Prognostic effects of the gastric mucosal microbiota in gastric cancer.

Gastric cancer (GC) is one of the most common malignant tumors with a high incidence and mortality. Microbiota play a significant role in human health and disease. We aimed to investigate the prognostic value of the gastric microbiota in different stomach microhabitats. We used our previously published 16S rRNA gene sequence data. We retrospectively enrolled a cohort of 132 patients with GC with complete prognostic information and selected 78 normal tissues, 49 peritumoral tissues, and 112 tumoral tissues for microbiota analysis. Patients with different prognoses showed different gastric microbiota compositions and diversity. The association network of the abundant gastric microbiota was more complicated in patients with poor prognoses. In the peritumoral microhabitat of patients with good prognoses, Helicobacter was significantly increased, whereas Halomonas and Shewanella were significantly decreased relative to that in the peritumoral microhabitat of patients with poor prognoses. PiCRUSt analysis revealed that the peritumoral microbiota had more different Kyoto Encyclopedia of Genes and Genomes pathways than did the tumoral and normal microbiota. This study evaluated the long-term prognostic value of the gastric mucosal microbiota in patients with GC. These findings suggested that the characteristic alterations of the gastric mucosal microbiota may be markers for clinical outcomes in these patients.

Prevalence of hyperhomocysteinemia and its associated factors in patients with primary hypertension in Chinese urban communities: A cross-sectional study from Nanjing.

OBJECTIVE: This study aimed to investigate the prevalence of hyperhomocysteinemia (Hhcy) and its major determinants in Chinese urban population with new-onset hypertension. METHODS: A total of 574 adults (404 men and 170 women) with newly diagnosed primary hypertension were recruited from seven communities in Nanjing, China. Data on lifestyle factors, such as physical activities, current smoking and drinking status, dietary habits, and familial factors were collected in interviews, and laboratory examinations were performed by well-trained personnel. Potential factors related to the prevalence of Hhcy in this population were analyzed using a logistic regression model. RESULTS: Of the 574 participants, 421 (73.3%) were diagnosed with Hhcy whereas the remainder were only hypertensive. The study highlighted a number of factors that were significantly correlated (p < 0.001) with the prevalence of Hhcy. Subjects with Hhcy were more likely to be male (odds ratio [OR] 3.007, 95% confidence interval [CI] 1.781-5.079), have a smoking history (OR 1.052, 95% CI 1.031-1.074), older (OR 1.052, 95% CI 1.031-1.074), have an elevated Body Mass Index BMI (OR 1.160, 95% CI 1.080-1.246) and higher levels of low density lipoprotein cholesterol (LDL-c) (OR 1.590, 95% CI 1.226-2.063). Regular and adequate physical activity was associated with normal homocysteine levels in both male and female groups (p < 0.05). For males only, having a higher BMI, higher LDL-c or being older significantly (p < 0.05) affected the chances of Hhcy. Whereas for females, lower levels of eGFR could be related to Hhcy (p < 0.05). CONCLUSIONS: Our present study reported a high prevalence of Hhcy in the Nanjing population with new-onset hypertension. Associated factors like physical activity, gender, smoking history, age, BMI, and LDL-c were important modifiers of plasma homocysteine concentration. Management and intervention of the above associated factors should be implicated to improve H type hypertension control.

Hepcidin in tumor-related iron deficiency anemia and tumor-related anemia of chronic disease: pathogenic mechanisms and diagnosis.

OBJECTIVES: Anemia is a common hematological abnormality in patients with cancer. Iron deficiency anemia (IDA) and anemia of chronic disease (ACD) are the most prevalent, both characterized by hypochromic microcytic anemia and low serum iron (SI). Their differential diagnosis is difficult in clinical practice, hampering their treatment. Our objective was to evaluate the use of hepcidin to discriminate tumor-related IDA and ACD and to investigate the mechanism of action of hepcidin in these anemias. METHODS: Blood samples were collected at Jiangsu Cancer Hospital. Patients were divided into IDA and ACD groups by Prussian blue staining of bone marrow smears. Serum hepcidin was measured by enzyme-linked immunosorbent assay. SI, total iron-binding capacity (TIBC), transferrin saturation (TSAT), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) also determined in this study. RESULTS: Areas under the curve on receiver operating characteristic analysis indicated the diagnostic sensitivity and specificity of hepcidin to be better than those of SI, TIBC, and TSAT. In ACD, hepcidin was correlated positively with IL-6 (r = 0.81, P < 0.01) and negatively with SI (r = -0.78, P < 0.01). In IDA, no significant relationship between IL-6 and hepcidin was found (r = -0.20, P = 0.17), but hepcidin decreased with decreasing quartiles of SI (r = 0.89, P < 0.01). SI was positively correlated with hemoglobin (r = 0.89, P < 0.01; r = 0.84, P < 0.01) in both groups. CONCLUSIONS: Hepcidin is a promising serological marker for the differential diagnosis of tumor-related ACD and IDA, clarifying the pathogenesis of these anemias and guiding corrective treatment.

B cells present a double-sided effect in digestive system tumors: a review for tumor microenvironment.

Over the past few years, there has been an increasing interest in investigating tumor-infiltrating lymphocytes. B lymphocytes (B cells) are extensively distributed within tertiary lymphoid structure (TLS) as multifaceted subgroups and are intimately linked to the anti-tumor properties of TLS, as well as the survival and prognostication of individuals. While the investigation of T lymphocytes in the TLS has advanced to the level of clinical practice, the study of B cells remains limited. The principal impediment to the utilization of B cells in immunotherapy is their notable dual impact on tumors. Compared with tumors in other parts and systems, the function of B cells in the microenvironment of digestive system tumors to promote tumors proliferation, differentiation and migration cannot be ignored. Therefore, this review collects the studies of B cell subsets in tumor microenvironments, particularly related single cell sequencing research. The multifaceted role and function of B cells are investigated in esophageal, liver, colorectal, gastric and pancreatic cancers. And through the identification of B cell subsets and specific markers, this review attempts to explain the reasons why B cells produce different tumor-promoting effects in those tumors. The insights gleaned from this review may provide potential help and support the development of B cell-based immunotherapies.

[Investigation of 1q21 amplification in patients with multiple myeloma using I-FISH and cIg-FISH].

OBJECTIVE: To investigate the prevlance of 1q21 amplification in patients with multiple myeloma (MM) and its correlation with the progression and prognosis of the disease. METHODS: 1q21 amplification was detected in 48 patients with MM using cytoplasmic light chain immunofluorescence with fluorescence in situ hybridization analysis (cIg-FISH) and interphase fluorescence in situ hybridization (I-FISH) analysis combined with CD138 immunomagnetic cell sorting (MACS). RESULTS: 1q21 amplification (≥ 3 red signals) was detected in 26/48(54.2%) cases by cIg-FISH and 31/48 (64.6%) cases by I-FISH combined with CD138 MACS. There was a good consistency between the two methods (P>0.05). The mortality of patients with 1q21 amplification was significantly higher than those without (P< 0.05). No significant difference was detected in terms of sex, age, Durie-Salmon stage, subgroup and international staging system (ISS) stage between patients with 1q21 amplification and those without (P>0.05). CONCLUSION: The frequency of 1q21 amplification in MM is high. There was also an association between the amplification and poor prognosis. cIg-FISH is consistent with CD138 MACS combined with I-FISH.

[Detection of complex chromosomal aberrations in patients with multiple myeloma using multiplex fluorescence in situ hybridization].

OBJECTIVE: To explore the value of multiplex fluorescence in situ hybridization (M-FISH) in the detection of the complex chromosomal aberrations (CCAs) in multiple myeloma (MM). METHODS: M-FISH was used in 10 MM patients with CCAs detected by conventional cytogenetics (CC) using R-banding to refine the rearrangement of CCAs and identify the characteristics of marker chromosome. RESULTS: M-FISH confirmed the 29 structural aberrations shown by CC analysis, and also confirmed the specific source of 21 types of chromosomal aberration, which were not detected by CC analysis. Among them, t(2;15)(q33;q22), t(6;7)(q23;q34), t(8;11) (q24;q23), t(1;14)(q10;q32) and t(X;1)(q26;q25) were new chromosomal aberrations. The median survival time of 9 MM patients with CCAs was 23 months and evidently shorter than that of MM patients without CCAs, with the mean survival time being 34 months. CONCLUSION: M-FISH could refine CCAs in MM patients, find or correct the missed or misidentified abnormalities analyzed by CC. It has provided one of the essential methods for the research of chromosomal aberrations in MM.

[Investigation of the molecular changes in patients with multiple myeloma by fluorescence in situ hybridization].

OBJECTIVE: To investigate the incidence and prognosis of 1q21 amplification, 13q14 deletion, TP53 gene deletion and IgH translocation in patients with multiple myeloma (MM). METHODS: Interphase fluorescence in situ hybridization (I-FISH) with four different specific probes for the regions containing 1q21, 13q14.3 (D13S319), 14q32 and TP53 gene were performed in 43 MM patients. RESULTS: Among the 43 MM patients, 1q21 amplification was observed in 28 (65.1%) cases, 13q14 deletion in 30 (69.7%) cases, TP53 gene deletion in 8 (18.6%) cases, and IgH translocation in 29 (67.4%) cases. The mortality of MM patients with 1q21 amplification, 13q14 deletion or TP53 gene deletion was higher than those without them. CONCLUSION: There is high frequency of 1q21 amplification, 13q14 deletion, TP53 gene deletion and IgH translocation in multiple myeloma, and 1q21 amplification, 13q14 deletion and TP53 gene deletion are poor prognosis factors for MM patients.

Promoter hypermethylation influences the suppressive role of long non-coding RNA MEG3 in the development of multiple myeloma.

Methylation is a fundamental regulator of gene transcription. Long non-coding RNA maternally expressed 3 (MEG3) inhibits cell proliferation in various types of cancer. However, the molecular mechanisms of MEG3 methylation in the regulation of multiple myeloma (MM) are unknown. In the present study, MEG3 upregulation was negatively associated with the International Staging System (ISS) status of the bone marrow samples of 39 patients with MM. MEG3 overexpression in an MM cell line resulted in elevated p53 expression. Furthermore, the results of methylation-specific PCR revealed that the abnormal methylation status of the MEG3 promoter region was present in eight of the 39 bone marrow samples collected. Treatment of the MM cell line with the DNA methylation inhibitor 5-Aza-2'-deoxycytidine (5-Aza-CdR) resulted in tumor cell proliferation inhibition, apoptosis induction and G0/G1 cell cycle arrest. Furthermore, 5-Aza-CdR decreased aberrant hypermethylation of the MEG3 promoter and increased the expression of MEG3. However, 5-Aza-CdR exerted no effect on p53 expression. To the best of our knowledge, the present study is the first to report that the demethylation reagent 5-Aza-CdR may serve as a therapeutic agent in MM by upregulating MEG3 expression. However, the mechanism of action was independent of p53 expression.

[Correlation between chromosome 13q14 deletion and 1q abnormality in multiple myeloma].

OBJECTIVE: To investigate the correlation between chromosome 13q14 deletion [del(13q14)] and chromosome 1q abnormality in multiple myeloma (MM). METHODS: The bone marrow plasma cells of 48 previously untreated MM patients were purified by CD138 and magnetic cell sorting system, and interphase fluorescence in situ hybridization (I-FISH) was applied to detect the del(13q14) with D13S319 probe and the abnormalities of chromosome 1q with CEP1 SpectrumOrange probe in sorted MM cells. RESULTS: Among the 48 MM patients, del(13q14) was observed in 22(45.8%) cases, the abnormalities of chromosome 1q were observed in 23 (47.9%) cases, among which 2 were 1q deletion and 21 were 1q duplication. The chromosome 1q abnormality was detected in 16 of the 22 cases of MM with del(13q14) and in 7 of the 26 cases of MM without del(13q14), and there was significant difference between the two groups (chi-square was 10.02, P was less than 0.01). CONCLUSION: There is high frequency of chromosome 13q14 deletion and 1q abnormality in multiple myeloma. The chromosome 1q abnormalities are highly associated with 13q14 deletion.

ROR1 associates unfavorable prognosis and promotes lymphoma growth in DLBCL by affecting PI3K/Akt/mTOR signaling pathway.

The receptor-tyrosine-kinase (RTK)-like orphan receptor 1 (ROR1) is a transmembrane glycoprotein regarded as a tumor-associated antigen. ROR1 plays an important role in cancer development, but the detailed function of ROR1 in diffuse large B-cell lymphoma (DLBCL) remains unclear. In this study, we first detected ROR1 expression and evaluated the relationship between ROR1 expression and the clinicopathological characteristics of DLBCL patients. Next we employed shRNA-mediated knockdown of ROR1 in DLBCL cell line to explore the characteristics of ROR1 in DLBCL development both in vitro and in vivo. The results showed a significantly higher level of ROR1 in DLBCL tissues than in lymphatic hyperplasia tissues. High ROR1 expression was correlated with unfavorable prognosis in DLBCL patients. Furthermore, ROR1 knockdown inhibited the growth and induced the apoptosis in DLBCL cells and xenografts. In addition, shROR1 inhibited activation of the PI3K/Akt/mTOR signaling pathway, both in vitro and in vivo. Taken together, our results suggest that ROR1 is a novel prognostic marker for DLBCL survival and ROR1 significantly promotes DLBCL tumorigenesis by regulating the PI3K/Akt/mTOR signaling pathway. Targeting ROR1 may provide a promising strategy for DLBCL treatment. © 2019 BioFactors, 45(3):416-426, 2019.

The Association Between Tea Consumption and Hyperhomocysteine in Chinese Hypertensive Patients.

BACKGROUND: There is no consistent evidence for the relationship between tea-drinking and hyperhomocysteine (hHcy). Because tea-drinking habit and hHcy have prevailed in Chinese hypertensive patients, this study aimed to investigate the association between hHcy and tea consumption in patients with hypertension. METHODS: A total of 335 hypertensive participants were recruited from 7 communities. Demographic characteristics of participants were collected through face-to-face interviews using a standard questionnaire, whereas laboratory data were obtained within 1 week after patient recruitment. Multiple logistic regression analysis was performed to examine the association between tea consumption and hHcy in hypertensive patients. RESULTS: Of the 335 patients, 245 had a tea-drinking habit, and 252 of them were detected with hHcy. A significant association was found between tea consumption and hHcy in hypertensive patients (adjusted odds ratio [OR] = 1.84, 95% confidence interval [CI] = 1.01-3.36, P = 0.048). Subgroup analyses showed that black tea drinking group (adjusted OR = 8.81, 95% CI = 2.74-28.33, P < 0.001) was significantly associated with the risk of hHcy, but not oolong and green tea drinking groups (P > 0.05). Furthermore, consuming a small amount (≤1 cup per day) of green tea was negatively associated with hHcy (adjusted OR = 0.19, 95% CI = 0.07-0.51, P = 0.001), whereas a large intake (>3 cups per day) of green tea was associated with high odds of hHcy (adjusted OR = 5.00, 95% CI = 1.33-18.79, P = 0.02). CONCLUSIONS: These data suggest a hypothesis that selecting green tea or limiting tea consumption might reduce risk of hHcy in hypertensive patients and that warrants further study.

High expression of NDRG3 associates with unfavorable overall survival in non-small cell lung cancer.

BACKGROUND AND OBJECTIVE: N-myc downstream-regulated gene 3 (NDRG3) is one of the important members of the NDRG family which crucially take part in cell proliferation, differentiation and other biological processes. METHODS: In this present study, western-blotting analysis was performed to evaluate NDRG3 expression in NSCLC cell lines. One-step quantitative reverse transcription-polymerase chain reaction (qPCR) with 16 fresh-frozen NSCLC samples and immunohistochemistry (IHC) analysis in 100 NSCLC cases were conducted to explore the relationship between NDRG3 expression and the clinicopathological characteristics of NSCLC. RESULTS: NDRG3 expression levels were statistically higher in NSCLC cell lines and tissue samples, compared with that of in non-cancerous cell line and tissue samples (p< 0.05). The IHC data demonstrated that the NDRG3 expression was significantly correlated with pathological grade (p= 0.038), N (p= 0.020) and TNM stage (p= 0.002). Survival analysis and Kaplan-Meier curve indicated that NDRG3 expression (p= 0.002) and T (p= 0.047) were independently associated with the unfavorable overall survival of patients with NSCLC. CONCLUSIONS: The data implied that NDRG3 expression may be identified as a new predictor in NSCLC prognosis.

Circulating microRNA 483-5p as a novel biomarker for diagnosis survival prediction in multiple myeloma.

Dysregulation of miRNA expression plays an important role in cancer development, and circulating miRNAs are biomarkers of several cancers. We explored whether the miRNAs in plasma could be useful clinical biomarkers for multiple myeloma. miRNA microarray was conducted to identify elevation of four miRNAs and reduced levels of eight miRNAs in the plasma of nine multiple myeloma patients and seven healthy controls. Increased miR-483-5p levels and decreased miR-20a were further validated in the plasma of 40 myeloma patients and 20 healthy controls using TaqMan quantitative real-time PCR. Receiver operating characteristic (ROC) analysis revealed that miR-483-5p and miR-20a had considerable diagnostic accuracy, yielding the areas under the ROC curve of 0.745 (sensitivity 58%, specificity 90%) and 0.74 (sensitivity 63%, specificity 85%), respectively. Plasma levels of miR-483-5p were associated with ISS staging. Within 14 months of diagnosis, the median progression-free survival of patients with high levels of plasma miR-483-5p was 15 months, in comparison with 21 months for patients with low levels of plasma miR-483-5p (p=0.025). However, miR-20a levels were not correlated with progression-free survival (p>0.05). miR-483-5p has the potential to be a predictor of myeloma survival.

Argonaute 2 promotes myeloma angiogenesis via microRNA dysregulation.

BACKGROUND: Dysregulated microRNA (miRNA) expression contributes to cancer cell proliferation, apoptosis and angiogenesis. Angiogenesis is a hallmark of multiple myeloma (MM) development and progression. Argonaute 2 (AGO2) protein, a core component of the RNA-induced silencing complex (RISC), can directly bind to miRNAs and mediate target messenger RNA (mRNA) degradation. A previous study showed that AGO2 knockdown suppressed human umbilical vein endothelial cell (HUVEC) growth and tube formation. However, the roles and molecular mechanisms of AGO2-induced myeloma angiogenesis are not yet fully understood. The aim of this study was to characterize these roles and effects and their associated mechanisms. RESULTS: Supernatants from AGO2-overexpressing MM lines induced HUVEC migration and accelerated tube formation. Conversely, supernatants from AGO2-knockdown MM lines suppressed HUVEC cell migration and tube formation. Moreover, a chick chorioallantoic membrane (CAM) assay was used to demonstrate that AGO2 could drive neovessel formation in MM lines in vivo. Using an miRNA microarray, we observed that 25 miRNAs were upregulated and 7 were downregulated in response to AGO2. Most let-7 family members and 2 miR-17/92 cluster members (miR-17a and miR-92-1), all known pro-angiogenic miRNAs, were positively regulated by AGO2 whereas anti-angiogenic miRNAs such as miR-145 and miR-361 were negatively regulated by AGO2. CONCLUSIONS: We conclude that AGO2 can drive neovessel formation in vitro and in vivo by dysregulating the expression of some angiogenic miRNAs. The pro-angiogenic miRNAs of the let-7 family and the miR-17/92 cluster, along with the anti-angiogenic miRNA miR-145, play crucial roles in AGO2-mediated angiogenesis by targeting angiogenesis-related genes.

TIMP-3 expression associates with malignant behaviors and predicts favorable survival in HCC.

The tissue inhibitors of metalloproteinases (TIMPs) are proteins that specifically inhibit the proteolytic activity of the matrix metalloproteinases (MMPs). TIMP-3, the only member of the TIMPs that can tightly bind to the extracellular matrix, has been identified as a unique tumor suppressor that demonstrates the ability to inhibit tumor angiogenesis, invasion, and metastasis. This study aimed to detect the expression of TIMP-3 in hepatocellular carcinoma (HCC) and investigate the association between TIMP-3 expression and its clinicopathological significance in HCC patients. In the current study, reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting of HCC cell lines and one-step quantitative reverse transcription PCR (qPCR) and immunohistochemistry (IHC) analyses in HCC tissues were performed, to characterize the TIMP-3 expression. Kaplan-Meier survival and Cox regression analyses were utilized to evaluate the prognosis of 101 HCC patients. The results showed that the expression of TIMP-3 in HCC was significantly decreased relative to that of non-cancerous cells and tissues. Furthermore, the TIMP-3 expression was statistically associated with malignant behaviors of HCC, including portal vein invasion (p = 0.036) and lymph node metastasis (p = 0.030). Cox regression analysis revealed that TIMP-3 expression was an independent prognostic factor for disease-free survival (p = 0.039) and overall survival (p = 0.049). These data indicate that TIMP-3 expression is a valuable prognostic biomarker for HCC and that TIMP-3 expression suggests a favorable prognosis for HCC patients.

[Expression level of microRNA-92a and its clinical significance in multiple myeloma patients].

OBJECTIVE: To investigate the relationship between the expression level of microRNA 92a (miR-92a) and del(13q14) and the prognosis of MM patients, and to explore the pathway that miR-92a involved. METHODS: Bone marrow samples from 53 newly diagnosed MM patients were collected, del(13q14) was analyzed by interphase fluorescence in situ hybridization in sorted CD138 positive plasma cell. The expression of miR-92a in plasma cells was measured by quantitative real-time PCR. The expression of c-jun was detected by Western blot in miR-92a transfected MM cell lines (LP-1, U266 and JJN3). RESULTS: Of the 53 MM patients, del(13q14) was detected in 31 (58.4%) patients. The median levels of miR-92a in MM patients with or without del(13q14) were 27.36±2.61 and 21.87±15.98, respectively (P>0.05). With the median follow-up of 13.5 (0.5-72.5) months, the median duration of progression-free survival of patients with high expression level of miR-92a was significantly shorter than those with low expression level of miR-92a (4.5 months vs 14.0 months, P=0.006). Overexpression of miR-92a in MM cell lines induces time-dependent down-regulation of c-jun. CONCLUSIONS: High expression of miR-92a was associated with poor prognosis in MM patients. The expression level of miR-92a was not associated with del(13q14), and the effect of miR-92a on the progress of MM might be involved in c-jun pathway.

IRF4 promotes cell proliferation by JNK pathway in multiple myeloma.

Interferon regulatory factor 4 (IRF4) is a member of the interferon regulatory family, which plays an important role in many lymphoid and myeloid malignancies. In the current study, using immunohistochemical staining, we found that IRF4 only expressed in plasma cells in bone marrow biopsy samples of multiple myeloma. IRF4-positive patients displayed increased disease stage (Durie-Salmon stage, p = 0.026; and International Staging System, p = 0.005). Silencing IRF4 in myeloma cell lines could inhibit myeloma cells proliferation and induce myeloma cell apoptosis, partly by JNK/Jun pathway. These results demonstrate that IRF4 plays important roles in myelomagenesis and disease progression.

[Clinical significance of serum free light chain in patients with multiple myeloma].

Multiple myeloma (MM) is a malignant disorder characterized by the proliferation of a single clone of plasma cells that can produce excessive amounts of serum free light chain (sFLC). sFLC plays an important role in MM diagnosis and disease monitoring. The quantitative immuno-nephelometric assay is sensitive and specific means for sFLC testing. The aim of this study was to investigate the levels of sFLC in multiple myeloma and the relationship between sFLC and serum total light chain (sTLC). sFLC in 45 newly diagnosed patients were detected by immuno-nephelometric assay, and then the ratio of free kappa to free lambda for every sample was calculated. Meanwhile, sTLC was also determined in these patients. The results showed that the difference of sFLC levels between MM patients and the normal controls was significant (tΚ = 8.86, P < 0.001; tλ = 15.48, P < 0.001;tΚ/λ = 5.54,P < 0.005). No correlation between sFLC and sTLC was found in MM patients. It is concluded that the level of sFLC in MM patients is significantly higher than that in normal controls. sFLC and its ratio may be served as a indicator for diagnosis of MM. sTLC can not replace the role of sFLC.