Clinical significance of LAG-3 on microvessel density in primary hepatocellular carcinoma.

Aims: We aimed to determine whether lymphocyte activation gene 3 (LAG-3), also known as CD223, is associated with microvessel density (MVD) in primary hepatocellular carcinoma (HCC), as well as their clinical significance in predicting survival. Materials and methods: One hundred and twenty-seven patients were enrolled in the study. Samples were obtained on resection at the Department of Hepatobiliary Surgery of the Qingdao Municipal Hospital from June 2014 to June 2016. Immunohistochemistry was used to determine vessel density and LAG-3 abundance. Statistical analyses were performed to test for correlation of LAG-3 density and other clinicopathological variables with overall survival (OS). Results: High LAG-3 abundance was significantly correlated with increased MVD in primary HCC (P < 0.05). The χ2 test revealed a significant association of LAG-3 with preoperative AFP level, tumor diameter, N stage, and the presence of HBV infection (P < 0.05). Patients with high LAG-3 expression had shorter OS compared to those with low LAG-3 expression (P < 0.05). The Cox proportional hazards model showed that both higher LAG-3 and MVD density, age, the number of tumors, preoperative AFP level, tissue differentiation, Child-Pugh grade, and lymph node metastasis correlated with survival. Conclusions: High expression of LAG-3 is associated with angiogenesis and poor prognosis in HCC patients. With the deepening of research, LAG-3 is likely to become a novel biomarker for clinical diagnosis and prognosis and can even be a therapeutic target of HCC.

LAG-3 is a promising inhibitory immune checkpoint for antitumor immunotherapy.

INTRODUCTION: Lymphocyte activation gene-3 (LAG-3) is a member of the immunoglobulin superfamily. Engagement of LAG-3 by its ligands to trigger downstream signaling can inhibit immune responses and regulate the pathogenesis of many diseases, including cancer and inflammatory diseases. AREAS COVERED: We used keywords to search for relevant publications in PubMed and information on websites. After systematic analysis, we discuss the biological characteristics of LAG-3 and its ligands, LAG-3 related signaling, its roles in the pathogenesis of tumors, and its blockages for the treatment of cancers, as well as current challenges and future directions of research. EXPERT OPINION: Although the mechanisms underlying the action of LAG-3/ligand-related signaling in tumor development are not fully understood, advances in scientific research and LAG-3-based immunotherapies are promising. Further studies to explore its biological roles and molecular mechanisms may aid in developing new LAG-3- and ligand-based therapeutic drugs to benefit patients with different types of cancers.

Correlation Analysis Among the Level of IL-35, Microvessel Density, Lymphatic Vessel Density, and Prognosis in Non-Small Cell Lung Cancer.

The aim of this study was to determine the expression of IL-35 and the lymphatic vessel density (LVD) and microvessel density (MVD) in the pathological tissues from patients with non-small cell lung cancer (NSCLC) and to analyze their correlation with other common clinical prognostic factors, as well as patients' overall survival and progression-free survival. We analyzed the pathological characteristics of 130 patients with NSCLC and determined the IL-35 expression, MVD, and LVD changes in the pathological tissues by immunohistochemistry. The results showed that IL-35 expression was significantly correlated with tumor differentiation, lymph node metastasis, T staging, LVD, and MVD (P < 0.05) but was not associated with age, sex, smoking, and other factors. Univariate analysis of risk models showed that age, lymph node metastasis, T stage, and high IL-35 expression, LVD, and MVD were significantly associated with NSCLC prognosis (P < 0.05), whereas sex, smoking, and high differentiation were not correlated with prognosis. Multivariate analysis of the proportional risk model showed that the IL-35 expression, lymph node metastasis, high LVD, and high MVD were significantly correlated with NSCLC prognosis (P < 0.05). In conclusion, IL-35, MVD, and LVD may be independent prognostic markers. In addition, IL-35 might represent a promising clinical drug target for the treatment of NSCLC.

Clinical significance of cancer-associated fibroblasts and their correlation with microvessel and lymphatic vessel density in lung adenocarcinoma.

BACKGROUND: To determine whether cancer-associated fibroblasts (CAFs) are associated with microvessel density (MVD) and lymphatic vessel density (LVD) in lung adenocarcinoma (ADC) or are not prognostic. METHODS: Ninety-three lung adenocarcinoma patients without adjuvant therapy between January 2010 and June 2011 were enrolled. CAFs, MVD, and LVD were identified by α-smooth muscle actin (α-SMA), CD34 and D2-40 staining via immunohistochemistry. Staining intensities were assessed and quantified. For statistics, Pearson's chi-square test, logistic regression, Kaplan-Meier, and log-rank tests were applied. In addition, the Cox proportional hazards model was used for multifactor analysis to predict survival. RESULTS: CAFs abundance in lung adenocarcinoma is associated with higher MVD and LVD. In addition, a correlation was demonstrated between MVD and LVD (P < 0.05). CAFs, MVD, and LVD are significantly correlating with age, tumor size, differentiation grade, clinical stage, and lymph node metastasis (P < 0.05), but not influenced by gender, tumor location, and smoking history. Three-year overall survival in the CAFs-poor group is 64.5%, which is significant higher than that in the CAFs-rich cohort (41.9%). Further, we found that age, clinical stage, α-SMA, CD34, D2-40 positivity, tumor size, differentiation grade, and lymph node metastasis significantly correlate with overall survival of patients with lung adenocarcinoma. However, sex, smoking history, and tumor location have no association with 3-year survival. The clinical stage is an independent prognostic factor in overall survival (P < 0.05). CONCLUSIONS: The density of CAFs identified by α-SMA staining is associated with progression and metastasis of lung adenocarcinoma and affects the patient's disease outcome.

Erratum: The association between human papillomavirus presence and epidermal growth factor receptor mutations in Asian patients with non-small cell lung cancer.

[This corrects the article DOI: 10.21037/tlcr.2018.03.16.].

The association between human papillomavirus presence and epidermal growth factor receptor mutations in Asian patients with non-small cell lung cancer.

BACKGROUND: The etiology of non-small cell lung cancer (NSCLC) in non-smoker patients remains largely unknown. It has been widely proved that human papillomavirus (HPV) participates in the development of various cancers. Epidermal growth factor receptor (EGFR) mutation patients represent a large portion of non-smokers with NSCLC. We performed this meta-analysis to determine whether HPV infection in NSCLC tissue is associated with EGFR mutations compared with HPV negative controls. METHODS: Online databases were searched up to June 30th 2017. We included studies in which HPV detection was based on polymerase chain reaction (PCR) methods. Random effects model was used in data synthesis and the relative effects were presented as odds ratio (OR) with 95% confidence intervals (CIs). RESULTS: Finally, four eligible studies with a total of 498 patients from Asian countries were identified and included. The general EFGR mutation positive rate was 38.2% among all patients, and the HPV DNA detection rate (HPV subtype being involved: 16, 18, 33 and 58) was 35.3%. The presence of EGFR mutation was significantly higher in HPV-positive patients compared with HPV-negative controls (52% vs. 31%; OR =2.41, 95% CI: 1.21 to 4.77; P=0.012), with moderate heterogeneity among studies (I2=59%; P=0.062). CONCLUSIONS: Our results suggest that HPV infection is associated with EGFR mutations in NSCLC, at least in Asian populations. Further efforts should be made on exploring the potential mechanism and the prognostic character of HPV/EGFR positive NSCLC patient.

The alteration of T790M between 19 del and L858R in NSCLC in the course of EGFR-TKIs therapy: a literature-based pooled analysis.

BACKGROUND: Treatment-naive epidermal growth factor receptor (EGFR) T790M mutation is more inclined to coexist with L858R than with 19 del in non-small cell lung cancer (NSCLC) patients. However, EGFR-tyrosine kinase inhibitors (EGFR-TKIs) might alter this status. We sought to compare the prevalence of T790M upon acquired resistance to EGFR-TKIs between 19 del and L858R by assembling all existing data. METHODS: Electronic databases were comprehensively searched for eligible studies. The primary endpoint was the odds ratio (OR) of T790M mutation in NSCLC co-existing with L858R mutation and 19 del upon resistance to first-generation EGFR-TKIs. A random effects model was used. Stratified analysis was performed based on study type (retrospective and prospective), race (Asians and Caucasians) and sample type (tissue and plasma). RESULTS: A total of 25 studies involving 1,770 patients were included. The overall T790M existent rate was 45.25%. Post-resistance T790M was more frequent in 19 del than in L858R mutated patients (53% vs. 36%; OR 1.87; P<0.001). All outcomes of subgroup and overall analyses were similar. In contrast, we re-analyzed the previous meta-analysis, finding that the pooled rate of pretreatment T790M was 14% and 22% in 19 del and L858R respectively (OR 0.59; P<0.001). The increase of T790M rate was 2.79-fold in 19 del and only 0.63-fold in L858R in the course of EGFR-TKIs therapy. CONCLUSIONS: Opposite to the situation of de novo T790M, it was observed that T790M was more frequent in exon 19 deletion than in L858R among patients with acquired resistance to EGFR-TKIs. The difference in T790M alteration between 19 del and L858R encourages development of detection or treatment strategies for the specific resistance mechanism.

Clinical significance of FAP-α on microvessel and lymphatic vessel density in lung squamous cell carcinoma.

AIMS: We aimed to determine whether cancer-associated fibroblasts (CAFs) are associated with microvessel density (MVD) and lymphatic vessel density (LVD) in lung squamous cell carcinoma, as well as their clinical significance in predicting survival. METHODS: 122 patients were enrolled in the study. Samples were obtained on resection at the Department of Thoracic Surgery of the Qingdao Municipal Hospital between January 2011 and December 2014. Immunohistochemistry was used to determine vessel and lymphatic vessel density, and CAF abundance (fibroblast activation protein α (FAP-α) positivity). Statistical analyses were performed on 85 patients to test for correlation of CAF density and other clinicopathological variables with 3-year overall survival (OS) and disease-free survival (DFS). RESULTS: High stromal CAF abundance significantly correlated with increased MVD and LVD in lung squamous cell carcinoma (p<0.05). χ2 test revealed a significant association of CAF density with lymph node metastasis. Cox proportional hazards model showed that both higher CAF density and lymph node metastasis negatively correlate with survival. CAF density or lymph node status can be used as an independent prognostic factor to predict 3-year OS and DFS. CONCLUSIONS: CAF density, identified by FAP-α staining pattern, should be considered as a novel biomarker for disease prognosis in patients with lung squamous cell carcinoma.

MicroRNA-432 functions as a tumor suppressor gene through targeting E2F3 and AXL in lung adenocarcinoma.

Abnormal proliferation and drug resistance are the hallmarks of lung adenocarcinoma (LAD). Dispite the advances in diagnosis and therapy, the 5-year survival remains low. Increasing studies regarding its pathological mechanism have been focused on microRNA (miRNA) due to its nodal regulatory properties. This study aims to characterize the expression of miR-432 in LAD and investigate its effects on the proliferation and sensitivity of lung cancer cells to cisplatin. Here, we report that downregulation of miR-432 in LAD tissues was correlated with a higher clinical stage (p = 0.03) and poor prognosis (p = 0.036). Additionally, miR-432 expression was negative correlated with high Ki67 labeling index (p = 0.016) in our cohorts. Functionally, over-expression of miR-432 inhibits cell proliferation through arresting cell cycle and sensitizes tumor cells to cisplatin. Mechanistically, miR-432 functions by directly targeting E2F3 and AXL, and they, in turn, mediate the regulation of miR-432 towards cell proliferation and cisplatin sensitivity. Importantly, miR-432 levels are negatively correlated with the levels of E2F3 and AXL in human LAD tissues. These results demonstrated that miR-432 functions as a tumor-suppressive miRNA and may represent a prognostic parameter and therapeutic target for LAD.

[Prevalence of acute coronary heart disease among farmers in Panyu, Guangzhou: a 20-year population-based study].

OBJECTIVE: To monitor the incidence change of acute coronary heart events in the all-ages farmers in Panyu District, Guangzhou City during 1991-2001 and 2010-2011. METHODS: The surveillance on the same defined population as that from the PRC-USA cooperative epidemiologic project on the cardiovascular and pulmonary diseases 30-year ago was carried out in Panyu, Guangzhou in 1991-2001 and 2010-2011. The crude incidence of acute coronary events and the age-standardized incidence rate were calculated by the year, gender and age, and standardized with the world standard population age distribution. Incidences at the two different periods were compared. The annual average changing rate of the incidence was obtained by the regression analysis methods. RESULTS: In the 11 consecutive years of 1991-2001, a upward trend on the incidence of acute coronary events among the farmers in women in Panyu District was found (P < 0.05). The incidence of the acute coronary events in the year of 2010-2011 was significantly higher than that in the year of 1991 to 2001 [34.06 per 100 000 (age-adjusted rate as 28.50 per 100 000) versus 16.14 per 100 000 (age-adjusted rate as 16.57 per 100 000), P < 0.05]. The age-adjusted rate increased by 83.04%. Peak incidence of acute coronary events in males was noticed in 75-79 age group, and in 80-84 age group in females. Comparing to the period of 1991-2001, the largest incidence increases appeared in the age groups of 35-39 and 75-79 years in males, while in the age groups of 50-54 and 65-69 years in females. Up to 47.37% (36/76) events occurred on the age group older than 75 years, raised by 40.44% comparing to that in 1991-2001 (33.73%, 56/166). CONCLUSIONS: The incidence of acute coronary events among farmers in Panyu District is at middle or low level of China but there is an increasing trend in acute coronary incidence from 1991 to 2011. Our results suggest that the prevention and treatment on acute coronary syndrome should be strengthened, and especially on the age group with the largest increase of disease incidence.

[Evaluation of induction chemotherapy with vinorelbine plus cisplatin (NP) or docetaxel plus cisplatin (TP) combined with concurrent chemoradiotherapy for patients with locally advanced nasopharyngeal carcinoma].

OBJECTIVE: To compare the efficacy and side effects of induction chemotherapy with vinorelbine plus cisplatin (NP) or docetaxel plus cisplatin (TP) combined with concurrent chemoradiotherapy in treating locally advanced nasopharyngeal carcinoma (NPC). METHODS: From January 2005 to December 2009, 146 patients with locally advanced nasopharyngeal carcinoma treated in our department were randomized into NP group (76 patients) or TP group (70 patients). Both groups received two cycles of induction chemotherapy and concurrent chemoradiotherapy. After three weeks of induction chemotherapy, the patients received concurrent chemoradiotherapy. The chemotherapy was recycled every three weeks. Two groups were treated with intensity-modulated radiation therapy. RESULTS: The short-term efficacy of NP group was similar to that of TP group. The 3-year overall survival rates, disease-free-survival rates, locoregional relapse-free survival rates and distant metastasis-free survival rates in the NP and TP groups were 84.2% and 82.9%, 71.1% and 74.3%, 89.5% and 91.4%, 81.6% and 77.1%, respectively (P > 0.05). The occurrence rates of leucopenia, anemia and acute mucositis were significantly higher in the TP group than those in the NP group (P < 0.05). The gastrointestinal toxicity, dermatitis and liver toxicity were similar in the two groups. CONCLUSIONS: The efficacy of NP regimen induction chemotherapy plus concurrent chemordiotherapy for advanced NPC is similar to that of TP regimen. The toxicity of the NP regimen is lower than that of NP regimen, tolerable, and with a good compliance.

A visual detection of protein content based on titration of moving reaction boundary electrophoresis.

A visual electrophoretic titration method was firstly developed from the concept of moving reaction boundary (MRB) for protein content analysis. In the developed method, when the voltage was applied, the hydroxide ions in the cathodic vessel moved towards the anode, and neutralized the carboxyl groups of protein immobilized via highly cross-linked polyacrylamide gel (PAG), generating a MRB between the alkali and the immobilized protein. The boundary moving velocity (V(MRB)) was as a function of protein content, and an acid-base indicator was used to denote the boundary displacement. As a proof of concept, standard model proteins and biological samples were chosen for the experiments to study the feasibility of the developed method. The experiments revealed that good linear calibration functions between V(MRB) and protein content (correlation coefficients R>0.98). The experiments further demonstrated the following merits of developed method: (1) weak influence of non-protein nitrogen additives (e.g., melamine) adulterated in protein samples, (2) good agreement with the classic Kjeldahl method (R=0.9945), (3) fast measuring speed in total protein analysis of large samples from the same source, and (4) low limit of detection (0.02-0.15 mg mL(-1) for protein content), good precision (R.S.D. of intra-day less than 1.7% and inter-day less than 2.7%), and high recoveries (105-107%).

Fast and selective determination of total protein in milk powder via titration of moving reaction boundary electrophoresis.

In this paper, moving reaction boundary titration (MRBT) was developed for rapid and accurate quantification of total protein in infant milk powder, from the concept of moving reaction boundary (MRB) electrophoresis. In the method, the MRB was formed by the hydroxide ions and the acidic residues of milk proteins immobilized via cross-linked polyacrylamide gel (PAG), an acid-base indicator was used to denote the boundary motion. As a proof of concept, we chose five brands of infant milk powders to study the feasibility of MRBT method. The calibration curve of MRB velocity versus logarithmic total protein content of infant milk powder sample was established based on the visual signal of MRB motion as a function of logarithmic milk protein content. Weak influence of nonprotein nitrogen (NPN) reagents (e.g., melamine and urea) on MRBT method was observed, due to the fact that MRB was formed with hydroxide ions and the acidic residues of captured milk proteins, rather than the alkaline residues or the NPN reagents added. The total protein contents in infant milk powder samples detected via the MRBT method were in good agreement with those achieved by the classic Kjeldahl method. In addition, the developed method had much faster measuring speed compared with the Kjeldahl method.

[Determination of total protein content in soya-bean milk via visual moving reaction boundary titration].

A visual, rapid and accurate moving reaction boundary titration (MRBT) method was used for the determination of the total protein in soya-bean milk. During the process, moving reaction boundary (MRB) was formed by hydroxyl ions in the catholyte and soya-bean milk proteins immobilized in polyacrylamide gel (PAG), and an acid-base indicator was used to denote the boundary motion. The velocity of MRB has a relationship with protein concentration, which was used to obtain a standard curve. By paired t-test, there was no significant difference of the protein content between MRBT and Kjeldahl method at 95% confidence interval. The procedure of MRBT method required about 10 min, and it had linearity in the range of 2.0-14.0 g/L, low limit of detection (0.05 g/L), good precision (RSD of intra-day < 1.90% and inter-day < 4.39%), and high recoveries (97.41%-99.91%). In addition, non-protein nitrogen (NPN) such as melamine added into the soya-bean milk had weak influence on MRBT results.

Combination chemotherapy with paclitaxel, cisplatin and fluorouracil for patients with advanced and metastatic gastric or esophagogastric junction adenocarcinoma: a multicenter prospective study.

OBJECTIVE: To evaluate the efficacy and toxicity of the combination regimen of paclitaxel, cisplatin and 5-FU (PCF) as first-line or second-line therapy in patients with advanced gastric and esophagogastric junction (EGJ) adenocarcinoma in China. METHODS: The patients were treated with paclitaxel 150 mg/m(2) on d1; fractionated cisplatin 15 mg/m(2) and continuous infusion 5-FU 600 mg/(m(2)·d) intravenously on d1-d5 of a 21-d cycle until disease progression or unacceptable toxicities. RESULTS: Seventy-five patients have been enrolled, among which, 41 received PCF regimen as the first-line therapy (group A) and 34 received the regimen as the second-line therapy (group B) with the median age of 59 years old and Karnofsky performance status (KPS) score ≥80. Toxicities were analyzed in all 75 patients. Seventy-one patients were evaluable for efficacy. The median overall survival (mOS) was 12.0 months (95% CI: 7.9-16.2 months) in group A and 7.3 months (95% CI: 4.3-10.3 months) in group B, respectively. The median progression-free survival (mPFS) was 5.7 months (95% CI: 4.1-7.2 months) and 5.0 months (95% CI: 3.1-6.9 months), respectively. The response rate (CR+PR) was 40% (16/40; 95% CI: 24.9-56.7%) in group A and 22.6% (7/31; 95% CI: 9.6-41.1%) in group B. Major grade 3 or 4 adverse events include neutropenia (41.3%), febrile neutropenia (9.3%), nausea/anorexia (10.7%), and vomiting (5.3%). There was no treatment-related death. CONCLUSIONS: The combination chemotherapy with PCF is active and tolerable as first-line and second-line therapy in Chinese patients with advanced gastric and EGJ adenocarcinoma. The response and survival of PCF are same as those of DCF, but the tolerance is much better.

Adverse trends of cardiovascular risk factors among low risk populations (1983-1994)--a cohort study of workers and farmers in Guangzhou, China.

BACKGROUND: The levels and trends of cardiovascular risk factors vary greatly throughout China. We examine 10-year trends of cardiovascular risk factors (1983-1994) and the factors related to these trends among low-risk cohorts of workers and farmers in Guangzhou, China. METHODS: This is a cohort study of 3,131 workers and 3,493 farmers aged 25-64 years at baseline with 10 years of follow-up. We performed a longitudinal analysis to account for the aging of the cohorts and the repeated measures of the same individual. RESULTS: At baseline the prevalence of overweight (including obese) ranged from 1.0% to 11.8%, hypertension ranged from 3.8% to 10.5%, and mean serum total cholesterol (TC) ranged from 155.4 mg/dl to 187.2 mg/dl. Although prevalence of smoking declined, blood pressure levels and body mass index (BMI) increased significantly, and lipid profiles changed unfavorably during the 10-year follow-ups. The prevalence of hypertension increased from 5.0 percentage points (female farmers) to 12.3 percentage points (male farmers). Mean TC increased significantly (e.g., +22.8 mg/dl and +17.0 mg/dl in male and female farmers, respectively). In the longitudinal data analyses, increase in BMI was associated with increase in blood pressure levels and TC. Significant adverse trends of risk factors persisted after adjustment for aging, education, BMI, smoking, and alcohol intake. CONCLUSION: Urgent action is needed to prevent and reverse the unhealthy trends occurring among these low risk Chinese workers and farmers.

[Epidemiological survey of rheumatic heart disease in schoolchildren in Guangdong and Xinjiang].

OBJECTIVE: To investigate the prevalence of rheumatic heart disease (RHD) among schoolchildren in Guangdong Province and Xinjiang Uygur Autonomous Region. METHODS: Using a cluster sampling method, an epidemiological survey of RHD was conducted in 16 682 primary and high school students by auscultation in Guangdong Province and Xinjiang Uygur Autonomous Region from 2005 to 2006. Review of the clinical records, RHD survey in adults, and examination of the positivity rate of group A beta-hemolytic Streptococcus (GAS) by throat swab cultures in the students aged between 9 and 12 years in the sampled schools were also carried out. RESULTS: No RHD patient was found in the sampled population. In Xinjiang, the prevalence of RHD was 12.9/1000 among adults, higher than that (2.2/1000) in Guangdong Province. The GAS-positive rate in the schoolchildren in Xinjiang ranged from 9.8% to 12.6%, higher than that in Guangdong (2.3%-3.9%). CONCLUSION: The GAS-positive rate among children and incidence of RHD in adults are higher in Xinjiang than in Guangdong. The prevalence of RHD among the schoolchildren shows a reduction compared with that in 1994.