Pooled analysis of NeoCARH and NeoCART trials: patient-reported outcomes in patients with early-stage breast cancer receiving platinum-based or anthracycline-based neoadjuvant chemotherapy.

PURPOSE: Anthracycline-based or platinum-based neoadjuvant chemotherapy belongs to the standard treatment for early-stage breast cancer (EBC) that is either triple-negative or human epidermal growth factor receptor 2 positive (HER2 +). Currently, there is a paucity of data comparing their impact on health-related quality of life (HRQoL). METHODS: Triple-negative or HER2 + EBC from our two prospective randomized controlled trials, neoCARH and neoCART, were divided into two groups based on the neoadjuvant chemotherapy regimens they received: anthracycline-based or platinum-based group. HRQoL was the exploratory endpoint in these two trials, which was assessed using the European Organization for Research and Treatment of Cancer Quality of Life-Core30 and Breast23 questionnaires. The primary variable of interest was the C30 summary score (C30-SumSc). Assessments were carried out at baseline, after neoadjuvant chemotherapy, and 1 year and 2 years after diagnosis. RESULTS: The mean questionnaires' compliance rate was 95.0%. After neoadjuvant chemotherapy, 210 patients had evaluable HRQoL data, the mean least square change from baseline for the platinum-based group was - 15.997 (95% confidence interval (CI): - 17.877 to - 14.117), and it was - 20.156 (95% CI: - 22.053 to - 18.258) for the anthracycline-based group (difference: 4.159, 95% CI: 1.462 to 6.855, P = 0.003, minimal important difference = 3). For the majority of the domains of interest assessed by the C30 and BR23 questionnaires, the platinum-based group demonstrated superior outcomes in comparison to the anthracycline-based group. CONCLUSION: Patients receiving platinum-based or anthracycline-based regimens both experienced worsened HRQoL after neoadjuvant chemotherapy; however, the former provided relatively better HRQoL compared with the latter. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov: NCT03140553. Registered 4 May 2017 (neoCARH). NCT03154749. Registered 16 May 2017 (neoCART).

Combined measurement of serum zinc with PSA ameliorates prostate cancer screening efficiency via support vector machine algorithms.

Background: Early screening of prostate cancer (PCa) is pivotal but challenging in the clinical scenario due to the phenomena of false positivity or false negativity of some serological evaluations, e.g. PSA testing. Decline of serum Zn2+ levels in PCa patients reportedly plays a crucial role in early screening of PCa. Accordingly, we combined 4 indices comprising the serum levels of total PSA (tPSA), free PSA (fPSA), Zn2+ and demographic information (especially age) in order to ameliorate the efficacies of PCa screening with support vector machine (SVM) algorithms. Methods: A total of 858 male patients with prostate disorders and 345 healthy male controls were enrolled. Patients' data included 4 variables and serum Zn2+ was quantified via a self-invented Zn2+ responsive AIE-based fluorescent probe as previously published. tPSA and fPSA were routinely determined by a chemiluminescent method. Mathematical simulations were conducted to establish a SVM model for the combined diagnostics with the four variables. Moreover, ROC and its characteristic AUC were also employed to evaluate the classification efficacy of the model. Sigmoid function was utilized to estimate corresponding probabilities of classifying the clinical subjects as per 5 grades, which were incorporated into our established prostate index (PI) stratification system. Results: In SVM model, the mean AUC of the ROC with the quartet of variables was approximately 84% for PCa diagnosis, whereas the mean AUC of the ROCs with tPSA, fPSA, [Zn2+] or age alone was 64%, 62%, 55% and 59%, respectively. We further established an integrated prostate index (PI) stratification system with 5 grades and a software package to support clinicians in predicting PCa, with the accuracy of our risk stratification system being 83.3%, 91.6% and 83.3% in predicting normal, benign and PCa cases in corresponding groups. Follow-up findings especially MRI results and PI-RADS scores supported the reliability of this stratification platform as well. Conclusion: Findings from our present study demonstrated that index combination via SVM algorithms may well facilitate clinicians in early differential screening of PCa. Meanwhile, our established PI stratification system based on SVM model and Sigmoid function provided substantial accuracy in preclinical risk prediction of developing prostate cancer.

The Genomic Characteristics of an Arthritis-Causing Salmonella pullorum.

Salmonella enterica subsp. enterica serovar Gallinarum biovar pullorum (Salmonella pullorum) is an avian-specific pathogen that has caused considerable economic losses to the poultry industry. High endemicity, poor implementation of hygiene measures, and lack of effective vaccines hinder the prevention and control of this disease in intensively maintained poultry flocks. In recent years, the incidence of arthritis in chicks caused by Salmonella pullorum infection has increased. In this study, four Salmonella pullorum strains were identified from the livers, spleens, and joint fluids of Qingjiaoma chicken breeders with arthritis clinical signs, and an arthritis model of chicks was successfully established using SP206-2. Whole genome sequencing of the SP206-2 strain showed that the genome was 4,730,579 bp, 52.16% GC content, and contained 5007 genes, including 4729 protein-coding regions. The genomic analysis of four arthritis-causing isolates and three diarrhea-causing isolates showed that the genome of arthritis-causing isolates was subject to nonsynonymous mutations, shift mutations, and gene copy deletions. An SNP phylogenetic tree analysis showed that arthritis-causing isolates are located in a different evolutionary branch from diarrhea-causing isolates. Further differential genes analysis showed that the genome of arthritis-causing isolates had missense mutations in genes related to substance metabolism and substance transport, as a result of adaptive evolution.

Research Note: Therapeutic effect of a Salmonella phage combination on chicks infected with Salmonella Typhimurium.

Antibiotic treatment failure is increasingly encountered for the emergence of pandrug-resistant isolates, including the prototypical broad-host-range Salmonella enterica serovar (S.) Typhimurium, which mainly transmitted to humans through poultry products. In this study we explored the therapeutic potential of a Salmonella phage composition containing a virulent phage and a nonproductive phage that does not produce progeny phage against chicks infected with a pandrug-resistant S. Typhimurium strain of avian origin. After approximately 107 CFU of S. Typhimurium strain ST149 were administrated to chicks by intraperitoneal injection, the phage combination (∼108 PFU) was gavaged at 8-h, 32-h, and 54-h postinfection. At d 10 postinfection, phage treatment completely protected chicks from Salmonella-induced death compared to 91.7% survival in the Salmonella challenge group. In addition, phage treatment also greatly reduced the bacterial load in various organs, with Salmonella colonization levels decreasing more significantly in spleen and bursa than in liver and cecal contents, possibly due to higher phage titers in these immune organs. However, phages could not alleviate the decreased body weight gain and the enlargement of spleen and bursa of infected chicks. Further examination of the bacterial flora in the cecal contents of chicks found that S. Typhimurium infection caused a remarkable decrease in abundance of Clostridia vadin BB60 group and Mollicutes RF39 (the dominant genus in chicks), making Lactobacillus the dominate genus. Although phage treatment partially restored the decline of Clostridia vadin BB60 group and Mollicutes RF39 and increased abundance of Lactobacillus caused by S. Typhimurium infection, Fournierella that may aggravate intestinal inflammation became the major genus, followed by increased Escherichia-Shigella as the second dominate bacterial genus. These results suggested that successive phage treatment modulated the structural composition and abundance of bacterial communities, but failed to normalize the intestinal microbiome disrupted by S. Typhimurium infection. Phages need to be combined with other means to control the spread of S. Typhimurium in poultry.

Development of a H2S-responsive NIR Fluorescent Probe for H2S Detection and H2S Releasing Monitoring From Prodrug.

H2S was deemed as a toxic gradient in the realm of food and environment but plays pivotal pathophysiological roles in organisms. H2S instabilities and disturbances are always responsible for multiple disorders. We fabricated a H2S-responsive NIR fluorescent probe (HT) for H2S detection and evaluation both in vitro and in vivo. HT exhibited rapid H2S response within 5 min, accompanied with visible color change and NIR fluorescence generation, and the fluorescent intensities were linearly correlated with corresponding H2S concentrations. When HT was incubated with A549 cells, the intracellular H2S and H2S fluctuations could be monitored ore rotundo via the responsive fluorescence. Meanwhile, when HT was co-administrated with H2S prodrug ADT-OH, the H2S release from ADT-OH could be visualized and monitored to evaluate its release efficacy.

Survival After Sentinel Lymph Node Biopsy Compared with Axillary Lymph Node Dissection for Female Patients with T3-4c Breast Cancer.

BACKGROUND: For patients with cN0 and T1-2 breast cancer, sentinel lymph node biopsy (SLNB) can provide survival results equivalent to axillary lymph node dissection (ALND). However, whether it can be performed on T3-4c patients is still controversial. MATERIALS AND METHODS: Female patients diagnosed with cN0, T3-4c, and M0 breast cancer from 2004 to 2019 were identified using the surveillance, epidemiology and end results (SEER) database and divided into 2 groups, the SLNB group (1-5 regional lymph nodes examined) and the ALND group (≥10 regional lymph nodes examined). Finally, only those with pN0 disease were included in the SLNB group. The baseline differences in clinicopathological characteristics between groups were eliminated by propensity score matching (PSM). We also conducted subgroup analyses according to age, overall TNM stage, breast cancer subtypes, surgical approaches, radiation therapy, and chemotherapy. The primary endpoint was survival. RESULTS: With a mean follow-up of 75 months, a total of 186 deaths were reported among 864 patients. The overall survival (OS) and breast cancer-specific survival (BCSS) in the SLNB group were 78.2% and 87.5%, respectively, and that in the ALND group were 78.7% and 87.3%, respectively. The unadjusted hazard ratio (HR) for OS and BCSS in the SLNB group (vs. the ALND group) was 0.922 (95% CI, 0.691-1.230, P = .580) and 0.874 (95% CI, 0.600-1.273, P = .481), respectively. Besides, the OS and BCSS between the 2 groups were also similar in all subgroup analyses. CONCLUSIONS: SLNB may be performed on female patients with cN0, T3-4c, and M0 breast cancer.

Survival differences between HER2-0 and HER2-low-expressing breast cancer - A meta-analysis of early breast cancer patients.

BACKGROUND: HER2-low (human epidermal growth factor receptor 2) breast cancer takes up 40-50% in all breast cancer subtypes. The survival difference between HER2-low and HER2-zero breast cancers remain uncertain. Therefore, the aim of this study was to compare survival outcome of the two subtypes and to explore the impact of hormone receptor status. METHODS: A comprehensive medical literature search was performed by searching PubMed, EMBASE, and the Cochrane Libraries up to August 2022. We included observational studies reporting hazard ratios (HRs) with corresponding 95% confidence intervals (CIs). The results of individual studies were pooled by random-effects models using Stata 16.0. Seventeen articles with a total of 78984 breast cancer patients were included in the meta-analysis. RESULTS: We observed a statistically significant association between low HER2 expression and better breast cancer survival outcomes (OS: HR: 0.83; 95% confidence interval: 0.75, 0.90; DFS/RFS: HR: 0.83; 95% confidence interval: 0.75, 0.91). In a subgroup analysis, we found that HER2-low patients had better survival outcomes relative to hormone receptor-positive breast cancer patients (OS: HR: 0.87; 95% confidence interval: 0.81, 0.93; DFS/RFS: HR: 0.91; 95% confidence interval: 0.85, 0.96). Similarly, in triple-negative breast cancer patients, we also observed a positive association between HER2 low expression and better survival (OS: HR: 0.85; 95% confidence interval: 0.71, 0.98; DFS/RFS: HR: 0.85; 95% confidence interval: 0.74, 0.95). CONCLUSIONS: Our study showed that HER2-low breast cancer had better survival outcomes compared to HER2 negative breast cancer in patients with early stage breast cancer, regardless of hormone receptor status. REGISTRATION: This meta-analysis was registered with PROSPERO (CRD42022335704) on June 10, 2022. AVAILABILITY OF DATA AND MATERIALS: All data generated or analysed during this study are included in this published article [and its supplementary information files].

Salmonella Phage CKT1 Effectively Controls the Vertical Transmission of Salmonella Pullorum in Adult Broiler Breeders.

Phage therapy is widely being reconsidered as an alternative to antibiotics for the treatment of multidrug-resistant bacterial infections, including salmonellosis caused by Salmonella. As facultative intracellular parasites, Salmonella could spread by vertical transmission and pose a great threat to both human and animal health; however, whether phage treatment might provide an optional strategy for controlling bacterial vertical infection remains unknown. Herein, we explored the effect of phage therapy on controlling the vertical transmission of Salmonella enterica serovar Gallinarum biovar Pullorum (S. Pullorum), a poultry pathogen that causes economic losses worldwide due to high mortality and morbidity. A Salmonella phage CKT1 with lysis ability against several S. enterica serovars was isolated and showed that it could inhibit the proliferation of S. Pullorum in vitro efficiently. We then evaluated the effect of phage CKT1 on controlling the vertical transmission of S. Pullorum in an adult broiler breeder model. The results demonstrated that phage CKT1 significantly alleviated hepatic injury and decreased bacterial load in the liver, spleen, heart, ovary, and oviduct of hens, implying that phage CKT1 played an active role in the elimination of Salmonella colonization in adult chickens. Additionally, phage CKT1 enabled a reduction in the Salmonella-specific IgG level in the serum of infected chickens. More importantly, the decrease in the S. Pullorum load on eggshells and in liquid whole eggs revealed that phage CKT1 effectively controlled the vertical transmission of S. Pullorum from hens to laid eggs, indicating the potential ability of phages to control bacterial vertical transmission.

Research Note: Effect of a live Salmonella Enteritidis vaccine against Salmonella Pullorum infection in breeder chickens.

Salmonella Pullorum is one of the most important avian pathogenic bacteria due to widespread outbreaks accompanied by high mortality. It has been demonstrated that the Salmonella Enteritidis live vaccine strain Sm24/Rif12/Ssq is able to induce cross-immunity protection against Salmonella Gallinarum and Salmonella Infantis, however, it is unknown whether this vaccine is effective against Salmonella Pullorum infection. In the present study, the Hubbard parent chickens were orally administrated this vaccine at 1-day-old, 40-day-old, and 131-day-old respectively, and challenged by Salmonella Pullorum at 157-day-old to evaluate the protective effect of the Salmonella Enteritidis live vaccine strain Sm24/Rif12/Ssq. After each vaccination, the vaccine strain could be recovered from cloacal swabs within a week, whereas no vaccine strain was re-isolated from environmental samples throughout the experiment. Vaccination for the breeder chickens with Salmonella Enteritidis Sm24/Rif12/Ssq could relieve swollen liver (P = 0.0066) caused by Salmonella Pullorum infection and decrease Salmonella Pullorum colonization level in spleen (P = 0.0035), whereas no significant difference was found in the bacterial counts of liver, ovary and oviduct of vaccinated chickens. These results suggested that the Salmonella Enteritidis live vaccine strain Sm24/Rif12/Ssq was high safety and effective against Salmonella Pullorum infection to a certain extent.

A Combination of Virulent and Non-Productive Phages Synergizes the Immune System against Salmonella Typhimurium Systemic Infection.

Effective phage cocktails consisting of multiple virus types are essential for successful phage therapy against pandrug-resistant pathogens, including Salmonella enterica serovar (S.) Typhimurium. Here we show that a Salmonella phage, F118P13, with non-productive infection and a lytic phage, PLL1, combined to inhibit pandrug-resistant S. Typhimurium growth and significantly limited resistance to phages in vitro. Further, intraperitoneal injection with this unique phage combination completely protected mice from Salmonella-induced death and inhibited bacterial proliferation rapidly in various organs. Furthermore, the phage combination treatment significantly attenuated the inflammatory response, restored the generation of CD4+ T cells repressed by Salmonella, and allowed macrophages and granulocytes to participate in immunophage synergy to promote bacterial clearance. Crucially, the non-productive phage F118P13 is less likely to be cleared by the immune system in vivo, thus providing an alternative to phage cocktail against bacterial infections.

Identification of Key Genes and Pathways Involved in Circulating Tumor Cells in Colorectal Cancer.

BACKGROUND: Characterization of the features associated with circulating tumor cells (CTCs) is one of major interests for predicting clinical outcome of colorectal cancer (CRC) patients. However, the molecular features of CTCs remain largely unclear. METHODS: For identification of key genes and pathways, GSE31023, contained CTCs from six metastatic CRC patients and three controls, was retrieved for differentially expressed gene (DEG) analysis. Protein-protein interaction networks of DEGs were constructed. Hub genes from the network were prognostic analyzed, as well as the association with tumor-infiltrating immune cells. RESULTS: 1353 DEGs were identified between the CTC and control groups, with 403 genes upregulated and 950 downregulated. 32 pathways were significantly enriched in KEGG, with ribosome pathway as top. The top 10 hub genes were included, including eukaryotic translation elongation factor 2 (EEF2), ribosomal protein S2 (RPS2), ribosomal protein S5 (RPS5), ribosomal protein L3 (RPL3), ribosomal protein S3 (RPS3), ribosomal protein S14 (RPS14), ribosomal protein SA (RPSA), eukaryotic translation elongation factor 1 alpha 1 (EEF1A1), ribosomal protein S15a (RPS15A), and ribosomal protein L4 (RPL4). The correlation between CD4+ T cells and RPS14 (correlation = -0.5) was the highest in colon cancer while CD8+ T and RPS2 (correlation = -0.53) was the highest in rectal cancer. CONCLUSION: This study identified potential role of ribosome pathway in CTC, providing further insightful therapeutic targets and biomarkers for CRC.

Salmonella phage CKT1 significantly relieves the body weight loss of chicks by normalizing the abnormal intestinal microbiome caused by hypervirulent Salmonella Pullorum.

Pullorum disease caused by Salmonella Pullorum remains an important disease for the poultry industry due to high morbidity and mortality in many countries. Phage therapy is becoming an alternative strategy to control multidrug-resistant Salmonella infections in young chicks. However, how bacteriophages affect the growth performance of chicks infected with S. Pullorum remains poorly understood. Herein, we assessed the therapeutic efficacy of Salmonella phage CKT1 against hypervirulent arthritis-causing S. Pullorum. The results showed that single phage treatment after hypervirulent S. Pullorum infection significantly improved body weight loss of chicks. Compared with enlarged liver and spleen in only Salmonella challenged group, phage administration substantially reduced the liver/body and spleen/body weight ratios, bacterial loads in organs and the degree of hepatic sinusoidal dilatation and congestion. Moreover, phage CKT1 can enter the organs of chicks and stay for at least 3 d in liver and spleen, and promote higher serum levels of IL-6 production within 6 d postinfection, indicating phage-induced bacterial lysis may be involved in inflammatory immune response to S. Pullorum infection. Analysis of the microbiome of gastrointestinal tract of chicks demonstrated that Salmonella challenge significantly reduced the relative abundances of Lachnoclostridium and Blautia, resulting in remarkably increased Escherichia-Shigella and Klebsiella becoming the predominant bacterial taxa. In contrast, the use of phage CKT1 significantly reduced Escherichia-Shigella and Klebsiella populations in intestine, permitting the proliferation of beneficial microbiota in Firmicutes including Lachnoclostridium, Ruminococcus, Lactobacillus, and Pseudoflavonifractor. In addition, phage alone treatments did not affect the normal gut microbiota structure of chicks, and phage therapy on Salmonella infected chicks increased bacteria species richness in the cecum. These results suggest that Salmonella phage CKT1 could improve growth performance of chicks challenged with S. Pullorum by normalizing the abnormal intestinal microbiome.

Synthesis and preliminary exploration of a NIR fluorescent probe for the evaluation of androgen dependence of prostate cancer.

PURPOSE: Castration resistance prostate cancer patients showing resistance to the androgen deprivation therapy always have low five-year survival rate and worse prognosis. A responsive NIR fluorescent probe was designed to report the androgen dependence and monitor the development of castration resistance for prostate cancer. METHODS: Intratumoral H2S in prostate cancer was closely related to castration resistance. A H2S-responsive NIR probe (HM) was developed as a dependent indicator to report the androgen dependence of prostate cancer. The specificity of HM to H2S and the influence of normal intracellular substrates to the response between H2S and HM were determined. Cell/in vivo animal imaging were performed on PC-3 and LnCAP cell/tumor bearing mice, which presented with androgen independence and androgen dependence, respectively. RESULTS: When HM responded to H2S, strong fluorescence at 770 nm could be rapidly turned on in 5 min with the stokes shift as large as 200 nm. The recognition between HM and H2S showed high specificity. Neither other common substrates showed capacity to turn on HM's fluorescence, nor their existence demonstrated competition. The fluorescence intensity was linearly dependent to the H2S concentration and the limited of detection was 0.15 µM. When HM was applied to PC-3/LNCaP prostate cancer cell and tumor, the intracellular and intratumoral H2S could be clearly imaged and monitored. CONCLUSION: HM showing obvious fluorescent behaviors in androgen dependence and independence prostate tumor, which could work as an indicator to reported the androgen dependence of prostate cancer and monitor the development of castration resistance.

Development and preliminary evaluation of an integrin α2ß1-targeted PET probe as a supplement and alternative of PSMA imaging for prostate cancer.

An integrin α2ß1-targeted PET probe (68Ga-IABtP) was developed to serve as a supplement and alternative of PSMA imaging for prostate cancer. 68Ga-IABtP was synthesized by labeling the precursor peptide with 68Ga with 93% labeling yield and 4.14 MBq/µg specific radioactivity. 68Ga-IABtP showed no specific uptake in LNCaP prostate cancer cell with low integrin α2ß1 expression but significantly increased uptake in PC-3 prostate cancer cell with high integrin α2ß1 expression, which could be specifically blocked by the integrin α2ß1 monoclonal antibody. The efflux experiments demonstrated that 68Ga-IABtP could rapidly penetrate into PC-3 cell after cell binding, thereby prolonging the residence time in the tumor and allow enough time for probe clearance from the circulation and non-specific organs. The biodistribution study indicated that 68Ga-IABtP showed no specific accumulation in non-target organs and was quickly cleared from the kidney. The in vivo PET-CT imaging demonstrated that 68Ga-IABtP showed no specific uptake in LNCaP tumor but could specifically accumulate in the PC-3 tumor, and was rapidly cleared from spleen, intestine, kidney and liver, resulting in excellent contrast effect with low background signal and high target to non-target ratios.

Research Note: Hypervirulent arthritis-causing Salmonella Pullorum isolated from Chinese native chicken breeds significantly decreased growth performance of chicks.

Pullorum disease is characterized by white diarrhea resulting from infection by S. Pullorum, but arthritis associated with S. Pullorum infection has become increasingly frequent recently, especially in Chinese native chicken flocks. In this study, we isolated and identified 4 S. Pullorum strains from the Qingjiaoma chicken breeders with arthritis symptoms. The LD50 of the isolate 20JS04 was 1.33 × 106 CFU, which was considered as a highly virulent strain in chicks. Reproducible arthritis symptoms were observed in the experimentally chickens infected with the isolate 20JS04, and the disease occurrence was 27.78% (5/18). In addition to the characteristics of high virulence and induced-arthritis, our results confirmed that the arthritis-causing isolate 20JS04 had greater negative impact on BW, ADFI, and ADG, compared with the white diarrhea-causing S. Pullorum standard strain CVCC526 (P < 0.05). These results suggest that the pathogenic diversity of S. Pullorum in China deserves more attention and stringent measures should be taken to control salmonellosis.

Development of a NIR fluorescent probe for the detection of intracellular cysteine and glutathione and the monitoring of the drug resistance.

Intracellular cysteine and glutathione was deemed as the most important reductants in the cell and played significant roles in the cellular homeostasis and redox adjustment. Here we developed a NIR fluorescent probe (HI) to detect and report the intracellular cysteine and glutathione, and monitor the development of the drug resistance of tumor. HI with both excited wavelength and emitting wavelength located within near infrared area showed no fluorescence in the normal physiological environment. However, when HI responded to cysteine and glutathione, strong NIR fluorescence could be turned on, which was linear dependent to the cysteine concentrations and the limited of detection was 0.18 µM. The response between HI and cysteine/glutathione demonstrated high specificity and no other amino acids showed influence or competition. The HPLC identification of the recognition results confirmed the response of acryloyloxy on the HI and active sulfhydryl on the cysteine/glutathione. DFT calculation of the HOMO and LUMO energy before and after response revealed the intramolecular charge transfer mechanism that induced the generation of the fluorescence. When HI was incubated with PATU-8988 and PATU-8988/Fu cell, the intracellular cysteine and glutathione could be clearly imaged and monitored by the enhanced fluorescence. Meanwhile, when HI was applied to the tumor-bearing mice, the drug resistance of tumor could be monitored and reported.

Detection and Clinical Value of Circulating Tumor Cells as an Assisted Prognostic Marker in Colorectal Cancer Patients.

BACKGROUND: Circulating tumor cells (CTCs) are cells that have been shed into the vasculature from a primary tumor and circulate in the bloodstream. It has been suggested that detecting CTCs could help the clinician to detect early metastasis or recurrence more effectively. This trial sets out to assess the detection and clinical value of CTCs as an assisted prognostic marker in patients with colon cancer and rectal cancer. METHODS: A prospective cohort of patients with colorectal cancer (CRC) was enrolled from July 2015 to February 2018 in Shanghai Minimally Invasive Surgery Center, Shanghai, China. In this study, 149 patients with CRC were enrolled and underwent surgical treatment. There were 79 cases of colon cancer and 70 cases of rectal cancer, including 93 males and 56 females. To investigate the correlativity and clinical value of CTCs, the patients were statistically analyzed in different subgroups: colon cancer group vs rectal cancer group, and left hemicolon cancer group vs right hemicolon cancer group. RESULTS: The results of analysis comparing CTC counts and clinical pathological features in colon and rectal cancer indicated that with increased tumor stage, the number of CTCs also increased, with significant statistical differences. CTC counts in patients with colon and rectal cancer showed positive correlations with TNM staging (P=0.001, 0.013, respectively), T staging (P=0.021, 0.001), N staging (P=0.014, 0.035) and M staging (P=0.018, 0.203). Detection of serum biomarkers in CTC-positive and CTC-negative groups indicated a significantly increasing expression in the CTC-positive group. To confirm the correlations between CTCs and histoembryological differences, analysis was conducted with the patients in two subgroups: left hemicolon cancer group and right hemicolon cancer group. The results showed that the positive rate of CTCs increased in both groups with the increase in tumor stage. The survival analysis indicated that there was a steep gradient in survival in the follow-up period, particularly in the CTC-positive group (P=0.000). Risk assessment curves showed that the change escalated more rapidly in the CTC-positive group. Furthermore, with the increase in T stage, changes in the survival curve and risk curve escalated more rapidly in the CTC-positive group. CONCLUSION: It was confirmed that in the left hemicolon cancer group, a much higher coincidence rate could be found on CTC-positive rate and clinicopathological features, than in the right hemicolon cancer group. The sensitivity of CTCs may be related to the histoembryological location of the tumor, lymphatic metastasis and the depth of infiltration. Monitoring CTCs may have value in evaluating clinical staging and estimating clinical prognosis.

Evaluation of a new magnetic bead as an integrated platform for systematic CTC recognition, capture and clinical analysis.

A novel form of magnetic bead, namely antibody-coated magnetic lipid nano-vehicle (AMLV), was synthesized by embedding Fe3O4 nanoparticles into an amphiphilic antibody-modified liposome as a high-performance circulating tumor cell (CTC) hunter. The CTC capture performance of AMLV was validated based on an enlarged patient sample (including 318 colorectal, 78 breast, 77 lung and 55 liver cancer patients) with high detection rate. The preliminary comparison with Cellsearch was also conducted, indicating that the cell membrane-semblance AMLVEpCAM showed higher capture performance for different kinds of EpCAM-expressed circulating tumor cells in the peripheral blood (4.4 ± 1.2-fold for AMLVEpCAM vs CellsearchTM, n=5, P<0.001). Moreover, the AMLVEpCAM-isolated CTCs could be used as a functional material to provide various clinical information for tumor patients and work as an alternative of tumor tissue to conduct gene analysis after conventional PCR amplification.

Ratiometric nanoprobe for circulating tumor cell detection and intracellular hydrogen peroxide evaluation in colorectal cancer patients.

The application of intensity-based H2O2-responsive fluorescence nanoprobe for circulating tumor cell detection was limited by the complex background and the nanoprobe uptake in each CTC. In this context, we developed a ratiometric fluorescence nanoprobe, on which a H2O2-responsive subunit and a stable subunit grafted working as a H2O2 detector and a reference, respectively. When responding to intracellular H2O2, the reference fluorescence (580 nm) maintained as a correction background while the detector fluorescence (450 nm) was turned on to conduct CTC enumeration and intracellular H2O2 evaluation. Two normal cells and three colon cancer cells were examined to evaluate their endogenous H2O2 with the ratiometric nanoprobe by flow cytometry and confocal laser scanning microscopy. CTC sample from colorectal cancer patients was used to validate the performance of the nanoprobe for CTC enumeration and H2O2 evaluation. The results indicated that not only CTC could be effectively identified based on the "turn on" fluorescence, but also the viability of the identified CTCs could be assessed with the intensity of the reference fluorescence to avoid the false-positive number. Moreover, the clinical results demonstrated that the viability CTC count combined with intracellular H2O2 content (described as I450/580)were related to the tumor TNM stage, which might provide significant guidance for clinical treatments.

Hydrogen peroxide-response nanoprobe for CD44-targeted circulating tumor cell detection and H2O2 analysis.

Circulating tumor cells (CTCs) represent the most common way of tumor metastasis and has been considered as a significant index for tumor diagnosis, staging and prognosis. However, CTC detection and analysis are always limited by the scarcity of CTC in the peripheral blood and the interference of blood cells. Therefore, here we presented with a hydrogen peroxide (H2O2)-response nanoprobes with CD44-targeted ability to reduce the interference of blood cells and improve the detection efficiency and accuracy and the pancreatic cancer cell was used to evaluate the feasibility of our probe. Shortly, hydrophobic H2O2-response naphthalimide-borate fluorophore was introduced onto the hydrophilic hyaluronic acid to form an amphiphilic complex, which could self-assemble into fluorescent nanoprobes in water. Our studies demonstrated that the nanoprobes were not only able to specifically recognize the pancreatic cancer cells with overexpressed CD44 proteins and reduce the influence of white blood cells in the peripheral blood, but also capable of semi-quantifying H2O2 content in CTCs, Which could be further used as a significant index for tumor clinical evaluation and therapy.