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.

Organic fertilizer application increases the soil respiration and net ecosystem carbon dioxide absorption of paddy fields under water-saving irrigation.

Quantifying carbon sequestration in paddy soil is necessary to understand the effect of agricultural practices on carbon cycles. The objective of this study was to assess the effect of organic fertilizer addition (MF) on the soil respiration and net ecosystem carbon dioxide (CO2) absorption of paddy fields under water-saving irrigation (CI) in the Taihu Lake Region of China during the 2014 and 2015 rice-growing seasons. Compared with the traditional fertilizer and water management (FC), the joint regulation of CI and MF (CM) significantly increased the rice yields and irrigation water use efficiencies of paddy fields by 4.02~5.08 and 83.54~109.97% (p < 0.05). The effects of organic fertilizer addition on soil respiration and net ecosystem CO2 absorption rates showed inter-annual differences. CM paddy fields showed a higher soil respiration and net CO2 absorption rates during some periods of the rice growth stage in the first year and during most periods of the rice growth stage in the second year. These fields also had significantly higher total CO2 emission through soil respiration (total Rsoil) and total net CO2 absorption compared with FC paddy fields (p < 0.05). The total Rsoil and net ecosystem CO2 absorption of CM paddy fields were 67.39~91.55 and 129.41~113.75 mol m-2, which were 27.66~135.52 and 12.96~31.66% higher than those of FC paddy fields. The interaction between water and fertilizer management had significant effects on total net ecosystem CO2 absorption. The frequent alternate wet-dry cycles of CI paddy fields increased the soil respiration and reduced the net CO2 absorption. Organic fertilizer promoted the soil respiration of paddy soil but also increased its net CO2 absorption and organic carbon content. Therefore, the joint regulation of water-saving irrigation and organic fertilizer is an effective measure for maintaining yield, increasing irrigation water use efficiency, mitigating CO2 emission, and promoting paddy soil fertility.

Quantification of endostar in rat plasma by LC-MS/MS and its application in a pharmacokinetic study.

LC-MS/MS is a promising analytical platform for the quantification of recombinant therapeutic proteins in biological fluids for pharmacokinetic (PK) studies. Herein, an absolute quantification method based on LC-MS/MS technique was developed to quantify endostar, which is modified from the recombinant human endostatin by adding a nine-amino acid sequence (MGGSHHHHH) at the N-terminal. A reproducible three-step analytical procedure was adopted: (1) Ni(2+) Sepharose was used to selectively extract endostar; (2) the signature peptide "TEAPSATGQASSLLGGR" (m/z 802.3(2+)-651.8(2+)) of endostar and a synthetic peptide "TEAPSATGQVSSLLGGR" (m/z 816.9(2+)-666.4(2+)) as internal standard (IS) were selected and analyzed in the multiple reaction monitoring (MRM) mode; (3) the proposed method was validated and applied to the pharmacokinetic study of endostar. The lower limit of quantification (LLOQ) for quantifying endostar was 50 ng/ml and this method is linear over 50-10,000 ng/ml. The accuracy was between 85% and 115%, and the intra-batch and inter-batch analytic precision and accuracy were below 15%. This LC-MS/MS approach was validated for the application to the pharmacokinetic study of endostar in rats.