Objective:To compare the human ??T lymphocytes expanded with HSP 70BCG and HSP 70 m-peptide complexes with those expanded with solid phase antibody in vitro.Methods:PBMCs were cultured with anti-TCR ?? antibody?HSP 70BCG and HSP 70 m -peptide complexes.The total cell number was counted.The flow cytometer was used to analyze the lymphocytes phenotypes and subtypes.RT-PCR was used to analyze the level of V? mRNA.The cytotoxitic activity of ??T cells against Daudi was determined using MTT calorimetric assay.Results:After 14 days of culture,in the antibody culture system,the total cell number increased about 30-40 fold,and the ratio of ??T cell reached to 86.3%;In the HSP 70BCG culture system,the total cell number increased about 7-8 fold,and the ratio of ??T cell reached to 71.23%;In the HSP 70-peptide complexes culture system,the total cell number increased about 4-5 fold,and the ratio of ??T cells reached to 27.26%.The antibody and HSP 70BCG activated ??T cells possessed a whole repertoire and mainly expressed V?9/V?2 subset and exhibited a strong cytotoxicity against Daudi.Conclusion:Anti-TCR ?? antibody and HSP 70BCG were able to proliferate purer ??T cells.The cells had a whole repertoire,and exhibited strong cytotoxicity against Daudi.

Objective To compare the effects of different spleen-strengthening recipes on water-electrolyte metabolism and water transport in rat model of spleen-deficiency syndrome. Methods The male Wistar rats were randomly assigned into the normal group and spleen-deficiency model group. The rat model of spleen-deficiency was established by overstrain plus disordered diet method. On the 15th modeling day, the model rats were randomly assigned into model group , Buzhong Yiqi Tang(BYT) group, Si Junzi Tang(SJT) group, Shenling Baizhu San(SBS) group, and the rats except for the model group were treated with gastric gavage of the corresponding medicine in the dosage of 4.05 g·kg-1·d-1. Rats of normal group and model group were given equal volume of normal saline. The medication lasted for 14 days. On the 29th experimental day, we detected serum Na+ and K+, plasma antidiuretic hormone(ADH) and aldosterone(ALD), aquaporin 3(AQP3) in the small intestine and colon, and renal AQP2. Results Compared with the normal group, plasma ADH and ALD levels, serum Na+ level, renal AQP2 content in the model rats were increased significantly, serum K+ and AQP3 content of small intestine and colon mucosa was decreased significantly(P<0.01). Compared with the model group, plasma ADH and ALD levels, serum Na+ level, renal AQP2 content in the three medication groups were decreased significantly, serum K+level and AQP3 in the small intestine and descending colon mucosa was increased significantly(P<0.05 or P<0.01). Of the three medication groups, SBS group had lower ADH, ALD, Na+levels, and higher serum K+level and AQP3 in the small intestine and descending colon mucosa than BYT group and SJT group(P<0.01); SBS group and BYT group had lower renal AQP2 than SJT group(P<0.05 or P<0.01). Conclusion Rat model with spleen-deficiency syndrome has disordered water-electrolyte metabolism and abnormal water transport. The spleen-strengthening recipes, which have the actions of strengthening spleen, strengthening spleen to drain dampness, strengthening spleen to elevate yang, can improve the disorder of water metabolism to some degrees, and SBS has the best effect.

Objective To investigate the expression and regulation of ASPP family mRNA in NSCLC cell lines.and study the diagnostic and therapeutic significances of apoptosis stimulating of ps3 expression in NSCLC Patients.Methods Real-time fluorescence quantitative PCR wag established and used to measure the expression Ievels of ASPP mRNA in NSCLC cell lines A549(wild-type p53)and NCI-H157 (mutant-type p53)with p53 different genetic background,and ASPP mRNA was also detected in NSCLC tissue,peripheral blood of 37 NSCLC patients.Western Bloting was used to examine the levels of ASPP1 and ASPP2 proteins of NSCLC cell Iines.Chemosensitivity of the cell lines to CDDP was analyzed by MTT assay,and detect the change of ASPP1 and ASPP2.Compared with the ASPPs mRNA expression between NSCLC patients and healthy controls.and monitoring ASPPs mRNA expression in patients with NSCLC chemotherapy.Results The slope rotes of standard curves were-3.249,-3.358 respectively.and correlation coefficients were more than 0.98.Amplification efficiency of standard curves were 1.156.1.028 respectively.The IC_(50) values of NCI-H157 and A549 cells were 3.70.10.48 μg/ml respectively.Mean ASPP1 ASPP2 mRNA levels were a statistically significant 2.66 folds and 6.98 folds higher in NCI-H157 cell compared to A549 cell The levels of ASPP1 mRNA in tumor tissues and peripheral blood were (4.27±0.57)×10~3,(2.49±0.32)×10~3 in patients with NSCLC respectively,and the levels of ASPP2 mRNA in tumor tissues and peripheral blood were(2.34±0.75)×10~3,(7.00±1.17)×10~3 in Patients with NSCLC respectively.The levels of ASPP1 and ASPP2 mRNA in tissues and peripheral blood were(1.32±0.21)×10~4,(1.46±0.31)×10~4 and(1.38±0.19)×10~4,(1.28±0.18)×10~4 in control groups respectively.The ASPP1,ASPP2 mRNA levels in patients with NSCLC were significantly lower than those of control groups(t=2.58,3.94,3.62,3.76,P＜0.05).Before chemotherapy the levels of ASPP1 and ASPP2 mRNA in patients with NSCLc were(2.34±0.56)×10~3 and(6.64±0.72)×10~3,and after 2 periods of postoperative chemotherapy the levels were(3.66±0.64)×10~3 and(9.42±0.44)×10~3,the expression was higher than those of ehemotherapy before(t=3.02,4.50,P＜0.05).Conclusions The expression of ASPP1.ASPP2 mRNA in p53 mutant-type cell lines is higher than that in p53 wild-type cell lines.The expression of ASPP maybe has close relationship with chemotherapeutic sensitivity of NSCLC.In NSCLC patient chemotherapy process,the change of ASPP1 and ASPP2 mRNA expression may be related with chemotherapy medicine function.Enhance the expression of ASPP1 and ASPP2 will be beneficial to the tumot treatment.

Objective To evaluate the diagnostic value of endoscopic ultrasonography (EUS) for unexplained bile duct expansion in patients before retrograde cholangiopancreatography (ERCP). Methods Sixty patients with unknown causes of bile duct dilatation were included in this study. Patients were examined by abdominal ultrasound (TUS), CT and (or) magnetic resonance imaging (MRCP) suggesting the dilatation of common bile duct, suspecting biliary pancreatic disease with unknown cause. EUS diagnosis was performed before ERCP surgery. The final diagnosis was confirmed by ERCP, pathology and follow-up diagnosis. Results Thirty-nine patients were diagnosed as distal bile duct stone by ERCP, 38 were diagnosed by EUS, and one case was diagnosed as common bile duct bottom tumors by EUS. There were 10 cases were diagnosed as common bile duct bottom tumors by ERCP and surgical pathology, 2 cases were diagnosed as biliary papillomatosis, 2 cases were diagnosed as periampullary carcinoma. There were 11, 0 and 3 cases were diagnosed by EUS. One case was diagnosed as distal bile duct stone, which was diagnosed as common bile duct bottom tumor by EUS. Two cases were diagnosed as papillary tumor of the bile duct, one of which was diagnosed as inflammatory stenosis, another one was diagnosed as periampullary carcinoma by EUS. Results of postoperative follow-up confirmed that 7 cases were duodenal papilla inflammatory stenosis. Eight cases were diagnosed by EUS, one of them was followed up and pathologically diagnosed as biliary papillomatosis by ERCP. The diagnostic accuray was 95%(57/60). Conclusion EUS has higher value in the diagnosis of unexplained bile duct expansion, which especially can improve the diagnostic rate of distal bile duct stone compared with that of MRCP detection, and can guide selectively ERCP, improve the therapeutic effect, and reduce its risk .

A total of 226 strains of organisms was isolated from the cultures of the subeschar unburnt tissues of the burn patients admitted to this institute in the period from April 1980 to April 1982. Among the organisms, gram-negative bacilli exceeded gram-positive cocci in number. The frequently seen gram-negative bacilli, in the order of frequency, were Pseudomonas, Serratia, Klebsialla, and E. coli. And the frequently seen gram-positive cocci were Staphylococcus aureus, Staphylococcus albus, Streptococcus fecalis, and Streptococcus hemolyticus.The quantitative culture of the biopsy specimen showed its value in our clinical application. In cases of multiple infections, after the identification and precise count of the bacterial colonies on the cultures were done, the percentage of the various organisms could be obtained and the main pathogen was revealed.It was pointed out that ordinary culture media were only favorable for rapid growth of bacteria but the existence of fungi was usually masked. A. modified method of fungus culture, tissue thread culture, was used for the early diagnosis of fungus infection. 38 specimens were studied simultaneously with three methods. The positive rate for fungus was 8% in ordinary cultures, 26% in his-tologic examinations, and 61% in tissue thread cultures.Anaerobic culture was performed for 102 swab specimens from the burn wounds and a positive rate of 14.7% was obtained. In addition, anaerobic blood culture was performed in 10 cases of severe burns with 2 positive cultures. It is suggested that anaerobic infections should not be neglected in burns.

AIM: To investigate the influence of microwave-assisted extraction on flavone contents of Pollen Tyhae micropowder. METHODS: Ultraviolet spectrophotometer and HPLC were applied to analyze Pollen Tyhae micropowder, total flavon and isorhamnetin-3-O-neohespridoside were adopted as the marker, respectively. RESULTS: Appropriate conditions of microwave-assisted extraction included: extraction time of 8min, ethanol concentration of 70%, Solid/liquid ratio of 1∶18 (g?mL -1) and the power of microwave oven of 540w. CONCLUSION: Compared with normal reflux method, microwave-assisted extraction of Pollen Typhae micropowder is more useful and can improve the extraction rate the reduce the extracting time.

To investigate the relationship between platelet-leukocyte aggregates (PLA) and serum high sensitive C-reactive protein (hs-CRP)in type 2 diabetic patients with retinopathy (DR).The blood levels of platelet-neutrophil aggregates( PNA )and platelet-monocyte aggregates ( PMA ) were determined by flow cytometry and serum hs-CRP by ELISA in 30 patients with proliferative diabetic retinopathy( PDR group),30 patients with no-proliferative diabetic retinopathy( NPDR group),25 diabetic patients without DR( DM group),and 30 normal controls.The levels of PNA,PMA,and hs-CRP in PDR group were significantly higher than those in normal control,NPDR,and DM groups.The levels of PNA,PMA,and hs-CRP in NPDR group were significantly higher than those in normal control and DM group( P＜0.01 or P＜0.05 ).The PNA and PMA were correlated positively with hs-CRP in patients with DR ( r =0.587,0.576 ; P ＜ 0.05 ).The data showed that the levels of PLA and hs-CRP were raised significantly in patients with DR,and the levels of PLA were positively correlated with inflammatory response and the seriousness of DR,suggesting that these findings may play an important role in the development of DR.

Objective To study the diagnostic value of endoscopic ultrasonography (EUS)for biliary stenosis.Methods Data of 83 patients with bile duct stenosis who underwent EUS from January 2010 to June 2012 at endoscopic center of Tianjin Nankai Hospital were reviewed.The EUS qualitative diagnosis and final diagnosis,and the accuracy of EUS for the lower malignant bile duct stenosis were analyzed.Results Malig-nant stenosis were confirmed histopathologically in 55 patients;benign stenosis were confirmed in 28 patients based on negative tissue sampling or extended clinical follow-up.Sensitivity,specificity,positive predictive val-ue,negative predictive value and accuracy of EUS for malignant bile duct stenosis were 98.2%(55 /56), 81.5%(22 /27),91.7%(55 /60),95.6%(22 /23)and 92.8%(77 /83),respectively.Conclusion The sensitivity and accuracy of EUS for the lower malignant bile duct stenosis is high,which is of clinical value.

Objective To explore the risk factors and targeted therapy for fungus infection in acute exacerbation of chronic obstructive pulmonary disease (COPD) pulmonary.Methods 83 acute exacerbations of COPD patients with pulmonary fungus infection were selected as the observation object(COPD fungal infection group),according to the time sequence of random month by month in acute exacerbations of COPD no secondary fungal infection of the 80 patients hospitalized patients as control group,analysis of risk factors lead to fungal infection.Results Two groups in age,live ICU time,antibiotic use time,hormone use time,albumin level mechanical ventilation people with diabetes mellitus,merger,as cor pulmonale had statistically significant differences (P ＜ 0.05) ; The conditional Logistic multiariate analysis,age,ICU patients live time,antibiotics use time,hormone use time,albumin level was acute exacerbations of COPD factors such as pulmonary fungus infection independent risk factors(P ＜0.01) ;Fungus infection were combined with antimycotic,cured 69 cases (83.13％) improved 12 cases (14.46％),there was no change in 1 case(1.20％),death 1 case (1.20％).Conclusion Patients older age,live long,long time ICU antibiotics and hormones and the low level of albumin is acute exacerbations of COPD pulmonary fungus infection independent risk factors,timely diagnosis and prognosis of patients with antifungal treatment is good.

Objective To construct an Apoptin prokaryotic vector,aiming to produce antigenic fusion protein Apoptin. Methods The Apoptin gene was amplified from the template of plasmid pSSCHG/NT4-Apoptin-HA2-TAT by PCR.The Apoptin was sub-cloned into the multiple clone sites of plasmid pET-28a(+) to get the prokaryotic vector of pET-28a(+)-Apoptin,which was transformed into E.coli BL21(DE3).Expression of E.coli BL21(DE3) was induced by IPTG.The specific protein expression was detected by SDS-PAGE. Results The fusion protein was expressed with high efficiency in E.coli BL21(DE3) transformed by pET-28a(+)-Apoptin after induction with IPTG.The specific fusion protein had an apparent related molecular weight of about 17 000 ku as indicated by SDA-PAGE analysis. Conclusion The Apoptin prokaryotic expression vector with pET-28a(+)-Apoptin can effectively express Apoptin fusion protein,laying a foundation for further study of Apoptin and preparation of antibodies against Apoptin.