GBP2 serves as a novel prognostic biomarker and potential immune microenvironment indicator in renal cell carcinoma.

Since the application of immune checkpoint therapy (ICT) has gradually become a new strategy for clear cell renal cell carcinoma (ccRCC) treatment, biomarkers that predict the individual response to ICT is needed. This study aimed to identify a new clinical indicator for postoperative surveillance of ccRCC and prediction of ICT response. We investigated the GBP2 expression and its relation with immune cell infiltration in tumor microenvironment using public databases, clinical specimens and ccRCC cell lines. Bioinformatic analysis using public database revealed that GBP2 expression is higher in cancer tissues than in adherent normal tissues among different cancer types including ccRCC, and the same results were acquired from clinical tissue samples tested by Western Blot and PCR. In ccRCC cell lines, CCk-8 proliferation assay and apoptosis assessment suggested GBP2 facilitates the malignancy of ccRCC. 286 ccRCC patients were randomly divided into a training or validation cohort, and immunohistochemistry (IHC) and Kaplan-Meier analysis revealed that higher GBP2 expression is related to worse prognosis. C-index analysis implied that integrating GBP2 expression with TNM stage improved the accuracy in predicting prognosis of ccRCC patients compared to the solitary use of either. Bioinformatic analysis implied a relation between GBP2 and immunity, and GBP2 expression is positively related with suppressive immune markers in ccRCC microenvironment. Taken together, our study demonstrated the potential of GBP2 to sever as a prognostic predictor of ccRCC, and an association between GBP2 and tumor-infiltrating lymphocytes in ccRCC was observed, making it a promising indicator of ICT response.

Circulating CXCR5+CD4+ T cells assist in the survival and growth of primary diffuse large B cell lymphoma cells through interleukin 10 pathway.

Diffuse large B cell lymphoma (DLBCL) is a common and aggressive cancer caused by the malignant transformation of B cells. Although it has been established that the follicular helper T (Tfh) cells play a central role in B cell development, little information is available on their involvement in DLBCL pathogenesis. We studied the role of the peripheral Tfh equivalent, the CXCR5+ CD4+ T cells, in DLBCL. Data showed that compared to CXCR5- CD4+ T cells, CXCR5+ CD4+ T cells were significantly more effective at promoting the proliferation as well as inhibiting the apoptosis of primary autologous DLBCL tumor cells. Surprisingly, we found that at equal cell numbers, CXCR5+ CD4+ T cells in DLBCL patients secreted significantly less interleukin (IL)-21 than CXCR5- CD4+ T cells, while the level of IL-10 secretion was significant elevated in the CXCR5+ compartment compared to the CXCR5- compartment. Neutralization of IL-10 in the primary DLBCL-CXCR5+ CD4+ T cell coculture compromised the CXCR5+ CD4+ T cell-mediated pro-tumor effects, in a manner that was dependent on the concentration of anti-IL-10 antibodies. The CXCR5+ compartment also contained significantly lower frequencies of cytotoxic CD4+ T cells than the CXCR5- compartment. In conclusion, our investigations discovered a previously unknown pro-tumor role of CXCR5-expressing circulating CD4+ T cells, which assisted the survival and proliferation of primary DLBCL cells through IL-10.

[Effects of Methanol Extracts from Atractylodes macrocephalae Rhizoma on Small Intestinal Epi- thelial Cell Proliferation and Migration, and Expression of Phospholipase C-γl].

OBJECTIVE: To observe the effects of methanol extracts from Atractylodes macro- cephalae Rhizoma (AMR) on the proliferation and migration of IEC-6 cell (small intestinal epithelial cells) and the expression of phospholipase C-γ1 (PLC-γ1) , and to explore the mechanism of AMR (a Chinese herb capable of invigorating Pi replenishing qi) for promoting repair of gastrointestinal mucosal injury. METHODS: IEC-6 cells were divided into the blank group, the positive control (spermidine, SPD; 5 µmol/L) group, AMR extracts groups (50, 100, and 200 mg/L). The alpha-difluoromethylornithine (DFMO, polyamines synthesis inhibitor) group, the SPD +DFMO group, AMR extracts (50, 100, and 200 mg/L) +DF- MO groups were set up in stress test. IEC-6 cells were cultured by adherence for 24 h,and then treated with AMR extracts for appropriate periods of time. Effects of IEC-6 cell proliferation after action of AMR extracts were detected by Real-time Cell Analyzer (RTCA). The effect of AMR extracts on IEC-6 cell migration number was detected using scratch method. mRNA and protein expressions of PLC-γ1 levels were detected by fluorescent quantitative polymerase chain reaction ( RT-qPCR) and Western blot respectively. RESULTS: Compared with the blank group, AMR extracts showed no obvious effect on IEC-6 cell proliferation (P >0. 05). But SPD and AMR extracts (100 and 200 mg/L) not only promoted IEC-6 cell migration (P <0. 01), but also improved mRNA and protein expressions of PLC-γl in the process of cell migration (P <0. 01). Compared with the DFMO group, SPD and AMR extracts (100 and 200 mg/L) could reverse inhibitory effects of DFMO on cell migration, and mRNA and protein expressions of PLC-γl (all P <0. 01). CONCLUSION: AMR extracts played roles in repairing gastrointestinal mucosal injury possibly by promoting polyamine mediated intestinal epithelial cell migration, and its effect on intestinal epithelial cell proliferation was not main potentcy.

[Effect of Sijunzi Decoction Polysaccharide on IEC-6 Cell Migration,Potassium Channel and Membrane Potential].

Objective: To investigate the effect of Sijunzi decoction polysaccharide( SJZDP) on intestinal epithelial cells( IEC-6)cell migration and polyamine signaling pathway potassium channel during intestinal epithelial cell migration, and to explore the mechanism of SJZDP on promoting gastrointestinal mucosal restitution after wounding. Methods: Cell migration model was established by scratch damage, and then the effect of SJZDP normal cultured or with difluoromethylornithine( DFMO) and 4-aminopyridine( 4-AP) on IEC-6 cell migration was observed and calculated on this wounding model. The effect of SJZDP on expression of IEC-6 cell kv1. 1 mRNA and protein levels were detected by RT-q PCR and Western blot analysis, respectively. The Effects of SJZDP on IEC-6 cell membrane potential were detected by flow cytometry. Results: The results showed that treatment with SJZDP( 40,80,160 mg / L) caused a promotion of IEC-6 cell migration,and increased of expression of in IEC-6 cell kv1. 1 mRNA and protein significantly( P < 0. 05 or P < 0. 01) compared with normal control group. In addition, SJZDP( 40,80,160 mg / L) increased cell membrane potential which resulted in cell membrane hyperpolarization compared with normal control group( P < 0. 05 or P < 0. 01). SJZDP( 40,80,160 mg / L) reversed the inhibition of cell migration was reduced,kv1. 1 mRNA,kv1. 1 protein expression, and cell membrane potential were decreased by polyamines synthesis inhibitor DFMO compared with DFMO model group( P < 0. 05 or P < 0. 01). SJZDP( 20,40,80 mg / L) reversed the inhibition of cell migration,kv1. 1 protein and mRNA levels expression were decreased by potassium channel inhibitor 4-AP compared with 4-AP model group( P < 0. 05 or P < 0. 01). Conclusion: These results indicate that the effect of SJZDP on promoting IEC-6 cell migration may be related to its influence on polyamine signaling pathway potassium channel and cell membrane potential.

Alterations of circulating follicular helper T cells and interleukin 21 in diffuse large B-cell lymphoma.

CD4+ CXCR5+ T cell in peripheral blood is known as circulating follicular helper T cell (Tfh), which can produce interleukin 21 (IL-21). In the current study, we investigated changes of circulating Tfh and its correlation with IL-21 in diffuse large B-cell lymphoma (DLBCL). Circulating Tfh and its subtypes were detected by flow cytometry in the peripheral blood of 32 healthy donors and 62 DLBCL cases. Data demonstrated that percentage of circulating Tfh in CD4+ T cells was significantly increased in DLBCL (11.3 %) than in controls (8.5 %) (p = 0.001). Studying the subtypes of Tfh revealed that the upregulation of circulating Tfh was contributed by Tfh-Th2 subtype and Tfh-Th17 subtype. Also, we identified that prevalence of Tfh was significantly elevated in cases with advanced stages (stages III and IV). Interestingly, the elevation of circulating Tfh was negatively correlated with serum IL-21 in DLBCL patients. In addition, a positive correlation between circulating Tfh and IL-21 receptor on CD + 8 T cells was observed in patients. This study suggests involvement of circulating Tfh and IL-21 in the pathogenesis and progression of DLBCL and provides a potential target for treating this disease.

Fibroblast growth factor receptor 4 polymorphisms and the prognosis of non-Hodgkin lymphoma.

Fibroblast growth factors and their receptors (FGFRs) play important roles in blood system. FGFR4 rs351855 (Gly388Arg) polymorphism has shown to be a risk factor for many diseases. The aim of this study was to investigate the association between FGFR4 polymorphisms and the susceptibility to non-Hodgkin lymphoma (NHL) in the Chinese population. We identified two polymorphisms in the FGFR4 gene, rs351855G/A (Gly388Arg), and rs147603016G/A, by polymerase chain reaction-restriction fragment length polymorphism in 412 NHL cases and 476 healthy controls. Results showed that frequencies of AA genotype and A allele in rs351855 (Gly388Arg) polymorphism were significantly higher in patients than in controls (odds ratio (OR) 2.12, 95 % confidence intervals (CI) 1.99-3.48, P < 0.0001; OR 1.45, 95 % CI 1.21-1.88, P < 0.0001, respectively; data were adjusted for age and sex). The rs147603016G/A polymorphism did not show any correlation with NHL. When analyzing the survival time of NHL patients with FGFR4 rs351855G/A polymorphism, cases with AA genotype had significantly shorter survival time compared to the patients with GG and GA genotypes (P = 0.002). These results suggested polymorphism in FGFR4 gene was associated with increased susceptibility to NHL and could be used as a prognostic marker for this malignancy.

Association of peripheral CD4+ CXCR5+ T cells with chronic lymphocytic leukemia.

Accumulating evidences indicate that immune dysregulation plays a key role in both lymphomagenesis and patient outcome of chronic lymphocytic leukemia (CLL). Peripheral blood CD4+ CXCR5+ T cells, known as circulating follicular helper T cells (Tfh), can induce B cell activation and production of specific antibody responses. The aim of the study was to investigate changes of circulating Tfh in CLL. Tfh and it subtypes were tested by measuring CD4, CXCR5, CXCR3, and CCR6 in 72 CLL cases and 86 healthy controls using flow cytometry. Data showed that the percentage of Tfh in the peripheral CD4+ T cells was significantly increased in CLL (25.1%) than in controls (8.4%) (p < 0.001). Further analysis revealed that the upregulation of Tfh was contributed by Tfh-th2 subtype and Tfh-th17 subtype. Investigating staging of the cases demonstrated that the prevalence of Tfh was significantly elevated in cases with Binet stage C (37.3%) than those with stage A (20.1 %) or stage B (23.9 %). In addition, we analyzed Tfh in patients with immunoglobulin variable heavy chain (IGHV) gene mutational status. Results presented that Tfh-th17 subtype had clearly higher frequency in patients with IGHV mutation compared to the unmutated cases (p = 0.035). This study suggested the involvement of Tfh in the pathogenesis and progression of CLL, and provided a potential target for treating this disease.

[Research progress of lower limb muscle strength training in the treatment of lliotibial band syndrome].

Iliotibial band syndrome (ITBS), as an overused injury of the lower extremities, has developed into a common cause of lateral knee pain. At present, the treatment of ITBS includes drug therapy, muscle strength training, physical therapy, and surgical treatment. Among these methods, physical therapy, drug therapy, and surgical treatment can only alleviate the symptoms of patients. As a safe and effective treatment, lower limb muscle strength training can improve patients' muscle strength, correct abnormal gait, and reduce the recurrence rate of the disease by paying attention to the dynamic changes of patients' recovery process. At present, the pathogenesis of ITBS remains unclear, and the treatment methods are not unified. It is necessary to further study the biomechanical factors related to the lower extremities and develop more scientific and comprehensive muscle strength training methods.

The 5-HT and PLC Signaling Pathways Regulate the Secretion of IL-1ß, TNF-α and BDNF from NG2 Cells.

The present study was clarified the relationship between NG2 glial cells and 5-hydroxytryptamine (5-HT) to further revealed a role in the regulation of cortical excitability. The co-localization of NG2 cells and 5-HT in rat prefrontal cortex was determined using immunofluorescence. Different concentrations of 5-HT were applied to cultured NG2 cells. Real-time PCR measured the expression of interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α) and brain-derived neurotrophic factor (BDNF). Changes in the expression of IL-1ß, TNF-α, and BDNF in NG2 cells were detected after the addition of 5-HT receptor specific blockers and phospholipase C (PLC) specific activators and inhibitors. The results confirmed that the NG2 protein and 5-HT co-localized in the prefrontal cortex. 5-HT treatment of NG2 cells significantly reduced the expression of IL-1ß and BDNF mRNA and increased the expression of TNF-α. The 5-HT receptor specific inhibitors alverine citrate, ketanserin, ondansetron and SB-399885 blocked the regulatory effects of 5-HT on NG2 cells. The PLC signal was linked to the secretion of IL-1ß, TNF-α and BDNF in NG2 cells. These results indicated that 5-HT affected IL-1ß, TNF-α, and BDNF secretion from NG2 cells via the 5-HT1A, 5-HT2A, 5-HT3, 5-HT6 receptors and the PLC signaling pathway.

Plasma Electrolytic Oxidation of Magnesium Alloy AZ31B in Electrolyte Containing Al2O3 Sol as Additives.

Plasma electrolytic oxidation (PEO) coatings were produced on AZ31B magnesium alloys in alkaline electrolytes with the addition of various concentrations of Al2O3 sols. Effects of Al2O3 sol concentrations on the microstructure, phase composition, corrosion resistance and hardness of PEO coatings were evaluated by scanning electron microscopy (SEM), X-ray diffraction (XRD), microhardness testing and potentiodynamic polarization measurements, respectively. It was revealed that the Al2O3 sol mostly participated in the formation of the ceramic coating and transferred into the MgAl2O4 phase. With the increase of the Al2O3 sol concentration in the range of 0⁻6 vol%, the coating performance in terms of the microstructure, diffraction peak intensity of the MgAl2O4 phase, corrosion resistance and microhardness was improved. Further increase of Al2O3 sol addition did not generate better results. This indicated that 6 vol% might be the proper Al2O3 sol concentration for the formation of PEO coatings.

The prevalence and function of CD4+CXCR5+Foxp3+ follicular regulatory T cells in diffuse large B cell lymphoma.

CD4+CXCR5+Foxp3+ follicular regulatory T (Tfr) cells possess critical roles in suppressing the germinal center reaction, B cell activation, and follicular helper T cell (Tfh) cytokine secretion. Since diffuse large B cell lymphoma (DLBCL) can arise from B cells undergoing germinal center reaction and/or differentiation, we hypothesized that Tfr cells might be involved in DLBCL. In the present study, we recruited thirty-five DLBCL patients and twenty-five healthy controls. Data showed that DLBCL patients presented an enrichment of circulating CD4+CXCR5+Foxp3+ Tfr cells compared to controls. In the primary tumor isolated from enlarged lymph nodes, Tfr cells made up of roughly 3% to 16% of infiltrating T cells. Higher levels of tumor-infiltrating Tfr cells were observed in patients with less advanced DLBCL stages, and in patients that stayed in remission 24 months after the initial R-CHOP treatment. High BCL6 and high FOXP3 expression was observed in Tfr cells ex vivo. After anti-CD3/CD28 and IL-2 stimulation, the Tfr cells more closely resembled Treg cells and presented high IL10 and TGFB1 expression. CD4+CD25+CXCR5+ Tfr cells and CD4+CD25+CXCR5- non-Tfr Treg cells could suppress CD4+CD25- Tconv cell and CD8+ T cell proliferation with similar capacity. However, Tfr cells were less capable of suppressing IFNG expression than Treg cells, and although both cell types supported CD19+ tumor cell proliferation, Tfr cells were less supportive than the non-Tfr Treg cells. Overall, this study suggested that Tfr cells were involved in intratumoral immunity, were likely beneficial to DLBCL patients, and were functionally distinctive from non-Tfr Treg cells. The distribution pattern and the prognostic value of Tfr cells in DLBCL should be examined in further studies.

Effect of interleukin 21 and its receptor on CD8+ T cells in the pathogenesis of diffuse large B-cell lymphoma.

Interleukin 21 (IL-21) and its receptor, IL-21R, play a key role in innate and adaptive immunity. In the present study, the effect of IL-21 and IL-21R on the pathogenesis of diffuse large B-cell lymphoma (DLBCL) was investigated. The serum levels of IL-21 were detected by enzyme-linked immunosorbent assay, and the expression of IL-21R on CD8+ T cells was examined through flow cytometry. The data showed that the serum level of IL-21 was significantly decreased in the patients with DLBCL compared with the healthy controls (P<0.001), whereas the expression of IL-21R was clearly elevated on the CD8+ T cells in the patients with DLBCL. Further analyses revealed that the downregulation of the IL-21 serum level was correlated with an increased tumor stage of DLBCL, while the expression of IL-21R on the CD8+ T cells was positively correlated with the tumor stage. Also, the serum level of IL-21 and the proportion of IL-21R on the CD8+ T cells were negatively correlated in the patients. Notably, it was identified that the proportion of IL-21R on the CD8+ T cells, but not the serum level of IL-21, was significantly upregulated in the patients with bone-marrow involvement and B symptoms. These results indicate that IL-21 and IL-21R may be involved in the pathogenesis of DLBCL, in which IL-21R may reflect the progression of the disease more accurately than the serum level of IL-21.

[Brain arousal dysfunction in severe craniocerebral injury treated with acupuncture].

OBJECTIVE: To explore the early rehabilitation effect of acupuncture on brain arousal in severe craniocerebral injury. METHODS: One hundred and two cases of severe craniocerebral injury were randomly divided into an observation group and a control group, 51 cases in each one. Based on the conventional nursing care in neurological external medicine, in observation group, acupuncture was applied at Shuigou (GV 26), Neiguan (PC 6) and Sanyinjiao (SP 6) mainly. In control group, functional electric stimulation was applied at stimulate the affected muscles of the upper limbs. Thirty days later, the lucid rate from coma, lucid interval and clinical efficacy were compared between two groups. RESULTS; The lucid rate from coma was 82.4% (42/51) in observation group, which was higher than 56.9% (29/51) in control group (P < 0.01). The lucid interval in observation group was shortened remarkably as compared with control group (P < 0.01), and the clinical efficacy was superior apparently to that in control group (P < 0.01). CONCLUSION: On the basis of conventional treatment, acupuncture intervention at early stage can accelerate the recovery of brain arousal function in patients with severe craniocerebral injury.