Objective:To investigate the effect of esculentoside (EsA) on apoptosis of murine thymocyte. Methods:Using electronic microscope, DNA agarose electrophoresis and flow cytometry analysis,the effect of EsA on apoptosis of murine thymocyte was examined. Results:The result showed that apoptosis of activated thymocyte by ConA was markedly promoted by 2.5, 5, 10 ?g/ml EsA in murine thymocyte culture for 3 h, but the spontaneous apoptosis was not affected by EsA. Conclusion:The results suggest that EsA has the positive effect on apoptosis of activated murine thymocyte.

Objective: To investigate the influence of esculentoside A（EsA） on production of IL-1 and TNF by rabbit synovial cells induced by LPS. Methods: levels of IL-1 and TNF in the supernatant of rabbit synovial cell were determined by examining proliferation of thymic cells and by bioassay L929 cells as target cells, respectively. Results: EsA in 5-40 μg/ml could significantly inhibit the production of IL-1 and TNF from rabbit synovial cells induced by LPS. Conclusion: EsA can inhibit the production of IL-1 and TNF from synovial cells. It suggests that EsA may play a role in improving the rheumatoid arthritis.

Objective To investigate and confirm the value of minimal invasion of separation between specific subcutancous layers of rats.Methods Thirty-two male SD rats were randomized into four groups,group 1:normal incision plus separation beneath superficial fascia; group 2:normal incision plus subdermal separation; group 3:normal incision plus extended separation beneath superficial fascia ; group 4:extended incision plus separation beneath superficial fascia.Blood sampling was taken preopeatively,2 h,12 h,24 h and 48 h postoperatively.The increase of serum IL-6 and neutrophil elastase levels were compared between different groups.Results Compared with each other,the increase of serum IL-6 and NE levels in group 1 vs 3 and group 2 vs 4 was not different significantly (P value was 0.074,0.096 and 0.747,0.897,respectively).However,the increase of serum levels in group 1 vs 2,1 vs 4,3 vs 2 and 3 vs 4 was different significantly (P ＜ 0.01).Conclusion Operation of separating beneath superficial fascia plus small skin incision in rats conduces to relatively minimal invasion.

AIM: To investigate the effects of interferon-alpha (IFN-?) on the development of dendritic cells (DCs) derived from bone marrow mononuclear cells in patients with chronic myeloid leukemia (CML). METHODS: Bone marrow mononuclear cells from 12 CML patients were cultured initially using cytokines as follows: recombined human granulocyte-macrophage colony stimulating factor (rhGM-CSF) plus IFN-? (IFN-?-DCs); rhGM-CSF plus recombined human interleukin-4 (rhIL-4) (IL-4-DCs); IFN-? alone; rhGM-CSF alone in 10% FBS RPMI-1640 medium for 7 days and then recombined human tumor necrosis factor-? (rhTNF-?) was added for another 3 days. The morphologic features were observed by Wright's staining under inverted microscope. CD_ 80,CD_ 86,CD_ 83,CD_ 1a and HLA-DR expression were assayed by flow cytometry. Cytogenetic analysis was performed for one CML patient by fluorescence in-situ hybridization (FISH), and the functions of antigen presenting were tested by mixed lymphocyte reaction (MLR). RESULTS: IFN-?-DCs displayed features in morphology that was similar to those of IL-4-DCs with delicate membrane projections. IFN-?-DCs showed an increase in expression of CD80, CD86, CD83, HLA-DR and more intense abilities of allogeneic antigen presentation with and without rhTNF-? stimulation, compared with the control groups of IL-4-DCs. FISH confirmed the DCs of both groups were leukemic origin. CONCLUSIONS: (1) IFN-? promoted the differentiation/activation of bone marrow mononuclear cells from patients with CML into activated dendritic cells. (2) The phenomenon of generation of activated DCs in vitro might contribute to therapeutic effect of IFN-? in CML. (3) IFN-? may be valuable for the generation of active bone marrow mononuclear cells-derived DCs to be as vaccination strategies of CML patients.

Objective To investigate the detection of monocyte chemotactic protein 1 (MCP-1),macrophage stimulating protein (MSP) and carcinoembryonic antigen (CEA) in differential diagnosis of pulmonary tuberculosis and lung cancer.Methods Thirty four patients with pulmonary tuberculosis,45 patients with pathologically confirmed lung cancer admitted in Quzhou People' s Hospital during December 2009 and December 2011,and 30 healthy controls were enrolled in the study.MCP-1 and MSP in serum and pleural effusion were determined by enzyme linked immunosorbent assay (ELISA),and CEA was detected by chemiluminescence method.Receiver operating characteristic method was used to determine the cut-off values of MCP-1,MSP and CEA in diagnosis of pulmonary tuberculosis or lung cancer.Results Serum MCP-1,MSP and CEA levels in pulmonary tuberculosis patients and lung cancer patients were higher than those in healthy controls.Compared with lung cancer patients,patients with pulmonary tuberculosis had higher serum MCP-1 and lower CEA levels (t =2.69 and 0.89,P ＜ 0.05),but there was no significant difference in serum MSP levels between two groups (t =2.89,P ＞ 0.05).While in pleural effusion,patients with pulmonary tuberculosis had higher MCP-1 level (t =3.54,P ＜ 0.05),lower MSP and CEA levels than those with lung cancer (t =3.47 and 3.48,P ＜ 0.05).Serum MCP-1 level was of the highest specificity (95.6％) with the cut-off value of 240 pg/mL in diagnosis of pulmonary tuberculosis,while MSP level in pleural effusion was of the highest specificity (94.1％) with the cut-off value of 1100 pg/mL in diagnosis of lung cancer.Conclusion Detection of MCP-1,MSP and CEA in serum and pleural effusion can be used for the differential diagnosis of pulmonary tuberculosis and lung cancer.

Objective To investigate the clinical application of electron-beam CT(EBCT)in the diagnosis of congenital cardiovascular diverticula. Methotis Retrospective analysis of 9 patients with congenital cardiovascular diverticula confirmed by operation and pathology was done.Of them,enhanced continuous volume scan was performed on 8 patients and enhanced single slice scan was performed on one patient with an Imatron C-150 scanner.Results The group of 9 pailents included one patient with diverticulum of the left ventricle.3 patients with diverticulum of the atria and 5 patients with diverticulum of the aorta.EBCT scan and three dimensional reconstruction could demonstrate not only the origin,size,shape,Location and adjacent structure of diverticula,but also other important complicated abnormalities such as ventrieuloarterial connection disorder,cardiac sepud defect,aortic coarctation and even dissection.Conclusion EBCT is an ideal noninvasive technique in the diagnosis of congenital cardiovascular diverticula

Objective To compare the effects of fundus-first laparoscopic cholecystectomy and laparoscopic subtotal cholecystectomy in complicated cholecystolithiasis cases. Methods The effects of fundus-first laparoscopic cholecystectomy (n = 21) and laparoscopic subtotal cholecystectomy (a = 18) in the 39 cases of complicated chole-cystolithiasis from our hospital within 2 years were analyzed retrospectively. Results The operation time in subtotal laparoscopic cholecystectomy group was shorter than in fundus-first laparoscopic cholecystectomy group (88.89±18.11) min vs. (109.52±21.79) min, P < 0.05). Less blood lose (82.78±44.96) ml and fluid replacement (847.22±169.32)ml during the operation were observed in the former group than those in the later group (116.67±53.23) ml and (964.29±147.60) ml, respectively, P < 0.05). However, the patients' postoperative recovery time and the duration of postoperative hospital staying were similar in the two groups(5.56±1.20) days vs. (5.29±1.38) days, P > 0.05). Conclusions Proper use of subtotal laparoscopic cholecystectomy in complicated cholecystolithiasis cases can simplify the operation and obligate the operation time, which will increase the safety of the operation with the outcome similar to fundus-first laparoscopic cholecystectomy.

OBJECTIVETo develop a simple, rapid, accurate, and cost-effective single-0tube multiplex polymerase chain reaction (PCR) assay, which could be used for molecular screening and prenatal diagnosis, for detection of three commonest deletional alpha-thalassemias (-- (SEA), -alpha (3.7) and -alpha (4.2)) in Chinese population.

METHODSFour groups of primers were designed on the basis of gap-PCR, and the PCR reaction condition was optimized systematically with the purpose of amplifying effectively specific DNA fragments that are indicative of the respective genotypes of these three deletional alpha thalassemias. In addition, a pair of primers was designed to amplify LIS1 3' untranslated region (UTR) fragment for use as a separate control for amplification running. A total of 72 blood and prenatal archival DNA samples with various known alpha thalassemia genes or normal alpha globin gene sequence that had been confirmed by Southern blotting analysis or DNA sequencing were collected to test the specificity of this assay by blind analysis. In addition, DNA samples from nine couples at high risk of alpha thalassemia were also analyzed to evaluate the reliability of this technique in prenatal implementation.

RESULTSHomozygote, heterozygote and double heterozygote of the three commonest deletional alpha thalassemias were well detected simultaneously by this established method. For normal allele, a 2.4 kb amplified band as a systematic control and an alpha (2) gene-specific amplicon of 1.8 kb were produced. Besides the two amplified fragments of normal allele, it was found that a 1.3 kb, a 2.0 kb or a 1.6 kb amplified band could be simultaneously shown for representing --(SEA), -alpha (3.7) and -alpha (4.2) alleles, respectively, in the heterozygous states. In a blind test, this technique accurately detected 100% of the DNA samples previously characterized by Southern blotting or DNA sequencing, and it was successfully applied to prenatal diagnosis of alpha thalassemia in nine at-risk families.

CONCLUSIONThe single-tube multiplex PCR protocol presented in this study is easy-to-handle, rapid, reliable and is cost-effective for detecting --(SEA), -alpha (3.7) and -alpha (4.2) chromosomes, and it is suitable for large-scale population screening and for rapid molecular genotyping in clinics.

Asian Continental Ancestry Group ; genetics ; China ; Female ; Heterozygote ; Homozygote ; Humans ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis ; Reproducibility of Results ; Sensitivity and Specificity ; alpha-Thalassemia ; diagnosis ; ethnology ; genetics