Dissociation Phenomenon of Erythrocyte Agglutination and Its Application to Assay of Functional Activity of the Complement System in Clinical Laboratory.

OBJECTIVE: The objective of the study was to validate the dissociation phenomenon of erythrocyte agglutination which is based on erythrocyte fragments and to apply it in the functional activity assay of the complement system. METHODS: The dissociation-agglutination effect of erythrocyte fragments was validated by detecting the number of free erythrocytes after the action of erythrocyte fragments on agglutinated erythrocytes. The number of free erythrocytes produced after hemolysis of agglutinated erythrocytes caused by complements and complement activators(CAs) was detected by auto hematology analyzer and the results were indicated by mean hemoglobin concentration of erythrocytes (MCHC). We optimized the test conditions and validated the inter-batch stability, explored the resolution of the assay method, and assayed for the total complement activity (AC) and the CAs activated complement activity (ACA) in serum from patients and healthy individual groups. RESULTS: Erythrocyte fragments have a dissociative effect on agglutinated erythrocytes. The auto hematology analyzer was able to detect AC and ACA, where AC showed an inverse correlation with MCHC, and ACA demonstrated a positive correlation with MCHC. The inter-batch CV of AC, ACA, and ACA/AC was found to be 5%, 9%, and 11.7%, respectively, with good stability. The study found that serum samples from acute phase reaction patients showed significant differences in ACA compared with healthy individuals, with a p value of 0.018; serum samples from patients with nephrotic syndrome showed significant differences in AC, ACA, and ACA/AC compared with healthy individuals, with p values of 0.014, 0.002, and 0.041, respectively. CONCLUSION: Erythrocyte fragments have dissociation-agglutination effect. The complement system immunological functional detection method, based on this effect, has potential clinical application value due to its sensitivity and accuracy.

The complement system testing in clinical laboratory.

With the advancement in research in the field of the complement system, a more comprehensive understanding developed about the complement system's role in the life process of an organism. It is a system of innate immune surveillance. This system plays a pivotal role in host defense against pathogens, inflammation, B and T cell homeostasis. Complement system analysis has a significant advantage in the assessment of the immune system status, diagnosis and prognosis of diseases, and medication guidelines. Currently, complement system testing is neither yet widely used across all clinical laboratoriesnor are the testing protocols yet systematic. Based on the current research, it is suggested that the analysis of complement activator-activated complement activity and total complement activity would be comprehensively assessed to evaluate the complement system's immunological function, and combine of the detection of its components to establish a systematic protocol for the complement system testing in the clinical laboratory. This article reviews the complement system's physiological role, disease relevance and the current testing status in clinical laboratories. Further more, some suggestions have also been provided for the preparation of complement standards i.e., the standardized preparation process for complement standards seems to be a feasible option given the easy inactivation of complement.

The value of noncontrast MRI in evaluating breast imaging reporting and data system category 0 lesions on digital mammograms.

Background: Benign and malignant diagnosis of nonpalpable breast imaging reporting and data system (BI-RADS) category 0 lesions on digital mammograms (DMs) is very important. We compared the diagnostic performance of non-contrast-enhanced magnetic resonance imaging (MRI) and dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) for them. We sought to evaluate BI-RADS category 0 lesions using 3 MRI sequences: short tau inversion recovery (STIR), STIR combined with high b value diffusion-weighted imaging (STIR-DWI), and DCE-MRI. Methods: We retrospectively reviewed 114 breast DMs rated as nonpalpable BI-RADS category 0 lesions in 112 patients from January 2014 to June 2019. STIR, high b value DWI, and DCE-MRI were performed for all patients. Two breast radiologists read individual sequences (STIR, DWI, DCE-MRI) and pairs of sequences (STIR-DWI) to detect BI-RADS category 0 lesions in DMs. Receiver operating characteristic (ROC) curve analysis was used to assess diagnostic performance according to a best valuable comparator that combined MRI imaging, clinical, and pathological data. Results: Among of 114 lesions (the median age of patients was 47 years; the median size of the lesion was 19 mm), 32 (48.5%) malignant lesions were missed by STIR, 9 (13.6%) malignant lesions were missed by STIR-DWI, and 3 (4.5%) malignant lesions were missed by DCE-MRI. The principal finding of our study was that STIR-DWI and DCE-MRI showed higher diagnostic accuracy than did STIR (P<0.01). STIR-DWI showed higher accuracy [area under the curve (AUC) =0.858; sensitivity =87.8%] for BI-RADS category 0 lesions in DMs than did STIR (AUC =0.754; sensitivity =51.5%), while the performance was comparable to that of DCE-MRI (AUC =0.884; sensitivity =95.5%). Conclusions: Using pairs of sequences (STIR-DWI) is a non-contrast-enhanced MRI technique and had an equal diagnostic performance in distinguishing benign from malignant lesions among nonpalpable BI-RADS category 0 lesions to that of DCE-MRI. As a result, STIR-DWI as having the potential to improve the safety and efficacy in of breast cancer screening, especially in nonpalpable BI-RADS category 0 lesions at in DMs.

Establishment and clinical application of an assay for the activity of circulating immune complexes using human O-erythrocytes as an indicator system.

OBJECTIVE: To develop a suitable clinical laboratory assay for detecting the activity of circulating immune complexes (CICs) that activate complement (ACIC). METHODS: CICs measured in serum were initially used to activate complement, and the remaining complement was activated through sensitized human O-erythrocytes. ACIC was quantified by the degree of hemolysis. Each serum sample was tested for ten consecutive days to determine its stability. Reference ranges are suggested. ACIC was measured in both healthy individuals and patients with autoimmune diseases as well. RESULTS: The OD values of the hemolysis degree index were inversely proportional to ACIC (r = -0.986, P = 0.002). A pooled serum was used to eliminate interference and optimize the experiment. The hemolysis degree (HD) was used to indicate the detection result. HD = (detection value OD/negative value OD)*100. Both the intra-batch and the inter-batch results showed good stability with a CV of 6.5% and 8.1%, respectively. HD differences between males and females were significant (P = 0.015) while the normal distribution for both genders was conformed. The HD recommended reference range for men is 56-88 while for women is 51-86. Serum HD of healthy subjects and autoimmune disease patients showed a significant difference (P = 0.001). Autoimmune disease patients have lower HD which was a result of having stronger ACIC. CONCLUSION: The ACIC assay while utilizing human O-erythrocytes as an indicator system is sensitive and accurate, and has potential in clinical applications.

Lipid metabolism contribute to the pathogenesis of IgA Vasculitis.

BACKGROUND AND OBJECTIVES: The underlying mechanism of IgA vasculitis (IgAV) and IgA vasculitis with nephritis (IgAVN) remains unclear. Therefore, there are no accurate diagnostic methods. Lipid metabolism is related to many immune related diseases, so this study set out to explore the relationship of lipids and IgAV and IgAVN. METHODS: Fifty-eighth patients with IgAV and 28 healthy controls were recruited, which were divided into six separate pools to investigate the alterations of serum lipids according to the clinical characteristics: healthy controls group (HCs) and IgAV group (IgAVs), IgAVN group (IgAV-N) and IgAV without nephritis group (IgAV-C), initial IgAV group (IgAV0) and IgAV in treatment with glucocorticoids group (IgAV1). RESULTS: 31 identified lipid ions significantly changed in IgAVs with p < 0.05, variable importance of the projection (VIP) > 1 and fold change (FC) > 1.5. All these 31 lipid ions belong to 6 classes: triacylglycerols (TG), phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylserine, ceramide, and lysophosphatidylcholine. TG (16:0/18:1/22:6) +NH4 over 888875609.05, PC (32:1) +H over 905307459.90 and PE (21:4)-H less than 32236196.59 increased the risk of IgAV significantly (OR>1). PC (38:6) +H was significantly decreased (p < 0.05, VIP>1 and FC>1.5) in IgAVN. PC (38:6) less than 4469726623 conferred greater risks of IgAV (OR=45.833, 95%CI: 6.689~341.070). CONCLUSION: We suggest that lipid metabolism may affect the pathogenesis of IgAV via cardiovascular disease, insulin resistance, cell apoptosis, and inflammation. The increase of TG(16:0/18:1/22:6) + NH4, and PC(32:1) + H as well as PE (21:4)-H allow a good prediction of IgAV. PE-to-PC conversion may participate in the damage of kidney in IgAV. PC (38:6) + H may be a potential biomarker for IgAVN.

A Flow Cytometry-Based Assay for the Measurement of Total Complement Activity in the Serum and Clinical Practice.

OBJECTIVE: To develop a novel sensitive and accurate assay suitable for high-volume testing of the total complement activity in the serum for clinical laboratories. METHODS: The total complement activity (TCA) to be measured was quantified by detecting the number of fragments produced by erythrocyte lysis and the erythrocyte fragmentation index (EFI), indicating TCA. EFI = M × M2/(M1 + M2), where M is the number of erythrocyte fragments (removed from the background), M1 is the number of unagglutinated red cells, M2 is the number of agglutinated red cell groups, and M2/(M1 + M2) is the agglutination coefficient indicating the degree of erythrocyte agglutination. Mild changes in hemolysin and erythrocyte concentrations were made to optimize the testing conditions. The same serum samples were tested for 10 consecutive days to determine the stability of the experimental results. Serum EFI was detected in both nephrotic syndrome patients and healthy subjects. RESULTS: There was a linear relationship between hemolysin and erythrocyte agglutination (r = 0.999, P < 0.001). A good linear relationship existed between EFI and TCA (r = 0.991, P < 0.001). The results were not affected by slight fluctuations in the concentrations of hemolysin or erythrocytes. The interbatch CV = 8.6% of the test results showed good stability. There was a significant difference in the EFI between nephrotic syndrome patients and healthy individuals, P < 0.001, and EFI was reduced in nephrotic syndrome patients compared to healthy individuals. CONCLUSION: The flow cytometry-based assay for TCA was sensitive and accurate and had potential value for clinical application.

Circadian rhythm disruption exacerbates Th2-like immune response in murine allergic airway inflammation.

BACKGROUND: Chronic jet lag (CJL)-induced circadian rhythm disruption (CRD) is positively correlated with an increased risk of allergic diseases. However, little is known about the mechanism involved in allergic rhinitis (AR). METHODS: Aberrant light/dark cycles-induced CRD mice were randomly divided into negative control (NC) group, AR group, CRD+NC group, and CRD+AR group (n = 8/group). After ovalbumin (OVA) challenge, nasal symptom scores were recorded. The expression of Occludin and ZO-1 in both nasal mucosa and lung tissues was detected by reverse transcription-quantitative polymerase chain reaction (RT-PCR) and immunohistochemical staining. The level of OVA-specific immunoglobulin E (sIgE) and T-helper (Th)-related cytokines in the plasma was measured by enzyme-linked immunosorbent assay (ELISA), and the proportion of Th1, Th2, Th17, and regulatory T cell (Treg) in splenocytes was evaluated by flow cytometry. RESULTS: The nasal symptom score in the CRD+AR group was significantly higher than those in the AR group with respect to eosinophil infiltration, mast cell degranulation, and goblet cell hyperplasia. The expression of ZO-1 and Occludin in the nasal mucosa and lung tissues in the CRD+AR group were significantly lower than those in the AR group. Furthermore, Th2 and Th17 cell counts from splenocytes and OVA-sIgE, interleukin 4 (IL-4), IL-6, IL-13, and IL-17A levels in plasma were significantly increased in the CRD+AR group than in the AR group, whereas Th1 and Treg cell count and interferon γ (IFN-γ) level were significantly decreased in the CRD+AR group. CONCLUSION: CRD experimentally mimicked CJL in human activities, could exacerbate local and systemic allergic reactions in AR mice, partially through decreasing Occludin and ZO-1 level in the respiratory mucosa and increasing Th2-like immune response in splenocytes.

Correlation Analysis between Gut Microbiota and Metabolites in Children with Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune-mediated diffuse connective tissue disease characterized by immune inflammation with an unclear aetiology and pathogenesis. This work profiled the intestinal flora and faecal metabolome of patients with SLE using 16S RNA sequencing and gas chromatography-mass spectrometry (GC-MS). We identified unchanged alpha diversity and partially altered beta diversity of the intestinal flora. Another important finding was the increase in Proteobacteria and Enterobacteriales and the decrease in Ruminococcaceae among SLE patients. For metabolites, amino acids and short-chain fatty acids were enriched when long-chain fatty acids were downregulated in SLE faecal samples. KEGG analysis showed the significance of the protein digestion and absorption pathway, and association analysis revealed the key role of 3-phenylpropanoic acid and Sphingomonas. Sphingomonas were reported to be less abundant in healthy periodontal sites of SLE patients than in those of HCs, indicating transmission of oral species to the gut. This study contributes to the understanding of the pathogenesis of SLE disease from the perspective of intestinal microorganisms, explains the pathogenesis of SLE, and serves as a basis for exploring potential treatments for the disease.

Intraperitoneal Chemotherapy Using Fluorouracil Implants Combined With Radical Resection and Postoperative Adjuvant Chemotherapy for Stage III Gastric Cancer: A Multi-Center, Randomized, Open-Label, Controlled Clinical Study.

BACKGROUND: Reducing peritoneal recurrence after radical surgery is an important choice to improve the prognosis of patients with advanced gastric cancer. Intraoperative intraperitoneal chemotherapy has the potential to be a promising treatment strategy. In the present study, we conducted a multi-center, randomized, controlled clinical study to evaluate the efficacy and safety of intraoperative intraperitoneal chemotherapy using sustained-release fluorouracil implants plus radical gastrectomy and adjuvant chemotherapy for cTNM stage III gastric cancer. METHODS: The patients were randomized into intraperitoneal chemotherapy group (sustained-release fluorouracil implants administration after standard D2 radical gastrectomy, and followed by XELOX adjuvant chemotherapy) and control group (standard D2 radical gastrectomy, and followed by XELOX adjuvant chemotherapy). A total of 122 patients from three centers were enrolled from September 2015 to February 2017. RESULTS: One hundred and two eligible patients completed the treatment course. The median follow-up time was 41.7 months (36.1-52.9 months). The 3-year progression-free survival rate and overall survival of patients in the intraperitoneal chemotherapy group were 43.9% and 49.1%, respectively, which were significantly better than those of the control group, 31.0% and 38.4%. In the intraperitoneal chemotherapy group, the number of cases with peritoneal recurrence was significantly less than that of the control group, 9 cases (17.3%) vs. 19 cases (44.2%). There were neither significant differences between the groups in the incidence of hematogenous metastasis, lymph node metastasis, nor local metastasis. CONCLUSION: For cTNM stage III gastric cancer, intraoperative sustained-release fluorouracil implants after radical resection combined with postoperative adjuvant chemotherapy, could significantly reduce the risk of peritoneal recurrence and prolong PFS.Clinical Trial Registration: https://clinicaltrials.gov/, identifier (NCT02269904).

Neutralizing interferon-α blocks inflammation-mediated vascular injury via PI3K and AMPK in systemic lupus erythematosus.

Plasmacytoid dendritic cells (pDCs) play a key role in the initiation and amplification of systemic lupus erythematosus (SLE)-associated vascular injury. In this study, we found that dsDNA induced dose- and time-dependent increase in IFN-α and Toll-like receptor 7 (TLR7), TLR9 and IRF7 expression in pDCs. Co-cultured circulating endothelial cells (ECs) with activated pDCs significantly decreased proliferation, tube formation and migration in ECs. The elevated level of cellular IFN-α increased cell adhesion, promoted cell apoptosis, induced cell senescence and arrested cells at G0/G1 phase of endothelial progenitor cells (EPCs). Additionally, the co-culture system activated MAPK and inactivated PI3K. Pristane was used to establish a in vivo SLE-like mouse model. Importantly, we showed that INF-α-neutralizing antibody (IFN-α-NA) rescued all the changes induced by IFN-α in vitro and prevented vascular injury in pristane-induced SLE model in vivo. In conclusion, we confirmed that activated pDCs promoted vascular damage and the dysfunction of ECs/EPCs via IFN-α production. IFN-α-neutralizing antibody may be a clinical implication for preventing vascular injury. PI3K signalling and AMPK signalling were associated with SLE-associated vascular functions.

IFN-I Mediates Lupus Nephritis From the Beginning to Renal Fibrosis.

Lupus nephritis (LN) is a common complication of systemic lupus erythematosus (SLE) and a major risk factor for morbidity and mortality. The abundant cell-free nucleic (DNA/RNA) in SLE patients, especially dsDNA, is a key substance in the pathogenesis of SLE and LN. The deposition of DNA/RNA-immune complexes (DNA/RNA-ICs) in the glomerulus causes a series of inflammatory reactions that lead to resident renal cell disturbance and eventually renal fibrosis. Cell-free DNA/RNA is the most effective inducer of type I interferons (IFN-I). Resident renal cells (rather than infiltrating immune cells) are the main source of IFN-I in the kidney. IFN-I in turn damages resident renal cells. Not only are resident renal cells victims, but also participants in this immunity war. However, the mechanism for generation of IFN-I in resident renal cells and the pathological mechanism of IFN-I promoting renal fibrosis have not been fully elucidated. This paper reviews the latest epidemiology of LN and its development process, discusses the mechanism for generation of IFN-I in resident renal cells and the role of IFN-I in the pathogenesis of LN, and may open a new perspective for the treatment of LN.

Alterations in mucosa-associated microbiota in the stomach of patients with gastric cancer.

PURPOSE: The purpose of this study was to characterize alterations in mucosa-associated microbiota in different anatomical locations of the stomach during gastric cancer progression and to identify associations between Helicobacter pylori infection and gastric microbial changes in patients with gastric cancer. METHODS: Twenty-five H. pylori negative subjects with chronic gastritis and thirty-four subjects with gastric cancer were recruited, including H. pylori negative and positive patients with tumors in the antrum and the corpus. Gastric mucosa-associated microbiota were determined by 16S ribosomal RNA gene sequencing using a 454 sequencing platform. RESULTS: We found that individuals with chronic gastritis from three different anatomical sites exhibited different microbiota compositions, although the microbial alpha diversity, richness and beta diversity were similar. Compared to patients with chronic gastritis, the gastric microbiota compositions were significantly different at the order level in the antrum and the corpus of patients with gastric cancer, which was dependent on the H. pylori infection status. Microbial alpha diversity and species richness, however, were similar between chronic gastritis and gastric cancer cases and independent of H. pylori status. The microbial community structure in patients with gastric cancer was distinct from that in patients with chronic gastritis. In addition, we found that the presence of H. pylori markedly altered the structure in gastric corpus cancer, but only mildly affected the antrum. CONCLUSION: Our data revealed distinct niche-specific microbiota alterations during the progression from gastritis to gastric cancer. These alterations may reflect adaptions of the microbiota to the diverse specific environmental habitats in the stomach, and may play an important, as yet undetermined, role in gastric carcinogenesis.

IFN-I Mediates Dysfunction of Endothelial Progenitor Cells in Atherosclerosis of Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a multi-system autoimmune disease including the cardiovascular system. Atherosclerosis is the most common cardiovascular complication of SLE and a significant risk factor for morbidity and mortality. Vascular damage/protection mechanism in SLE patients is out of balance, caused by the cascade reaction among oxidative stress, proinflammatory cytokines, Neutrophil Extracellular Traps, activation of B cells and autoantibodies and abnormal T cells. As a precursor cell repairing vascular endothelium, endothelial progenitor cells (EPCs) belong to the protective mechanism and show the reduced number and impaired function in SLE. However, the pathological mechanism of EPCs dysfunction in SLE remains ill-defined. This paper reviews the latest SLE epidemiology and pathogenesis, discusses the changes in the number and function of EPCs in SLE, expounds the role of EPCs in SLE atherosclerosis, and provides new guidance and theoretical basis for exploring novel targets for SLE treatment.

[Sinus balloon dilation for treatment of chronic sinusitis: a systematic review and Meta-analysis].

Objective:To systematically assess the efficacy of sinus balloon dilation（SBCD） for patients with chronic rhinosinusitis（CRS）. Method:To research the references whose the contents were related to the randomized controlled trials of sinus balloon dilation for treatment of chronic sinusitis from PubMed, Embase, Cochrane Library, WanFang, CNKI and other databases from January 2000 to June 2019. Assessed the quality and extracted the data of these included references and made Meta analysis by RevMan5.3 software. Result:A total of 11 references in English and Chinese were included. Result:Of Meta analysis showed that the difference had no statistical significance between the SNOT-20 score which were based on the statistics measured 3 months later and 6 months later [3 months later, SMD=-0.30, 95%CI（-1.01-0.41）, P=0.40, 6 months later, SMD=-0.53, 95%CI（-3.22-2.16）, P=0.70]and the score in the control group. Lund-Mackay CT score [SMD＝-0.69, 95%CI（-0.72--0.66, P<0.00001） ]and VAS symptom score [SMD＝-1.67, 95%CI（-1.91--1.44）, P<0.00001]following SBCD versus FESS in the treatment of CRS had statistical significance. Besides, SBCD had a lower incidence [OR=0.29, 95%CI（0.14-0.59）, P=0.0007]to develop postoperative complications compared with that in the control group. Conclusion:Balloon dilation of sinuses can significantly relieve the symptoms of patients with chronic sinusitis, this effect is equal or even better. Moreover, the incidence of developing the postoperative complications is lower, which SBCD of CRS is worthy of clinical popularization.

Abnormalities of Serum Fatty Acids in Children With Henoch-Schönlein Purpura by GC-MS Analysis.

Purpose: The objectives of this work were to test the levels of serum medium- and long- chain fatty acids (MLCFAs) in children and to discover their possible relationship with Henoch-Schönlein Purpura (HSP), also known as Immunoglobulin A vasculitis. Methods: A total of 57 children with HSP (HSP group) and 28 healthy children (CON group) were recruited for this study. Serum specimens were collected to detect the compositions and contents of MLCFAs by gas chromatography with mass spectrometry (GC-MS) analysis. Results: The contents of all detected 37 MLCFAs in the HSP group were higher than the healthy group. Thirty-one species of MLCFAs were discovered to have a significant difference (p < 0.05) in two groups. Comparing to healthy controls, there were 31, 31, 18 fatty acids showed a statistical difference in the untreated group, regular treated group, and withdrawal group of HSP, respectively. The trend of fatty acids in the three HSP groups was similar to the healthy controls, as well as the untreated group and regular treated group changed more obviously than the withdrawal group. Almitate (C16:0) and 18 carbon atoms (C18) of fatty acids were abundant in all three HSP groups, divided according to the treatment of glucocorticoid. Some fatty acids were found having considerable differences (p < 0.05) in three groups. Monounsaturated fatty acids (MUFAs), including elaidate (C18:1T), cis-11,14,17-eicosatrienoic acid ester (C20:1), and cis-15-tetracosenoate (C24:1), were distinctly higher in HSP children with renal damage. Conclusion: Our study revealed that the abnormalities in MLCFA may be associated with the development of HSP. Another interesting finding was that fatty acids contents were changing during the glucocorticoid treatment. Meanwhile, long-chain MUFAs may have an impact on renal damage in HSP patients. Further studies need to be carried out in order to explore the specific mechanism of fatty acids in the course of HSP.

Prognostic significance of lymphovascular infiltration in overall survival of gastric cancer patients after surgery with curative intent.

OBJECTIVE: Lymphovascular infiltration (LVI) is frequently detected in gastric cancer (GC) specimens. Studies have revealed that GC patients with LVI have a poorer prognosis than those without LVI. METHODS: In total, 1,007 patients with curatively resected GC at Department of Gastric Cancer, Tianjin Medical University Cancer Institute and Hospital were retrospectively enrolled. The patients were categorized into two groups based on the LVI status: a positive group (PG; presence of LVI) and a negative group (NG; absence of LVI). The clinicopathological factors corrected with LVI and prognostic variables were analyzed. Additionally, a pathological lymphovascular-node (lvN) classification system was proposed to evaluate the superiority of its prognostic prediction of GC patients compared with that of the eighth edition of the N staging system. RESULTS: Two hundred twenty-four patients (22.2%) had LVI. The depth of invasion and lymph node metastasis were independently associated with the presence of LVI. GC patients with LVI demonstrated a significantly lower overall survival (OS) rate than those without LVI (42.8% vs. 68.9%, respectively; P<0.001). In multivariate analysis, LVI was identified as an independent prognostic factor for GC patients (hazard ratio: 1.370; 95% confidence interval: 1.094-1.717; P=0.006). Using strata analysis, significant prognostic differences between the groups were only observed in patients at stage I-IIIa or N0-2. The lvN classification was found to be more appropriate to predict the OS of GC patients after curative surgery than the pN staging system. The -2 log-likelihood of lvN classification (4,746.922) was smaller than the value of pN (4,765.196), and the difference was statistically significant (χ2=18.434, P<0.001). CONCLUSIONS: The presence of LVI influences the OS of GC patients at stage I-IIIa or N0-2. LVI should be incorporated into the pN staging system to enhance the accuracy of the prognostic prediction of GC patients.

Impact of extranodal tumor deposits on prognosis and N stage in gastric cancer.

BACKGROUND: Extranodal tumor deposits have been reported to be associated with a poor prognosis in many malignancies and are also included in the tumor, node, and metastasis staging system for colorectal cancer. METHODS: We reviewed retrospectively a total of 2,344 gastric cancer patients who underwent gastrectomy with curative intent at the Tianjin Medical University Cancer Institute and Hospital (Hexi District, Tianjin, China) and the First Affiliated Hospital of Hainan Medical University (Longhua District, Haikou, China). Patients were categorized into 2 groups based on extranodal tumor deposit status: a positive group, including those with extranodal tumor deposits, and a negative group composed of those with no extranodal tumor deposits. Clinicopathologic factors were correlated with extranodal tumor deposits, and their individual prognoses were analyzed. In addition, a pathologically modified node classification system was proposed by incorporating the extranodal tumor deposit status into the 8th ed of the N staging system. The superiority of prognostic prediction between the modified node classification and node stage was compared. RESULTS: A total of 645 (27.5%) patients had extranodal tumor deposits. The presence of extranodal tumor deposits was associated with a larger tumor size, Borrmann type III and IV, a deeper depth of invasion, and an advanced node stage. In the multivariate analysis, extranodal tumor deposits were an independent prognostic factor for gastric cancer patients after curative resection. Gastric cancer patients with extranodal tumor deposits demonstrated a lesser 5-year overall survival than those with no extranodal tumor deposits (31.9% vs 61.4%, P < .001). With the strata analysis, statistically significant prognostic differences between the two groups were only observed in patients at the N0-N2 stage. The modified node classification was found to be more appropriate for predicting the overall survival of gastric cancer patients after curative resection than node stage, and the -2 log likelihood of the modified node classification (16,042.890) was smaller than the value of node stage (16,150.811). CONCLUSION: Extranodal tumor deposits in gastric cancer patients indicate aggressive characteristics and a poorer prognosis of gastric cancer. We maintain that extranodal tumor deposits should be incorporated into the N staging system to enhance the accuracy of the prognostic prediction of patients with gastric cancer.

Upregulation of microRNA-140-3p inhibits epithelial-mesenchymal transition, invasion, and metastasis of hepatocellular carcinoma through inactivation of the MAPK signaling pathway by targeting GRN.

Invasion and metastasis in hepatocellular carcinoma (HCC) results in poor prognosis. Human intervention in these pathological processes may benefit the treatment of HCC. The aim of the present study is to elucidate the mechanism of miR-140-3p affecting epithelial-mesenchymal transition (EMT), invasion, and metastasis in HCC. Microarray analysis was performed for differentially expressed genes screening. The target relationship between miR-140-3p and GRN was analyzed. Small interfering RNA (siRNA) against granulin (GRN) was synthesized. EMT markers were detected, and invasion and migration were evaluated in HCC cells introduced with a miR-140-3p inhibitor or mimic, or siRNA against GRN. A mechanistic investigation was conducted for the determination of mitogen-activated protein kinase (MAPK) signaling pathway-related genes and EMT markers (E-cadherin, N-cadherin, and Vimentin). GRN was highlighted as an upregulated gene in HCC. GRN was a target gene of miR-140-3p. Elevation of miR-140-3p or inhibition of GRN restrained the EMT process and suppressed the HCC cell migration and invasion. HCC cells treated with the miR-140-3p mimic or siRNA-GRN exhibited decreased GRN expression and downregulated the expressions of the MAPK signaling pathway-related genes, N-cadherin, and Vimentin but upregulated the expression of E-cadherin. GRN silencing can reverse the activation of the MAPK signaling pathway and induction of EMT mediated by miR-140-3p inhibition. Taken together, the results show that miR-140-3p confers suppression of the MAPK signaling pathway by targeting GRN, thus inhibiting EMT, invasion, and metastasis in HCC.

Elevated preoperative plasma D-dimer dose not adversely affect survival of gastric cancer after gastrectomy with curative intent: A propensity score analysis.

OBJECTIVE: Elevated plasma D-dimer has been reported to be associated with advanced tumor stage and poor survival in several types of malignancies. The purpose of this study was to assess the potential impact of preoperative plasma D-dimer level (PDL) on overall survival (OS) of gastric cancer (GC) patients undergoing curative surgery by applying propensity score analysis. METHODS: A total of 1,025 curatively resected GC patients in Tianjin Medical University Cancer Institute & Hospital were enrolled. Patients were categorized into two groups based on preoperative PDL: the elevated group (EG) and the normal group (NG). To overcome bias due to the different distribution of covariates for the two groups, a one-to-one match was applied using propensity score analysis, after matching, prognostic factors were analyzed. RESULTS: In analysis for the whole study series, patients in the EG were more likely to have a larger proportion of tumor size ≥5 cm (67.5% vs. 55.8%, P=0.006), elder mean age (64.0±10.8 years vs. 60.5±11.6 years, P<0.001) and advanced tumor (T), node (N), and TNM stage. Patients with elevated PDL demonstrated a significantly lower 5-year OS than those with normal PDL (27.0%vs. 42.6%, P<0.001), however, the PDL was not an independent prognostic factor for OS in multivariate analysis [hazard ratio: 1.13, 95% confidence interval (95% CI): 0.92-1.39, P=0.236]. After matching, 163 patients in the EG and 163 patients in the NG had the same characteristics. The 5-year OS rate for patients in the EG was 27.0% compared with 25.8% for those in the NG (P=0.809, log-rank). CONCLUSIONS: The poor prognosis of GC patients with elevated preoperative PDL was due to the advanced tumor stage and elder age rather than the elevated D-dimer itself.

Risk factors for metastasis to No.14v lymph node and prognostic value of 14v status for gastric cancer patients after surgery.

BACKGROUND: D2 procedure has been accepted as the standard lymphadenectomy for advanced GC, while the role of No.14v lymph node (14v) dissection for distal GC is still controversial. METHODS: A total of 284 GC patients receiving D2 plus 14v dissection in our center were enrolled. Patients were categorized into two groups based on 14v status: positive group (PG) and negative group (NG). Clinicopathological factors correlated with 14v metastasis and prognostic variables were respectively analyzed. RESULTS: Thirty-five patients (12.3%) had 14v metastasis. Metastasis to No.4d and No.6 lymph node were independent variables affecting 14v metastasis. Patients with positive 14v had a significant lower 3-year overall survival (OS) rate than those without (3-year OS: 42.9% vs. 70.3%, P < 0.001). Multivariable analysis demonstrated that 14v status was an independent prognostic factor for III stage GC (hazard ratio 1.462, 95% confident interval: 1.182-2.309, P = 0.027). The prognosis of 14v positive patients correlated with tumor size and No.6 lymph node status in univariate analysis. CONCLUSION: GC patients with No.4d and No.6 lymph node metastasis were more likely to have positive 14v. Status of 14v was an independent prognostic factor for III stage GC. Patients with 14v metastasis usually had a poorer prognosis, while survival in such patients after curative surgery was similar to that of patients staged IIIc without 14v metastasis.