BACKGROUNDAdoptive transfer of allogeneic tumor-specific T cells often results in severe graft-versus-host disease (GVHD). Here, we sought to maximize graft-versus-tumor and minimize GVHD by using haploidentical T cells in pre-irradiated B16-melanoma bearing mice.

METHODSC57BL/6 mice bearing B16-melanoma tumors were irradiated with 0, 5, or 7 Gy total body irradiation (TBI), or 7 Gy TBI plus bone marrow transplantation. Tumor areas were measured every 3 days to assess the influence of irradiation treatment on tumor regression. B16-melanoma bearing mice were irradiated with 7 Gy TBI; sera and spleens were harvested at days 1, 3, 5, 7, 9, 11, and 13 after irradiation. White blood cell levels were measured and transforming growth factor β1 (TGF-b1) and interleukin 10 (IL-10) levels in serum were detected using enzyme-linked immunosorbent assay (ELISA) kits. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry were performed to test TGF-b1, IL-10 and Foxp3 mRNA levels and the proportion of CD4+CD25+Foxp3+ T-regulatory cells (Tregs) in spleens. B16-melanoma bearing C57BL/6 mice were irradiated with 7 Gy TBI followed by syngeneic (Syn1/Syn2) or haploidentical (Hap1/Hap2), dendritic cell-induced cytotoxic T lymphocytes (DC-CTLs) treatment, tumor areas and system GVHD were observed every 3 days. Mice were killed 21 days after the DC-CTLs adoptive transfer; histologic analyses of eyes, skin, liver, lungs, and intestine were then performed.

RESULTSIrradiation with 7 Gy TBI on the B16-melanoma-bearing mice did not influence tumor regression compared to the control group; however, it down-regulated the proportion of Tregs in spleens and the TGF-b1 and IL-10 levels in sera and spleens, suggesting inhibition of autoimmunity and intervention of tumor microenvironment. Adoptive transfer of haploidentical DC-CTLs significantly inhibited B16-melanoma growth. GVHD assessment and histology analysis showed no significant difference among the groups.

CONCLUSIONAdoptive transfer of haploidentical tumor-specific T cells in irradiation-pretreated B16-melanoma bearing mice preserved antitumor capacity without causing a GVHD response.

Animals ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Graft vs Host Disease ; Immunotherapy, Adoptive ; methods ; Male ; Melanoma, Experimental ; metabolism ; therapy ; Mice ; Mice, Inbred C57BL ; Real-Time Polymerase Chain Reaction ; T-Lymphocytes ; immunology ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo develop an HPLC method for determination of saponin I in bulb extracts of Allium macrostemon.

METHODThe extract was refluxed by methanol and then filtrated. The filtrate was concentrated and the bioactive saponin was measured by HPLC-RID.

RESULTThe calibration curve was in good linearity over the range of 5-100 microg with the regression equation of Y= 9.35 x 10(4) X - 2.78 x 10(4) (r = 0.9998) and the average recovery was 101.1% (n = 5, RSD 1.6%).

CONCLUSIONThe method is simple, quick and reproducible and can be applied in the quality control of A. macrostemon.

Allium ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Plant Extracts ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Saponins ; analysis ; Sterols ; analysis

Objective To evaluate the correlation of lens density (LD) measured by opacity lensmeter (OLM) and Pentacam with phacodynamics in patients with age-related nuclear cataract.Methods A prospective study was conducted in 78 patients (78 eyes) with age-related nuclear cataract.The lens density was measured with OLM and Pentacam respectively before operation,and the phacoemulsification kinetic parameters in phacoemulsification,including ultrasound time (UST),cumulated dissipated energy (CDE) and balanced salt solution (BSS) usage,were recorded for the analysis of the correlations of LD values with phacodynamics.Results LD measured by OLM and Pentacam was 32.83 ± 14.41 and 15.26 ±5.82,respectively.LD from OLM and Pentacam was positively correlated with both UST (r =0.645,0.807,respectively;both P ＜ 0.05)and CDE (r =0.669,0.828,respectively;both P ＜ 0.05).UST and CDE were more strongly correlated with LD from Pentacam than from OLM (Z =2.153,2.283;both P ＜0.05).Conclusion LD from both OLM and Pentacam are correlated with UST and CDE,and a stronger correlation is observed in Pentacam than OLM.

Jiangsu province is rich in intertidal marine resources,especially gastropod and bivalve shellfish.In this study,we performed shotgun proteomic analysis on shellfish from Lüsi,Jiangsu to identify protein content.In total,265 proteins were identified in nine species of shellfish using nano LC-MS/MS.The highest number of proteins were identified in the gastropod Hemifusus tuba (HF) (105 proteins),whereas the bivalve Scapharca subcrenata (SSC) produced the fewest (39 proteins).The most abundant proteins are constituents of cytoskeleton,including actin,tubulin,tropomyosin,myosin heavy chain,as well as histone proteins.Additional proteins were also identified,including ribosomal proteins,enzymes such as arginine kinase and ATP synthase,and housekeeping proteins such as elongation factor and heat shock protein.Proteins were classified based on cellular components and molecular function using the principal component analysis (PCA).Results of PCA analysis revealed that Sinonovacula constrzcta (SC),Bullacta exarata (BE) and Hemifusus tuba (HF) showed similar protein profiles,whileSolen strictus (SS),Meretrix meretrix (MM),Ruditapes philippinarum (RP),and Mactra veneriformis (MV) also showed similar protein profiles.Hierarchical clustering results of heat map studies showed that RP and Mactra antiquate (MA) were classified into one group.SC,BE,HF,MM,SSC,and MV were classified into another group.SS showed the furthest distance from this group.We established a method for analyzing and characterizing the protein composition characteristics of predominant shellfish species in Jiangsu providence's intertidal zone area,which provides basic data for the development of shellfish resources.

Our work focuses on the quality control and structural identification of Photocyanine as a cancer therapeutic photosensitizer. Photocyanine is a mixture which contains four ZnPcS2P2 type substituted Phthalocyanine isomers. In order to obtain the single component from Photocyanine, the mixture of four isomers possessing the similar structures and chemical property had been isolated and purified. An HPLC method with a mixture of methanol-acetonitrile-ion-pair buffer as the mobile phase was applied to isolate the four isomers by means of a semi-preparative C18 column. To remove the salts which were mixed in the preparative product, a SPE C18 column was used to separate the salts by elution with water and then the marker component was eluted by methanol. Subsequently, a column of Sephadex LH-20 gel was applied to elute the crudes with methanol to desalination. The purity of the isolated compound was measured by TLC and four different isomers of phthalocyanine were obtained. The chemical structures of them were elucidated by 1H NMR spectra, gCOSY and NOE1D. An HPLC-DAD method was developed for simultaneously determination of four major isomers in Photocyanine with a C18 column (Grace Smart, 150 mm x 4.6 mm ID, 5 microm). The separation was carried out with a gradient program at a flow rate of 1.0 mL x min(-1). The mobile phase was a mixture of acetonitrile and ion-pair buffer (0.01 mol x L(-1) hexadecyl trimethyl ammonium bromide and 0.01 mol x L(-1) potassium dihydrogen phosphate, adjusted the pH value to 6.8 with potassium hydroxide solution). The resolution values of four isomers were 2.5, 1.20, 1.33, and 1.8. Linear regression analysis for four compounds was performed by the external standard method. Four constituents were linear in the concentration range of 0.005 to 10 microg. The values of relative standard deviation (RSD) of intra-day were 0.12%, 0.66%, 0.99%, and 1.21%, respectively. The limits of detection for four compounds were 15 ng, 20 ng, 12 ng, and 25 ng, respectively. This method was simple, accurate and reproducible. The developed method can be successfully applied to analyze isomers in Photocyanine.
Antineoplastic Agents ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Indoles ; analysis ; chemistry ; Isomerism ; Molecular Structure ; Organometallic Compounds ; analysis ; chemistry ; Photochemotherapy ; Photosensitizing Agents ; analysis ; chemistry ; Quality Control

BACKGROUNDFractional flow reserve (FFR)-guided percutaneous coronary intervention (PCI) is associated with fewer unfavorable events. However, the hemodynamic change in FFR after different stenting approaches for bifurcation lesions is still not fully studied. The aim of this study was to analyze the hemodynamic changes in FFR after double kissing (DK) crush and provisional side branch (SB) stenting (PS) for true coronary bifurcation lesions.

METHODSSeventy-five patients with true bifurcated lesions were randomly divided into DK (n = 38) and PS (n = 37) groups. Additional SB stenting in the PS group was required if there was any pinched SB ostium > 70% stenosis, or ≥ type B dissection, or TIMI flow < grade 3. FFR at hyperemia in the main vessel (MV) and SB was measured prior- and post-stenting, and at 8 months follow-up.

RESULTSBaseline clinical, angiographic and lesion characteristics were matched well between the two groups, with the exception of the final kissing balloon inflation (FKBI, 100.0% in the DK vs. 83.8% in the PS group, P < 0.001). Baseline FFR was comparable between the DK and the PS groups, however, the acute gain and late loss of SB FFR at 8-month follow-up in the DK group were 0.18 ± 0.15 and -0.06 ± 0.11, compared to 0.12 ± 0.18 (P = 0.044) and -0.002 ± 0.07 (P = 0.037) in the PS group, respectively. MV FFR post-stenting > 0.94 was seen in about 40% of patients. There was no significant difference in the clinical events at 1-year follow-up between the two groups.

CONCLUSIONSDK crush was associated with improved acute gain and late loss of SB FFR. The lower rate of FFR > 0.94 after stenting underscored the further improvement of stenting quality.

Adolescent ; Adult ; Aged ; Angioplasty, Balloon, Coronary ; Coronary Artery Disease ; therapy ; Coronary Stenosis ; therapy ; Drug-Eluting Stents ; Female ; Hemodynamics ; physiology ; Humans ; Male ; Middle Aged ; Percutaneous Coronary Intervention ; methods ; Treatment Outcome ; Young Adult

BACKGROUNDStenting strategies and clinical outcomes of bifurcation lesions in a chronic total occlusion (CTO) vessel after successful recanalization remain to be unknown.

METHODSBetween January 2001 and December 2009, 195 (41.1%) patients with 254 (47.0%) bifurcation lesions in CTO vessels from a pool of 564 patients with 659 CTO lesions were included and divided into proximal (n = 134) and distal (n = 120) groups, according to the location of the bifurcation lesions. The primary endpoint was the occurrence of major adverse cardiac events (MACE) at the end of clinical follow-up, including cardiac death, myocardial infarction, or target vessel revascularization (TVR).

RESULTSCollaterals with Rentrop class 3 were seen more in distal group (100% and 68.3%), compared to proximal group (76.9% and 45.6%). Two-stent technique for proximal bifurcation lesions was used in 24.6%, significantly different from the distal group (6.7%, P < 0.001), without significant difference in composite MACE between proximal and distal groups, or between one- and two-stent subgroups in proximal group. The composite MACE after 1-year in complete revascularization subgroup was 17.9% relative to 29.6% in the incomplete revascularization group (P = 0.044). Stents in long false lumen in main vessel were mainly attributive to decreased TIMI grade flow, with resultant increased in-stent restenosis, total occlusion, TVR and coronary aneurysms. Imcomplete revasculzarization (HR 2.028, P = 0.049, 95%CI 1.002 - 4.105) and post-stenting TIMI flow (HR 6.122, P = 0.020, 95%CI 1.334 - 28.092) were two independent predictors of composite MACE at the 1-year follow-up.

CONCLUSIONSTwo-stent was more used for proximal bifurcation lesions. No significant difference was observed in MACE between proximal and distal, or between one- and two-stent subgroups in the proximal group. Placement of a safety wire was critical for proximal bifurcation lesions. Complete revascularization was mandatory to improve clinical outcomes.

Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; Coronary Stenosis ; therapy ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Prospective Studies ; Registries ; Stents ; adverse effects ; Treatment Outcome

Aged ; Angina, Unstable ; pathology ; therapy ; Angioplasty, Balloon, Coronary ; methods ; Coronary Angiography ; Coronary Disease ; pathology ; therapy ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Prospective Studies ; Treatment Outcome

OBJECTIVETo investigate the value of intensity modulated radiation therapy (IMRT) for patient with gynecological malignancies after treatment of hysterectomy and chemotherapy/radiotherapy.

METHODSAll 32 patients with cervical or endometrial cancer after hysterectomy received full course IMRT after 1 to 3 cycles of chemotherapy (Karnofsky performance status(KPS) > or =70). Seventeen of these patients underwent postoperative preventive irradiation and the other 15 patients were pelvic wall recurrence and/or retroperitoneal lymph node metastasis, though postoperative radiotherapy and/or chemotherapy had been given after operation.

RESULTSThe median dose delivered to the PTV was 56.8 Gy for preventive irradiation, and 60.6 Gy for pelvic wall recurrence or retroperioneal lymph node metastasis irradiation. It was required that 90% of iso-dose curve could covere more than 99% of GTV. However, The mean dose irradiated to small intestine, bladder, rectum, kidney and spinal cord was 21.3 Gy, 37.8 Gy, 35.3 Gy, 8.5 Gy, 22.1 Gy, respectively. Fourteen patients presented grade I (11 patients) or II (3 patients) digestive tract side-effects, Five patients developed grade I or II bone marrow depression. Twelve patients had grade I skin reaction. The overall 1-year survival rate was 100%. The 2- and 3- year survival rate for preventive irradiation were both 100%, but which was 5/7 and 3/6 for the patients with pelvic wall recurrence or retroperioneal lymph node metastasis.

CONCLUSIONIntensity modulated radiation therapy can provide a better dose distribution than traditional radiotherapy for both prevention and pelvic wall recurrence or retroperioneal lymph node metastasis. The toxicity is tolerable. The adjacent organs at risk can well be protected.

Adult ; Aged ; Combined Modality Therapy ; Diarrhea ; etiology ; Endometrial Neoplasms ; drug therapy ; radiotherapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; methods ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Recurrence, Local ; Radiotherapy Dosage ; Radiotherapy, Intensity-Modulated ; adverse effects ; methods ; Survival Analysis ; Uterine Cervical Neoplasms ; drug therapy ; radiotherapy ; surgery

OBJECTIVETo investigate the distribution of metastatic pelvic lymph nodes in the women with early stage cervical carcinoma, and the feasibility of dividing these nodes into three stations in those patients.

METHODS(99m)Tc-DX of 2 ml was injected into the cervix to a depth of 5 to 10 mm at 3, 6, 9, 12 o'clock positions preoperatively in 196 patients with early stage cervical cancer. Pelvic lymphadenectomy and radical hysterectomy were performed in all patients. Pelvic lymph nodes were detected by gamma-probe. The sentinel lymph nodes (SLN) were determined if the radioactivity reached 5 times higher than that in the ipsilateral nodes. All resected pelvic lymph nodes were examined by histopathology with HE stained serial sections.

RESULTSOf the 196 patients, 41 were found to have metastasis in 83 lymph nodes. The metastatic rate was 78.3% (65/83) in the parametrial and obturator lymph nodes, 20.5% (17/83)in the internal and external iliac lymph nodes, 1.2% (1/83) in the commmon iliac lymph nodes. Of the 22 patients with metastatic parametrial lymph nodes, metastatic external iliac lymph nodes were detected in 5 patients, and metastatic internal iliac lymph nodes in 3 patients. Among the 19 patients with metastatic obturator lymph nodes, metastatic external iliac lymph nodes were found in 4 patients, and metastatic internal iliac lymph nodes in 3 cases. It was shown by Chi-sqare test that the metastases in parametrial and/or obturator lymph nodes were positively correlated with lymph node metastases in other pelvic sites. Eighty-one SLN were found to have metastasis. The metastatic rate of parametrial and obturator SLN was 79.0% (64/81) versus 21.0% (17/81) of internal and external iliac SLN. No statistically significant difference in 1- and 3-yr survival was observed between the groups with and without metastasis in parametrial and obturator lymph nodes, while the 5-yr survival rate in the patients without metastatic lymph node was 93.2%, significantly higher than that of patients with lymphatic metastasis (65.1%).

CONCLUSIONIt is feasible for cervical cancer to divide the pelvic lymph nodes into three levels. The level I lymph nodes consist of parametrial and obturator lymph nodes. Internal and external iliac lymph nodes can be considered as level II lymph nodes, and the common iliac and inguinal lymph nodes as level III nodes. A rational treatment plan can be made according to the distribution of metastatic pelvic lymph nodes.

Adenocarcinoma ; pathology ; surgery ; Adult ; Carcinoma, Squamous Cell ; pathology ; surgery ; Dextrans ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; methods ; Lymph Node Excision ; Lymph Nodes ; diagnostic imaging ; pathology ; surgery ; Lymphatic Metastasis ; pathology ; Middle Aged ; Neoplasm Staging ; Organotechnetium Compounds ; Pelvis ; Radionuclide Imaging ; Sentinel Lymph Node Biopsy ; Survival Rate ; Uterine Cervical Neoplasms ; pathology ; surgery ; Young Adult