Objective To study the influence of microenvironment created by dissoluble cytokines derived from hepatocellular carcinoma cells (HCCs) on the functional status of mitochondria in dendritic cells (DCs) in order to investigate the mechanisms of tumor immune escape.Methods CD14+ monocytes,which were isolated with immune magnetic beads from fresh peripheral blood of healthy human,were then cultured in RPMI1640/10% FBS supplemented with 100 ng/ml rhGM-CSF and 100 ng/ml rhIL-4 for 7 d to develop into immature DCs (imDCs).Maturation was induced by addition of 20 ng/ml rhTNF-? to imDCs for another 3 d of culture.Acquired imDCs and mDCs were respectively co-cultured with human HCCs for 48 h in Transwell chamber.These treated cells were investigated for mitochondrial membrane potential,enzyme activity and intracellular Ca2+ concentration ([Ca2+]in) in the cytoplasma by immune fluorescence and MTT assay.Those untreated cells served as control.Results After co-cultured with HCCs,flow cytometry showed that the mitochondrial membrane potentials of imDCs and mDCs were respectively decreased from (659.991?17.052)and(473.741?11.676)to(482.681?7.935)and(407.189?5.051)(P

Objective To study the influence of microenvironment created by dissoluble cytokines derived from hepatocellular carcinoma cells (HCCs) on the expression of IL-12 of dendritic cells (DCs) at different differentiating stage. Methods The CD14+ monocytes,isolated with immune magnetic beads from fresh peripheral blood of healthy human,were cultured in RPMI 1640/10% FBS supplemented with 100 ng/ml rhGM-CSF and 100 ng/ml rhIL-4 for 7 d to develop into immature DCs (imDCs). Maturation was induced by addition of 20 ng/ml rhTNF-? to imDCs for another 3 days' culture. Acquired imDCs and mDCs were respectively co-cultured with human HCC Bel7402 cells for 48 h in Transwell chamber. The concentrations of IL-10,IL-12,TGF-?1 and VEGF in culture supernatant were investigated by ELISA,imDCs and mDCs cultured alone served as control. Results After co-cultured with Bel7402 cells,IL-12 were decreased from (115.076?8.129) pg/ml to (17.599?1.757) pg/ml in imDCs,and from (258.346?3.609) pg/ml to (6.787?1.123) pg/ml in mDCs (P

Objective To observe the clinical effects of Shenqifuzheng injections combined with PTX and DDP on advanced non small cell lung cancers. Methods Fifty cases of advanced non small cell lung cancers were randomly divided into pure chemotherapy and chemotherapy combined with traditional Chinese medicine groups. The former was treated with 30 mg/m2 DDP from day 1 to day 3, 135 mg/m2 PTX only at the first day. The latter was treated with 250 ml Shenqifuzheng injections every day until the twentieth-first one, three days before the beginning of chemotherapy. After two periods of treatments, the therapeutic effects were evaluated, respectively. Results The recent effective ratios of chemotherapy combined with traditional Chinese medicine and pure chemotherapy groups were 48 % and 40 %, respectively. The former could ameliorate the physical status, the numbers of peripheral leucocytes, the decreases of blood platelet, the gastrointestinal reactions of subjects. Conclusion The Shenqifuzheng injections could increase the recent treatment effect of PTX combined with DDP on advanced non small cell lung cancers, effectively meliorate the clinical symptoms and alleviate the side effects of chemotherapy leading to improve the life qualities of subjects.

To establish reference values of various immunophenotypic markers in B lymphocyte population in healthy Chinese adults and build background information for accurate interpretation of B cell immunophenotyping data in clinical practice, peripheral blood from 41 healthy adults were collected separately into test tubes containing EDTA-K(2) and stored in room temperature no more than 24 hours before analysis. Whole blood lysis technique and multiparameter flow cytometry were applied to immunophenotype B cells gated on CD19/SSC dot-plot. The results showed that CD22, CD20, CD62L, CD40, CD24, CD79b, CD79a, and FMC-7 were almost positive in the circulating B cell population, whereas CD11a, CD80, CD103, CD10, CD40L, CD54, CD95L, CD86, and CD95 were almost negative in the peripheral blood B lymphocytes. CD18, CD44, CD23, CD5, CD11c and CD43 were positive in different B cell subpopulations. 78% of B cells were IgD positive and ratio kappa/lambda was 1.26. The significance of all these markers in the differential diagnosis of lymphoproliferative diseases was discussed. The conclusion is that it is necessary to consider the qualitative and quantitative levels of expression of various markers in normal B cell population in order to accurately interpret the pathological immunophenotypic data in clinical practice. It is also important to note the immunotypic differences of B cells between Chinese and Western populations.
Adult ; Aged ; B-Lymphocytes ; immunology ; CD5 Antigens ; analysis ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Intercellular Adhesion Molecule-1 ; analysis ; Leukemia, Lymphocytic, Chronic, B-Cell ; immunology ; Lymphoma ; immunology ; Male ; Middle Aged ; Receptors, IgE ; analysis

Objective To investigate the protective effect of c-jun N-terminal kinase (JNK)inhibitor SP600125 against acute liver failure in mice.Methods Fifty-five male C57/BL6 mice were divided into control group (n =30) and SP600125 group (n =25).The animals were given an intraperitoneal injection of D-galactosamine (D-GalN,400 mg/kg body weight)/lipopolysaccharide (LPS,30 μg/kg body weight).The control group and SP600125 group were given 10％ dimethyl sulfoxide (15 mL/kg body weight) or SP600125 (75 mg/kg body weight) subcutaneously 12 h and 1 h before D-GalN/LPS administration,respectively.D GalN/LPS induced mouse JNK activation was detected by immunohistochemistry for phospho JNK (p-JNK).D-GalN/LPS induced mouse liver cell apoptosis was detected by immunohistochemistry for Caspase-3 and TdT-mediated-dUTP nick endlabeling (TUNEL).Serum alanine transaminase (ALT) level was tested to assess liver injury.Survival rate of mice within 24 h after D-GalN/LPS administration was observed.The comparison between groups was done by t test and survival rate was analyzed by Kaplan-Meier method.Results JNK activity in liver tissues,as indicated by observation of p-JNK positive cells by immunohistochemistry,was diminished 4 h after D-GalN/LPS administration in SP600125 group.Reduced Caspase-3 activity was observed 6 h after D-GalN/LPS administration in SP600125 group (as indicated in immunohistochemistry by Caspase-3 positive cells).Mice in SP600125 group showed significantly lower TUNEL-positive cell count than control group (43.0±24.5 vs 194.7±73.8; t=9.743,P=0.000).Serum ALT level 6 h after D-GalN/LPS administration was (24.0±54.7) U/L in SP600125 group,which was significantly lower than that in control group [(1234.4±478.4) U/L; t=4.734,P=0.0015].SP600125 also significantly improved the survival rate within 24 h after D-GalN/LPS administration (4/5 vs 1/10； x2=5.225,P=0.0223).Conclusions JNK inhibitor SP600125 exerts protective effects against D-GalN/LPS induced acute liver failure in mice by suppressing JNK activation and hepatocyte apoptosis.

Background Phakic posterior chamber implantable contact lens (PPC-ICL) or phakic posterior chamber Toric implantable contact lens (PPC-TICL) implantation is an effective way for the correction of high myopia or high myopia with astigmia,but it often has residual myopic power.Excimer laser-assisted subepithelial keratectomy (LASEK) can correct the residual myopia following PPC-ICL or PPC-TICL,but its effectiveness and safety deserve attention.Objective This study was to analyze the clinical effectiveness and safety of LASEK for residual myopia after PPC-ICL implantation for extreme high myopia.Methods A prospective cases-observational study was performed,and written informed consent was obtained from each patient before any surgery.Fourteen eyes of 9 patients with residual myopia following PPC-ICL or PPC-TICL for the eyes with spherical equivalent refraction of ≥-20.00 D were collected in the Affiliated Hospital of Guizhou Medical University from July 2010 to March 2015,including PPCICL implantation in 8 eyes and PPC-TICL implantation in 6 eyes.LASEK were performed on the eyes to correct the residual myopic power.Uncorrected visual acuity (UCVA),best corrected visual acuity (BCVA),haze,the distance of intraocular lens to lens,corneal thickness,corneal topography,corneal endothelial cell counting,intraocular pressure (IOP) and fundus were examined and compared before and after surgery.The effectiveness and safety of the surgery were evaluated.Results The operation was smooth and no complication was found after surgery in all of the eyes.The UCVA and BCVA were significantly different in the eyes among before surgery,6 months after PPC-ICL implantation and 12 months after LASEK (F =31.360,1.778;both at P＜0.05),and the UCVA after LASEK was higher than BCVA before LASEK.The refractive powers were (-22.27-±4.29),(-3.75±2.25) and (-0.42±0.63) D before surgery,6 months after PPC-ICL implantation and 12 months after LASEK,showing a significant difference among them (F=46.370,P＜0.05),and the refractive power was considerably lower after LASEK than that before surgery and after PPC-ICL implantation (both at P＜0.05).No significant difference was found in IOP or corneal endothelial cell counting in operated eyes among before surgery,6 months after PPC-ICL implantation and 12 months after LASEK (F=1.663,1.055;both at P＞0.05).The distance of intraocular lens to lens was (0.69±0.26)mm in the eyes after LASEK and (0.71 ±0.29)mm in the eyes after PPC-ICL implantation,with no significant difference between them (t =0.192,P＞0.05).Conclusions PPC-ICL or PPC-TICL implantation for the correction extreme high myopia often remains a certain degree of myopia,and LASEK for the correction of residual refractive power is safe and effective.

The paper discusses the experimental teaching reform of biochemistry by referring to the characteristics of biochemical development and special feature.The reform increases effectually the interests of medical students,and may contribute to their creative competence and the abilities of scientific research.

ObjectiveTo prospectively investigate the impact of short-time response on survival of concurrent chemotherapy and thoracic three-dimensional radiotherapy (CCTTRT) for stage Ⅳ non-small cell lung cancer (NSCLC). Methods From Jan.2003 to Oct.2010,201 patients with pathologically or cytologically proven stage Ⅳ NSCLC were included.All patients received platinum-based chemotherapy.Of the 167 patients eligible for analysis,the median number of chemotherapy were 4 cycles.The median dose for planning target volume (PTV) of thoracic primary tumor was 63 Gy.Response was scored according to WHO criteria. Survival was calculated by Kaplan-Meier method and compared using the Logrank. Cox regression model were used to examine the effect of response on overall survival.ResultsThe follow-up rate of 201 patients was 97.5％.with 201,170 and 134 patients finished ＜ 1,1 -2 and ≥3 years' follow-up.For the 167 patients eligible for analysis,the CR,PR,NC and PD rate of primary tumor was 5.4％,65.9％,21.0％ and 7.7％,respectively.The effective group ( CR + PR) and ineffective group ( NC + PD) was 71.3％ and 28.7％,respectively.The median survival time (MST) for patients with CR,PR,NC and PD was 22.6,13.4,8.8 and 4.8 months,respectively ( χ2 =44.79,P =0.000).The MST for effective and ineffective group was 13.9 and 7.6 months,respectively in the whole group ( χ2 =8.3 0,P =0.004 ),12.1months and 7.3 months in those treated with 2 - 3 cycles chemotherapy ( χ2 =7.71,P =0.007 ),and 13.9months and 7.9 months in those treated with 2 -5 cycles chemotherapy and radiation dose to PTV ≥36 Gy ( χ2 =4.00,P =0.045 ).No significant MST difference was detected between patients of effective group and ineffective group treated with 4 -5 cycles chemotherapy ( χ2 =0.67,P =0.413),or those treated with 4 -5 cycles of chemotherapy and radiation dose to primary lesion ≥36 Gy (χ2 =0.00,P =0.956).Multivariate analysis showed that 4-5 cycles of chemotherapy and CR and PR achieved in primary tumor (β =0.182,P=0.041 ) were independent favorable factors for survival. Conclusion CCTTRT can improve local control,and prolong the survival time for Stage Ⅳ NSCLC.

ObjectiveTo investigate the delineation of gross tumor volume (GTV) in locally advanced nasopharyngeal carcinoma (LANC) according to imageological changes before and after induction chemotherapy (IC) in order to decrease high dose area and protect normal tissue better.MethodsBetween Mar 2010 to Jan 2011,11 patients with LANC were enrolled and treated with TPF regimen followed by intensity-modulated radiotherapy (IMRT) with concurrent chemotherapy,target volumes were delineated based on fused CT imaging before and after IC following project determination.Tumor target volumes after and before IC were respectively delineated according to imaging tumor residues and were overlaid by CTVnx in order to ensure radical doses for the imaging tumor volume before IC,the resulting differences of tumor target volumes of IC before and after were measured and analyzed by paired t-test.ResultsBefore and after IC,the average volumes of GTVnx were respectively 44.72 cm3 and 28.87 ( t =3.89,P =0.003 ),the average volumes of GTVnd were respectively 32.76 cm3 and 19.82 cm3 ( t =2.47,P =0.033 ),the volumes of maximum dose area in brainstem and spinal cord as well as eyeball decreased ( t =2.93-4.59,all P ＜0.05).ConclusionsLANC treated by 3 cycle TPF regimen followed by IMRT with concurrent chemotherapy showes significant shrinkage of tumor volume.The volume of high dose region which caused by normally recovered tissues were decreased by re-delineation of target volume in brainstem and spinal cord as well as eyeball of CT images after IC.

Objective To evaluate the importance of three-dimensional radiotherapy for elderly patients of stage Ⅳ non-small cell lung cancer (NSCLC).Methods Comparing with treatment outcome of ≥65 years 67 patients and ＜ 65 years 134 patients using concurrent chemotherapy and thoracic threedimensional radiotherapy during 2003 to 2010 years.Survival analysis was taken by Kaplan-Meier method.The multivariate prognosis was analyzed by Cox model.Results The follow-up was 97.8％.The percentage of ≥65 years and ＜ 65 years patients accepted with concurrent 4-5 cycles chemotherapy were 30％ and 55％,and with 42％ and 49％ patients with radiotherapy ≥63 Gy.The median survival time (MST) were 17 months and 14 months (x2 =0.76,P =0.384) for ≥65 years and ＜ 65 years patients accepted with concurrent 4-5 cycles chemotherapy concurrent ≥63 Gy radiotherapy respectively.The MST and 1-,2-,3year overall survival rate were 17 months and 8 months,65％ and 23％,30％ and 13％,24％ and 9％(x2 =7.90,P =0.005) for whole groups patients treated with chemotherapy concurrent ≥63 Gy and ＜ 63 Gy radiotherapy.And the MST of patients ≥ 63 Gy was significantly longer than those with ＜ 63 Gy either concurrent chemotherapy any cycles (x2 =9.54,P =0.023).The MST were 14 months and 8 months (x2 =1.82,P=0.178),17 months and 17 months (x2 =0.47,P=0.492) for ≥ 65 years and ≥ 63 Gy radiotherapy patients accepted with concurrent 4-5 cycles and 2-3 cycles chemotherapy concurrent respectively.Multivariate analysis showed local response (β =0.600,P =0.003) and numbers of tumor metastasis (β =0.670,P =0.040) were independent factors for survival.Conclusions For a part of elderly patients of stage Ⅳ NSCLC,concurrent chemotherapy and thoracic three-dimensional radiotherapy can prolong survival time with acceptable toxicity.Perhaps radiotherapy is more important.