on days 1-3, and capecitabine at 1 250 mg/m² on days 1-14, and the treatment cycle was repeated ever 3 weeks.

RESULTSOf the 22 patients, 14 (63%) achieved partial remission, 2 (9%) had complete remission, and 5 (23%) showed stable disease. The overall clinical response rate of the patients was 72% with a 1-year survival rate of 68%, median progression-free survival of 8 months, and overall survival of 14 months. The main toxicity was myelosuppression; 7 (32%) patients experienced grade 3/4 neutropenia, and 5 (23%) had grade 3/4 anemia. All the other adverse effects were tolerable and reversible.

CONCLUSIONThe TXP regimen is safe and effective for treatment of metastatic NPC, and the results are comparable with those of the reports in recent literatures.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Neoplasms ; drug therapy ; secondary ; Capecitabine ; Carcinoma, Squamous Cell ; drug therapy ; pathology ; Cisplatin ; administration & dosage ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Female ; Fluorouracil ; administration & dosage ; analogs & derivatives ; Humans ; Lung Neoplasms ; drug therapy ; secondary ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; drug therapy ; pathology ; Retrospective Studies ; Taxoids ; administration & dosage

OBJECTIVETo investigate the correlation between the change of liver enzymes and the severity of acute graft-versus-host disease (aGVHD).

METHODSThe liver enzyme and severity of aGVHD in 82 patients were analyzed retrospectively.

RESULTSAmong the 82 aGVHD patients, 7 developed grade I (18.3%), 47 grade II (57.3%), 13 grade III (15.9%) and 7 grade IV (8.5%) aGVHD. The elevation of ALT, AST and ALP was found in 49 (59.8%), 36(43.9%) and 8(9.8%)patients respectively. Among these patients, 34 (69.4%) were ALT > 40 U/L, 18 (50%) AST > 40 U/L and 4 (50%) ALP > 132 U/L before the onset of aGVHD. The elevation of ALT and AST was more common and the peak value was significantly higher in patients with grade III-IV aGVHD than with grade I-II aGVHD, but those of ALP showed no difference between the two groups. Compared with the patients without ALT elevation, the proportion of severe aGVHD was higher in patients with ALT elevation.

CONCLUSIONSThe alteration of liver enzymology is common in aGVHD and its level is correlated with the severity of aGVHD. Liver enzymes, especially the ALT, may be used as a predictor of aGVHD.

Graft vs Host Disease ; Humans ; Liver ; Retrospective Studies

OBJECTIVETo compare the therapeutic effect and adverse effects of two regimens, namely cisplatin and docetaxel (DC) regimen and fluorouracil (PF) regimen, both with concurrent radiotherapy, in the treatment of advanced esophageal squamous cancer.

METHODSForty-eight patients with esophageal squamous cancer were randomly assigned in DC regimen and PF regimen groups. All the patients received conventional radiotherapy at a total dose of 60 Gy (in 30 fractions) for 6 weeks. In DC regimen group, the patients received intravenous infusion of docetaxel (75 mg/m(2)) for 1 h on day 1 and DDP (25 mg/m(2) daily) on days 1-3, with every 28 days as one cycle. PF regimen consisted of cisplatin (25 mg/m(2)) on days 1-3 and continuous intravenous infusion of fluorouracil (500 mg/m(2)) for 5 days, with every 28 days as one cycle. All the patients were suggested to have no less than 2 cycles.

RESULTSThe 3-year median survival time in DC regimen was slightly longer than that in PF regimen group (26 vs 23 months, Χ2=3.4041, P=0.065). The same result was also found in the short-term effect and adverse reactions including ?myelosuppression and gastrointestinal reactions. Only the adverse reaction of radiotherapy-induced esophagitis showed a significant difference between the two groups (P=0.049).

CONCLUSIONDC regimen with synchronous radiotherapy is effective and safe for treating advanced esophageal squamous cancer.

Adult ; Aged ; Antineoplastic Protocols ; Carcinoma, Squamous Cell ; drug therapy ; radiotherapy ; therapy ; Combined Modality Therapy ; Esophageal Neoplasms ; drug therapy ; radiotherapy ; therapy ; Female ; Humans ; Male ; Middle Aged

Objective: To assess the prognostic significance of immunophenotype complete remission (ICR) and hematological complete remission (HCR) before human-leukocyte antigen (HLA)-matched sibling donor transplantation (MSDT) in acute myeloid leukemia (AML) patients. Methods: A cohort of 182 AML (non-APL) patients undergoing MSDT in HCR was retrospectively studied [including complete remission with ANC and PLT recovery (CR), CR with incomplete PLT recovery (CRp), CR with inconplete ANC and PLT recovery (CRi)]; ICR was determined as undetective minimal resudial disease (MRD) by multi-parameter flow cytometer. Results: ①Of the 182 patients, 97 were male, 85 female, and the median age was 41(4-62) years. ②The CR and CRi+CRp rates were 80.8% (147/182) and 19.2%(35/182), respectively; The 4-year cumulative incidence of relapse[CIR, (11.0±4.3)% vs (16.0±7.1)%, χ(2)=0.274, P=0.600], non-relapse mortality[NRM, (14.0±4.3)% vs (9.0±6.3)%, χ(2)=0.913, P=0.339], leukemia-free survival[LFS, (75.0±5.1)% vs (75.0±8.3)%, χ(2)=0.256, P=0.613], and overall survial [OS, (77.0±5.2)% vs (80.0±8.1)%, χ(2)=0.140, P=0.708] were comparable between the CRp+CRi and CR groups. ③Compared with the non-ICR group (n=35), the ICR group (n=147) showed lower 4-year CIR [(11.3±3.4) % vs (55.2±8.8) %, χ(2)=32.687, P<0.001], better 4-year LFS [(76.2±4.7)% vs (32.8±8.7)%, χ(2)=26.234, P<0.001] and OS[(79.0±4.7)% vs (39.0±9.1)%, χ(2)=25.253, P<0.001], and comparable NRM[(12.5±4.1)% vs (12.0±7.1)%, χ(2)=1.002, P=0.656]. ④Mulitvariate analysis confirmed the independent prognostic value of ICR in lower CIR [HR=11.026(95%CI 4.685-25.949), P<0.001], higher LFS [HR=5.785 (95% CI 2.974-11.254), P<0.001] and OS[HR=5.578 (95% CI 2.575-27.565), P<0.001]. Conclusion: The results indicated that ICR instead of HCR pre-transplantation had a significant prognostic value in AML patients undergoing MSDT.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; Male ; Middle Aged ; Retrospective Studies ; Siblings ; Young Adult

BACKGROUNDChronic graft-versus-host disease (GVHD), the commonest long-term complication after allogeneic hematopoietic stem cell transplantation (HSCT), has a negative impact on patients' health related quality of life (HRQoL). This study was designed to investigate the HRQoL in patients with chronic GVHD in China.

METHODSTwo hundred and sixty-four patients with chronic GVHD who were ≥ 24 months post-HSCT and had been in continuous complete remission since HSCT were enrolled in this retrospective study. HRQoL was evaluated using an SF-36 questionnaire. Multivariate analysis was used to identify the factors that affect HRQoL in patients with chronic GVHD.

RESULTSHRQoL in patients categorized as having mild and moderate chronic GVHD was significantly better than in those in the severe category. In the moderate chronic GVHD category, markedly poorer HRQoL was observed in patients with both multiple organ involvement and more severe organ impairment than in those without these factors. According to multivariate analysis, chronic GVHD severity had the greatest significant negative impact on patients' HRQoL; whereas being female was associated with a negative impact on psychological health.

CONCLUSIONChronic GVHD severity strongly correlates with negative impacts on patients' HRQoL.

Adult ; Chronic Disease ; Female ; Graft vs Host Disease ; psychology ; Humans ; Male ; Middle Aged ; Quality of Life ; Retrospective Studies ; Severity of Illness Index

BACKGROUNDThe occurrence of bronchiolitis obliterans syndrome (BOS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is rare but severe. We examine the role of pre-HSCT chemotherapeutic exposure, pre-HSCT comorbidities, and transplant-related complications in the development of BOS after allo-HSCT.

METHODSA nested case-control study was designed. Cases with BOS and controls matched for the year of allo-HSCT and length of the follow-up were identified from a cohort of 1646 patients who underwent allo-HSCT for treatment of hematologic malignancies between 2006 and 2011. Antithymocyte globulin was used in the partial matched related and unrelated matched donor HSCT, or patients with severe aplastic anemia.

RESULTSThirty-six patients suffered from BOS; the mean age at the time of presentation was (32.7 ± 12.4) years, and the mean time to presentation was (474 ± 350) days post-HSCT. A pre-HSCT cyclophosphamide dose of ≥ 3.2 g/m(2)(OR = 8.74, P = 0.025), chronic graft-versus-host disease (moderate to severe) (OR = 12.02, P = 0.000), and conditioning regimens without antithymocyte globulin (OR = 2.79, P = 0.031) were independently associated with BOS.

CONCLUSIONSWe found that higher pre-HSCT cyclophosphamide exposure, a conditioning regimen without antithymocyte globulin, and moderate to severe chronic graft-versus-host disease are significantly and independently associated with BOS. Based on these results, we can identify patients who are at a higher risk of developing BOS after allo-HSCT, select a more appropriate therapeutic strategy, and improve the outcome of HSCT recipients.

Adult ; Bronchiolitis Obliterans ; etiology ; Case-Control Studies ; Female ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Risk Factors ; Transplantation Conditioning ; Transplantation, Homologous