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Nitrate is the main form of inorganic nitrogen that crop absorbs, and nitrate transporter 2 (NRT2) is a high affinity transporter using nitrate as a specific substrate. When the available nitrate is limited, the high affinity transport systems are activated and play an important role in the process of nitrate absorption and transport. Most NRT2 cannot transport nitrates alone and require the assistance of a helper protein belonging to nitrate assimilation related family (NAR2) to complete the absorption or transport of nitrates. Crop nitrogen utilization efficiency is affected by environmental conditions, and there are differences between varieties, so it is of great significance to develop varieties with high nitrogen utilization efficiency. Sorghum bicolor has high stress tolerance and is more efficient in soil nitrogen uptake and utilization. The S. bicolor genome database was scanned to systematically analyze the gene structure, chromosomal localization, physicochemical properties, secondary structure and transmembrane domain, signal peptide and subcellular localization, promoter region cis-acting elements, phylogenetic evolution, single nucleotide polymorphism (SNP) recognition and annotation, and selection pressure of the gene family members. Through bioinformatics analysis, 5 NRT2 gene members (designated as SbNRT2-1a, SbNRT2-1b, SbNRT2-2, SbNRT2-3, and SbNRT2-4) and 2 NAR2 gene members (designated as SbNRT3-1 and SbNRT3-2) were identified, the number of which was less than that of foxtail millet. SbNRT2/3 were distributed on 3 chromosomes, and could be divided into four subfamilies. The genetic structure of the same subfamilies was highly similar. The average value of SbNRT2/3 hydrophilicity was positive, indicating that they were all hydrophobic proteins, whereas α-helix and random coil accounted for more than 70% of the total secondary structure. Subcellular localization occurred on plasma membrane, where SbNRT2 proteins did not contain signal peptides, but SbNRT3 proteins contained signal peptides. Further analysis revealed that the number of transmembrane domains of the SbNRT2s family members was greater than 10, while that of the SbNRT3s were 2. There was a close collinearity between NRT2/3s of S. bicolor and Zea mays. Protein domains analysis showed the presence of MFS_1 and NAR2 protein domains, which supported executing high affinity nitrate transport. Phylogenetic tree analysis showed that SbNRT2/3 were more closely related to those of Z. mays and Setaria italic. Analysis of gene promoter cis-acting elements indicated that the promoter region of SbNRT2/3 had several plant hormones and stress response elements, which might respond to growth and environmental cues. Gene expression heat map showed that SbNRT2-3 and SbNRT3-1 were induced by nitrate in the root and stem, respectively, and SbNRT2-4 and SbNRT2-3 were induced by low nitrogen in the root and stem. Non-synonymous SNP variants were found in SbNRT2-4 and SbNRT2-1a. Selection pressure analysis showed that the SbNRT2/3 were subject to purification and selection during evolution. The expression of SbNRT2/3 gene and the effect of aphid infection were consistent with the expression analysis results of genes in different tissues, and SbNRT2-1b and SbNRT3-1 were significantly expressed in the roots of aphid lines 5-27sug, and the expression levels of SbNRT2-3, SbNRT2-4 and SbNRT3-2 were significantly reduced in sorghum aphid infested leaves. Overall, genome-wide identification, expression and DNA variation analysis of NRT2/3 gene family of Sorghum bicolor provided a basis for elucidating the high efficiency of sorghum in nitrogen utilization.
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Nitrate Transporters , Nitrates/metabolism , Sorghum/metabolism , Anion Transport Proteins/metabolism , Phylogeny , Protein Sorting Signals/genetics , Nitrogen/metabolism , DNA , Gene Expression Regulation, Plant , Plant Proteins/metabolismABSTRACT
Objective:To systematically evaluate the efficacy and safety of precision thoracic radiotherapy (TRT) in the limited-stage small cell lung cancer (LS-SCLC) patients by network meta-analysis.Methods:Randomized controlled trials (RCTs) of TRT regimes in the LS-SCLC were electronically searched from PubMed, Web of Science, The Cochrane Library, CNKI and Wanfang Data from inception to September 1 st, 2021. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Statistical analysis was performed by Stata 17 and R 4.1.1 software. Results:A total of 6 RCTs involving 1730 patients with six radiation regimens including hyperfractionated radiotherapy (HFRT): HFRT 45(45 Gy/30 F) and HFRT 60(60 Gy/40 F); conventional fractionated radiotherapy (CFRT): CFRT 70(70 Gy/35 F) and CFRT 66(66 Gy/33 F); moderately hypofractionated radiotherapy (MHFRT): MHFRT 65(65 Gy/26 F) and MHFRT 42(42 Gy/15 F) were included. The network meta-analysis showed that: in terms of improving progression-free survival and overall survival, there was no statistically significant difference among the six radiotherapy regimens. The probabilistic ranking results were: MHFRT 65> HFRT 60>CFRT 66>CFRT 70>MHFRT 42>HFRT 45, and HFRT 60>MHFRT 65>CFRT 66>CFRT 70>HFRT 45>MHFRT 42, respectively. The HFRT 60 regimen was superior to other regimens in reducing the incidence of grade ≥3 pneumonia, and there was no difference between the regimens in causing grade ≥3 radiation esophagitis, and the results of ranking probability were: HFRT 60> MHFRT 42>CFRT 66>CFRT 70>HFRT 45>MHFRT 65, and HFRT 60>CFRT 70>CFRT 66>HFRT 45>MHFRT 42>MHFRT 65, respectively. Conclusions:HFRT 60 radiotherapy regimen may be more effective and safer in the treatment of LS-SCLC patients as a priority choice for LS-SCLC TRT. Limited by the number and quality of included studies, the above conclusions need to be verified by more high-quality studies.
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Objective:To determine the protective effect of fasudil on acute lung injury in septic mice.Methods:Forty-five 4-6-week-old male C57BL mice were randomly(random number) assigned to three groups ( n=15 each group): control group, lipopolysaccharide (LPS) group and Fasudil intervention group (FAS+LPS). Acute lung injury model of septic mice was established with an intraperitoneal injection and intratracheal infusion of LPS. The mice in the FAS+LPS group were injected with fasudil hydrochloride intraperitoneally 30 min before intraperitoneal LPS injection and 1 h after intratracheal LPS infusion, respectively. All mice were sacrificed at 4 h after modeling, and lung tissues were collected. Hematoxylin-eosin staining was preformed to observe the morphological changes in the lung tissue. The wet /dry weight (W/D) ratio, malondialdehyde (MDA) content and the activity of myeloperoxidase (MPO) in the lung tissues were detected. Caspase-3 expression was examined by immunohistochemical (IHC) staining. Western blot was employed to detect the expression of RhoA, ROCK1, endothelial nitric oxide synthase (eNOS), and p-eNOS. Results:Inflammatory cell infiltration and erythrocyte exudation were significantly reduced, and the degree of interstitial oedema and derangement of alveolar structure appeared in a decreasing degree after FAS intervention. Compared with the LPS group, the W/D ratio, MDA content, MPO activity and the expression of Caspase-3 in the FAS+LPS group were significantly reduced (all P<0.01). Meanwhile, the expression of RhoA and ROCK1 of the LPS group were obviously higher than those in the control group ( P<0.05), and p-eNOS was obviously lower than that in the control group ( P<0.05). Furthermore, the expression of RhoA and ROCK1 of the FAS+LPS group were obviously lower than those in the LPS group, and p-eNOS was obviously higher than that in the LPS group. There was no significant difference on the expression of eNOS among the three groups. Conclusions:Fasudil can alleviate the degree of inflammatory cell infiltration, reduce apoptosis in lung tissue, inhibit the RhoA/ROCK1 signaling activity, and promote the phosphorylation expression of eNOS in septic mice.
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RNA N6-methyladenosine (m6A) modification is an important gene expression regulation mechanism of eukaryotes. The m6A modification mainly mediates the methylation of adenosine N6. It is a reversible epigenetic modification that not only occurs in messenger RNA (mRNA), but also occurs in non-coding RNA (ncRNA). In addition, RNA m6A modification participates in many physiological and pathological processes, and also plays an important role in the occurrence and development of tumors. This article reviews the role of RNA m6A modification in malignant tumors of the digestive system.
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Objective@#To explore the association between gastrointestinal problems and autism spectrum disorders(ASD) related symptoms, in order to give some clues to the management of ASD children.@*Methods@#Three hundred and thirty-six ASD children aged from 3-8 years old were recruited for the study according to the inclusion and exclusion criteria.General information about the children, including birth date, gender, food allergy history, gastrointestinal problems in the recent 3 months and social psychological scales, were completed by parents or caretakers.The association between gastrointestinal problems and ASD related symptoms (repetitive and stereotypic behaviors, sensory processing problems, emotional and behavioral problems) were analyzed.@*Results@#Among 336 ASD individuals, gastrointestinal problems were detected in 85 children.General gastrointestinal problem detection rate was 25.3%.ASD children with gastrointestinal problems were more severe in sensory over responsitivity(t=3.172, P<0.05), as well as emotional problems(t=-3.215, P<0.05). According to the regression analysis, gastrointestinal problem was significantly associated with sensory over responsitivity[β(SE)=-0.041, P<0.05, Exp(β)=-0.959], as well as emotional problems[β(SE)=0.375, P<0.05, Exp(β)=1.456]. Specifically, vomit was significantly associated with sensory over responsitivity(B=-14.446, P<0.05), and constipation was significantly associated with emotional problems(B=1.555, P<0.05).@*Conclusions@#Gastrointestinal problems are often seen in Chinese ASD children.Gas-trointestinal problems are significantly associated with sensory over responsitivity, as well as emotional problems, implying that resolving gastrointestinal problems may be helpful to remit these problems.
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Objective To explore the association between gastrointestinal problems and autism spectrum disorders(ASD) related symptoms,in order to give some clues to the management of ASD children.Methods Three hundred and thirty-six ASD children aged from 3-8 years old were recruited for the study according to the inclusion and exclusion criteria.General information about the children,including birth date,gender,food allergy history,gastrointestinal problems in the recent 3 months and social psychological scales,were completed by parents or caretakers.The association between gastrointestinal problems and ASD related symptoms (repetitive and stereotypic behaviors,sensory processing problems,emotional and behavioral problems) were analyzed.Results Among 336 ASD individuals,gastrointestinal problems were detected in 85 children.General gastrointestinal problem detection rate was 25.3％.ASD children with gastrointestinal problems were more severe in sensory over responsitivity (t =3.172,P ＜ 0.05),as well as emotional problems(t =-3.215,P ＜0.05).According to the regression analysis,gastrointestinal problem was significantly associated with sensory over responsitivity[β(SE) =-0.041,P ＜ 0.05,Exp(β) =-0.959],as well as emotional problems[β(SE) =0.375,P ＜0.05,Exp(β) =1.456].Specifically,vomit was significantly associated with sensory over responsitivity(B =-14.446,P ＜ 0.05),and constipation was significantly associated with emotional problems(B =1.555,P ＜ 0.05).Conclusions Gastrointestinal problems are often seen in Chinese ASD children.Gastrointestinal problems are significantly associated with sensory over responsitivity,as well as emotional problems,implying that resolving gastrointestinal problems may be helpful to remit these problems.
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Chromosome microarray analysis (CMA) is a cost-effective molecular cytogenetic technique that has been used as a first-line diagnostic test in neurodevelopmental disorders in the USA since 2011. The impact of CMA results on clinical practice in China is not yet well studied, so we aimed to better evaluate this phenomenon. We analyzed the CMA results from 434 patients in our clinic, and characterized their molecular diagnoses, clinical features, and follow-up clinical actions based on these results. The overall diagnostic yield for our patients was 13.6% (59 out of 434). This gave a detection rate of 14.7% for developmental delay/intellectual disability (DD/ID, 38/259) and 12% for autism spectrum disorders (ASDs, 21/175). Thirty-three recurrent (n ≥ 2) variants were found, distributed at six chromosomal loci involving known chromosome syndromes (such as DiGeorge, Williams Beuren, and Angelman/Prader-Willi syndromes). The spectrum of positive copy number variants in our study was comparable to that reported in Caucasian populations, but with specific characteristics. Parental origin tests indicated an effect involving a significant maternal transmission bias to sons. The majority of patients with positive results (94.9%) had benefits, allowing earlier diagnosis (36/59), prioritized full clinical management (28/59), medication changes (7/59), a changed prognosis (30/59), and prenatal genetic counseling (15/59). Our results provide information on de novo mutations in Chinese children with DD/ID and/or ASDs. Our data showed that microarray testing provides immediate clinical utility for patients. It is expected that the personalized medical care of children with developmental disabilities will lead to improved outcomes in long-term developmental potential. We advocate using the diagnostic yield of clinically actionable results to evaluate CMA as it provides information of both clinical validity and clinical utility.
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Child , Child, Preschool , Female , Humans , Infant , Male , Age Factors , China , Epidemiology , Ethnology , Chromosome Disorders , Genetics , Chromosomes , Genetics , DNA Copy Number Variations , Genetics , Disease Management , Microarray Analysis , Methods , Neurodevelopmental Disorders , Diagnosis , Ethnology , GeneticsABSTRACT
Objective To explore the clinical feature,treatment and prognosis of incidental prostate cancer(IPC) after transurethral resection of the prostate (TURP) for benign prostatic hyperplasia (BPH).Methods From January 2009 to April 2017,24 cases undergoing TURP for benign prostatic hyperplasia and being diagnosed with prostate cancer(T1a-T1b) was retrospectivey analysed,who aged from 62 to 84 years (mean 71.8 years).Digital rectal examination (DRE) showed prostate medium texture,smooth surface,and no nodules.Ultrasound presented no low echo nodules in the prostate.Prostate volumes were 19.2-93.4 ml,with median of 40.1 ml.PSA were 1.81-9.11 ng/ml,with median of 4.12 ng/ml.The patients with PSA between 6-10 ng/ml accepted prostate biopsy,and pathological results were negative.Results The The pathology of TURP specimens in 24 cases were diagnosed prostate cancer (21 cases of T1a,3 cases of T1b).According to the new WHO/ISUP classification group,there were 18 cases of hierarchical group 1,3 cases of hierarchical group 2,1 case of hierarchical group 3,2 cases in hierarchical group 4.All patients were treated with hormonal therapy,and 7 cases (5 cases of hierarchical group 1,and 2 cases of hierarchical group 2) underwent laparoscopic radical prostatectomy (LRP) after 3 months of hormonal therapy.The specimens of prostatectomy were examined by whole-mount serial,showing 3 cases of prostate cancer (T1a) with negative margin,and 4 cases of benign prostate cells.They were followed up for 5-82 months with median of 43.5 months.No biological progression or tumor progression was found,and,1 case died of colon cancer after 26 months of follow-up.The patients' age and Gleason score of stage T1b were higher than that of stage T1a.Prostate volume and preoperative PSA had no statistically significant difference between the two stages.Conclusions The patients' age and Gleason score of stage T1b were higher than that of stage T1b.The proportion of residual tumor following TURP was high.The prognosis of incidental prostate cancer was good by hormonal therapy or radical prostatectomy.
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Epigenetics consists of DNA methylation,post-translational modification of histon,microRNA and long non-coding RNA,which regulate angiogenesis by acting on different targets.Epigenetics has become potential targets of anti-angiogenesis in tumor.
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Objective To investigate the effect of signal transducer and activator of transcription-3(STAT-3)-mediated non-dependent vascular endothelial growth factor receptor-2(VEGFR-2) pathway on the radiosensitivity of non-small cell lung cancer (NSCLC) cells and its possible mechanism.Methods NSCLC cells were divided into control group, apatinib (VEGFR-2 inhibitor) group, apatinib+S3I-201(STAT-3 inhibitor) group, radiation (RT) group, RT+apatinib group, and RT+apatinib+S3I-201 group.Q-PCR and Western blot were used to measure the mRNA and protein expression of VEGFR-2, STAT-3, other proteins involved in the signaling pathway, hypoxia-inducible factor 1α(HIF-1α), and cyclin D1.The cellular radiosensitivity was determined by colony-forming assay and cell apoptosis and cell cycle distribution were evaluated by flow cytometry.Results For Calu-1 cells, there was no significant difference in the expression of VEGFR-2 mRNA and STAT-3 mRNA between the apatinib group and the control group (P>0.05);the apatinib group had significantly lower expression of HIF-1α mRNA and cyclin D1 mRNA than the control group (P<0.05);the apatinib+S3I-201 group had significantly lower mRNA and phosphorylated protein expression of VEGFR-2, STAT-3, and related downstream target proteins than the control group (F=304.54, P<0.01;F=118.99, P<0.01, F=144.34, P<0.01;F=529.66, P<0.01);compared with the control group, the apatinib+S3I-201 group and the apatinib group showed increases in apoptosis rate and proportion of cells in G2+M phase, and the apatinib+S3I-201 group had significantly greater increases than the apatinib group (F=72.37, P<0.01).Compared with Calu-1 cells, the radiosensitizing effect of apatinib on A549 cells was limited (SER[sensitizer enhancement ratio]=1.39).The radiosensitizing effect of apatinib+S3I-201 on A549 cells was significantly higher than that of apatinib (SER:1.72 vs.1.39, P=0.000).Conclusions The inhibition of STAT-3 can enhance the radiosensitivity of NSCLC cells.When VEGFR-2 is inhibited by apatinib, activated STAT-3 regulates the expression of cyclin D1 directly or indirectly to affect the radiosensitivity of NSCLC cells.The inhibition of VEGFR-2 and STAT-3 has a good radiosensitizing effect on NSCLC cells.
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Objective: To evaluate the effect and safety of molecular targeted therapy of apatinib mesylate combined with multiple antigen stimulatiing cellular therapy in treatment of osteosarcoma and soft tissue sarcoma. Methods:Six patients with sarcoma were collected by the failure of surgery, radiation and chemotherapy treatment or refusal surgery, radiation and chemotherapy, and at least one month from the last treatment of surgery, radiation and chemotherapy. All of the patients at least received three cycle MASCTTM . From Day 1,everyone were given Apatinib 500 mg,po,qd ,until the disease progression. To measure the patient’s quality of life depending on EORTC QLQ-C30,meanwhile,detecting the cellular immunity function and circulating tumor cells(CTCs) of patients before treatment and one month after 3 cycle MASCTTM . At last, monitoring the cellular immune responses by the Enzyme-linked immuno spot ( ELISPOT) assay. Results: All of the four patients completed the treatment of 3 cycle MASCTTM . Only one patient reduced apatinib from 500 mg to 250 mg because of palmar-plantar erythrodyses-thesia. The response rates of the four patients received MASCTTM and apatinib mesylate after treatment were 1 for complete response (CR),3 for partial response (PR). The life quality and cellular immunity function were improved in all of the patients. ELISPOT assay suggested that the majority of antigen peptides could induce specific cytotoxic T lymphocytes( CTLs) response. The Progression-Free-Survival ( PFS) of four patients received MASCTTM and apatinib mesylate was 7,6,9 and 4 months ,while the response rates of the two patients received apatinib mesylate were 1 for ( Stable disease) SD,one for ( Progression disease) PD. And PFS of the two patients were one month and two months. Conclusion:Combination of MASCTTM and apatinib mesylate is safe,effective and were good prospects for application.
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Objective@#To explore radiosensitivity-associated genes in esophageal squamous cell carcinoma by targeted sequencing panel.@*Methods@#The peripheral blood from 22 esophageal squamous cell carcinoma (ESCC) patients received radiotherapy alone were collected, respectively. The genomic DNA (gDNA) of peripheral blood was extracted and used to create a library of gDNA restriction fragments. The gDNA restriction fragments were hybridized to the HaloPlex probe capture library, which comprises 356 cancer genes selected from the Catalogue of Somatic Mutations in Cancer (Cosmic) database of 2011 updated edition. The sequencing data were aligned by the Genome Analysis Toolkit GATK (version 3.0) and Picar. The single nucleotide polymorphism and inserted-deletion (SNP/InDel) variations were annotated by online database. The pathway enrichment was analyzed by Ingenuity Pathway analysis (IPA). Moreover, according to the short-period curative effect, 22 patients were divided into two groups: the radiation- sensitivity group (CR+ PR) and the radiation-resistant group (PD+ SD). The nonsynonymous mutation sites were statistically analyzed and the genes associated with radiosensitivity of ESCC were screened.@*Results@#More than 97% sequencing reads were aligned to human genome reference sequence and more than 90% sequencing reads were the target sequences. SNP/InDel database annotation results showed that the mutations of 22 cases mainly distributed in exons, and the mutant types were mainly missense and synonymous single nucleotide variant (SNV). There were 23 genes of high-frequency mutation associated with esophageal cancer. Pathway enrichment by IPA showed that 3 pathways were associated with the development of esophageal cancer, which were roles of BRCA1 in DNA damage response pathway, DNA double-strand break repair by non-homologous end joining pathway and ATM signaling pathway. According to the curative effect, five genes including mismatch repair system component (PMS1), fibronectin 1(FN1), mutL homolog 1 (MLH1), B-Raf proto-oncogene, serine/threonine kinase (BRAF), patched 1 (PTCH1) and cytochrome P450 family 2 subfamily C member 19 (CYP2C19) were associated with radiosensitivity of ESCC patients.Moreover, the PTCH1 was mutated in all of 22 ESCC patients, while the variations of rs199476092 and rs202111971 sites of PTCH1 were only identified in the radiation-resistant group.@*Conclusions@#We find that the variations of rs199476092 and rs202111971 in the encoding region of PTCH1 gene are significantly associated with radiosensitivity of ESCC patients.
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Objective@#To investigate the impact of allergic airway diseases on the risk of attention deficit hyperactivity disorder (ADHD) in school-age children.@*Method@#Used stratified cluster sampling method, school-age children in first to sixth grade in primary schools in 9 randomly selected cities including Shanghai, Guangzhou, Xi′an, and Wuhan were enrolled in the study. Interview of parents with questionnaires, which included school-age individual and family social environment questionnaire (including history of diagnosed ADHD, allergic rhinitis, and bronchial asthma) and Children′s Sleep Habits Questionnaire (CSHQ), were finished and collected during November to December in 2005.Diagnosed allergic rhinitis and asthma by specialist were independent variables and divided into following three categories as no allergic diseases (neither allergic rhinitis nor asthma), single allergic disease (allergic rhinitis or asthma), and combined allergic diseases (allergic rhinitis and asthma). Diagnosed ADHD as dependent variable, binary logistic regress model was used to analyze the risks of ADHD in school-age children.@*Result@#Totally 23 791 questionnaires were handed out, while 22 018 were collected. The children had an average age of (8.8±1.8) years, within which 10 869 were male, and 11 021 were female. The risk ratios of ADHD were 2.197 (95%CI: 1.823-2.648) and 3.150 (95%CI: 2.082-4.760) in children with single allergic disease and combined allergic diseases separately. There was no significant difference after adjusting for the factor of sleep habits, as the risk ratios were 2.055 (95%CI: 1.683-2.508) and 3.140 (95%CI: 2.061-4.784) in children with single and combined allergic airway disease separately.@*Conclusion@#Allergic rhinitis and bronchial asthma increased the risk of ADHD, not depending on sleep habits. Hence, allergic airway diseases could be independent risk factors of ADHD.
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Hypoxia results from long-term anti-angiogenic therapy and can stimulate hypoxia-inducible factors (HIFs). HIF-induced hy-poxia signaling is involved in various steps in tumor invasive-metastatic cascade. On the one hand, HIFs regulate epithelial-mesenchy-mal transition. On the other hand, the characteristics of pericytes around vessels and the links among endothelial cells can change;thus, tumor cells can more easily intravasate into blood vessels, survive in peripheral blood, and then reach specific organs, ultimately resulting in metastasis. This review discusses the emerging mechanisms of long-term anti-angiogenic therapy and the occurrence of metastasis.
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Malignant tumor therapy has entered a new era ofprecise treatment.Nowadays, targeted anti-angiogenic agents have become a popular research topic that continues to attract increasing interest. Tumor immune escape plays an indispensable role in therapeutic resistance. Anti-angiogenic therapies not only prevent the tumor angiogenesis and suppress tumor growth but also neu-tralize tumor escape from a host's immune system by reducing the immunosuppressive cells and increasing the number of tumor-infil-trating lymphocyte (TIL) and cytotoxic lymphocte (CTL). This paper aims to review the mechanism underlying the manner by which an-ti-angiogenesis enhances immunity by influencing tumor microenvironment.
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Objective To investigate the value of X-ray guided desmopressin (DDAVP) stimulated bilateral inferior petrosal sinus sampling (IPSS) in diagnosing the recurrence of ACTH dependent Cushing disease or ineffectiveness after surgery or radiotherapy. Methods Retrospective analyses of patients with recurrent ACTH dependent Cushing disease (31 cases) or ineffective (3 cases) treatment after surgery or radiotherapy from January 2013 to October 2014 in our hospital was conducted. Bilateral inferior petrosal sinus angiography showed the same side of the cavernous sinus to prove successful intubation. The cases with discontinuous of the inferior petrosal sinus and cavernous sinus were excluded by this study. Finally, there were 34 cases of the patients in this study. Diagnosis was based on the ratio of ACTH level in IPS to peripheral vein after desmopressin test.The gradient≥2 at baseline or gradient≥3 after desmopressin test suggested the sources were in the pituitary. Diagnosis was confirmed by gold standard to investigate the value of X-ray guided desmopressin (DDAVP) stimulated bilateral IPSS. Results The IPS gradient≥2 at baseline or gradient≥3 after desmopressin test suggested the sources were in the pituitary in 30 patients. A total of 22 (22/30) patients underwent surgery with a final diagnosis of ACTH adenoma. The symptoms were obviously relieved in 8 (8/30) cases after sellar area gamma knife treatment and lesions were confirmed in the pituitary. IPS gradient<2 at baseline or gradient<3 after desmopressin test was found in 4 cases. One case (1/4) was found to have for ACTH adenoma after pituitary surgery. The other 3 cases (3/4) were confirmed to have lung carcinoid and clinical symptom alleviated after surgery. The sensitivity of desmopressin stimulated IPSS was 96.8%, the specificity was 100%, and the accuracy was 97.1%. Conclusion Desmopressin stimulated IPSS is an effective diagnostic procedure in diagnosing ACTH dependent Cushing disease recurrence or ineffectiveness after surgery or radiotherapy.
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To observe whether HMGB1could enhance the paracrine effect of MSCs when the Mesenchymal stem cells [Mesenchymal stem cells, MSCs] are pre-proccessed by High Mobility Group Box-1 [High Mobility Group Box-1, HMGB1]. And to observe whether it can further increase the quantity of local angiogenesis in myocardial infarcts on the rat model with acute myocardial infarction, HMGB1 was combined with MSCs transplantation. MSCs in rats were cultivated with adherence and centrifugation method. Receptors of TLR4and RAGE in HMGB1 were tested. The MSCs were interfered by HMGB1 with different concentration gradient respectively, then the expression of VEGF was tested with ELISA method. SD male rats were divided into four groups: the model group, the MSCs transplantation group, the HMGB1 injection group, the HMGB1 injection plus MSCs transplantation group [n = 24], preparing rat model with acute myocardial infarction. The serum VEGF concentration levels were detected on the 3[rd] day, 7th and 28th day with ELISA method. On the 28th day after post operation the density of angiogenesis in infarction area was detected by immunohistochemal. [1] MSCs owned the expression of TLR4 and RAGE. [2] the secretion of VEGF increased significantly after the intervention of HMGB1 with concentration of 12.5 ng/mL, 25 ng/mL, 50 ng/mL, 100 ng/mL and 200ng/ml on MSCs compared with the control group. While the concentration was 400ng/ml or 800ng/ml, the secretion of VEGF decreased compared with the control group [P < 0.05]. [3] detection of the serum VEGF on the 3[rd] or7th day after post operation was arranged: The results showed that: HMGB1 injection plus MSCs transplantation group > MSCs transplantation group >HMGB1 injection group >model group [P < 0.05]. [4] the quantity of CD31 stained angiogenesis in HMGB1 injection plus MSCs transplantation group increased obviously. Combining MSCs transplantation, contributed to new angiogenesis of rats with acute myocardial infarction in myocardial infarction area and its near area in rats with acute myocardial infarction
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Objective To investigate the effects of vascular endothelial growth factor receptor?2 ( VEGFR?2) on proliferation, migration, invasion, and apoptosis after radiotherapy in lung cancer cell line Calu?1, and to explore the probable mechanisms. Methods Small interference RNA ( siRNA )?mediated silencing of VEGFR?2 gene was performed on Calu?1 cells, and the mRNA and protein expression of VEGFR?2 was determined by quantitative real?time PCR and Western blot, respectively. The cells were divided into control group, vascular endothelial growth factor ( VEGF ) group, VEGFR?2 specific siRNA (siKDR) group, and siKDR+VEGF group. The changes in proliferation, migration, and invasion were evaluated by the CCK8 assay, cell scratch wound?healing assay, and transwell migration assay, respectively. The protein expression of VEGFR?2 and proteins in the related downstream signaling pathway was measured by Western blot. Apoptosis in each group was determined after radiotherapy. Results After RNA interference?mediated silencing of VEGFR?2, the mRNA and protein expression of VEGFR?2 was significantly reduced ( P=0. 001,P=0. 000);the proliferation, migration, and invasion of Calu?1 cells were also significantly reduced ( P=0. 000,P=0. 000,P=0. 000);the phosphorylation levels of AKT, ERK 1/2, and p38 were significantly reduced in Calu?1 cells ( P=0. 336,P=0. 986,P=0. 553);the apoptosis in Calu?1 cells was significantly elevated ( P=0. 0012);the protein expression of HIF?1α was significantly inhibited ( P= 0. 016 ) . Conclusions The VEGFR?2 gene silencing significantly inhibits several physiological functions of Calu?1 cells and elevates the apoptosis rate after radiotherapy.
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Objective To investigate the radiosensitivity of stem cells in pancreatic adenocarcinoma PANC‐1 cell line and the possible mechanism of its radiation resistance .Methods Using flow cytometry ,the cells were isolated and sorted into CD44+CD24+ ,CD44-CD24+ ,CD44+CD24- ,and CD44-CD24- .Multi‐target click model was used to fit cell survival curves and deter‐mine the sensitizer enhancement ratio .The apoptosis and cycle distribution of the four cell subsets were determined using flow cy‐tometry ,and the level of ROS was determined by DCFH‐DA probe .Results The ratio of CD44+ and CD24+ in the sorted PANC‐1 cell line was 92 .0% and 4 .7% respectively .Before radiation ,there was no statistically significant difference between four groups (P>0 .05);After treated with 6MV‐X ray ,The ratio of apoptosis was the lowest in CD44+CD24+ (P<0 .01);The percentage G0/G1 cell was the highest in CD44+CD24+ (P<0 .01) ,the sensitizer enhancement ratio of CD44+ CD24- ,CD44-CD24+ and CD44-CD24- were 1 .61 ,1 .81 ,1 .94 ,respectively .The level of ROS in CD44+CD24+ was lower (P<0 .01) .Conclusion Tumor stem cells of pancreatic adenocarcinoma have properties of a lower level of ROS and relative stationary that maybe the reasons of radio resist‐ance .
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Aim To study modulation of NMDA recep-tors on inward rectifying potassium channels through investigation of retinal horizontal cells. Methods Whole cell recording was conducted on isolated retinal horizontal cells for observation of inward-rectifying po-tassium currents before and after application of NMDA. Modulation of inward-rectifying currents by NMDA re-ceptor was also tested under calcium-free and chelation of intracellular calcium condition. Results Reduction of inward rectifying potassium currents was induced by activation of NMDA receptors. The effect was attenua-ted when calcium was removed. Conclusion Inhibi-tion of inward rectifying potassium channels can be in-duced by activation of NMDA receptors via intracellular calcium signals.