1.The expression of CD14 in rat Kupffer cells
Junming FENG ; Jingquan SHI ; Yousheng LIU ; Yusong YIN
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: To explore the expression of CD14 in rat Kupffer cells (KCs). METHODS: In rat KCs induced by LPS or the mediators from KCs induced by LPS,the changes of CD14 expression were measured by RT-PCR and immunohistochemistry.The expressions of TNF? mRNA?IL-6 mRNA or the concentrations of TNF??IL-6 were estimated by in situ hybridization and radioimmunoassay,respectively. RESULTS: LPS increased the expression of CD14 in KCs in a dose-dependent fashion (LPS,1 ?g/L-10 mg/L) and in a time-dependent fashion(0.5 h-24 h,peaked at 3-6 hours). While the expression of CD14 in KCs stimulated by the active mediators from KCs which had been exposed to LPS 1 hour were obviously increased. CONCLUSIONS: There was a close relationship between LPS or the active mediators from KCs induced by LPS and the expressions of CD14. It is implied that the increase in CD14 expression may be induced by LPS and the cytokines produced by KCs,it also reveals that there is a auto-regulated loop in CD14 expression.
2.The roles of thymic stromal lymphopoietin receptor and its antibody in airway inflammatory response in asthmatic mice
Liyun SHI ; Hongping YIN ; Xielai ZHOU ; Feng XU ; Jie YAN
Chinese Journal of Microbiology and Immunology 2008;28(6):492-496
Objective To determine the roles of thymic stromal lymphopoietin receptor (TSLPR) and its antibody in airway inflammatory response in asthmatic mice, and to promote maturation and activation of dendritic cells (DCs) in mouse airway. Methods BALB/c mice were randomly divided into group A, B and C. The mice in group B and C were intraperitoneally injected with OVA for allergization while the mice in group A were intraperitoneally injected with PBS as the normal control. The mice in group B and C were treated by inhalation of non-specific IgG and TSLPR IgG respectively, before provocation of asthma using OVA. The bronchoalveolar lavage fluid (BALF) of the mice in different groups were collected for cell differential counts and quantitative detection of IL-4, IL-5, IFN-γand IL-10 levels by ELISA. Moreover, the pulmonary tissue specimens of the mice were collected for pathological examination, and the numbers and phenotypes of DCs from the local lymph nodes and pulmonary tissue were determined by flow cytometry. Results The levels of all the tested cytokines in the BALF from mice in group B and C were remarkably higher compared to those from mice in group A (P<0.01). However, both the IL-4 and IL-5 levels in the BALF from group C mice that pre-blocked with TSLPR IgG were lower than those from group B (P<0.05, P<0.01), whereas both the IFN-γ and IL-10 levels in the BALF from group C mice were higher than those from group B (P<0.05, P<0.01). Furthermore, the numbers of total cells, eosinophils and lymphocytes in the BALF from group C mice were also lower than those from group B (P<0.01). A large number of inflammatory cell infiltration around the bronchus, beaker cell proliferation and mucous secretion reinforcement could be found in the samples from group B mice, while slight inflammatory cell infiltration and beaker cell proliferation in the samples from group C mice. The numbers of DCs in mediastinal lymph node and the levels of I-Ad, CD40, CD80 and CD86 expression of pulmonary DCs from group B mice were higher than those from group C mice (P<0.05). Conclusion TSLP/TSLPR have an effect on promoting asthma, which is closely relative to its regulation of DCs activation. And the interference of TSLPR antibody can decrease the effect of TSLP/TSLPR which indicating a potential of the antibody as a novel anti-asthma drug.
3.Effects of inducible co-stimulator gene on the cytotoxic activity of cytokine-induced killer cells against cholangiocarcinoma cells
Jian WANG ; Min HE ; Yin WANG ; Huifang SHA ; Jiuxian FENG ; Yuqian SHI ; Tao CHEN ; Weijin SHI
Chinese Journal of Digestive Surgery 2008;7(3):213-217
Objective To explore the effects of inducible co-stimulator (ICOS) gene on the cytotoxic activity of cytokine-induced killer (CIK) cells against cholangiocarcinoma cells. Methods CIK-ICOS cells were obtained by stable transfecting ICOS genes into CIK cells through the adenovirus vector whereas untransfected and EGFP-transfected CIK cells were treated as controls. The proliferation and apoptosis of different CIK cells, as well as their cytotoxicity against cholangiocarcinoma cells in the three groups were detected. The expressions of IFN-T, IL-2 and TNF-α in the supernatant of different CIK cells were measured by ELISA. SCID mice with cholangiocarcinoma were randomly divided into CIK group, CIK-EGFP group, CIK-ICOS group and normal saline group. The cytotoxic activity of CIK-ICOS cells against cholangiocarcinoma cells in vivo was observed. Results CIK-ICOS cells displayed better proliferation than CIK cells and CIK-EGFP cells. At day 20 and 23 of culture, the apoptosis rate of CIK-ICOS cells was 0.69% and 0.89%, respectively, while that of the CIK cells was 2.90% and 4.92%. The cytotoxic effect of CIK-ICOS cells at different E: T ratio against cholangiocarcinoma cells was significantly stronger than that of CIK cells and CIK-EGFP cells (F=13.37, 6.46, 25.51, P<0.05). The concentration of IFN-γ in CIK-ICOS cultured supernatant was (49.50±4.73)μg/L, which was significantly higher than that in the cultured supernatant of CIK cells [(30.53±3.73)μg/L] and CIK-EGFP cells [(30.12±2.64)μg/L](F=38.89, P<0.05). The growth of cholangiocarcinoma was significantly slower in CIK-ICOS group than that in CIK group and CIK-EGFP group, whereas the necrosis area of tumor was larger and the CIK cells in CIK-ICOS group was more than those in the other two groups. Conclusions CIK cells had the function of killing cholangiocarcinoma cells in vitro and in vivo. After ICOS genes were transfected into CIK cells, the survival time of CIK cells in vitro was prolonged and the proliferation of CIK cells was enhanced, as well as the secretion of IFN-γ was increased so that the cytotoxicity of CIK cells against cholangiocarcinoma cells in vitro and in vivo was enhanced.
5.Effects of laparoscopic and open D2 gastrectomy on the expression of interleukin-6 and interleukin-10 : a prospective analysis
Peng YIN ; Yan SHI ; Peiwu YU ; Feng QIAN ; Yongliang ZHAO ; Bo TANG ; Yingxue HAO
Chinese Journal of Digestive Surgery 2013;(5):358-361
Objective To compare the effects and significance of laparoscopic and open D2 gastrectomy on the expression of interleukin (IL)-6 and IL-10.Methods The clinical data of 146 patients with gastric cancer who were admitted to the Southwest Hospital from November 2010 to October 2011 were prospectively analyzed.All the patients were randomly divided into the laparoscopic group (75 patients) and open group (71 patients)according to the sealed envelope method.Laparoscopic or open D2 gastrectomy were performed according to the 14th edition of gastric cancer treatment guidelines of Japan Gastric Cancer Association.Peritoneal lavage fluid was collected at the beginning and the end of operation,and the concentrations of IL-6 and IL-10 in the peritoneal lavage fluid were detected by enzyme linked immunosorbent assay.The measurement data were analyzed using the t test,and the count data were analyzed using the chi-square test.Results The preoperative concentrations of IL-6 in the laparoscopic group and the open group were (34 ± 13)μg/L and (35 ± 12)μg/L,respectively,with no significant difference between the 2 groups (t =-5.110,P > 0.05).The postoperative concentrations of IL-6 in the laparoscopic group and the open group were (4015 ± 1592)μg/L and (6724 ± 2112)μg/L,respectively.The postoperative concentration of IL-6 in the laparoscopic group was significantly lower than that of the open group (t =-8.367,P < 0.05),and the postoperative concentrations of IL-6 were significantly higher than those before operation in the laparoscopic group and open group (t =-59.065,-87.123,P <0.05).The preoperative concentrations of IL-10 in the laparoscopic group and the open group were (43 ±9) μg/L and (42 ± 10) μL,respectively,with no significant difference between the 2 groups (t =1.190,P >0.05).The postoperative concentrations of IL-10 in the laparoscopic group and the open group were (92 ± 32)μg/L and (62 ± 23)μg/L,respectively.The postoperative concentration of IL-10 was significantly higher than that of the open group (t =6.408,P < 0.05),and the postoperative concentrations of IL-10 were significantly higher than those before operation in the laparoscopic group and the open group (t =-12.680,-6.802,P < 0.05).Conclusion Peritoneal inflammatory reaction is relatively lighter after laparoscopic D2 gastrectomy when compared with open D2 gastrectomy,which might prevent the peritoneal metastasis of gastric cancer mediated by IL-6.
6.Determination of Bioequivalence of Cefixime by Three-way Crossover Design
Aixin SHI ; Yufei FENG ; Kexin LI ; Lei LIU ; Qi YIN ; Chunhua SUN
China Pharmacy 1991;0(02):-
OBJECTIVE:To study the bioequivalence of suspension formulation of cefixime(A),capsule formulation of ce-fixime(B) and reference preparation(C: Cefixime Capsules or Cefspan) in human body.METHODS: The study was conducted as a 3- way crossover design in 18 healthy volunteers whose plasma concentrations of cefixime were determined by HPLC after receiving a single oral dose of 200 mg trial preparations or reference preparation.RESULTS:The main pharmacokinetics of the three preparations(A、B、C) were as follows after undergoing BIO3 program fitting:AUC0-1 were(18.54?6.31)mg?h-1?L-1, (16.10?5.51)mg?h-1?L-1 and (17.16?5.96)mg?h-1?L-1, Cmax were(2.63?0.76) mg?L-1, (2.43?0.78)mg?L-1 and (2.57?0.90)mg?L-1;tmax were(4.11?0.58)h,(4.56?0.51)h and (4.56?0.70)h,respectively .The relative bioavailability of cefixime suspensions(A) and cefixime capsules(B) were (108.8?12.3)% and (95.7?15.9)% ,respectively as against reference preparation(C) .CONCLUSION:The test formulations(A and B) were found bioequivalent to the reference formulation(C).
7.Effect of emodin in attenuating endoplasmic reticulum stress of pancreatic acinar AR42J cells.
Li WU ; Feng ZHANG ; Shi-zhong ZHENG ; Yin LU ; Bao-chang CAI
China Journal of Chinese Materia Medica 2015;40(3):501-505
OBJECTIVETo explore the effect of emodin on endoplasmic reticulum (ER) stress of pancreatic acinar AR42J cells.
METHODRat pancreatic acinar AR42J cells were cultured in 6-well plates, and divided into the normal control group, the model group (with the final concentration at 1 x 10(-7) mol · L(-1) for cerulean and lipopolysaccharide at 10 mg · L(-1)) and the emodin group (10, 20, 40 μmol · L(-1)). Cells in each group were cultured in three multiple pores for 24 h, and their supernate was removed after cell attachment. The normal control group was added with haploids, the model group was added with the modeling liquid for haploids, and the treatment groups were added with different concentrations of emodin at 15-20 min before the modeling liquid. The cells were continuously cultured for 3 h under 37 °C and 5% CO2. Their intracellular protease and lipase expressions were detected with kits. The cellular morphology was observed under optical microscope. The level of calcium in endoplasmic reticulum was measured under laser confocal microscopy. Western blot assay were used to determine the protein expression of ER-related signaling molecules.
RESULTEmodin could significantly inhibit levels of amylase, lipase and intracellular calcium and ER.
CONCLUSIONEmodin could reduce pancreatic acinar cell injury induced by the combination of cerulean and lipopolysaccharide. Its action mechanism is correlated with the inhibition of intracellular calcium overload and ER stress.
Animals ; Calcium ; metabolism ; Cell Line, Tumor ; Emodin ; pharmacology ; Endoplasmic Reticulum Stress ; drug effects ; Pancreatic Neoplasms ; metabolism ; pathology ; Rats ; Unfolded Protein Response ; drug effects
8.Comparison among the immune effects of DNA-or protein-FimH of UPEC type 1 pilus
Xiaolin YIN ; Xinli SHI ; Lin WEI ; Cuiqing MA ; Xiurong WANG ; Huidong FENG
Chinese Journal of Immunology 2000;0(08):-
Objective:To observe cellular,humoral and mucosal immune responses induced by DNA-or protein-based of FimH of UPEC type 1.Methods:After mice were immunized respectively with recombinant plasmid pcDNA3.1/fimH or pcDNA3.1/fimC,and the combinant FimH and FimC protein,the anti-FimH protein IgG of sera and SIgA in bladders were detected by ELISA.The lymphocyte phenotypes of CD3,CD4 and CD8 were analyzed by FCM.Results:The titers of IgG in sera and SIgA in the bladders were all low in the group immunized by recombinant FimH plamid,but the percentage of CD4+T cells in spleen was high,which revealed that recombinant FimH plamid was able to trigger better cellular immune response.The titers of IgG were very high in the group immunized by FimH protein,which suggested that the FimH protein was able to trigger better humoral immune response,but SIgA in the bladders was not detectable.Conclusion:The DNA for FimH can induce humoral,mucosal and cellular immune response.FimH protein can only induce humoral immune response.FimC protein is able to enhance the immunogenicity of FimH protein.
9.Effects of Low Dose Ultrashort Wave Therapy on Inflammation and Edema after Spinal Cord Injury in Rats
Yucen WAN ; Shi SUN ; Lina ZHAO ; Yanmei YIN ; Zhiping FENG ; Yuxin ZHOU ; Zhiqiang ZHANG ; Lixin ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2016;22(2):150-155
Objective To investigate the effect of ultrashort wave therapy on edema and inflammatory reaction after spinal cord injury (SCI) in rats. Methods 56 Sprague-Dawley rats were divided into sham-operated group (n=8), model group (n=24) and ultrashort wave group (n=24). The model was established with Allen's method. The sham-operated group was exposed endorhachis without hit. The ultra-short wave group was exposed to ultrashort wave radiation 7 minutes, once a day, 24 hours after modeling until the animals were sacrificed. Locomotors functional recovery was assessed every week post operation period by BBB score. Immunohistochemical staining was per-formed to observe the expression of the aquaporin-4(AQP-4) and ED-1. Results The BBB scores increased in the model group and the ultra-short wave group after treatment, and was higher in the ultrashort wave group than in the model group from 1 week after treatment (t>3.368, P<0.01). The expressions of AQP-4 and ED-1 were higher in the model group and ultrashort wave group than in the sham-operated group, and decreased as time went on. They were lower in the ultrashort wave group than in the model group(t>3.156, t>4.466, P<0.05). Conclu-sion Ultrashort wave therapy can alleviate the edema after SCI, reduce the activation and infiltration of inflammatory cells, and promote the recovery of neurological function.
10.An experimental study on the removal of thrombi by Fogarty thrombectomy in different time phases
Yi GU ; De SHI ; Yu ZHAO ; Leping YIN ; Chao FENG ; Tong LIU
Chinese Journal of General Surgery 1997;0(06):-
Objective To clarify the effect of Fogarty balloon catheter thrombectomy on venous wall integraty when performed on different time phases.MethodsA murine model of inferior vena caval thrombosis was established. Collagen of venous wall was measured by Van Gieson staining and this was used as the criteria of venous wall injury. The thrombus residue was determined after Fogarty balloon catheter thrombectomy in each individual time phase. Results Collagen deposit in the adventitia of venous wall increased every day,to an amount of (5 902?399) ?m2 on the third day which was significantly different from that of controls (5 333?454) ?m2(P