Targeting DKK1 enhances the antitumor activity of paclitaxel and alleviates chemotherapy-induced peripheral neuropathy in breast cancer.

Chemotherapy in combination with immunotherapy has gradually shown substantial promise to increase T cell infiltration and antitumor efficacy. However, paclitaxel in combination with immune checkpoint inhibitor targeting PD-1/PD-L1 was only used to treat a small proportion of metastatic triple-negative breast cancer (TNBC), and the clinical outcomes was very limited. In addition, this regimen cannot prevent paclitaxel-induced peripheral neuropathy. Therefore, there was an urgent need for a novel target to enhance the antitumor activity of paclitaxel and alleviate chemotherapy-induced peripheral neuropathy in breast cancer. Here, we found that Dickkopf-1 (DKK1) expression was upregulated in multiply subtypes of human breast cancer specimens after paclitaxel-based chemotherapy. Mechanistic studies revealed that paclitaxel promoted DKK1 expression by inducing EGFR signaling in breast cancer cells, and the upregulation of DKK1 could hinder the therapeutic efficacy of paclitaxel by suppressing the infiltration and activity of CD8+ T cells in tumor microenvironment. Moreover, paclitaxel treatment in tumor-bearing mice also increased DKK1 expression through the activation of EGFR signaling in the primary sensory dorsal root ganglion (DRG) neurons, leading to the development of peripheral neuropathy, which is charactered by myelin damage in the sciatic nerve, neuropathic pain, and loss of cutaneous innervation in hindpaw skin. The addition of an anti-DKK1 antibody not only improved therapeutic efficacy of paclitaxel in two murine subtype models of breast cancer but also alleviated paclitaxel-induced peripheral neuropathy. Taken together, our findings providing a potential chemoimmunotherapy strategy with low neurotoxicity that can benefit multiple subtypes of breast cancer patients.

Expression of the small cysteine-rich protein SCR96 from Phytophthora cactorum in mammalian cells: phytotoxicity and exploitation of its polyclonal antibody.

OBJECTIVE: We aimed to investigate the expression of a novel small cysteine-rich (SCR) effector protein SCR96 from the phytopathogenic oomycete Phytophthora cactorum in mammalian cells, its bioactivity and to exploit its polyclonal antibody. RESULTS: The gene encoding the SCR effector protein SCR96 was codon-optimized, custom-synthesized, cloned into pcDNA3.1(-) and overexpressed in human embryonic kidney (HEK) 293-6E cells. The recombinant protein SCR96 was prone to aggregation and purified with its monomer to homogeneity with a predicted molecular weight of 8.9 kDa. SCR96 exhibited strong phytotoxic activity on tomato seedlings at 24 h post treatment with 4.2 µg of the purified protein. An anti-SCR96 polyclonal antibody was prepared by immunization of New Zealand white rabbits. The good-titer antibody had a detection sensitivity at 6.25-ng level and could specifically detect the SCR96 protein expressed either in yeast, or in tomato leaves. CONCLUSIONS: Transient production of the SCR effector protein SCR96 in mammalian cells is reliable, providing sufficient recombinant protein that can be utilized for analysis of its phytotoxic activity and preparation of its polyclonal antibody.

The RXLR Effector PcAvh1 Is Required for Full Virulence of Phytophthora capsici.

Plant pathogens employ diverse secreted effector proteins to manipulate host physiology and defense in order to foster diseases. The destructive Phytophthora pathogens encode hundreds of cytoplasmic effectors, which are believed to function inside the plant cells. Many of these cytoplasmic effectors contain the conserved N-terminal RXLR motif. Understanding the virulence function of RXLR effectors will provide important knowledge of Phytophthora pathogenesis. Here, we report the characterization of RXLR effector PcAvh1 from the broad-host range pathogen Phytophthora capsici. Only expressed during infection, PcAvh1 is quickly induced at the early infection stages. CRISPR/Cas9-knockout of PcAvh1 in P. capsici severely impairs virulence while overexpression enhances disease development in Nicotiana benthamiana and bell pepper, demonstrating that PcAvh1 is an essential virulence factor. Ectopic expression of PcAvh1 induces cell death in N. benthamiana, tomato, and bell pepper. Using yeast two-hybrid screening, we found that PcAvh1 interacts with the scaffolding subunit of the protein phosphatase 2A (PP2Aa) in plant cells. Virus-induced gene silencing of PP2Aa in N. benthamiana attenuates resistance to P. capsici and results in dwarfism, suggesting that PP2Aa regulates plant immunity and growth. Collectively, these results suggest that PcAvh1 contributes to P. capsici infection, probably through its interaction with host PP2Aa.

Sterol metabolism and protein metabolism are differentially correlated with sarcopenia in Asian Chinese men and women.

OBJECTIVES: Our aim was to investigate the prevalence and predictive variables of sarcopenia. METHODS: We recruited participants from the Peking Union Medical College Hospital Multicenter Prospective Longitudinal Sarcopenia Study (PPLSS). Muscle mass was quantified using bioimpedance, and muscle function was quantified using grip strength and gait speed. Logistic regression revealed the relationships between sarcopenia and nutritional, lifestyle, disease, psychosocial and physical variables. RESULTS: The prevalence of sarcopenia and sarcopenic obesity was 9.2%-16.2% and 0.26%-9.1%, respectively. Old age, single status, undernourishment, higher income, smoking, low physical activity, poor appetite and low protein diets were significantly associated with sarcopenia. Multiple logistic regression analysis showed that age was a risk factor for all stages of sarcopenia, and participants above 80 years were greater than fivefold more susceptible to sarcopenia, while lower physical activity was an independent risk factor. The optimal cut-off value for age was 71 years, which departs from the commonly accepted cut-off of 60 years. Female participants were greater than twofold less susceptible to sarcopenia than male participants. The sterol derivative 25-hydroxyvitamin D was associated with fourfold lower odds of sarcopenia in male participants. Several protein intake variables were also correlated with sarcopenia. Based on these parameters, we defined a highly predictive index for sarcopenia. CONCLUSIONS: Our findings support a predictive index of sarcopenia, which agglomerates the complex influences that sterol metabolism and nutrition exert on male vs female participants.

The correlations of coronary artery disease with epicardial adipose tissue, insulin resistance and C-reactive protein in different body mass index patients / 中国综合临床

Objective To evaluate the influence of C-reactive protein (CRP),insulin resistance (IR) and epicardial fat volume (EFV) on the extent of coronary atherosclerosis in patients with different body mass index(BMl).Methods One hundred and three patients with coronary artery disease were involved in current study who underwent 64-slice dual source CT and percutaneous coronary angiography.Measurements of height,weight,waist circumference (WC) were recorded,and BMI was calculated.All patients were divided into obesity group (n =45) and non-obesity group (n =58) based on BMI.EFV were calculated through 64-slice dual source CT.Blood samples were collected for biochemical examination.Gensini score were adopted to quantify the severity of coronary artery stenosis.The relationship between Gensini score and EFV,CRP and homeostasis model assessment-insulin resistance(HOMA-IR) index were statistical analyzed by SPSS16.0 software.Results The level of CRP,WC,EFV and BMI in obesity group were (11.0 ± 5.8) mg/L,(96.1 ± 7.0) cm,(122.7 ± 43.3) cm3,(27.9 ± 2.9) kg/m2 respectively,significantly higher than those in non-obesity group ((6.5 ± 3.4) mg/L,(86.4 ± 7.6) cm,(92.9 ± 39.5) cm3,(22.4 ± 1.9) kg/m2) and the differences were significant (t =2.24,6.74,3.64,11.74,and P ＜ 0.05).CRP were positively correlated with EFV (r =0.404,0.364,P ＜0.05) in both obesity and non-obesity group,While HOMA-IR were only associated with BMI in obese group(r =0.322,P ＜0.05).Gensini score in non-obesity groups were positively related with EFV and CRP (r =0.358,0.315,P ＜ 0.05),while in obesity groups were positively related with EFV,CRP and HOMA-IR(r =0.348,0.297,0.384; P ＜ 0.05).The associations between Gensini score and CRP were not significant in obesity group after adjusting BMI and WC.Multiple linear regression analysis showed that EFV and diabetes mellitus were independent risk factors of patient Gensini score.Conclusion Coronary atherosclerosis is positively related with EFV and CRP in all patients.While,coronary atherosclerosis is influenced by BMI,WC and HOMA-IR in obese group.EFV is an independent risk factor of coronary atherosclerosis.

The effect of epicardial fat volume in patients with metabolic syndrome on insulin resistance and coronary heart disease / 医学研究生学报

Objective Epicardial fat volume ( EFV) was a risk factor for coronary heart disease ( CHD) , but there is little research regarding the relationship of EFV with insulin resistance ( IR) and CHD in patients with metabolic syndrome ( MS) .The aim of the article was to explore the effect of EFV in patients with MS on CHD and IR . Methods Patients with MS treated by percutane-ous coronary angiography ( CAG) in our hospital from February 2013 to August 2013 were recruited in this study .The data of height , weight, waist circumference(WC) and hip circumference(HP) were recorded.EFV were measured by MSCT.Fasting blood samples were collected for blood biochemical test . Results EFV in patients with MS was in positive relation with IR index (IRI)(r=0.335, P<0.001) and CHD (r=0.321, P<0.05), and the correlation still remained when the influences of WC and body mass index (BMI) were excluded.Logistic regression analysis showed that EFV was an independent risk factor for CHD (P<0.05), while linear regression analysis indicated EFV , BMI and LDL-C were the risk factors for IRI .ROC curves analysis proved EFV and BMI had diag-nosis value for IRI, and the areas under curve of EFV were 0.755 and 0.679 (P<0.05) respectively. Conclusion EFV is an in-dependent risk factor for CHD and IRI in patients with MS , and EFV has an advantage over BMI in the diagnosis value of IRI .

Automatic synthesis of 18F-FB-RGD and evaluation of its biodistribution / 中华核医学杂志

Objective To evaluate the automatic synthesis of 18F-labeled cyclic RGD peptide c(RGDyK)and its biological distribution in the tumor-bearing mice. Methods N-succinimidyl-4-18 F-fluorobenzoate (18F-SFB) was automatically synthesized and then re-dissolved in acetonitrile (MeCN). The cyclic RGD peptide c(RGDyK) was mixed with an hydrous DMSO and N, N-diisopropyl ethylamine (DIPEA). 18F-FBRGD was obtained by the reaction of peptide solution with 18 F-SFB. The final product was purified by HPLC gradient separation system and solid-phase extraction method. The biodistribution study and competition test of N-4-18F- fluorobenzoyl-RGD (18F-FB-RGD) in the tumor-bearing mice was performed. Results The labeling yield of 18 F-FB-RGD was (33.6 ± 3.5)%. The synthesis time was 110 min. The radiochemical purity was more than 98%. The tumor uptake of 18F-FB-RGD was (3.43 ±0.15), (2.61 ±0.14), (2.11 ±0.13), and (1.79 ±0.18) %ID/g, respectively, at 30, 60, 90 and 120 min after injection. The ratio of tumor to muscle activity ranged from 4.26 ±0.69 to 5.80 ±0.78. The tumor uptake decreased dramatically after RGD blockage. The uptake was (0.46 ±0.21) %ID/g and (2.87 ±0.59) %ID/g in the blocked and unblocked mice, respectively, at 60 min after blockage. Conclusions 18 F-FB-RGD can be automatically synthesized and it may become a promising tumor imaging agent.

Resistance mutation patterns of hepatitis B virus in patients with suboptimal response to adefovir dipivoxil therapy after lamivudine resistance / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the resistance mutation patterns of hepatitis B virus(HBV) during adefovir dipivoxil (ADV) monotherapy or combination therapy after lamivudine(LAM) resistance.</p><p><b>METHODS</b>Serum samples from fifteen patients with suboptimal viral response to ADV therapy after LAM resistance were collected. The RT region of HBV P gene was PCR-applied, cloned and sequenced, and the mutation patterns in relation to resistance were analyzed.</p><p><b>RESULTS</b>The ADV resistance mutation patterns of A181T+N236T, A181V, A181T were selected in ADV monotherapy group. The LAM resistance mutation patterns of M204V+L180M, M204V+L180M+L229V, M204I+L80I, M204V+L180M+V207I were detected in the combination therapy group. 20% of clones from three serum samples were detected double resistance to LAM and entecavir (ETV) in the combination therapy group, the resistance patterns were M204I+L80I+T184I (2/10), M204V+L180M+T184S (2/10), and M204V+L180M+G173L+S202G (2/10) respectively. I269L clones were detected in two serum samples from both two groups and P109S clones also detected in the one from monotherapy group.</p><p><b>CONCLUSIONS</b>In the patients with suboptimal viral response to ADV therapy after LAM resistance, the ADV resistance mutation patterns of A181T+N236T, A181V and A181T could easily be selected during ADV monotherapy; while in the patients with combination therapy, the LAM resistance mutation patterns of M204V+L180M, M204V+L180M+L229V, M204I+L80I, and M204V+L180M+V207I were predominant, the ETV resistance mutation T184I/S and S202G could be selected. The mutation patterns of I269L and P109S may impact the responses to ADV therapy in some patients.</p>

Transient ischemia and reperfusion induce the expression of glucose transporters GLUT1 and GLUT4 in rat myocardium / 医学研究生学报

Objective : To investigate the dynamic changes of the expression of GLUT1 mRNA and GLUT4 mRNA in rats myocardium with transient ischemia and reperfusion, and the relationship between the dynamic changes and the time during reperfusion. Methods : In rats, the left anterior descending coronary artery was occluded for 20 min followed by reperfusion for 4 hours, 1, 3 or 7 days as ischemia reperfusion model. The relative content of GLUT1 mRNA and GLUT4 mRNA in myocardium was detected by RT-PCR and gel electrophoresis imaging. Results: During myocardial post-ischemia and reperfusion, the levels of GLUT1 mRNA got up to the peak at 4th hour[ (0.666?0. 003 ) vs (0. 509?0.002) controls , P 0.05). Conclusion; Transient ischemia and reperfusion induce the expression of glucose transporters GLUT1 and GLUT4 genes in rat myocardi- um, which contribute to promote glucose utilization during ischemia, protect ischemic myocardium and improve functional recovery on reperfusion.

Expression and function of the glucose transporter and their correlation with heart failure / 医学研究生学报

The glucose transporter(GLUT) is an energy-related carrier protein located on the cell membrane.Most researches have shown that changes of energy metabolism play an important role in the development of heart failure.This review summarizes recent advances in the understanding of the relationship between GLUT and heart failure.