[A retrospective analysis of estimating postmortem interval in 256 murder cases].

OBJECTIVE: To discuss the method of reducing error in estimating postmortem interval (PMI). METHODS: Two hundred and fifty-six solved murder cases from 2003 January to 2013 January in Changzhou and Nanjing City were collected, The PMI of all cases was estimated by traditional method and then compared with the real PMI obtained after the cases were solved. The cases were grouped according to the PMI, the accuracy was calculated, and the reasons of suboptimal PMI were analyzed. RESULTS: The accuracies of early PMI (less than 12h and 13-24 h) were 90% and 89%, respectively; while the accuracies of late PMI (1-7 d, 1-2 weeks, 3-4 weeks, 1-6 months, 7-12 months and 1-5 years) decreased over time, being 79%, 76%, 83%, 79%, 60% and 50%, respectively. The common reasons of estimating error included improper inference methods, water submerged body, extreme temperature, lack of objective evidence, intentionally abandoned body, and changed or destroyed scene, etc. CONCLUSION: The multiple index data can reduce the error in estimating PMI.

Efficient chain elongation synthesis of n-caproate from shunting fermentation of food waste.

Food waste was used to produce ethanol by yeast fermentation and volatile fatty acids (VFAs) by hydrolytic acidogenesis for chain elongation. Effectiveness of mole ratio of ethanol in yeast fermentation effluent (YFE) to VFAs in hydrolytic acidification effluent (HAE) on chain elongation was examined. The ideal YFE to HAE ratio for chain elongation was 2:1, the highest n-caproate production was 169.76 mg COD/g vS and the food waste utilization was 65.43 %. Electron transfer and carbon distribution did not completely correspond to n-caproate production, suggesting timely product extraction. The abundance of Romboutsia and Clostridium_sensu_stricto_12 increased as chain elongation progressed, which was critical for the chain elongation to n-caproate. The food waste shunting ratio of yeast fermentation to hydrolytic acidogenesis was 6:5, and 572.6 CNY can be created through chain elongation from shunting fermentation of 1 t food waste. This study proposed a new approach for efficient producing n-caproate from food waste.

Metformin Use and Risk of All-Cause Mortality and Cardiovascular Events in Patients With Chronic Kidney Disease-A Systematic Review and Meta-Analysis.

Background: To evaluate whether metformin use assuredly alters overall all-cause death in patients with type 2 diabetes mellitus (T2DM) and chronic kidney disease (CKD). Methods: Pubmed, Web of Science, Embase, and Cochrane Central Register of Controlled Trials were systematically searched from inception to Feb. 29, 2020 with no language restriction. All related articles comparing all-cause death of T2DM and CKD patients after metformin use (monotherapy or combination) versus non-metformin treatment were identified. Pooled risk ratios (RR) and 95% confidence intervals (CI) were computed using random-effects models regardless of the heterogeneity quantified by Cochrane χ2 and I2 statistics. Results: Totally 13 studies (9 cohort studies [CSs], 3 subanalyses or post-hoc analyses of randomized controlled trials [RCTs], and 1 nested case-control article) involving 303,540 patients were included. Metformin-based treatments relative to any other measure displayed significantly lower risks of all-cause mortality (Pooled RRs 0.71, 95%CI 0.61 to 0.84; I2 = 79.0%) and cardiovascular events (Pooled RRs 0.76, 95%CI 0.60 to 0.97; I2 = 87.0%) in CKD patients at stage G1-3, with substantial heterogeneity. Metformin use was not significantly related with these end points in advanced CKD patients. Conclusions: Metformin use is connected with significantly less risks of all-cause mortality and cardiovascular events in patients with T2DM and mild/moderate CKD. However, RCTs with large sample sizes are warranted in the future to assess whether these key benefits extend to later stages of CKD by dose adjustment.

Assessing the binding properties of the anti-PD-1 antibody landscape using label-free biosensors.

Here we describe an industry-wide collaboration aimed at assessing the binding properties of a comprehensive panel of monoclonal antibodies (mAbs) against programmed cell death protein 1 (PD-1), an important checkpoint protein in cancer immunotherapy and validated therapeutic target, with well over thirty unique mAbs either in clinical development or market-approved in the United States, the European Union or China. The binding kinetics of the PD-1/mAb interactions were measured by surface plasmon resonance (SPR) using a Carterra LSA instrument and the results were compared to data collected on a Biacore 8K. The effect of chip type on the SPR-derived binding rate constants and affinities were explored and the results compared with solution affinities from Meso Scale Discovery (MSD) and Kinetic Exclusion Assay (KinExA) experiments. When using flat chip types, the LSA and 8K platforms yielded near-identical kinetic rate and affinity constants that matched solution phase values more closely than those produced on 3D-hydrogels. Of the anti-PD-1 mAbs tested, which included a portion of those known to be in clinical development or approved, the affinities spanned from single digit picomolar to nearly 425 nM, challenging the dynamic range of our methods. The LSA instrument was also used to perform epitope binning and ligand competition studies which revealed over ten unique competitive binding profiles within this group of mAbs.

Purification of common light chain IgG-like bispecific antibodies using highly linear pH gradients.

Monovalent bispecific antibodies (BsAbs) are projected to have broad clinical applications due to their ability to bind two different targets simultaneously. Although they can be produced using recombinant technologies, the correct pairing of heavy and light chains is a significant manufacturing problem. Various approaches exploit mutations or linkers to favor the formation of the desired BsAb, but a format using a single common light chain has the advantage that no other modification to the antibody is required. This strategy reduces the number of formed molecules to three (the BsAb and the two parent mAbs), but the separation of the BsAb from the two monovalent parent molecules still poses a potentially difficult purification challenge. Current methods employ ion exchange chromatography and linear salt gradients, but are only successful if the difference in the observed isoelectric points (pIs) of two parent molecules is relatively large. Here, we describe the use of highly linear pH gradients for the facile purification of common light chain BsAbs. The method is effective at separating molecules with differences in pI as little as 0.10, and differing in their sequence by only a single charged amino acid. We also demonstrate that purification resins validated for manufacturing are compatible with this approach.

VEGF gene expression is regulated post-transcriptionally in macrophages.

The macrophage is critical to the innate immune response and contributes to human diseases, including inflammatory arthritis and plaque formation in atherosclerosis. Vascular endothelial growth factor (VEGF) is an angiogenic cytokine that is produced by macrophages. To study the regulation of VEGF production in macrophages we show that stimulation of monocyte-macrophage-like RAW-264.7 cells by lipopolysaccharide (LPS) increases expression of VEGF mRNA and protein. Three alternative splicing VEGF mRNA isoforms are produced, and the stability of VEGF mRNA increases following cellular activation. To study post-transcriptional regulation of the VEGF gene the 3'-untranslated region (3' UTR) was introduced into the 3' UTR of the luciferase gene in a reporter construct. In both RAW-264.7 cells and thioglycollate-elicited macrophages, the 3' UTR sequence dramatically reduces reporter expression. Treatment with activators of macrophages, including LPS, lipoteichoic acid, and VEGF protein, stimulates expression of 3' UTR reporters. Finally, mapping studies of the 3' UTR of VEGF mRNA show that deletion of the heterogeneous nuclear ribonucleoprotein l binding site affects basal reporter expression in RAW-264.7 cells, but does not affect reporter activation with LPS. Together these results demonstrate that a post-transcriptional mechanism contributes to VEGF gene expression in activated macrophage cells.

[Study of zinc in protecting sperm from sodium nitroprusside damage].

OBJECTIVE: To study the function of zinc in preventing human sperm from being damaged by sodium nitroprusside (SNP), an external NO donor. METHODS: Analyses were made of the function of zinc in protecting sperm from being influenced by SNP in such aspects as sperm motility, head-tail connection and the breakage of sperm DNA chain by using phase-contrast microscope and single cell gel electrophoresis (SCGE). RESULTS: Sperm motility was obviously inhibited by SNP. The percentage of comet cells increased significantly but the stability of sperm head-tail connection decreased. Zinc could promote sperm motility, protect the DNA chain and prevent the sperm head-tail connection from breaking. CONCLUSION: Zinc can protect sperm from being damaged by NO. Its mechanism may be related to the mercaptol group of sperm chromatin.

AUF1/hnRNP D represses expression of VEGF in macrophages.

Vascular endothelial growth factor (VEGF) is a regulator of vascularization in development and is a key growth factor in tissue repair. In disease, VEGF contributes to vascularization of solid tumors and arthritic joints. This study examines the role of the mRNA-binding protein AUF1/heterogeneous nuclear ribonucleoprotein D (AUF1) in VEGF gene expression. We show that overexpression of AUF1 in mouse macrophage-like RAW-264.7 cells suppresses endogenous VEGF protein levels. To study 3' untranslated region (UTR)-mediated regulation, we introduced the 3' UTR of VEGF mRNA into a luciferase reporter gene. Coexpression of AUF1 represses VEGF-3' UTR reporter expression in RAW-264.7 cells and in mouse bone marrow-derived macrophages. The C-terminus of AUF1 contains arginine-glycine-glycine (RGG) repeat motifs that are dimethylated. Deletion of the RGG domain of AUF1 eliminated the repressive effects of AUF1. Surprisingly, expression of an AUF1-RGG peptide reduced endogenous VEGF protein levels and repressed VEGF-3' UTR reporter activity in RAW-264.7 cells. These findings demonstrate that AUF1 regulates VEGF expression, and this study identifies an RGG peptide that suppresses VEGF gene expression.