Ureaplasma urealyticum GrpE promotes mouse dendritic cell maturation and elicits Th1 immune responses / 中华微生物学和免疫学杂志

Objective:To investigate the effects of Ureaplasma urealyticum GrpE ( Uu-GrpE) on the maturation of dendritic cells and the polarization of T cells. Methods:Uu-GrpE was expressed and purified, and then identified by Western blot. The cytotoxicity of Uu-GrpE to mouse bone marrow-derived dendritic cells (BMDCs) was analyzed by LDH kit. After stimulating BMDCs with Uu-GrpE, the expression of costimulatory molecules, CD80, CD86 and major histocompatibility complex Ⅱ (MHCⅡ), on the surface of BMDCs was detected by flow cytometry, and ELISA was used to detect the cytokines such as IL-12p70, TNF-α, IL-1β and IL-6. CD4 + Na?ve T cells were isolated from mouse spleen tissues by magnetic beads. A co-culture system of BMDCs and Na?ve T cells was constructed to analyze the effects of GrpE-stimulated mature BMDCs (GrpE-BMDCs) on T cell proliferation and polarization towards Th1/Th2. Mice were immunized with GrpE-BMDCs through the tail vein, and the induced humoral and cellular immune responses were detected by ELISA and flow cytometry. Results:Uu-GrpE was successfully express and high purity BMDCs were isolated. Uu-GrpE could stimulate BMDCs to secrete cytokines such as IL-12p70, TNF-α, IL-1β and IL-6 without having cytotoxicity. Uu-GrpE significantly increased the expression of CD80 [mean flourscence indensity (MFI): (324.00±22.11) vs (91.03±10.95), P<0.01], CD86 [MFI: (1 176.00±51.39) vs (217.00±14.93), P<0.01] and MHCⅡ [MFI: (708.70±56.32) vs (185.70±16.77), P<0.01] on BMDCs. Compared to the GrpE-BMDCs only group and GrpE (boiled)-BMDCs+ T cell group, the GrpE-BMDCs+ T cell group showed significantly increased T cell proliferation [stimulation index: (7.25±0.21) vs(6.55±0.23) and (6.09±0.35), both P<0.05], and dramatically promoted T cell secretion of IL-2 and IFN-γ [IL-2: (145.60±14.67) pg/ml vs(55.92±3.12) pg/ml and (26.05±2.40) pg/ml, P<0.05 and P<0.01; IFN-γ: (267.20±37.80) pg/ml vs(146.70±20.65) pg/ml and(27.84±6.69) pg/ml, both P<0.05]. However, no significant change was observed in the expression of Th2-type cytokines. Moreover, the adoptive transfer of GrpE-BMDCs induced a Th1-type immune response. Conclusions:Uu-GrpE could stimulate the maturation and polarization of BMDCs. Moreover, it could induce Th1 immune response as a candidate protein vaccine for Ureaplasma urealyticum.

The changes of serum iron markers in pregnant women with hepatitis B virus infection / 国际检验医学杂志

Objective To investigate the effect of hepatitis B virus(HBV)infection on serum iron markers in pregnant women.Methods A total of 188 pregnant women with HBV infection and 157 normal pregnant women were recruited in this study.Serum levels of iron,ferritin,transferrin,HBV DNA,alanine aminotrans-ferase(ALT)and aspartate aminotransferase(AST)were measured and compared.Results At the same stage of pregnancy,the serum ferritin level in pregnant women with HBV infection was higher than that of normal pregnant women(Z= -1.72,P=0.04;Z= -2.33,P=0.01;Z= -4.42,P=0.01),while the serum transfer-rin concentration increased in normal pregnant women in the second and third trimesters,and the differences between the two groups were statistically significant(Z= -3.26,P<0.01;Z= -2.25,P=0.01).In pregnant women with HBV infection,the serum ferritin levels in patients with positive results of ALT and AST were higher than those in patients with negative results(ALT:P=0.01;AST:P=0.02),however,there was no change in healthy pregnant women.Conclusion There is iron metabolism imbalance in pregnant women with HBV infection.We should monitor serum iron markers to guide pregnant women for appropriate iron supple-mentation in gestation period.

Preliminary research on the establishment of reference interval of serum pepsinogen in healthy people / 国际检验医学杂志

Objective To investigate the distribution characteristics of serum pepsinogen (PG) in healthy people and its reference interval establishment .Methods 3 753 healthy people were enrolled and divided into 0 .05) .In the same gender ,pairwise comparison of PGⅠlevels was conducted in different age groups ,and the difference showed no statistical sig-nificance(P>0 .05) .PGⅡlevel increased with age increasing (P<0 .01) while PGⅠ /PGⅡlevel increased with age reducing (P<0 .05) .Percentile method was adopted to determine the 95% reference interval ,the bilateral reference intervals (P2 .5 - P97 .5 ) was taken for PGⅠ ,unilateral upper limit(≤ P95 ) for PGⅡ and unilateral limit (≥ P5 ) for PGⅠ /PGⅡ .Conclusion The establishment of serum PG Ⅰ ,PG Ⅱ ,PG Ⅰ /PG Ⅱ reference intervals of healthy people provides a basis for the prevention and treatment for stomach disease .

Study on inhibitory effect of baicalein on mycoplasma pneumonia and protection mechanism of pulmonary epithelial cells of mice / 中国医师杂志

Objective To investigate the inhibitory effect of baicalein on mycoplasma pneumonia and its the protective mecha -nism in body , and to provide scientific experimental basis for prevention and treatment of mycoplasma pneumoniae infection .Methods The mycoplasma pneumoniae and baicalein treated BALB /c mice lung tissues were stained with hematoxylin eosin (HE), and his-topathological grading .Minimum inhibitory concentration ( MIC) of baicalein on mycoplasma pneumonia was determined by real-time quantitative polymerase chain reaction ( PCR) .The expression of P 1 adhesion molecules mRNA and protein in lung tissue of BALB /c mice was determined with reverse transcription-PCR and Western blot .The expression of epidermal growth factor ( EGF) mRNA and protein in lung tissue was detected by quantitative RT-PCR and immunofluorescence .Results Baicalein significantly reduced the my-coplasma treated mice's lung tissue pathological score .The minimal inhibitory concentration of baicalein was 32 μg/ml.Baicalein sig-nificantly downregulated P 1 gene transcription and protein translation , and upregulated EGF gene transcription and protein expression . Conclusions Baicalein shows significant resistance to mycoplasma pneumoniae , and can protect the body against mycoplasma damage by inhibiting the expression of P 1 protein and promoting the expression of EGF protein .

Serodiagnosis of Chlamydia pneumoniae infection using three inclusion membrane proteins.

The Chlamydia pneumoniae genome-encoded open reading frames Cpn0146, Cpn0147, and Cpn0308 were expressed as recombinant proteins for detecting C. pneumoniae-specific antibodies in samples from three groups of individuals including 183 with C. pneumoniae-associated respiratory infection (group I), 60 healthy blood donors (group II), and 32 with no known respiratory infection (group III). The recombinant Cpn0146 was recognized by 71 (38.8% positive recognition rate), 15 (25%) and 1 (3.1%), Cpn0147 by 75 (40.9%), 14 (23.3%), and 2 (6.3%), and Cpn0308 by 82 (44.8%), 16 (26.7%), and 0 (0%) samples from groups I, II, and III, respectively. The positive recognition rates with any of the three antigens were significantly higher in group I than those in groups II and III, suggesting that more individuals from group I were likely infected with C. pneumoniae. This conclusion was confirmed with a commercially available whole organism-based ELISA kit (Savyon Diagnostics Ltd., Ashdod, Israel), which detected C. pneumoniae antibodies in 98 (64.1%), 26 (43.3%), and 4 (12.5%) samples from group I, II, and III, respectively. Comparing to the commercial kit, the recombinant antigen-based detection assays displayed >97% of detection specificity and >87% of sensitivity, suggesting that these recombinant antigens can be considered alternative tools for aiding in serodiagnosis of C. pneumoniae infection.

Comparison of three different novel Chlamydophila pneumonia recombinant antigens for serodiagnosis / 中华检验医学杂志

Objective To evaluate three different Chlamydophila pneumoniae recombinant antigens for use in Chlamydophila pneumoniae serodiagnosis. Methods The recombinant plasmids pGEX6p-2/ Cpn0146,Cpn0147 and Cpn0308 were constructed and expressed as GST fusion proteins. The immunogenicity and the immunocompetence of these recombinant protein were analyzed by Western-blot and indirect ELISA. A total of 183 sera samples of patients with respiratory tract infection and 32 sera samples of patients with Chlamydia trachomatis infection were detected with indirect ELISA coated microwell plates with the purified recombinant proteins comparing with SeroCP-TM IgG ELISA kits. The positive recognition rate, sensitivity and specificity of each method were analyzed. Results GST-Cpn0146, Cpn0147 and Cpn0308 were obtained after expression and purification. The titers of the specific IgG antibodies against Cpn0146, Cpn0147 and Cpn0308 were higher than 1:6 400, 1:128 00 and 1:128 00, respectively. When the indirect ELISA was developed to detect the IgG antibody against Chlamydophila pneumoniae in 183 samples, the concordance rate between the indirect ELISA test and SeroCP-TM IgG ELISA kits were 92. 3% (Cpn0146) , 94.5% (Cpn0147) and 96.7% (Cpn0308), respectively. The recombinant Cpn0146, Cpn0147 and Cpn0308 were recognized by 71 (38.8% positive recognition rate), 75 (40.9%), and 82 (44.8%) samples, respectively. The recombinant antigen-based detection assays displayed > 97% of detection specificity and>87%of sensitivity.Condusion GST-Cpn0308 shows a better sensitivity and specificity,which suggests it could be used for developing serodiagnosis kits of Chlamydophila pneumoniae infection.