Objective To obtain and identify dendritic cell (DC) from mouse bone marrow in vitro. Methods Recombinant mouse granulocyte and macrophage colony stimulus factor (GM-CSF) induction was used to induce mouse bone marrow cells to form dendritic cells in vitro. The morphological changes were observed with light inverted microscope and scanning electron microscope (SEM). CD11c, MHCⅡ, and CD86 were identified with flow cytometry. The biological function was studied with antigen phagocytosis test and mixed lymphocyte reaction (MLR). Result The cultured DC from mouse bone marrow displayed the typical morphological characteristics of DC. Immature DC, which had high expression in CD11c and low expression in MHCⅡ and CD86, could phagocytize antigen. Mature DC, which had high expression in CD11c, MHCⅡ and CD86, could stimulate allogenic mixed lymphocyte proliferation. Conclusion DC can be generated from mouse bone marrow cells through cytokine induction in vitro and be used for further study associated with DC.

Objective To investigate the diagnostic value of carotid artery ultrasonography,CT angiography (CTA)and digital subtraction angiography (DSA)for carotid artery dissection. Methods The image data of carotid artery ultrasonography,CTA,and DSA of 24 patients with carotid artery dissection were analyzed retrospectively. Results Twenty-four,16,and 21 patients were examined with DSA,CTA,and carotid artery ultrasonography respectively. The detection rates of carotid artery dissection with DSA,CTA, and carotid artery ultrasonography were 95. 8%,75.0%,and 71. 4% respectively. The DSA mostly showed the line-like sign (n=12,50 %). CTA and carotid artery ultrasonography mostly showed the double lumen sign;they were 37. 5%(n=6)and 52. 4%(n=11)respectively. Compared with DSA,the concordance rates of carotid artery ultrasonography and CTA were 66. 7% and 81. 3% respectively. There was no significant difference (Kappa=0. 39,P=0. 08 and Kappa=0. 43,P =0. 22 respectively). The concordance rate of ultrasonography in combination with CTA and DSA reached 87. 5%(n=15,Kappa=0. 67,P =0.047). There was significant difference. Conclusion DSA is a gold standard for the diagnosis of carotid artery dissection,and it is irreplaceable. Carotid artery ultrasonography in combination with CTA can improve the diagnostic rate. Carotid artery ultrasonography can be used as a screening method for carotid artery dissection.

Purpose To investigate the clinicopathologic changes of the lung epithelial tumor with epithelioid granuloma. Methods E-leven cases of lung malignant tumor with epithelial epithelioid granuloma reaction were studied according to the histological changes of lung epithelial tumor, immunophenotype, acid-fast staining, and tuberculosis PCR test results combined with follow-up data. Results The mean age was 62. 3 years, and male to female ratio was 10∶ 1. The main clinical manifestations displayed a cough, chest pain, fever and hemoptysis or bloody sputum. Nine cases were malignant tumors, tuberculosis or inflammatory lesions was 1 case each in pre-vious surgery. The tumor with epithelioid granuloma formation of the 11 cases displayed in the same site ( mixed) . The histopathologic changes of the lesions showed a epithelioid or typical tuberculous granulomas formation, including 4 cases of adenocarcinoma, 5 cases of squamous cell carcinoma and 2 cases of neuroendocrine tumors. The acid-fast bacilli staining was positive in 3 cases;6 cases were positive for PCR detection of mycobacterium tuberculosis. 5 patients died within 2 years due to various causes. Conclusions The lung epithelial tumors with epithelioid granuloma is more commonly found in the middle-aged, The signs and symptoms of the patients are similar to primary epithelial malignant tumors of the lung . The biological behavior was progressive with poor prognosis. The diagnosis of the lesions depends on pathological examination. It is very important to pay more attention to the pathological examination, especially in lung puncture samples, and the differential diagnosis for a definate diagnosis and treatment.

Objective To explore the distribution of specific allergens for children in Sichuan Province and provide the evidences of clinical diagnosis and treatment.Methods Totally 451 cases in West China Second University Hospital,from April to August of 2016,were detected for 20 kinds of allergen-specific immunoglobulin E (IgE) by a new reverse enzyme-allergosorbent test,SPSS 18.0 statistical analysis was used to evaluate allergen-specific IgE levels in different groups of age,gender and diverse population distribution of disease.Results A total of 282 cases among 451 subjects were detected as IgE positive and the positive rate was 62.53%,among which 248 cases were positive for inhaled allergen-specific IgE,and the positive rate was 54.99%.There were 261 positive cases in food allergen-specific IgE,and the positive rate was 57.87%.For inhaled allergens,the highest positive rate was dermatophagoides pteronyssinus (n=111,24.61%),then dermatophagoides farina (n=110,24.39%),and house dust (n=108,23.95%);For food allergens,the highest positive rate was milk (n=159,35.25%),then egg (n=111,24.61%),and house dust (n=108,23.95%).Conclusions Both Inhaled allergens and food allergens were the important cause of allergic disease for children in Sichuan area.Serum allergen specific IgE determination was helpful to understand the patient′s allergy status,and diagnose the allergic disease.

Standerdized patiets oan serve as patients, teachers and evaluators. Establishment of standardized patients' teacher-identity is helpful to improving their teaching passion, and it also makes sure all teaching steps may develop successfully and promote accuracy and fairness of evaluation in Standerdized patiets examinations.

Objective:To observe the effects of auto-dendritic cell vaccine on T cells of patients with condyloma acuminatum(CA) related human papilloma virus(HPV) and its clinical outcome.Methods:73 patients with condyloma acuminata(CA) in the present study were positive of HPV-DNA and have been unsuccessfully treated with freezing,laser,or electric therapy combined with interferon anti-virus treatment for half a year.The peripheral blood samples were harvested from patients and the monouclear cells were extracted to cultivate DCs in presence of cytokines.Auto DCs were sensitized with corresponding HPV virus antigen of a patient to prepare the auto-dendritic cell vaccine.Then the vaccine were cultivated with auto T-cells and the proliferation of T cells were examined.Meanwhile,the auto-DC vaccine(1-4)?107 was also injected into the patients' inguinal lymph glands and its therapeutic effectiveness was observed.Results:The prepared DC-vaccine significantly promoted the proliferation of auto T cells compared with control group(P

Objective To construct an eukaryotic expression vector pcDNA-UCA1a(CUDR)and to observe its expression in bladder cancer UM-UC-2 cells, and to provide experimental basis for study on the relationship between UCA1a(CUDR)gene and bladder cancer.Methods Human total length of UCA1a(CUDR)gene was obtained from the 5′-RACE-Ready cDNA of bladder cancer BLZ-2 1 1 cells by PCR and was inserted into pcDNA3.1 (+)vector.pcDNA-UCA1a(CUDR)was identified by digestion with EcoRⅠ and BamHⅠ.The bladder cancer UM-UC-2 cells were transfected stably with the constructed eukaryotic expression vector pcDNA-UCA1a(CUDR). The expressions of UCA1a(CUDR)gene in the UM-UC-2 cells transfected with pcDNA-UCA1a(CUDR)and the UM-UC-2 cells transfected with pcDNA3.1(+)(control vector)were detected by RT-PCR.Results The inserted fragment with 2 200 bp was successfully amplified, which was in accordance with the expected results. The eukaryotic expression vector pcDNA-UCA1a(CUDR)was constructed successfully after identified by double enzyme digestion and sequencing.The RT-PCR results showed that the expression of UCA1a(CUDR)gene in the cells transfected with pcDNA-UCA1 a (CUDR ) was significantly increased compared with the cells transfected with pcDNA3.1 (+). Conclusion The eukaryotic expression vector pcDNA-UCA1a (CUDR ) is successfully constructed.The UCA1a(CUDR)gene highly expresses in the UM-UC-2 cells transfected with the expression vector.

Objective To investigate the clinical significance of serum CEA , CA19-9, CA72-4, CA242, CA50, PGⅠand PGR ( PGⅠ/PGⅡ) in the Diagnosis of preoperative and postoperative gastric cancer.Methods Retrospective study.The levels of CEA , CA19-9, CA72-4, CA242, CA50, PGⅠand PGⅡin serum of 41 patients with gastric cancer preoperative and postoperative and 60 healthy people were detected by AutoLumo A2000 chemiluminescence immunoassay and compared.Statistical analysis was performed using Rank-sum test by SPSS 17.0.Results The median of CEA, CA19-9, CA72-4, CA242, CA50, PGI, PGII and PGR in postoperative gastric cancer group were 3.79 ng/ml, 17.85 U/ml, 3.50 U/ml, 14.52 U/ml, 17.62 U/ml, 32.81 ng/ml, 11.48 ng/ml, 3.35.The postoperative gastric cancer group were 1.67 ng/ml, 7.76 U ml, 1.73 U/ml, 6.30 U/ml, 7.57 U/ml, 20.56 ng/ml, 5.71 ng/ml, 2.94.The healthy group were 1.53 ng /ml, 7.59 U/ml, 1.47 U/ml, 6.08 U/ml, 5.68 U/ml, 90.86 ng/ml, 14.85 ng/ml, 6.67.There were statistical differences in the serum levels of CEA , CA19-9, CA72-4, CA242, CA50, PGⅠ, PGⅡand PGR among different groups (chi-squared values were 79.108, 20.678, 20.374, 7.252, 56.73, 131.212, 20.38, 86.37, P<0.05).By the Mann-Whitney rank sum test,the serum levels of CEA , CA19-9, CA72-4, CA242 and CA50 in patients with preoperative gastric cancer were significantly higher than those in healthy controls (Z values were -8.598, -4.425, -4.365, -2.000,-7.420, P<0.05).The level of postoperative group was significantly lower than that of preoperative group (Z value were -4.641, -2.383, -2.459, -2.399, -2.903, P<0.05).The serum PGⅠ, PGⅡand PGR levels in patients with preoperative gastric cancer were significantly lower than those in healthy controls (Z values were -10.309, -2.695, 8.637, P<0.05).The PGⅠlevel in the postoperative group was significantly lower than that in the preoperative group (Z value was -2.109, P<0.05).PGⅡ,PGR levels of postoperative group were lower than those of preoperative group , but the difference were not statistically significant.(Z values were -1.506,-0.838, P values were 0.132,0.402).Conclusion The detection of the seven serum markers can help to preoperative diagnosis and postoperative monitoring of gastric cancer .

BACKGROUND: According to present theories and our clinical experience, immune ablative and tolerance inducing theory is proposed. Immune ablative means to clear out mutate cell clones and without transfusion of hemopoietic stem cells afterwards; intolerance inducing means to induce animal models not to react to mutate somatic cells, which avoids relapse or new occurrence of autoimmune disease. OBJECTIVE: To explore the effects of immune-ablative and tolerance inducing therapy in treating animal model of immune polymyositis (PM). DESIGN, TIME AND SETTING: Randomized, controlled animal experiment was performed at the Animal Experimental Center of Nanfang Hospital from December 2008 to April 2009. MATERIALS: One New Zealand rabbit, female, weighing 4.1 kg and 36 England guinea pigs, female, weighing 400-500 g, were used. METHODS: New Zealand rabbit's muscle tissue homogenate and complete Freund's adjuvant (CFA) were injected into guinea pigs to make PM animal models. The 28 animal models were randomly divided into intense immune-ablative and tolerance inducing group (Busulfan 1 mg/kg, every 12 hours, totally 8 doses; followed by CTX 40 mg/kg per day for 4 days; then cyclosporine A (CsA) 3 mg/kg per day was given till animals were dead); cyclophosphamide (CTX) group: CTX was given, 10 mg/kg per day for 3days; immune-ablative and tolerance inducing group: Busulfan 0.8 mg/kg, CTX 30 mg/kg, CsA 3 mg/kg; the administration time and dose were the same as group 1. Control group was not treated.MAIN OUTCOME MEASURES: Full blood count (FBC) and biochemical index were tested before and after treatment, and surviving time was recorded. In addition, muscle pathological changes were observed.RESULTS: Compared with control group, number of white cells was significantly decreased in the other groups, and hematopoiesis function gradually restored after administration. The number of white cells in the immune-ablative and tolerance inducing group was the most, and striated muscle pathology showed PM. Following administration, the glutamic oxaloacetic transaminase and creatine kinase of intense immune-ablative and tolerance inducing and immune-ablative and tolerance inducing groups were significantly reduced (P < 0.05, P < 0.01), but no obvious striated muscle pathological changes were found. The glutamic oxaloacetic transaminase, lactic dehydrogenase and creatine kinase in the CTX and control groups remained unchanged. Survival time of intense immune-ablative and tolerance inducing group was the shortest among all groups, and there was no significant difference between CTX and control groups. The animals in immune-ablative and tolerance inducing group survived for the longest time. CONCLUSION: Immune-ablative and tolerance inducing therapy has preferable effect on treating animal models of PM, and its prognosis is better than intense immune-ablative and tolerance inducing therapy and regular CTX therapy.

Objective To study the serum levels of progastrin-releasing peptide (pro-GRP) and neuron-specific enolase (NSE) for the clinical diagnosis,therapy monitoring and survival time analysis of small cell lung cancer (SCLC).Methods All 41 SCLC samples (30 males,11 females,age range from 46 to 78 years),95 NSCLC samples (55 males,40 females,age range from 42 to 88 years),and 127 normal individuals samples (80 males,47 females,age range from 35 to 78 years) which were diagnosed by People's Hospital of Zhengzhou from May 1,2008 to April 30,2011 were collected.Serum levels of pro-GRP,NSE and their changes in SCLC patients before and after therapy were evaluated.ANOVA analysis,randomized block design analysis of variance and the log-rank test were collected SPSS 16.0 to evaluate the survival time.Results The serum levels of pro-GRP (median 357.8 ng/L) and NSE (median 89.5 μg/L) in SCLC group were significantly higher than those in the NSCLC group (pro-GRP:39.9 ng/L;NSE:11.43 μg/L) and normal individuals group (pro-GRP:12.7 ng/L;NSE:10.03 μg/L) (P=0.000).The sensitivity of pro-GRP and NSE for the diagnosis of SCLC were 80.4％ and 78.0％,while the specificity were 92％ and 87％,respectively.There is a poor correlation between pro-GRP and NSE serum levels,but when combined the sensitivity can be 95％ and specificity can be 85％.Significantly statistical difference of pro-GRP levels was observed in the different stages of treatment (before and after therapy) in SCLC-LD patients (F =3.53,P =0.038),and significant statistical difference of NSE levels was also observed in SCLC-ED patients in different stages (F =16.049,P =0.000).In partied response SCLC patients,the group with NSE level lower than cut-off value had longer survival time than the other group with NSE level higher than cut-off value (P =0.001).Conclusions The sensitivity of the combined analysis of pro-GRP and NSE is better than single marker for the diagnosis of SCLC.The serum level of pro-GRP has better correlation with therapeutic effect of SCLC-LD patient than NSE.The serum level of NSE are well correlated with therapeutic effect in SCLC-ED patients.There are some certain value of NSE level for evaluation the survival time of SCLC patients who were in partial response.