Citrus tristeza virus is a member of the genus Closterovirus in the family Closteroviridae. The p23 of citrus tristeza virus (CTV) is a multifunctional protein and RNA silencing suppressor. In this study, we identified a p23 interacting partner, FK506-binding protein (FKBP) 17-2, from Citrus aurantifolia (CaFKBP17-2), a susceptible host, and Nicotiana benthamiana (NbFKBP17-2), an experimental host for CTV. The interaction of p23 with CaFKBP17-2 and NbFKBP17-2 were individually confirmed by yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. Subcellular localization tests showed that the viral p23 translocated FKBP17-2 from chloroplasts to the plasmodesmata of epidermal cells of N. benthamiana leaves. The knocked-down expression level of NbFKBP17-2 mRNA resulted in a decreased CTV titer in N. benthamiana plants. Further, BiFC and Y2H assays showed that NbFKBP17-2 also interacted with the coat protein (CP) of CTV, and the complexes of CP/NbFKBP17-2 rapidly moved in the cytoplasm. Moreover, p23 guided the CP/NbFKBP17-2 complexes to move along the cell wall. To the best of our knowledge, this is the first report of viral proteins interacting with FKBP17-2 encoded by plants. Our results provide insights for further revealing the mechanism of the CTV CP protein movement.
Objective
To evaluate the clinical value of combined detection of T cell receptor rearrangement excision circles (TRECs) and CD31+ regulatory T (Treg) cells for accessing the recent thymic output in patients with chronic hepatitis B.
Methods
Four groups involving 135 subjects were set up in this study as follows: mild chronic hepatitis B (Mild CHB, n=35), moderate chronic hepatitis B (Moderate CHB, n=35), severe chronic hepatitis B (Severe CHB, n=35) and healthy control (HCs, n=30) groups. CD4+ CD25+ Treg cells in these subjects were sorted out using magnetic cell separation. The ratio of peripheral CD31+ Treg cells to Treg cells in each group was analyzed by flow cytometry. Real-time PCR was performed to detect TRECs in CD4+ CD25+ Treg cells. The percentages of CD3+ , CD4+ and CD8+ T cell subsets were also measured.
Results
The ratios of CD31+ Treg/Treg cells and the numbers of TRECs in peripheral blood of the Moderate CHB and Severe CHB groups were significantly lower than those of the Mild CHB and HCs groups (P 0.05). No significant difference in the percentages of CD3+ , CD4+ or CD8+ T cell subsets was observed between the four groups (P>0.05). CD31+ Treg/Treg cell ratio had a positive correlation with the number of TRECs (r=0.551, P=0.014).
Conclusions
Both CD31+ Treg/Treg cell ratio and the number of TRECs were reduced in the peripheral blood of patients with moderate or severe CHB. CD31+ Treg/Treg cell ratio and the number of TRECs were positively correlated and could be used as new indices to evaluate recent thymus output.
Key words:
Chronic hepatitis B; CD4+ CD25+ regulatory T cell; CD31+ regulatory T cell; T cell receptor rearrangement excision circles
To discuss the significance of testing hepatitis B virus (HBV) from saliva in HBV patients.HBV DNA content in serum and saliva of 200 HBV patients and 20 healthy subjects were detected by fluorescence quantitative polymerase chain reaction. According to the serum level of HBV content, four groups were divided: control group A, group B negative, low virus C (1 x 10(3) - 1 x 10(5) copies/ml) and high-group D ( > 1 x 10(5) copies/ml). The relationship of serum and virus content in saliva was analysed.Of 200 HBV cases, 180 were found HBV DNA in serum with positive rate of 90.0%; while 145 were found HBV DNA in saliva with positive rate of 72.5%, and there was no significant difference (chi2 = 1.35, P > 0.05). The significant difference was observed in testing serum and saliva in Group C (100.0% vs. 38.5%; Z = 14.11, P < 0.01). In group D, there was no significant difference found either (100.0% vs. 83.8%; chi2 = 1.05, P > 0.05). Group D virus serum had a high average level of (6.63 +/- 1.55) log copies/ml virus and in the saliva had an average level of (5.21 +/- 1.85) log copies/ml; saliva had serum viral load lower than an order of magnitude average. No HBV DNA was found in serum or saliva from 20 health subjects.When the serum contains a high content of HBV DNA virus, the content of saliva HBV DNA virus should be likely high, which might pose a threat of source of infection. A precise quantitative detection of HBV DNA in saliva might be used as evaluation of the level of virus in the body copy for judgment of infection.
Background/aims The accurate diagnosis of lymph node metastasis is necessary in gastric cancer. This study was carried out to identify the risk factors of lymph node metastasis by extreme analysis. Methodology Data from 77 patients with lymph node-positive small gastric cancer and 128 patients with lymph node-negative large gastric cancer were collected. The relationship between the two groups was compared, based on the clinical, pathologic, and preoperative clinical laboratory analysis findings. The independent risk factors influencing lymph node metastasis were determined by multiple logistic regression analysis. Results Rural residence, introversive personality, low third gastric cancer, low serum fibrinogen content, serosal invasion, and high serum CEA level were found to be associated with lymph node metastasis. The rural residence, introversive personality, serosal invasion, and high serum CEA level were the independent risk factors for lymph node metastasis in the multivariate logistic regression model. Conclusions Rural residence, introversive personality, serosal invasion, and high serum CEA level were significantly and independently related to lymph node metastasis. The limited surgery with potentially incomplete lymph node dissection is not recommended in gastric cancer patients with these risk factors.
Aim:To study the effects of high dose methylprednisolone on expression of BMP-2 in acetabulum,caput ossis femoris and proximal femurs bone of rabbit.Methods:A total of 27 adult rabbits were randomly allocated into three groups,9 in each group.The rabbits in group A and group B were injected with methylprednisolone sodium succinate at 80 mg/(kg·d) for 3 and 6 days,respectively.The rabbits in group C were injected with normal saline.At 2,4,and 8 weeks after steroid administration,the bone tissue of acetabulum,caput ossis femoris and proximal femurs were collected to observe histopathological changes,and examine the expression of BMP-2 by immunohistochemical method.Results:The number of bone empty lacunae in caput ossis femoris and proximal femurs in group A and B were more than that of group C(Fgroup=116.782 and 110.112,Ftime=57.207 and 61.092,P0.001),especially in group B(P0.05).The average optical density of BMP-2 in acetabulum,caput ossis femoris and proximal femurs of group A and B were higher than those of group C(Fgroup=232.623,239.894,and 338.533;Ftime=273.064,198.320,and 230.326;P0.001),especially in group B(P0.05).Conclusion:High dose methylprednisolone can reduce the expression of BMP-2 in bone around hip.
In the complex tumor microenvironment, TGFβ is a pleiotropic cytokine involved in regulating cellular processes such as cancer cell proliferation, apoptosis and metastasis. TGFβ defines three subtypes (TGFβ1, TGFβ2, and TGFβ3), of which TGFβ is highly expressed in many cancers, especially those showing high dissemination potential. In addition, increased expression of TGFβ in multiple cancers is usually positively correlated with epithelial mesenchymal transition (EMT) and coordinated with the expression of genes driving EMT-related genes. TGFβ signaling in the tumor microenvironment inhibits the antitumor function of multiple immune cell populations, including T cells and natural killer cells, and the resulting immunosuppression severely limits the efficacy of immune checkpoint inhibitors and other immunotherapeutic approaches. As a major pathway to enhance the efficacy of cancer immunotherapy effects, the role of TGFβ signaling inhibitors have been evaluated in many clinical trials. However, the potential functions and mechanisms of TGFβ1, TGFβ2 and TGFβ3 in gastric cancer progression and tumor immunology are unclear. In this study, we comprehensively analyzed TGFβ1, TGFβ2 and TGFβ3 and gastric cancer microenvironmental features, including immune cell infiltration, EMT, hypoxia, mutation, immunotherapy and drug treatment, based on HMUCH sequencing data (GSE184336) and public databases. We also validated the protein expression levels of TGFβ in gastric cancer tissues as well as the role of TGFβ factor in cytology experiments. This report reveals the important role of the TGFβ gene family in gastric cancer and provides possible relationships and potential mechanisms of TGFβ in gastric cancer.
Objective To study the clinic laboratory characters of AIDS with cryptococcus neoformans meningitis coinfection patients. Methods 58 cases of AIDS patients in our hospital were selected. All the cases were divided into two groups, 26 AIDS patients were infection group, while those with meningitis were coinfection group. T lymphncytes,liver function, blood routine test were detected by the immunological, biochemical methods. Results Compared with the infection group, cerebral protein in coinfection group was high[(601.28±545.24) mmol/L, t =0.494, P＞ 0.05], CL and GLU in coinfection group were low[( 121.571 ± 5.255) mmol/L and ( 1.944 ± 0.678) mg/L, t = 0.129, 1.984,P＞0.05]. In coinfection group, the counts of lymphocyte, CD3, CD4, and CD8 were (495.0± 141.1), (479.9±137.4), (35.1 ± 32.2), (413.9 ± 104.8) colls/μL separately. They were all lower than those in control group ( t=2.236, 2.007,2.767,1.997, P ＜ 0.05 ). Conclusiions The counts of CD4 in AID Spatients with cryptococcus neoformans meningitis is significantly decreased, so in the clinical, pathogen detection should be consided to the patients with CD4 ＜ 100/μL, and the CD3 or CD8 decreased obviously.
Key words:
Acquired immunodeficiency syndrome; Cryptococcus neofotmans meningitis; CD4 lymphocyte
// Yi-Xun Liu 1, * , Guo-Dong Wang 1, * , Xiao Wang 1 , Yong-Le Zhang 1 and Tian-Lun Zhang 2 1 Department of Orthopedic, Huaihe Hospital of Henan University, Kaifeng, China 2 School of Aerospace Engineering, University of Electronic Science and Technology of China, Chengdu, China * These authors have contributed equally to this work Correspondence to: Xiao Wang, email: wangxiaokf@yeah.net Keywords: degenerative osteoarthritis, toll-like receptor 2, nuclear transcription factor-kappa B, TLR-2/NF-κB signaling pathway, matrix metalloprotease-13 Received: November 04, 2016 Accepted: February 06, 2017 Published: March 15, 2017 ABSTRACT The study aims to explore the effects of TLR-2/NF-κB signaling pathway on the occurrence of degenerative knee osteoarthritis (OA). Degenerative knee OA and normal cartilage samples were collected from patients with degenerative knee OA receiving total knee arthroplasty and amputation. Expressions of TLR-2, NF-κB and MMP-13 were determined by qRT-PCR and immunochemistry. The chondrocytes were divided into control, IL-1β, IL-1β + anti-TLR-2 and IL-1β + PDTC groups. MTT assay and flow cytometry were performed to determine proliferation and apoptosis of the chondrocytes. Expressions of TLR-2, NF-κB and MMP-13 were measured by Western blotting. ELISA was conducted to detect the expressions of related inflammatory factors. The positive expressions of TLR, NF-κB and MMP13 were associated with body mass index (BMI), family history, exercise, and WOMAC scores of OA patients. Logistic regression analysis showed that OA influencing factors were TLR, NF-κB, MMP13, BMI, family history and exercise. Compared with normal chondrocytes, the expressions of TLR-2, NF-κB, MMP-13 and related inflammatory factors increased in degenerative knee OA. The chondrocytes in the IL-1β + anti-TLR-2 and IL-1β + PDTC groups showed lower apoptosis rates than those in the IL-1β group. Compared with the control group, increased expressions of TLR-2, NF-κB, phosphorylated-NF-κB (p-NF-κB), MMP-13, IL-1, IL-6 and TNF-α were found in the IL-1β group. In the IL-1β + anti-TLR-2 and IL-1β + PDTC groups, decreased expressions of NF-κB, p-NF-κB, MMP-13, IL-1, IL-6 and TNF-α were found compared with those in the IL-1β group. TLR-2/NF-κB signaling pathway contributes to the occurrence of degenerative knee OA.
Objective To analyse the clinical value of HBV large surface protein(LHBs)in the therapeutic observation of adefovir dipivoxil on chronic hepatitis B.Methods The levels of serum LHBs、HBV DNA、 HBeAg of 30 chronic hepatitis B cases with anti-viral of adefovir dipivoxil were detected at the time of 0,6,12,18,24 months.Results There was no statistical difference between the positive rate of LHBs(90.00%) and HBV DNA(100%) (χ~2=3.16,P > 0.05).LHBs levels were correlated with the logarithm of HBV DNA copies (r=0.912).There was no statistical difference between the positive rate of HBeAg(80.00%)and LHBs(χ~2=1.18,P > 0.05).Conclusions The serum LHBs has a good consistency with HBV DNA.It can be used as a new indicator of viral replication.The long LHBs positive duration indicates the high risk of invalid anti-viral treatment.
Key words:
Hepatitis B; chronic; Anti-viral treatment; Large surface protein; Adefovir dipivoxil
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