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Objective:To explore the clinical features, diagnosis and differential diagnosis of splenic marginal zone lymphoma.Methods:The clinical diagnosis and differential diagnosis processes of 3 cases of CD5 - CD10 - B cell non-Hodgkin lymphoma with splenomegaly and cytopenia who were admitted to Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology in 2019 were retrospectively analyzed, and the relevant literature was reviewed. Results:The 3 cases were all elderly patients with varying degrees of splenomegaly and cytopenia. CD5 - CD10 - monoclonal B lymphocytes were found in the bone marrow or lymph nodes. Based on the patient's clinical characteristics, peripheral blood and bone marrow morphology, immunophenotype and genetic characteristics, 2 patients were diagnosed as splenic marginal zone lymphoma, and 1 patient was diagnosed as diffuse large B-cell lymphoma. Conclusions:The diagnosis of splenic marginal zone lymphoma requires comprehensive analysis of clinical characteristics, peripheral blood and bone marrow morphology, immunophenotype and genetic characteristics. Careful differentiation from other CD5 - CD10 - small B-cell lymphomas is also needed. The next-generation gene mutation high-throughput sequencing and mutational spectrum analysis will help the accurate diagnosis of atypical and difficult cases.
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Purpose@#Accumulating evidence has suggested that toll-like receptor 4 (TLR4) is critically involved in the pathogenesis of asthma. The aim of this study was to investigate the role of TLR4 in toluene diisocyanate (TDI)-induced allergic airway inflammation. @*Methods@#TLR4−/− and wild-type (WT) C57BL/10J mice were sensitized and challenged with TDI to generate a TDI-induced asthma model. B-cell lymphoma 2 (Bcl-2) inhibitors, ABT-199 (4 mg/kg) and ABT-737 (4 mg/kg), were intranasally given to TDI-exposed TLR4−/− mice after each challenge. @*Results@#TDI exposure led to increased airway hyperresponsiveness (AHR), granulocyte flux, bronchial epithelial shedding and extensive submucosal collagen deposition, which were unexpectedly aggravated by TLR4 deficiency. Following TDI challenge, TLR4−/− mice exhibited down-regulated interleukin-17A and increased colony-stimulating factor 3 in bronchoalveolar lavage fluid (BALF), while WT mice did not. In addition, TLR4 deficiency robustly suppressed the expression of NOD-like receptor family pyrin domain containing 3 and NLR family CARD domain containing 4, decreased caspase-1 activity in TDI-exposed mice, but had no effect on the level of high mobility group box 1 in BALF. Flow cytometry revealed that TDI hampered both neutrophil and eosinophil apoptosis, of which neutrophil apoptosis was further inhibited in TDI-exposed TLR4−/− mice, with marked up-regulation of Bcl-2. Moreover, inhibition of Bcl-2 with either ABT-199 or ABT-737 significantly alleviated neutrophil recruitment by promoting apoptosis. @*Conclusions@#These data indicated that TLR4 deficiency promoted neutrophil infiltration by impairing its apoptosis via up-regulation of Bcl-2, thereby resulting in deteriorated AHR and airway inflammation, which suggests that TLR4 could be a negative regulator of TDI-induced neutrophilic inflammation.
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Purpose@#Accumulating evidence has suggested that toll-like receptor 4 (TLR4) is critically involved in the pathogenesis of asthma. The aim of this study was to investigate the role of TLR4 in toluene diisocyanate (TDI)-induced allergic airway inflammation. @*Methods@#TLR4−/− and wild-type (WT) C57BL/10J mice were sensitized and challenged with TDI to generate a TDI-induced asthma model. B-cell lymphoma 2 (Bcl-2) inhibitors, ABT-199 (4 mg/kg) and ABT-737 (4 mg/kg), were intranasally given to TDI-exposed TLR4−/− mice after each challenge. @*Results@#TDI exposure led to increased airway hyperresponsiveness (AHR), granulocyte flux, bronchial epithelial shedding and extensive submucosal collagen deposition, which were unexpectedly aggravated by TLR4 deficiency. Following TDI challenge, TLR4−/− mice exhibited down-regulated interleukin-17A and increased colony-stimulating factor 3 in bronchoalveolar lavage fluid (BALF), while WT mice did not. In addition, TLR4 deficiency robustly suppressed the expression of NOD-like receptor family pyrin domain containing 3 and NLR family CARD domain containing 4, decreased caspase-1 activity in TDI-exposed mice, but had no effect on the level of high mobility group box 1 in BALF. Flow cytometry revealed that TDI hampered both neutrophil and eosinophil apoptosis, of which neutrophil apoptosis was further inhibited in TDI-exposed TLR4−/− mice, with marked up-regulation of Bcl-2. Moreover, inhibition of Bcl-2 with either ABT-199 or ABT-737 significantly alleviated neutrophil recruitment by promoting apoptosis. @*Conclusions@#These data indicated that TLR4 deficiency promoted neutrophil infiltration by impairing its apoptosis via up-regulation of Bcl-2, thereby resulting in deteriorated AHR and airway inflammation, which suggests that TLR4 could be a negative regulator of TDI-induced neutrophilic inflammation.
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Objective To study effect of rubescens on inflammatory cytokines and liver function of alcoholic liver injury rat model.Methods 120 SD rats were chosed for studied, randomly divided into control group, model group and treatment group,40 rats per group.Alcoholic liver injury rat model was made by ethanol feeding approach, rubescens were given for treatment.Then serum AST, ALT, α-GST, GLDH content and liver tissue NF-κB, TNF-α, IL-6, IL-8 levels were detected by electrochemiluminescence and fluorescent quantitation PCR,respectively.Results Model group serum AST, ALT,α-GST, GLDH contents and NF-κB, TNF-α, IL-6, IL-8 contents in liver tissue were higher than control group(t=12.030~37.417, P<0.05). Treatment group serum AST,ALT, α-GST, GLDH contents and NF-κB, TNF-α, IL-6, IL-8 contents in liver tissue were lower than model group (t=5.128~32.325, P<0.05).Conclusion Alcohol feeding can cause liver damage, rubescens therapy can reduce liver injury, relieve inflammation.
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Background Researches indicated that etiology and epidemiology of pertussis toxin (PTX)dependent experimental autoimmune uveoretinitis(EAU)model are very different with human uveoretinitis owing to the influence of PTX on immune.Our previous study has established lipopolysaccharide (LPS),an endotoxin,which instesad of PTX,mediated EAU model.However,the exact roles of LPS and PTX in EAU still remained unclear.Objective This study was to investigate the roles of LPS and PTX in EAU model.Methods Twenty SPF C57BL/6(H-2b) mice were assigned to 0 d-PTX-EAU group,7 d-PTX-EAU group,0 d-LPS-EAU group and 7 d-LPS-EAU group using random number table method.The mice were immunized with interphotoreceptor retinoid-binding protein 1-20(IRBP 1-20) emulsified in complete Freund adjuvant (CFA),and concurrently with or on day 7 postimmunization,LPS or PTX was injected in the footpad or intraperitoneally respectively.Delayed-type hypersensitivity (DTH) of the mice was evaluated by measuring the ear thickness 48 hours after IRBP was injected into the ear pinna,and lymphocyte proliferation was assessed by tritiated thymidine uptake.Retinal histopathological examination was performed and scored based on criteria of Caspi.The use and care of experimental animals complied with Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Serious infiltration of inflammatory cells,disorder of entire retinal structure and retinal folds were seen in the mice of the 0 d-PTX-EAU group and 7 d-LPS-EAU group on 21 days after injection of PTX or 14 days after injection of LPS,and severe vitritis and a few granuloma-like lesions were found in the 0 d-PTX-EAU group.However,only mild vasodilatation or less retinal folds were found in the 7 d-PTX-EAU group and 0 d-LPS-EAU group.The pathological scores in the mice of the 0 d-PTX-EAU group and 7 d-LPS-EAU group were higher than those of the 7 d-PTX-EAU group and 0 d-LPS-EAU group (all at P ＜ 0.05).The ear thickness was (62.600 ± 3.362) μm,(60.000±2.345) μm,(30.400± 1.817) μm and (32.800 ± 1.643) μm in the 0 d-PTX-EAU group,7 d-PTX-EAU group,0 d-LPS-EAU group and 7 d-LPS-EAU group,showing a significantly difference among the 4 groups (Fgroup =259.751,P=0.000),and the ear thicknesses of 0 d-PTX-EAU group and 7 d-PTX-EAU group were significantly higher than those of the 0 d-LPS-EAU group and 7 d-LPS-EAU group (all at P＜0.05).The lymphocyte proliferation was strongly enhanced in PTX-EAU groups,and the radiation count per minute (cpm) was (16 150.000±799.218)/min and (16 120.000±729.383)/min in the 0 d-PTX EAU group and 7 d-PTX EAU group,and (8 348.000±258.979)/min and (8 540.000±81.548)/min in the 0 d-LPS EAU group and 7 d-LPS EAU group respectively,with a significant difference among the PTX-EAU groups and LPS-EAU groups (Fgroup =316.978,P=0.000).Conclusions LPS and PTX play different roles during the EAU formation.LPS may be involved in the breakdown of blood-retina barriers (BRB).
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Background The pathogenesis and management of human autoimmunity uveoretinitis is a focus in ophthalmology.For decades,a traditional experimental autoimmunity uveoretinitis (EAU) induced by pertussis toxin (PTX) was used for the basic investigation,which was thought to have a large deviation from the natural course of human autoimmunity uveoretinitis.Objective This study was to establish a new mice model of autoimmunity uveoretinitis which mimics the human autoimmunity uveoretinitis pathogenesis and offer a basis for the investigation and treatment of uveoretinitis.Methods Twenty 6-8 weeks old specific pathogen-free female C57BL/6 (H-2b) mice were randomized into normal control group,only endotoxin (lipopolysaccharide,LPS) induced uveitis group (endotoxin induced uveitis [EIU] group),interphotoreceptor retinoid-binding protein (IRBP1-20) +complete Freund adjuvant (CFA) induced uveoretinitis group (EAU group) and IRBP+CFA+LPS induced uveoretinitis group (LPS-EAU group).The mice of the EAU were only immunized with IRBP emulsified in CFA,and LPS-EAU group firstly were immunized with IRBP emulsified in CFA and then LPS was injected in the footpad of the mice on 7 days following immunization.The ocular pathological examination,histopathological scoring,delayed-type hypersensitivity and specific lymphocyte proliferating response were evaluated and compared with the EIU models,traditional EAU models without PTX and LPS-EAU models.The use and care of experimental animals complied with Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results No inflammatory response was found in the iris,cilliary body and retina of mice in the normal control group.However,mild blood vessels dilation and fibrin exudation were seen in the iris and cilliary body of mice in the EIU group.In the EAU group,mild vasculitis and swelling of nerve fiber layer were exhibited in the retinas; while in the LPS-EAU group,severe disorder of retinal structure,infiltration of inflammatory cells and damage of photoreceptor were found under the optical microscope.The pathological score was 0 in the models of the normal control group and EIU group,0.5 score in the EAU group and 3.0 scores in the LPS-EAU group,with a significant difference in the pathological scores between the EAU group and the LPS-EAU group (U=16.246,P =0.001).The earthickness of the mice was (35.60±0.55) μm in the LPS-EAU group,and this value was significantly higher than (12.60±0.55) μm of the EIU group (q =23.003,P＜0.01),but closed to (34.80±0.84) μm of the EAU group (t =0.820,P＞0.05).The obvious cloning were seen and theradiation count per minute was (8 540.00 ±54.77)/min in the model mice of the LPS-EAU group,and that in the EAU group was (8 484.00±47.75)/min,without significant difference between them (q =56.634,P =0.069).Compared with the β particle number (2 050.00±50.00)/min in the EIU group,that of the LPS-EAU group was significantly elevated (q =195.683,P =0.000).Conclusions LPS injection can induce EAU in mice,and this model can better imitate the pathogenesis of human autoimmunity uveoretinitis.
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0.70,cluster in two pairs of medicines are similar. Conclusion:It's reliable to cluster analysis the TCM by regarding the element contents as the character of these medicines. The larger the correlation coefficient is,the more similar the nature,flavor and efficacy of the medicines are,which suggested that trace element contents were closely related to(have much to do with) the flavor,nature and efficacy. The study provides method for the cluster relationship among TCM.
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OBJECTIVE:To establish an HPLC-CSP (chiral stationary phase) for the determination of plasma levels of enantiomers of fluoxetine. METHODS: Enantiomers of fluoxetine were separated on a Chirobiotic V column. The mobile phase consisted of methanol-acetic acid-triethylamine (100∶0.04∶0.04) with a flow rate of 1 mL?min-1. The detection wavelength was set at 226 nm.Clozapine was used as the internal standard. RESULTS: Under this chromatographic condition, both R-norfluoxetine and S-norfluoxetine had no interference on the enantiomers of fluoxetine and the internal standard. The calibration curve was linear in the range of 8.1～432 ng?mL-1(r=0.999 8) for R-enantiomer and in the range of 10.8～432 ng?mL-1 (r=0.999 2) for S-enantiomer. The relative recovery of R-fluoxetine and S-fluoxetine in plasma were 97.2%～101.9% and 98.7%～102.2%, respectively. RSD of intraday and interday ranged from 2.6% to 5.3% and from 7.7% to 8.9% for R-fluoxetine, and from 2.1% to 6.7% and from 8.0% to 10.6% for S-fuoxetine, respectively. CONCLUSION: The method is simple, reliable and sensitive, and applicable for the determination and pharmacokinetic study of the enantiomers of fluoxetine in plasma.
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OBJECTIVE:To promote rational drug use and improve medical quality.METHODS:A comprehensive analysis was performed on the contents of drug counseling problems presented by 483 patients and medical staff.RESULTS:The use of cardio-cerebral-vascular drugs,anti-bacterial drugs and digestive system drugs were the most common drugs inquired by patients,and their questions focused on the action and clinical use,adverse reactions,administration and dosage etc of drugs.However,anti-bacterial drugs were the leading drugs inquired by medical staff,and their questions focused on drugs available for choice and combined use of drugs etc.CONCLUSIONS:Drug counseling can help promote rational drug use,improve medical quality and patients' awareness toward medical care,besides,it is conducive to the publicity for hospital pharmacists.
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Objective: To study the expression and activity changes of methionine adenosyltransferase (MAT) in human peripheral T lymphocytes. Methods: The expression of MAT mRNA was detected by RT-PCR and the activity of MAT was measured. Results: After stimulated by IL-2, PHA and anti-CD3 antibody, MAT-Ⅱ gene expression increased by (8.9? 2.1), (7.7?1.9) and (8.0?1.8) times, respectively, and the expression peak was at 8, 4 and 8 h,respectively; MAT activity continuously increased in 48 h. S-adenosylmethioinie (SAM) moderately induced IL-2 and IFN secretion by human T cells. SAM(0.1 mg/ml) downregulated the expression and activity of MAT-Ⅱ and the secretion of IL-2 and IFN induced by PHA or anti-CD3 antibody in human T cells. Conclusion:MAT is involved in the activation of T lymphocytes, and high dose of SAM may also inhibit its activation through PHA and anti-CD3 antibody.
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Objective: To study antibody dependent cell-mediated cytotoxicity (ADCC) on hepatocytes during immune response in halothane hepatitis of guinea pigs. Methods: The model of halothane hepatitis of guinea pigs was used. Mixed lymphocyte hepatocytes were cultured and supernatants containing specific antigens and cytokines were added. The changes of hepatic function in immune response of halothane hepatitis were analyzed. Results: The cytotoxicity on hepatocytes induced by halothane 3 times included ADCC mediated by specific antigens. ADCC on hepatocytes induced by halothane once required the second induction. There was no cytotoxicity on hepatocytes of specific antigens and cytokines produced by proliferative lymphocytes induced by Kupffer cells. Conclusion: Humoral immunity is mainly ADCC. Specific antibodies and cytokines can not separately kill hepatocytes. The extent of immune response is related to induction by halothane.
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Objective: To prepare the monoclonal antibodies against polycystin 1 intracellular region and to study the distribution and expression of polycystin 1 in kidney tissues. Methods: Using the recombinant fusion protein containing polycystin 1 intracellular region as antigen, the hybrid cells secreting the monoclonal antibodies against polycystin 1 intracellular region were established by hybridoma technique. The distribution and expression of polycystin 1 in polycystic kidney, fetal kidney and adult kidney were investigated by immunohistochemical methods(standard EnVision method) with the monoclonal antibodies. Results: Four cell lines of hybrids steadily secreting the monoclonal antibodies against polycystin 1 intracellular region were established. The antibody titers were 1∶10 6. The 50th generation of these cell lines of hybrids still can secret the monoclonal antibodies and the titers remain similar. Polycystin 1 was weakly expressed in tubules and collecting ducts of normal kidney, the strong staining was seen at tubules of fetal kidney, and very strong staining of cyst lining epithelium of polycystic kidney was observed. Conclusion: The monoclonal antibodies against polycystin 1 intracellular region will be a useful tool in the studies of polycystin 1 structure and function.
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AIM: To investigate the effect of lipopolysaccharide (LPS) priming on macrophage(M?). METHODS: Macrophage cell line RAW264.7 were pretreated with or without LPS for 1 h, then challenged with PMA, or LPS, muramyl dipeptide(MDP), Zymosan, formyl-methionyl-leucyl-phenylalanine(FMLP) for 1 h . O 2 production in supernatants and intracellular free calcium([Ca 2+ ]i ) were measured, and changes in [Ca 2+ ]i and LPS induced O 2 production were compared. RESULTS: LPS pretreatment significantly increased O 2 production in RAW264.7 cells challenged with the stimuli, and in a certain extent, both O 2 production and increase of resting intracellular [Ca 2+ ]i were dose- and time-dependent on LPS pretreatment.Furthermore,the peak [Ca 2+ ]i was significantly higher in LPS pretreated groups than that of LPS unpretreated groups when challenged with PMA. Pretreatment with Ca 2+ inophore A23187 mimicked the LPS priming effects on O 2 production, but pretreatment with Ca 2+ chelator BAPTA and EGTA blocked this priming effect. CONCLUSION: LPS induced M? priming effect on O 2 production is dependent on elevation of resting intracellular [Ca 2+ ]i .
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Objective: To investigate the effect of a long-term CpG-ODN stimulation on the maturation of murine bone-marrow derived dendritic cells (BMDCs). Methods: Murine bone-marrow cells were cultured in GM-CSF alone or with CpG-ODN for 7 d or for last 36 h (days 6, 7). Cell phenotypes and antigens uptake by BMDCs were analyzed by flow cytometry. Cytokines released by BMDCs were detected by ELISA. The antigen presenting capability by BMDCs was evaluated by mixed lymphocyte responses.Results:Compared to those of the short-term CpG-ODN stimulation group, the expression of MHCⅡ, CD86, CD40, and secretion of IL-12(p70) by BMDCs in long-term stimulation group were not increased. The phagocytosis of FITC-OVA by BMDCs in long-term CpG-ODN stimulation group was strengthened, but the activation of allogenic and homogenic lymphocyte cells proliferation was impaired. Conclusion:Long-term CpG-ODN stimulation can suppress the maturation of DCs, which may explain the low adaptive immunity in sepsis patients.
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Objective:To study the antigen presenting of Kupffer cells in immune response of Guinea pig halothane hepatitis. Methods:Sixteen male Guinea pigs were randomly divided into 2 groups. Experimental group:1% halothane and 40% O 2 were inhaled for 4 h, re inhaled on the 42nd and 84th day; Control group:only inhaled 40% O 2. Lymphocytes and Kupffer cells were separated and the culture was mixed on the 21st day after the last inhalation. 3H TdR was added 18 h before the end of culture. Immune response and the antigen presenting action of Kupffer cells were analysis by lymphocyte transformation test (LTT). Results:Halothane had significant pro proliferative effects on autologous lymphocytes( P
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Objective:To construct a mutispecific internal control for competitive RT-PCR analysis of Fas、FasL、GB、P、TIA-1 and ?-actin.Methods:Invitro synthesized fragments were amplified by PCR,there are two products,of which 145 and 147 bp.The 145 bp one contained 5′ primer sequences of Fas、FasL、GB、P、TIA-1 and ?-actin,another contained 3′ primer sequences of the same genes.The products with restriction sites were inserted into the vector PKF_3.Results:Restriction enzyme analysis and DNA sequencing were used to identify the recombinant plasmid,corresponding internal control was obtained by the amplification of the recombinant plasmid with each primer pair.The mutispecific internal control was then used for quantitative detection of TIA-1 in peripheral blood leukocytes from a patient with acutely rejecting allograft.Our study on Fas、FasL、GB、P、TIA-1 and ?-actin showed that the coamplified templates accumulated in a parallel manner throughout not only the exponential phase.Conclusion:The mutispecific internal control can be used for quantitative detection of the six genes. [
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Interleukin 24(IL 24),also called the melanoma differentiation associated gene 7(MDA 7),is stably expressed in human tissues associated with the immune system such as the spleen,thymus,peripheral blood leukocytes and normal melanocytes. IL 24 binds to IL 20 and IL 22 receptor complexes(IL 22R1/IL 20R2 and IL 20R1/IL 20R2),and induces secretion of high levels of IL 6,TNF alpha,and IFN gamma and low levels of IL 1beta,IL 12,and GM CSF from human PBMC. Adenoviral IL 24(Ad IL 24) induces growth suppression and apoptosis selectively in diverse human cancers and tumor cell lines without producing any apparent harmful effect in normal cells. The effects of Ad IL 24 are associated to the ratio of pro apoptotic(BAX,BAK) to anti apoptotic(Bcl 2) proteins,and the up regulation of p38 MAPK and a family of growth arrest and DNA damage(GADD) inducible genes. These demonstrate the potential therapeutic effects of Ad IL 24 in human cancer.