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.

Objective To prospectively evaluate the survival of different metastasis organs with concurrent chemotherapy and thoracic three-dimensional radiotherapy (CCTTRT) for stage Ⅳ non-small cell lung cancer (NSCLC).Methods Two hundred and one patients of stage Ⅳ NSCLC were enrolled from January,2003 to July,2010.Of the 182 patients eligible for analysis,The number of patients with single-organ metastasis or multiple-organ metastasis was 107 and 75,respectively.Patients were treated by platinum-based chemotherapy,the median number of cycle was 4.The median dose to planning target volume of primary tumor (DTPTv) was 63 Gy.Survival was calculated by Kaplan-Meier method and compared using the Logrank.Results The follow-up rate of 201 patients was 97.5％.with 201,170 and 134 patients finished ＜ 1,1 -2 and ≥3 years'follow up.Of 182 patients,the 1-,2-,and 3-year overall survival (OS) rate and median survival time (MST) was 41.0％,17.0％,10.0％ and 10.5 months,respectively ;with single-organ metastasis and multi-organ metastasis were 50％,20％,14％ and 13 months and 29％,12％,0％ and 8.5 months ( x2 =10.10,P =0.001 ),respectively; compared with multi-organ metastasis,the 1-,2-,and 3-year OS arte and MST of patients with bone,lung metastasis only was 58％,25％,16％ and 14 months (x2 =10.42,P=0.001 ) and 49％,21％,21％ and 11 months (x2 =6.39,P=0.011 ) respectively;patients with brain metastasis only did not show advantage of survival comparing with patients with multi-organ metastasis (49％,8％,0％ and 12 months and 29％,12％,0％ and 8 months,respectively;x2 =0.71,P =0.401 ) ;the 1-,2-,and 3-year OS rate and MST was 63％,23％,19％ and 15 months and 42％,15％,0％ and 10 months,respectively for patients with single-organ metastasis and multi-organ metastasis patients who accepted 4 - 5 cycles of chemotherapy ( x2 =6.47,P =0.011 ) ; for patients under the same metastasis and 4 - 5 cycles of chemotherapy,no matter whether single-organ or multiple-organ metastases,the 1 -,2-,3-year OS rate and MST of patients with enough radiotherapy on DTPTV ≥63 Gy were better than patients without enough radiotherapy ( DTPTV ＜ 63 Gy ) ( 71％,25 ％,25％ and 16.8 months and 33％,17％,0％ and 10.5 months,respectively;x2 ＝4.73,P =0.030 ;54％,21％,0％ and 14.3 months and 29％,10％,0％ and 7.6 months,respectively,x2 =8.16,P =0.004).The MST of liver metastases was 6 months,there was significantly difference when comparing with non liver matastasis ( x2 =17.21,P =0.000).Conclusions It is very important to treat stage Ⅳ NSCLC with CCTTRT,especially patients with single-organ metastasis.Liver metastases is a unfavorable prognostic factor.

Objective To evaluate the overall survival and safety among patients for stage Ⅳ non-small cell lung cancer (NSCLC) treated with concurrent chemotherapy and thoracic three-dimensional radiotherapy (CCTTRT).Methods From Jan.2003 to July 2010,201 patients with stage Ⅳ NSCLC were included.All patients were treated with CCTTRT.Those patients who received only one cycle chemotherapy were not included in survival analysis,but analysis of toxicity.One hundred and eighty-two patients were eligible for survival analysis.All patients received platinum-based two-drug chemotherapy.The median number of cycles was 4.The median dose to planning target volume of primary tumor ( DTPTV ) was 63 Gy.Treatment-related gastrointestinal and hematological toxicity were scored according to WHO criteria.Radiation-related pneumonitis and esophagitis were evaluated according to the National Cancer Institute's Common Terminology Criteria for Adverse Events (CTC) version 3.0.Survival was calculated by Kaplan-Meier method and compared using the Logrank.Cox regression model was used to examine the effect of CCTTRT on overall survival.Results The follow-up rate of 201 patients was 97.5％.with 201,170 and 134 patients finished ＜ 1,1 -2 and ≥3 years' follow-up,respectively.Of the 182 patients eligible for survival analysis,further stratified analysis showed that the 1-,2-and 3-year overall survival rate and median survival time (MST) was 54％,20％,13％ and 14.3 months,respectively for patients treated with concurrent 4 -5 cycles chemotherapy and CCTTRT,and 66％,23％,19％ and 16.1 months,respectively for those treated with 4 -5 cycles chemotherapy and DTPTV ≥ 63 Gy.Under similar chemoradiotherapy intensity,the MST of patients with single organ metastasis was significantly longer than that with multiple organ metastases ( 13.0 months versus 8.5 months,x2 =10.10,P =0.001 ).For patients eligible for survival analysis and received 4 - 5 cycles of systemic chemotherapy,MST of patients treated with DTPTV≥63 Gy was significantly longer than those treated with DTPTV ＜63 Gy[14.9 months vs.8.4 months (x2 ＝20.48,P =0.000) and 16.1 months vs.8.8 months ( x2 =11.75,P =0.001 )].For patients with single organ metastasis,MST was 16 months for those treated with DTPTV ≥63 Gy and 9 months for those with DTPTV ＜63 Gy (x2 =10.51,P=0.000) ;for patients with multiple organ metastasis,it was 11 months and 7 months,respectively ( x2 =7.90,P =0.005 ).Multivariate analysis showed that concurrent 4 - 5 cycles chemotherapy and DTPTV ≥63 Gy (β =0.243,P=0.019) and improved KPS (β =1.268,P=0.000) were independent factors for survival.For the whole group,45％ patients had Grade 2 -3 gastrointestinal toxicity,35.0％ grade 3- 4 leukopenia,18％ grade 3- 4 thrombocytopenia.15.0％ grade 3- 4 anemia,9.5％ Grade 2 - 3 radiation pneumonia and 13.4％ radiation esophagitis,respectively.Conclusions For stage Ⅳ NSCLC,CCTTRT can prolong survival time with acceptable toxicity.Radiotherapy to thoracic primary tumor should be under consideration.

Background and purpose:When the patients with nasopharyngeal carcinoma (NPC) receive radiotherapy, their thyroids are inevitably involved. As a result, thyroid damage occurs. This study aimed to explore the effects of intensity modulated radiation therapy (IMRT) on dynamics of thyroid blood flow in patients with NPC.Methods:A total number of 68 patients with NPC were enrolled in the study who received primary treatment of radical radiation and chemotherapy from Jul. 2012 to Oct. 2013. And the TMN stage was fromⅡ toⅣc according to UICC 2010. The treatment method consisted of 2 cycles of TPF induction treatment, concurrent radiation therapy (IMRT) with 2 cycles of DDP and 2 cycles of adjuvant therapy sequentially. Before radiotherapy, at the end of radiotherapy, 3 and 6 months after radiotherapy, serum free triiodothyronine (FT3), free thyroxin (FT4) and thyroid-stimulating hormone (TSH) concentrations of all cases were detected by electrochemiluminescence. The highest systolic velocity, mean velocity, minimum diastolic velocity, resistance index, and the value of all thyroid diameter lines were measured by type-B ultrasound.Results:All the patients were followed up for 6 months. Hypothyroidism: the incidence of immediate clinical hypothyroidism after radiotherapy was 5.9%; 3 months later, the incidence was 13.2%; and 6 months later, the incidence was 26.5%. The difference in volume change between before radiotherapy and at the end of radiotherapy had no statistical signiifcance (P>0.05). The difference in volume change between 3 and 6 months after radiotherapy had statistical signiifcance (P<0.05). The difference in FT3, FT4 and FSH between the end of radiotherapy and before radiotherapy had no statistical signiifcance, while there was statistically signiifcant difference between at the end of radiotherapy and 3 months after radiotherapy. The thyroid volume correlated with the average dose at the end of radiotherapy, 3 and 6 months after radiotherapy as shown by the single factor correlation analysis (P<0.05). The results of sinlge factor correlation analysis also showed that the occurrence of hypothyroidism correlated with thyroid dose-volume parameter V40 at the end of radiotherapy (P<0.05). The correlation between hypothyroidism and the average dose on thyroid 6 months after radiotherapy was demonstrated by independentt test (P<0.05). Hypothyroidism had no correlation with thyroid artery systolic maximum velocity and resistance index at the end of radiotherapy, 3 and 6 months after radiotherapy (P>0.05).Conclusion:The incidence of hypothyroidism may increase with time after radiotherapy. The volume may decrease with the increased dose of radiotherapy and the follow-up time. The patients with NPC after radiotherapy should be tested for thyroid lesions routinely. The thyroid dose-volume parameter V40 may be a predictor for acute radioactive thyroid lesions. The study did not reveal temporarily that hypothyroidism was associated with thyroid ultrasound blood lfow velocity.