Objective To open out new method,we probe into immune function of??ST and NKT cells in NSCLC.Methods The peripheral blood cells were stained with antibodies labeled with fluorescence in NSCLC,??T,NKT and their set group cells were counted with flow cytometry.Results The absolute counts of??T,CD~+_(56)??T,CD~-_(56)??T,NKT and??T~+ NKT cells in NSCLC was significantly lower than that of normal controls.The relative counts of NKT,??T~+ NKT,??T~-NKT and CD~+_56??T cells in NSCLC was not sig- nificantly lower than that of normal control.The absolute counts of??T cells in NSCLC positively correlated to the number of NKT cells,(r=0.426,P=0.009).Conclusion The absolute counts of??T,NKT and their set ground cells in NSCLC was significantly lower than that of normal controls.In NSCLC the relation of the??T and NKT had positive correlation.

Objective To investigation the pathological characteristics of esophageal squamous cell carcinoma to provide reference criteria for delineating the target area in radiotherapy.Methods Fifty-two patients from the Fourth Hospital of HeBei Medical University underwent resection whom all had been proved to have esophageal squamous cell carcinoma before operation.Chest CT was scanned and transmitted to the 3- dimensional conformal planning system for radiotherapy by VRX-16 scanner.The lesion of esophageal carcinoma was delineated in the 3-dimensional rebuild CT image and the lesion volume was computed by digital rebuild program.Every surgically resected specimen was made into pathologic giant section.The actual size of the specimen was obtained by calculating the size under the microscope with the shrinkage ratio.Multicentric carcinomatous lesion,severe dysplasia and direct intramural infiltration were observed in the giant section with a microscope and the order of such pathological characteristics were analysed statistically.Results 1.The tumor length by different method of preparation of operated specimens differed obviously.The longest was shown by CT. 2.Multicentric carcinomatous lesion was found in 15(29%)cases out of 52 patients.Proximal to the tumor,the mean distance between the multicentric carcinomatous lesion and the main lesion plus the length of the multicentric carcinomatous lesion was 3.02?1.45cm.Distal to the tumor,it was 2.60?2.44 cm.Severe dysplasia was found in 28 patients.Proximally,the mean distance between the severe dysplasia and the main lesion plus the length of the severe dysplasia was 2.45?1.30 cm.Distal to the tumor,it was 3.24?2.19 cm.Direct intramural infiltration was found in 41 patients,of which the mean length being 2.80?1.52 cm proximally and 2.02?1.51 cm distally. 3.Tumor thrombus was found in 6 patients and lymphoduct infiltration in 36 patients.Direct intramural infiltration was found at higher incidence in specimens complicated with lymphoduct infiltration(86%)and those complicated with tumor thrombus(91%).There were no apparent factors affecting severe dysplasia.The proximal distance to direct intra- mural infiltration was much longer than distally.Conclusions Multicentric carcinomatous lesion,severe dysplasia and direct intramural infiltration may be observed in esophageal squamous cell carcinoma.Multicentric carcinomatous lesion and direct intramural infiltration are obviously correlated with lymphoduct infiltration.To cover 95% of the microscopic extension,a margin of 5.0 cm is needed proximal to the base of gross tumor volume,and 7.5 cm distal to it.To cover 90% of the microscopic extension,a margin of 4.5 cm is needed proximally,and 5.0 cm distally.

Objective:To study the antimalarial mechanism of benflumetol (B). Methods: Flow cytometry (FCM) was used to analyze the effects of B and chloroquine (CQ) on DNA content of Plasmodium berghei and pH value of the lysosome of malarial parasites. Results: DNA content of the plasmodia not treated with any drugs was not changed in 24 hours,while benflumetol could decrease the DNA content: the DNA content began to decrease 2 h after the drug administration and reached the minimum by 16 h, but somewhat increased at 24 h after administration. The pH in the lysosome increased 1 h and restored premedication level 4 h after benflumetol administration. Chloroquine had the same effects on DNA and lysosome pH of malarial parasites.Conclusions: The antimalarial mechanism of benflumetol is directly related to its effect to inhibit the synthesis of DNA.

Background Allograft rejection is a main cause of failure of penetrating keratoplasty,especially in the patient with high risk of rejection condition.Previous study on allograft rejection mechanism focused on limbal and corneal neovascularization,but these factors did not explain all the phenomena of allograft rejection.Research found that immune cells appeared in iris and ciliary body when rejection occurred,but the relationship between these immune cells and allograft rejection is unclear Objective This study was to evaluate the relationship between diversity of vascular permeability in the iris and ciliary body and allograft rejection after penetrating keratoplasty.Methods Seventy clean eight-week-old BALB/c mice were divided into allogeneic corneal transplantation group (60 mice) and blank control group (10 mice).Allogeneic corneal transplantation was performed with the same age of C57BL/6 mice as donor and BALB/c mice as the recipients.The grafts were examined under the slit lamp microscope and scored based on the criteria of Hegde.The mice were sacrificed and iris and ciliary tissue were obtained 5,10 days and rejection after surgery.Immunohistochemistry and reverse transcription PCR (RT-PCR) was used respectively to detect the expression diversities of occludin,zonula occludens protein-1 (ZO-1),matrix metalloproteinase-9(MMP-9),major histocompatibility complex-Ⅱ (MHC-Ⅱ),and CCR5,CCR7 and their mRNA in iris and ciliary body.Image-J image analysis software was used to calculate the quantity of positive cells on iris wholemount,and absorbance of target genes (A values).The use and care of the experimental animals complied the ARVO Resolution on the Use of Animals in Research.Results The mean survival time of corneal gratts was (17±3) days after operation.The mean score was 0.6 in 5 days and 0.5 in 10 days,and 3.3 in 18 days after operation.Expression of ZO-1 reduced significantly,and that of MMP-9 increased obviously at the time of rejection.MHC Ⅱ + cells were scattered in iris and ciliary body in normal mice,and the number of the positive cells (cells/field) was increased after operation with a peak value when rejection occurred.A significant difference was seen between normal mice and rejection mice (1559.67±350.29 vs.4021.83±495.18) (P=0.000).The expressions of occludin mRNA and ZO-1 mRNA in the iris and ciliary body decreased obviously in the rejection mice.Compared with normal mice,theA value of ZO-1 and occluding were 36.74±3.13 vs.110.11±11.88 and 57.54±3.41 vs.59.90±3.50respectively,with significant differences between them (all P＜0.05).The expressions of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA in the iris and ciliary body increased gradually with the time lapse after operation and peaked when the rejection appeared.The A value of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA were significantly higher than those of normal mice (20.29±1.19 vs.2.77±0.85 for MMP-9 mRNA; 35.43±2.56 vs.9.11±0.29 for CCR5 mRNA,and 60.83±0.87 vs.0.89 ±0.95 for CCR7 mRNA) respectively (all P＜0.05).Conclusions The permeability of vascules in the iris and ciliary body increase during the allograft rejection after penetrating keratoplasty.Increased antigen presenting cells were also detected.

This retrospective analysis compared standard regimen of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) with the dose-dense ABVD regimen (ABVD-21) in terms of efficacy and toxicity. Patients who had early-stage unfavorable or advanced Hodgkin's lymphoma (HL) according to German Hodgkin Study Group criteria from March 1999 to February 2011 were analyzed for treatment response, long-term survival and hematological toxicity. There were 85 patients in the ABVD-21 group and 118 patients in the ABVD group respectively. The complete remission rates after completion of treatment were 92.9% and 90.7% for ABVD-21 and ABVD, respectively. During a median follow-up period of 62 months, no significant difference was found in projected 10-year progression-free survival (PFS) and overall survival (OS) rates (84.7% and 94.1% respectively for ABVD-21; 81.4% and 91.5% for ABVD). Subgroup analyses showed that ABVD-21 was significantly better than ABVD for patients with IPS≥3 in terms of PFS and OS rates. Grade 3 to 4 leukopenia (51.8% vs. 28.8%, P=0.001) and neutropenia (57.6% vs. 39.0%, P=0.009) were more common with ABVD-21. We were led to conclude that dose-dense ABVD did not result in better tumor control and overall survival than did ABVD for early-stage unfavorable HL. However, patients at high risk, for example, with IPS≥3, may benefit from dose-dense ABVD.

OBJECTIVETo explore the inhibition and molecular mechanism of icaritin (ICT) combined doxorubicin (DOX) on human osteosarcoma MG-63 cells in vitro.

METHODSThe control group, ICT groups (10, 20, 40, 80, and 160 µmol/L), DOX groups (1, 2, 4, 8, and 16 µg/mL), and combination groups (20 µmol/ L ICT +1 µg/mL DOX, 20 µmol/L ICT +2 µg/mL DOX, 20 µmol/L ICT +4 µg/mL DOX, 40 µmol/L ICT +1 µg/mL DOX, 40 µmol/L ICT +2 µg/mL DOX, 40 µmol/L ICT +4 µg/mL DOX, 80 µmol/L ICT +1 µg/mL DOX, 80 µmol/L ICT +2 µg/mL DOX, 80 µmol/L ICT +4 µg/mL DOX) were set up. Human osteosarcoma MG-63 cells were respectively cultured and their effects on morphological changes were observed using inverted phase contrast microscope after 24-and 48-h intervention. The cell proliferation inhibition rate of each group was de- termined using CCK-8, and IC50 calculated. The MG-63 apoptosis rate was detected using Annexin V-FITC/ PI double dye flow cytometry. Expression levels of bcl-2, caspase-3, and p21 were detected using RT-PCR.

RESULTSICT and DOX could obviously inhibit the proliferation of MG-63 cell. Along with ICT concentration increasing from 10 µmol/L to 160 µmol/L, the cell proliferation inhibition rate also increased gradually from 9.67% ± 3.62% to 89.18% ± 9.66%. The IC50 was 46.93 µmol/L and 3.87 µg/mL respectively. ICT and DOX could cause either early or late stage apoptosis, down-regulate Bcl-2 gene expression, and up-regulate gene expressions of Caspase-3 and p21 respectively (P < 0.05). Aforesaid changes were more obviously seen in combination groups than in lCT groups and DOX groups (P < 0.05).

CONCLUSIONCT combined DOX had additive or synergistic inhibition effect for the proliferation of osteosarcoma MG-63 cells, which might be related with regulating gene expressions of bcl-2, caspase-3, and p21.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Down-Regulation ; Doxorubicin ; pharmacology ; Drug Synergism ; Flavonoids ; pharmacology ; Humans ; Osteosarcoma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

The chemical constituents were isolated and purified by various chromatographic techniques indluding silica gel, reverse phase silica gel, sephadex LH-20 and pre-HPLC and identified by their physicochemical properties and spectral data. Sixteen phenolic compounds had been isolated and n-butanol extracts which were fractionated from the ethanol extract of Oplopanax horridus roots bark. Their structures were identified as below, including 7 phenylpropanoid compounds, ferulic acid (1), 3-acetylcaffeic acid (2), caffeic acid (3), homovanillyl alcohol 4-O-beta-D-glucopyranoside (4), 3-hydroxyphenethyl alcohol 4-O-beta-D-glucopyranoside (5), 3, 5-dimethoxycinnamyl alcohol 4-O-beta-D-glucopyranoside (6), and 3-dimethoxycinnamyl alcohol 4-O-beta-D-glucopyranoside (7). Three coumarins, scopoletin (8), esculetin (9) and 3'-angeloyl-4'-acetyl-cis-knellactone (10). And 6 lignan compounds, (+)-isolaricires-inol-9'-O-beta-D-glucopyranoside (11), 3, 3'-dimethoxy-4, 9, 9'-trihydroxy-4', 7-epoxy-5', 8-lignan-4, 9-bis-O-beta-D-glucopyranoside (12), (+)-5, 5'-dimethoxylariciresinol 4'-O-beta-D-glucopyranoside (13), (-)-5,5'-dimethoxylariciresinol 4'-O-beta-D-glucopyranoside (14), (-)-pinoresinol 4'-O-beta-D-glucopyranoside (15), and (+)-5, 5'-dimethoxylariciresinol 9'-O-beta-D-glucopyranoside (16). All compounds were isolated and identified for the first time from this plant All the constituents except compounds 4, 6, 12 and 13 were obtained for the first time from the genus Oplopanax.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Oplopanax ; chemistry ; Phenols ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo study the therapeutic effect of chitosan-coated basic fibroblast growth factor (bFGF) slow-releasing microspheres on the knee osteoarthritis in the rabbit.

METHODSFrom November 2008 to July 2009, 54 New Zealand rabbits were divided into 6 groups at random, which were the control group, the model group, the PBS-M group, the bFGF-S group, the 10-bFGF-M group and the 100-bFGF-M group, respectively. The model of knee osteoarthritis was induced by the injection of papain in the rabbit. Except the control and model groups, all the experimental groups were implanted 1 ml intervention solution at the third and sixth weeks, including the PBS microspheres, bFGF solution, 10 µg bFGF microspheres and 100 µg bFGF microspheres, respectively. The rabbits were sacrificed at the ninth week after operation, and then articular cartilage was conducted the morphological and histopathological evaluation.

RESULTSThe damage of articular cartilage in the model group was more serious than that in the control group, with statistical differences according to the Ink score (t = 8.22, P = 0.00) and Mankin score (t = 17.20, P = 0.00). The damage of articular cartilage in the PBS-M and bFGF-S groups were similar with that in the model group, according to the Ink score (t = 0.26, P = 0.79; t = 0.80, P = 0.45) and Mankin score (t = 1.51, P = 0.17; t = 0.56, P = 0.60). The Ink and Mankin scores in the 10-bFGF-M and 100-bFGF-M groups were better than that in the model group (Ink score: t = 3.58, P = 0.01; t = 6.82, P = 0.00; Mankin score: t = 3.41, P = 0.01; t = 5.00, P = 0.00), with the 100-bFGF-M group much better (t = 5.29, P = 0.00; t = 2.80, P = 0.02).

CONCLUSIONSThe bFGF slow-releasing microsphere can keep its effective intra-articular concentration, which may accelerate the synthesis of proteoglycan and inhibit its decomposition to reverse the damage of articular cartilage.

Animals ; Drug Carriers ; administration & dosage ; therapeutic use ; Fibroblast Growth Factor 2 ; administration & dosage ; therapeutic use ; Injections, Intra-Articular ; Microspheres ; Osteoarthritis, Knee ; therapy ; Rabbits

In order to establish HPLC fingerprint of Liuwei Dihuang soft capsule, and to provide certain reference for an quality control of it, the HPLC method was performed on an Agilent C18 (4.6 mm x 250 mm, 5 μm) column with acetonitrile-0.02% trifluoroacetic acid as mobile phase, gradient elution volume flow of 1.0 mL x min(-1), column temperature was 30 degrees C, detection wavelength: 0-60 min, 238 nm, 60-70 min, 210 nm. The software for chromatographic fingerprint was applied to analysis different batches of Liuwei Dihuang soft capsule samples. Sixteen mutual peaks were selected as the fingerprint peaks in 12 samples with loganin as the reference peak, and all of the detected peaks were separated effectively. Cluster analysis (HCA) and similarity analysis (SA) were done based on data of 12 samples clustering analysis of 12 batches of samples were divided into 2 categories. Including 7 for the first class, the rest was second, similarities calculated by SA were all above 0.92, indicating a good similarity between the reference and twelve batches of samples, also, the analysis results of HCA and SA basically the same. This method is simple with good precision, repeatability and stability, and provides the basis for Liuwei Dihuang soft capsule quality control.
Capsules ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Quality Control

OBJECTIVETo assess bone health in epileptic children who have been treated with topiramate (TPM) or carbamazepine (CBZ).

METHODSSixty-three epileptic children who received TPM or CBZ treatment and 36 eileptic children who did not receive any drug treatment (control group) were enrolled. Bone mineral density (BMD) at lumbar vertebrae (L1-L4) and radius-ulna was evaluated by the dual-energy X-ray absorptiometry method. Biochemical indices of bone metabolism, including serum calcium, phosphorus and alkaline phosphatase contents were measured.

RESULTSThe serum calcium content was higher in the TPM group (2.41+/-0.17 mmol/L), but it was lower in the CBZ group (2.15+/-0.26 mmol/L) than that (2.26+/-0.11 mmol/L) in the control group (p<0.05). The serum phosphorus content in both the TPM (1.55+/-0.17 mmol/L) and the CBZ groups (1.52+/-0.26 mmol/L) was significantly lower than that in the control group (1.70+/-0.30 mmol/L) (p<0.05). There were no significant differences in the serum content of alkaline phosphatase between three groups. BMD was significantly reduced in both the TPM and the CBZ groups when compared to the control group (p<0.05).

CONCLUSIONSTPM and CBZ may result in alterations in serum contents of calcium, phosphorus and alkaline phosphatase as well as BMD reduction.

Adolescent ; Alkaline Phosphatase ; blood ; Anticonvulsants ; adverse effects ; Bone Density ; drug effects ; Bone and Bones ; drug effects ; metabolism ; Calcium ; blood ; Carbamazepine ; adverse effects ; Child ; Child, Preschool ; Epilepsy ; drug therapy ; metabolism ; Female ; Fructose ; adverse effects ; analogs & derivatives ; Humans ; Male ; Phosphorus ; blood