Electrical Characterization and Analysis of Single Cells and Related Applications.

Biological parameters extracted from electrical signals from various body parts have been used for many years to analyze the human body and its behavior. In addition, electrical signals from cancer cell lines, normal cells, and viruses, among others, have been widely used for the detection of various diseases. Single-cell parameters such as cell and cytoplasmic conductivity, relaxation frequency, and membrane capacitance are important. There are many techniques available to characterize biomaterials, such as nanotechnology, microstrip cavity resonance measurement, etc. This article reviews single-cell isolation and sorting techniques, such as the micropipette separation method, separation and sorting system (dual electrophoretic array system), DEPArray sorting system (dielectrophoretic array system), cell selector sorting system, and microfluidic and valve devices, and discusses their respective advantages and disadvantages. Furthermore, it summarizes common single-cell electrical manipulations, such as single-cell amperometry (SCA), electrical impedance sensing (EIS), impedance flow cytometry (IFC), cell-based electrical impedance (CEI), microelectromechanical systems (MEMS), and integrated microelectrode array (IMA). The article also enumerates the application and significance of single-cell electrochemical analysis from the perspectives of CTC liquid biopsy, recombinant adenovirus, tumor cells like lung cancer DTCs (LC-DTCs), and single-cell metabolomics analysis. The paper concludes with a discussion of the current limitations faced by single-cell analysis techniques along with future directions and potential application scenarios.

Aptamer-conjugated gold nanoparticles and their diagnostic and therapeutic roles in cancer.

The burden of incidence rate and mortality of cancer is increasing rapidly, and the development of precise intervention measures for cancer detection and treatment will help reduce the burden and pain of cancer. At present, the sensitivity and specificity of tumor markers such as CEA and CA-125 used clinically are low, while PET, SPECT, and other imaging diagnoses with high sensitivity possess shortcomings, including long durations to obtain formal reports and the inability to identify the molecular pathological type of cancer. Cancer surgery is limited by stage and easy to recur. Radiotherapy and chemotherapy often cause damage to normal tissues, leading to evident side effects. Aptamers can selectively and exclusively bind to biomarkers and have, therefore, gained attention as ligands to be targeted for cancer detection and treatment. Gold nanoparticles (AuNPs) are considered as promising nano carriers for cancer diagnosis and treatment due to their strong light scattering characteristics, effective biocompatibility, and easy surface modification with targeted agents. The aptamer-gold nanoparticles targeting delivery system developed herein can combine the advantages of aptamers and gold nanoparticles, and shows excellent targeting, high specificity, low immunogenicity, minor side effects, etc., which builds a bridge for cancer markers to be used in early and efficient diagnosis and precise treatment. In this review, we summarize the latest progress in the application of aptamer-modified gold nanoparticles in cancer targeted diagnosis and delivery of therapeutic agents to cancer cells and emphasize the prospects and challenges of transforming these studies into clinical applications.

Multi-omics characterization of the unsaturated fatty acid biosynthesis pathway in colon cancer.

The biosynthesis of unsaturated fatty acids is involved in the initiation and progression of colon adenocarcinoma (COAD). In this study, we aimed to investigate the multi-omics characteristics of unsaturated fatty acid biosynthesis-related genes and explore their prognostic value in colon cancer by analyzing the data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. An unsaturated fatty acid biosynthesis pathway related-genes enrichment score (BUFAS) was constructed utilizing the single sample gene set enrichment analysis (ssGSEA). We discovered that a high BUFAS was associated with longer overall survival (OS) in both the training and the validation sets. Multivariable analysis including the clinical characteristics further verified the independent prognostic value of the BUFAS in both the TCGA-COAD and the GSE39582 datasets. In addition, GSEA analysis revealed that BUFAS was positively associated with several signaling pathways, including MTORC1, peroxisome, and pathways related to fatty acid metabolism, while was negatively associated with other signaling pathways, such as hedgehog, NOTCH, and Wnt/beta-catenin pathway. Furthermore, in the COAD cell lines of the Genomics of Drug Sensitivity in Cancer (GDSC) database, we found that BUFAS was positively correlated with the drug sensitivities of cisplatin, gemcitabine, camptothecin, lapatinib, and afatinib, while was negatively correlated with that of ponatinib. Moreover, in the COAD single-cell transcriptomic dataset (GSE146771), the BUFAS varied among different cell types and was enriched in mast cells and fibroblasts. Taken together, the BUFAS we constructed could be used as an independent prognostic signature in predicting the OS and drug resistance of colon cancer. Unsaturated fatty acid biosynthesis pathway might serve as potential therapeutic targets for cancer treatment.

Risk Factors for Synchronous Peritoneal Metastases in Colorectal Cancer: A Systematic Review and Meta-Analysis.

Background: Early detection of synchronous colorectal peritoneal metastases (CPMs) is difficult due to the absence of typical symptoms and the low accuracy of imaging examinations. Increasing the knowledge of the risk factors for synchronous CPM may be essential for early diagnosis and improving their management. This study aimed to identify the risk factors for synchronous CPM. Method: The study was registered at PROSPERO (CRD42020198548). The PubMed, Embase and Cochrane Library databases were searched for studies comparing the clinicopathological and molecular features between patients with or without synchronous CPM. The pooled data were assessed by a random-effects model. Results: Twenty-five studies were included. A synchronous CPM was positively associated with female sex (OR 1.299; 1.118 to 1.509; P = 0.001), PROK1/PROKR2-positivity (OR 2.244; 1.031 to 4.884; P = 0.042), right-sided colon cancer (OR 2.468; 2.050 to 2.970; P < 0.001), poorly differentiated grade (OR 2.560; 1.537 to 4.265; P < 0.001), BRAF mutation (OR 2.586; 1.674 to 3.994; P < 0.001), mucinous adenocarcinoma (OR 3.565; 2.095 to 6.064; P < 0.001), signet-ring cell carcinoma (OR 4.480; 1.836 to 10.933; P = 0.001), N1-2 (OR 5.665; 3.628 to 8.848; P < 0.001), T4 (OR 12.331; 7.734 to 19.660; P < 0.001) and elevated serum CA19-9 (OR 12.868; 5.196 to 31.867; P < 0.001). Conclusions: These evidence-based risk factors are indicators that could predict the presence of synchronous CPMs and can improve their management. Systematic Review Registration: www.crd.york.ac.uk/prospero, identifier: CRD42020198548.

Comprehensive Analysis of Microsatellite-Related Transcriptomic Signature and Identify Its Clinical Value in Colon Cancer.

Background: Microsatellite has been proved to be an important prognostic factor and a treatment reference in colon cancer. The transcriptome profile and tumor microenvironment of different microsatellite statuses are different. Metastatic colon cancer patients with microsatellite instability-high (MSI-H) are sensitive to immune checkpoint inhibitors (ICIs), but not fluorouracil. Efforts have been devoted to identify the predictive factors of immunotherapy. Methods: We analyzed the transcriptome profile of different microsatellite statuses in colon cancer by using single-cell and bulk transcriptome data from publicly available databases. The immune cells in the tumor microenvironment were analyzed by the ESTIMATION algorithm. The microsatellite-related gene signature (MSRS) was constructed by the least absolute shrinkage and selection operator (LASSO) Cox regression based on the differentially expressed genes (DEGs) and its prognostic value and predictive value of response to immunotherapy were assessed. The prognostic value of the MSRS was also validated in another cohort. Results: The MSI-H cancers cells were clustered differentially in the dimension reduction plot. Most of the immune cells have a higher proportion in the tumor immune microenvironment, except for CD56 bright natural killer cells. A total of 238 DEGs were identified. Based on the 238 DEGs, a neural network was constructed with a Kappa coefficient of 0.706 in the testing cohort. The MSRS is a favorable prognostic factor of overall survival, which was also validated in another cohort (GSE39582). Besides, MSRS is correlated with tumor mutation burden in MSI-H colon cancer. However, the MSRS is a barely satisfactory factor in predicting immunotherapy with the area under the curve (AUC) of 0.624. Conclusion: We developed the MSRS, which is a robust prognostic factor of overall survival in spite of a barely satisfactory immunotherapy predictor. Further studies may need to improve the predictive ability.

Association of minimal residual disease levels with clinical outcomes in patients with mantle cell lymphoma: A meta-analysis.

Some studies have elucidated that Minimal residual disease (MRD) in patient with Mantle Cell Lymphoma (MCL) was a significant prognostic factor, with potential value in assessing overall survival (OS) and progression-free survival (PFS). However, most studies were widely varied in included population, sample sources and MRD detection time points. Some studies even have conflicting results. In view of this, a meta-analysis was performed to evaluate association of MRD levels with clinical outcomes in patients with MCL. We identified 7 included articles, which were published in recent 20 years. Then, we extracted or calculated hazard ratios (HRs) and their 95 % confidence intervals (CIs). Our results reveal that patients with MRD negativity have improved OS (HR = 0.63; 95 % CI: 0.50-0.79) and PFS (HR = 0.40, 95 % CI: 0.21-0.76), comparing with patients with MRD positivity. There are also consistent results in subgroups based on sample sources and MRD detection time points. Our study also demonstrates that MRD level is a strong prognostic factor of clinical outcomes. Thus, MRD is expected to be an effective clinical indicator for assessing prognosis and guide treatment decisions in MCL patients.

Hyperhomocysteinemia exaggerates adventitial inflammation and angiotensin II-induced abdominal aortic aneurysm in mice.

RATIONALE: A number of epidemiological studies have suggested an association of hyperhomocysteinemia (HHcy) and abdominal aortic aneurysm (AAA), but discrepancies exist. In addition, we lack direct evidence supporting a causal role. OBJECTIVE: We determined the association and contribution of HHcy to AAA formation. METHODS AND RESULTS: We first performed a meta-analysis of studies involving 1489 subjects and found a strong association of HHcy and AAA (odds ratio, 7.39). Next, we used angiotensin II-infused male apolipoprotein E-deficient mice and tested whether HHcy contributes to AAA pathogenesis. Homocysteine (Hcy) supplement (1.8 g/L) in drinking water resulted in mild HHcy. Intriguingly, HHcy greatly increased the incidence of angiotensin II-induced AAA and aortic dissection in apolipoprotein E-deficient mice (vehicle versus Hcy: 50% versus 100%; P<0.05). Histology indicated HHcy markedly exaggerated aortic adventitial inflammation. Increased levels of proinflammatory interleukin-6 and monocyte chemoattractant protein-1 were preferentially colocalized within adventitial fibroblasts in HHcy plus angiotensin II mice, which suggested the importance of adventitial fibroblasts activation in Hcy-aggravated AAA. Hcy sequentially stimulated adventitial fibroblasts transformation into myofibroblasts, secretion of interleukin-6 and monocyte chemoattractant protein-1, and consequent recruitment of monocytes/macrophages to adventitial fibroblasts, which was abolished by the NADPH oxidase inhibitor diphenyliodonium. NADPH oxidase 4, but not other homologs of NADPH oxidase, was significantly upregulated by Hcy in adventitial fibroblasts, whereas NADPH oxidase 4 small interfering RNA silencing diminished Hcy-induced adventitial fibroblasts activation. Finally, folic acid supplement (0.071 µg/g per day) markedly reduced HHcy-aggravated angiotensin II-induced AAA formation in apolipoprotein E-deficient mice. CONCLUSIONS: HHcy may aggravate AAA formation at least partially via activating adventitial fibroblast NADPH oxidase 4.

[Expression and clinical significance of matrilysin (MMP-7) in human rectal cancer].

OBJECTIVE: To detect the matrilysin (MMP-7) expression in human rectal cancer and to investigate whether it is correlated with invasion and metastasis of human rectal cancer. METHODS: The paired samples obtained from 100 inpatients were allocated into cancer group and control group. By Quantitative-Real-Time RT-PCR, immunohistochemical staining and computerized image analysis, the mRNA and protein expressions of MMP-7 in human rectal cancer were measured and further analyzed for the relationship between MMP-7 expression and clinicopathologic characters. RESULTS: MMP-7 mRNA expression in cancer group was higher than that in control group (P = 0.001), the mRNA expression ratio of 67 (7%) samples was over 1. The mRNA expression level of MMP-7 was correlated with age, Dukes's Staging, lymph node metastasis and histological differentiation grade. The positive degree of immunohistochemical staining for MMP-7 in cancer group (1.94 +/- 0.21) was higher than that in control group (1.15 +/- 0.20, P = 0.002). The protein expression of MMP-7 was correlated with age, Dukes's Staging and lymph node metastasis. CONCLUSION: MMP-7 expression in human rectal cancer increases significantly and plays a key role in the invasion and metastasis of human rectal cancer.

Overexpression of connective tissue growth factor WISP-1 in Chinese primary rectal cancer patients.

AIM: To clarify the expression change of Wnt-induced secreted protein-1 (WISP-1) in human rectal cancer and to determine whether it is correlated with invasion and metastasis of human rectal cancer. METHODS: Eighty-six paired samples of rectal cancer and surgically resected distant normal rectal tissue were collected and allocated into cancer group and control group respectively. WISP-1 mRNA was detected by relative quantitative real-time RT-PCR and WISP-1 protein was examined by immunohistochemical staining. RESULTS: WISP-1 gene overexpression was found in 65% (56/86) primary rectal cancers, 2-30 times that of the level in normal matched rectal tissues (P = 0.001). The mRNA expression level was correlated with Duke's staging, histological differentiation grade and lymph node status. The WISP-1 protein expression was in accordance with mRNA expression level. The positive degree of immunohistochemical staining in the cancer group (1.40 +/- 0.35) was different from that in control group (1.04 +/- 0.08, P < 0.001). Moreover, in cancer group the positive staining degree in high-level mRNA cancers (1.46 +/- 0.37, n = 56) was higher than that in low-level mRNA (1.28 +/- 0.28, n = 30, P = 0.018). CONCLUSION: Aberrant levels of WISP-1 expression may play a role in rectal tumorigenesis. WISP-1 may be used as a specific clinical diagnosis and prognosis marker in rectal cancer.

Clinicopathological characteristics of 15 patients with combined hepatocellular carcinoma and cholangiocarcinoma.

BACKGROUND: Combined hepatocellular carcinoma and cholangiocarcinoma (cHCC-CC) is a rare subtype of primary liver cancer, and clinicopathological features of cHCC-CC have seldom been reported in detail. This study was undertaken to explore the diagnosis and clinicopathological characteristics of cHCC-CC in comparison with hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC), respectively. METHODS: The clinical data from 15 patients with cHCC-CC, 132 patients with HCC and 44 patients with CC who had undergone hepatic resection were analyzed retrospectively. Clinicopathological characteristics of cHCC-CC, HCC and CC such as hepatitis B viral infection, serum hepatitis C virus (HCV) antibody, serum alpha-fetoprotein (AFP) level, cirrhosis, vascular invasion, lymph node metastasis, surgical procedure and adjuvant treatment were also analyzed. Follow up was carried out in the patients, and their 1-, 3-, and 5-year survival rates were calculated. RESULTS: Two patients with cHCC-CC were correctly diagnosed by enhanced CT before operation, the other 13 patients were diagnosed by histology and immunohistochemistry after operation. Radical (8/15) and conservative hepatectomy (7/15) for cHCC-CC was similar to that for HCC and CC (P>0.05). Pathologically cHCC-CC showed more significantly vascular invasion and lymph node metastasis than HCC (P<0.05), and a similarity to CC (P>0.05). Hepatitis B viral infection, serum HCV antibody, cirrhosis, and serum AFP level of cHCC-CC patients were similar to those of HCC patients (P>0.05) but different from CC patients (P<0.05). The cumulative 1-, 3-, and 5-year survival rates in patients with cHCC-CC were poorer than in patients with HCC or CC (P<0.05). CONCLUSIONS: Patients with cHCC-CC are seldom diagnosed before operation. The progression of cHCC-CC is more rapid than that of HCC or CC. Survival rate of patients with cHCC-CC after hepatic resection is poorer than that of patients with HCC or CC.

Improved early diagnosis of cystadenocarcinoma of the pancreas.

BACKGROUND: Cystadenocarcinoma of the pancreas is insensitive to radiotherapy and chemotherapy, and surgery is at present the definitive treatment. Early and accurate diagnosis of cystadenocarcinoma is crucial for increasing the five-year survival rate and the resectable rate. There is no definitive and effective method of early diagnosis of cystadenocarcinoma of the pancreas in China and other countries. METHODS: We compared endoscopic ultrasonography-guided (EUS-guided) fine needle aspiration biopsy combined with cyst fluid carcinoembryonic antigen (CEA), CA19-9 examination with computed tomography (CT), B-ultrasonography (B-US) and serum CEA and CA19-9, to explore methods of early diagnosis of cystadenocarcinoma of the pancreas. Retrospective analysis was made on the clinical data of 126 cases of benign pancreatic lesion (90 cases) and cystadenocarcinoma (36). RESULTS: The sensitivity of B-US and CT for cystadeno-carcinoma was 52.8% and 77.8%, while the specificity was 78.9% and 86.7%, respectively. When measurement of CEA and CA19-9 of cyst fluid was combined with EUS-guided fine needle aspiration biopsy, the sensitivity was 94.4%, higher than that of B-US and CT (P<0.05). The sensitivity of cyst fluid CEA, CA19-9 examinations was considerably higher than that of serum CEA, CA19-9 (P<0.05). Upper gastrointestinal barium meal and endoscopic retrograde cholangiopancreatography (ERCP) had low sensitivity and specificity. CONCLUSIONS: EUS-guided fine needle aspiration biopsy combined with examination of cyst fluid CEA, CA19-9 is a credible means for early diagnosis of cystadenocarcinoma of the pancreas. B-US, CT and serum CEA, CA19-9 measurements are in common use, their findings are also very important.

[Quantification of PPARdelta mRNA by real-time RT-PCR in rectal cancer tissues].

OBJECTIVE: To clarify the expression change of PPARdelta gene in human rectal cancer tissues and determine the correlation of PPARdelta expression with the clinical and pathological parameters of rectal cancer. METHODS: Applying real-time RT-PCR, we quantified PPARdelta mRNA in 86 tissues from excised primary rectal cancers. In each case, accompanying normal mucosa was collected for comparison. RESULTS: Among the 86 rectal cancer tissues, 48 (55.8%) cases showed PPARdelta overexpression: 39 (81.3%) tumors gave an expression level 1.5 to 5.0 times, 5 (10.4%) tumors 10 to 20 times, and 4 (8.3%) tumors more than 20 times relative to normal mucosa. However, the general level of PPARdelta mRNA in rectal cancer tissues is not statistically different from that in normal mucosa. There was no evidence for the relationships of PPARdelta expression with cell differentiation, pathological categories and Dukes stages. CONCLUSION: The expression of PPARdelta gene in rectal cancers is not statistically different from that in normal mucosa, and it is not correlated with cell differentiation, pathological categories and Dukes stages.

Association of E1AF mRNA expression with tumor progression and matrilysin in human rectal cancer.

OBJECTIVE: To examine E1AF mRNA expression and to determine whether it is correlated with tumor progression and matrilysin in human rectal cancer. METHODS: Real-time RT-PCR was used to determine E1AF and matrilysin expression in 100 matched rectal cancers and normal tissues. RESULTS: Among the 100 rectal cancers, 69 cases of E1AF mRNA overexpression were observed. E1AF mRNA overexpression correlated well with matrilysin. In carcinomas, E1AF mRNA overexpression correlated significantly with depth of invasion, lymph node metastasis, venous involvement and advanced pTNM stage. CONCLUSIONS: E1AF was correlated significantly with tumor progression of human rectal cancer and may be an important factor in rectal cancer progression.

Caudate lobectomy by the third porta hepatis anatomical method: a study of 16 cases.

BACKGROUND: The treatment for primary tumor in the caudate lobe of the liver is difficult because of its unique anatomical location. This study was undertaken to improve operative techniques and results by a new anatomical method of caudate lobectomy. METHODS: Clinical data of 16 patients who had had caudate lobectomy for the liver from January 1996 to November 2004 were retrospectively analyzed. The third porta hepatis anatomical method was performed in all 16 patients. Operative time, intraoperative blood loss, postoperative complications were recorded. The 1-, 3-, and 5-year survival rates of 13 patients with caudate lobe carcinoma were followed up. Anatomical status, operative routes, operative procedures, liver blood supply were evaluated. RESULTS: The operation was successful in the 16 patients. The operative time was 255+/-70 minutes and blood loss 740+/-402 ml. None of the patients died from massive bleeding during the operation, nor did complications such as biliary fistula and liver failure occurred. In 13 patients with malignant tumor, 7 died from recurrence and metastasis of the tumor and the other 6 are still alive at the end of follow-up. One patient has survived for 6 years. The 1-, 3-, and 5-year survival rates in the 13 patients were 83.9%, 58.7% and 39.2%, respectively. CONCLUSION: Caudate lobectomy by the third porta hepatis anatomical method can improve operative effect and increase the resection probability for solitary tumor in the caudate lobe.

Clinicopathologic and prognostic significance of MMP-7 (matrilysin) expression in human rectal cancer.

OBJECTIVE: There is increasing evidence to indicate that MMP-7 plays a more important role in tumor progression than other MMPs. The aim of this study was to detect MMP-7 expression in human rectal cancer and normal rectal tissue and to determine whether it is correlated with invasion and metastasis of human rectal cancer. METHODS: Eighty-six paired samples of rectal cancer and distant normal rectal tissue obtained from 100 inpatients were allocated into two groups (cancer group and control group). MMP-7 mRNA was detected by relative quantitative real-time RT-PCR and MMP-7 protein was examined by immunohistochemical staining and computerized image analysis. RESULTS: MMP-7 mRNA expression in cancer group was higher than that in control group (P = 0.006), the expression ratios of 31 samples (37.35%) were <1 and 52 (62.65%) were >1. The mRNA expression level was correlated with Dukes Staging, histological differentiation grade and CEA level. The MMP-7 protein expression was in accordance with mRNA expression level. The positive degree of immunohistochemical staining in cancer group (1.82 +/- 0.03) was different from that in control group (1.17 +/- 0.13, P = 0.002). Moreover, in cancer group the positive staining degree in high-level mRNA cancers (2.04 +/- 0.18, n = 52) was higher than that in low-level mRNA ones (1.58 +/- 0.23, n = 31, P = 0.008). CONCLUSIONS: Our results suggest that MMP-7 plays an important role in the progression of human rectal cancer. MMP-7 may be selected as a clinical diagnosis and prognosis index in rectal cancer.