Objective To express human inducible eostimulator (ICOS) extracellular region and IgG Fc fusion protein, and analyze their function in allogenie lymphocyte proliferation in vitro and in vivo. Methods Human ICOS extraeellular region and IgG Fc fragment were cloned into a soluble expression vector. ICOS-Ig fusion protein was expressed and purified in CHO cells. To monitor primary MLR, Balb/c spleen T cells were isolated as responder cells, and irradiated C57BL/6 spleen cells as stimulator cells. 50 μg/ml ICOS-Ig or IgG was added to primary MLR cultures. The cells responsive rates were detected by 3 H-TdR methods. ELISA tested supernatants for eytokines (IL-2,IL-4, IL-10 and IFN-γ). T cells of each group in primary MLR were cultured as responder cells for secondary MLR, and irradiated C57BL/6 (donor) or C3H (third party) spleen cells as stimulator cells. Similar indexes were detected in secondary MLR. Then vital dye CFSE was used to study alloreactive T cell proliferation in vivo. CFSE-labeled C57BL/6 spleen cells were transferred to irradiated Balb/c mice. Mice were then intraperitoneally injected with 0. 2 mg IgG, ICOS-Ig or CsA each day.At the 3rd day after transfection, the spleen cells of the mice were harvested to detect CD4+ CFSE+ and CD8+ CFSE+ by FACS. Results In primary MLR, ICOS-Ig inhibited allogenic T-cell proliferation with inhibition rate being (58 ± 8)% in 50 μg/ml, and increased IFN-γ secretion. In secondary MLR, ICOS-Ig specifically inhibited the proliferation of donor spleen cells with inhibition rate being (42±8)%, and in ICOS-Ig group the levels of IL-4 and IL-10 were lower and the level of IFN-γ higher than in IgG group. However, ICOS-Ig didn't inhibit the proliferation of third-party spleen cells. In the CFSE dye assay, CFSE intensity of CD4+ and CD8+ T cells in ICOS-Ig and CsA groups was stronger than that in control group (P < 0. 05), while CFSE intensity in combined treatment group were even stronger than that in ICOS-Ig and CsA groups (P<0. 05). Conclusion ICOS-Ig could inhibit allo-reactive T cell proliferation in vitro and in vivo, and induce donor-specific T cell hyporesponsiveness specifically.

Objective To understand laboratory diagnosis and clinical characteristics of human immunodeficiency virus (HIV ) infection and AIDS patients in three comprehensive hospitals .Methods Laboratory diagnosis and clinical characteristics dates of people living with HIV/AIDS patients consulted in Chongqing Emergency Medical Center between 2007 to 2013 were retrospective‐ly analyzed .Results Totally 47 355 cases were carried HIV antibody screening during 7 years ,179 cases of HIV antibody were positive in preliminary screening ,171 cases were confirmed as positive .Among 5 cases of HIV antibodies result unconfirmed ,2 ca‐ses were followed up ,1 case was ruled out HIV infection ,1 case was converted to HIV antibody positive .People living with HIV al‐ways merging double or multiple infection with hepatitis b virus (HBV) ,hepatitis c virus (HCV) and treponema pallidum (TP) and so on .People living with HIV aged from 18 to 86 years old ,9 .36% was over 60 years old .Most patient has two or more clinical manifestations when consulted a doctor .Conclusion There were false‐positive of HIV antibody preliminary screening ,HIV anti‐body positive results must be confirmed by Western blot confirmatory test .Uncertainty of HIV antibody results should be judged by regularly follow‐up or combining with other detection methods ,epidemiological data .Routine HIV antibody screening should be adopt for HBV ,HCV and TP infection .Elder patients should not be ignored .Clinical specificity of HIV/AIDS is not strong ,it is need to be valued and identified from other cause similar symptoms of diseases caused by phase identification ,in order to reduce missed diagnosis and misdiagnosis .

Objective:To investigate the etiololgy and pathogenesis of cholesterol granuloma of the marillary sinus. Method:Five patients with cholesterol granuloma of the maxillary sinus were analysed retrospectively. Result: The cholesterol granuloma was a particular form of tissue reactions of granulation. The clinical symptoms of the disease was short of specificity. Conclusion; The pathogenesis of cholesterol granuloma includes hyperlipemia, obstruction of airfilled cavity ventilation and impaired drainage. The final diagnosis was based on pathologic examination. The treatment should begin with the intervention and treatment of hyperilpemia. Radical cure can be a-chieved by operation.

Humans ; Sarcoma ; diagnosis ; Skin Neoplasms ; diagnosis

Objective To investigate residual hearing of children with sensorineural hearing loss in whom wave V was not found in ABR testing and to emphasize the importance of behavioral audiometry in determining the residual hearing.Methods Residual hearing obtained by behavioral audiometry of 101 children with SNHL was studied in relation to the absence of wave V in both click-ABR and tone burst-ABR tests.Results All children have residual hearing of different degrees at different frequencies.There appeared to be a higher percentage of lowfrequency residual hearing than middle and high frequencies.Also,the residual hearing at low frequencies appeared to be better than those in the middle and high frequencies.Average residual hearing thresholds in the right ears from 500 to 4 000 Hz were 106.81±7.13,110.00±7.90,111.78±5.22,112.06±7.08 dB HL and those in the left ears were 98.01±3.98,111.30±7.18,112.06±7.08,108.33±7.23 dB HL.Conclusion The absence of wave V in ABR does not mean total deafness.For those children with no wave V in ABR,behavioral audiometry must be conducted to determine children's behavioral hearing thresholds in order to know their residual hearing.

Objective To investigate the therapeutic effect and safety of the clip with the flossmethod during endoscopic submucosal dissection for early gastric angle cancer. Methods A total of 27 gastric angle lesions diagnosed as early gastric cancer were treated by ESD. They were randomized to two groups, routine ESD group and clip with the flossgroup. The procedure time, complication events, en-block resection rate and complete resection rate were compared between the two groups. Also,the learning time was divided to two stages and the learning curve was studied according to the resected specimen areas per minute. Results The en block rate was 85. 7%(12/14) in the routine ESD group and 100. 0%(13/13) in the clip with the floss group. The procedure time in the clip with the flossgroup was significantly less than that in the routine ESD group (the median time 30 min VS 40 min, P =0. 011) . Perforation and the post operative bleeding did not occur in either group. The ESD learning curving during the first learning period and the mean resected specimen (area/min) in theclip with the floss group were larger than routine ESD group(30±6 mm2/min VS 20±5 mm2/min,P=0. 01). However, no difference presented during the second learning period between the two groups. Conclusion Clip with the flossmethod during endoscopic submucosal dissection for early gastric angle cancer as a novel procedure is safe, efficacious and worthy to recommend to beginning learners.

OBJECTIVETo study the inhibitory effect and mechanism of Ganoderma lipsiense extract (GLE) on the growth of triple-negative breast cancer (TNBC) cell line MDA-MB-231-HM in a mouse model.

METHODSThe mouse model of TNBC was established by subcutaneous injection of 1.5 x 10(6) of MDA-MB-231-HM cells into BALB/c-nu mouse. Twenty successfully modeled mice were divided into the GLE group and the negative control group according to random digit table, 10 in each group. GLE (0.2 mL 100 mg/mL) was peritoneally injected to mice in the GLE group, while equal dose of normal saline was peritoneally injected to mice in the negative control group. The medication was administered once per 3 days and discontinued after 45 days. The CD34 expression was detected using immunohistochemical assay for counting microvessels. Meanwhile, expressions of thrombospondin 1 (TSP-1) and cyclin D1 were detected using immunohistochemical assay.

RESULTSThe average weight was obviously lower in the GLE group than in the negative control group [(0.33 ± 0.16) g vs (0.68 ± 0.37)g, P < 0.05]. The tumor inhibition rate was 51.4% in the GLE group. The volume of transplanted tumor was obviously lesser in the GLE group than in the negative control group (P < 0.05). Results of immunohistochemical staining showed, the microvessel density (MVD) under every field was (20.7 ± 2.1), TSP-1 positive cell count was (66.2 ± 9.2), cyclin D1 positive cell count was (33.8 ± 16.4) in the GLE group, and they were 34.0 ± 2.0, 24.0 ± 6.6, and 168.2 ± 32.6, respectively in the negative control group. There was statistical difference in all indices between the two groups (P < 0.05).

CONCLUSIONGLE could inhibit malignant proliferation of tumor cells by suppressing angiogenesis of blood vessels in tumor tissues and regulating cell cycles, thereby inhibiting TNBC.

Animals ; Biological Products ; pharmacology ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Disease Models, Animal ; Ganoderma ; chemistry ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Random Allocation ; Thrombospondin 1 ; metabolism ; Triple Negative Breast Neoplasms ; drug therapy

24 strains obtained from root nodules of Psoralea corylifolia, Pueraria lobata, and Campylotropis macrocarpa of Yunnan province were studied with numerical taxonomy and 16S rDNA PCR-RFLP. Results of numerical taxonomy indicated that all strains included 10 reference strains were divided into 3 groups at 84% similarity. Group III is an unknown group with no reference strains. Group I is slow-growing kind, and group II fast and middle-slow-grower. The dendrogram derived from 16S rDNA PCR-RFLP showed that all strains divided into five phylogenetic branches at the similarity of 70%. They are branches I and V with no reference strains, Agrobacterium-Sinorhizobium-Rhizobium, Mesorhizobium and Bradyrhizobium. Not all results of numerical taxonomy are accord with 16S rDNA PCR-RFLP, and 2 strains at the same group with A. tumefaciens IAM13129T.

Human ScFv against botulinum neurotoxin serotype A(BoNTa) was modified by fusing human IgG1 Fc to C terminal of ScFv. ScFv-Fc fusion protein was expressed at high level over 30% of total host cell proteins in E.coli. Recombinant protein existed in inclusion body form. Renatured ScFv-Fc was purified to 90%~95% by Protein G Sepharose column. In vitro ScFv-Fc could bind specific to toxiod BoNTa in ELISA. Recombinant ScFv-Fc had similar relative affinity to parent ScFv and had improved stability.

Objective To determine if Shenfu injectio(SFI)has any analeptic action in patients emerging from general anesthesia.Methods Eighty-six ASAⅠorⅡadult patients undergoing elective abdominal surgery under general anesthesia were randomly divided into 2 groups(n=43 each):SFI group and control group.The patients were premedicated with intramuscular phenobarbital 0.1g and atropine 0.5mg.Anesthesia was induced with propofol 2 mg?kg~(-1),fentanyl 4-5?g?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with propofol infusion(2-4 mg?kg~(-1)?h~(-1)),0.5%-1.0% isoflurane inhalation and intermittent i.v.boluses of fentanyl and vecuronium.The patients were intubated and mechanically ventilated.The propofol infusion and isoflurane inhalation were stopped during skin closure.The patients were still unconscious and on mechanical ventilation at the end of surgery and transferred to PACU with a tube in trachea.As soon as the patients reached the PACU,SFI 1 ml?kg~(-1) in Ringer's solution 100 ml was infused over 10 min.In control group the patients received Ringer's solution 100 ml without SFI.The following times were recorded:(1)the time when the patients opened their eyes on command;(2)the time when mechanical ventilation was stopped;(3)the time when oxygen inhalation was stopped;(4)the extubation time;(5)the time of staying in PACU.Venous blood samples were taken before(T_0) and 5,15 and 45 min(T_(1,2,3))after SFI infusion for determination of plasma?-endorphin concentration.Results The awakening time,the mechanical ventilation time,oxygen inhalation time,extubation time and duration of PACU stay were significantly shorter in SFI group than in control group.There were no significant differences in MAP and HR after SFI between the two groups.The plasma?-endorphin concentration was significantly higher in group SFI than in control group.Conclusion Shenfu injectio can make patients emerging from general anesthesia faster.