锁阳多糖提取工艺的研究

以浸膏得率和浸膏中多糖质量分数综合评价,利用L9(34)正交实验从水提和醇沉两方面优选锁阳中水溶性多糖最佳提取工艺条件,为锁阳活性多糖的开发提供科学依据和实验基础。最佳水提工艺条件为:m(锁阳):v(水)=1:12,回流提取0.5 h,提取2次;最佳醇沉工艺条件为:在m(生药):v(水提液)=1 g:2 mL溶液中加入乙醇,使φ(乙醇)=70%,静置6 h。     

Resistance outside the substrate envelope: hepatitis C NS3/4A protease inhibitors

Direct acting antivirals have dramatically increased the efficacy and tolerability of hepatitis C treatment, but drug resistance has emerged with some of these inhibitors, including nonstructural protein 3/4?A protease inhibitors (PIs). Although many co-crystal structures of PIs with the NS3/4A protease have been reported, a systematic review of these crystal structures in the context of the rapidly emerging drug resistance especially for early PIs has not been performed. To provide a framework for designing better inhibitors with higher barriers to resistance, we performed a quantitative structural analysis using co-crystal structures and models of HCV NS3/4A protease in complex with natural substrates and inhibitors. By comparing substrate structural motifs and active site interactions with inhibitor recognition, we observed that the selection of drug resistance mutations correlates with how inhibitors deviate from viral substrates in molecular recognition. Based on this observation, we conclude that guiding the design process with native substrate recognition features is likely to lead to more robust small molecule inhibitors with decreased susceptibility to resistance.     

Optimization and mathematical modeling of the transtubular bioreactor for the production of monoclonal antibodies from a hybridoma cell line

This report describes the use of a transtubular bioreactor to study the relative effects of diffusion versus perfusion of medium on antibody production by a hybridoma cell line. The study was performed with a high-density cell culture maintained in a serum-free, low-protein medium for 77 days. It was determined that the reactor possessed a macro-mixing pattern residence time distribution similar to a continuous stirred tank reactor (CSTR). However, due to the arrangement of the medium lines in the reactor, the flow patterns for nutrient distribution consist of largely independent medium path lengths ranging from short to long. When operated with cyclic, reversing, transtubular medium flow, some regions of the reactor (with short residence times) are more accessible to medium than others (with long residence times). From this standpoint, the reactor can be divided into three regions: a captive volume, which consists of medium primarily delivered via diffusion; a lapped volume, which provides nutrients through unilateral convection; and a swept volume, which operates through bilateral convection. The relative sizes of these three volumes were modified experimentally by changing the period over which the direction of medium flow was reversed from 15 min (larger captive volume) to 9 h (larger swept volume). The results suggest that antibody concentration increases as the size of the diffusion-limited (captive) volume is increased to a maximum at around 30 min with a sharp decrease thereafter. As reflected by changes in measured consumption of glucose and production of lactate, no significant difference in cellular metabolism occurred as the reactor was moved between these different states. These results indicate that the mode of operation of the transtubular bioreactor may influence antibody productivity under serum-free, low-protein conditions with minimal effects on cellular metabolism.     

区域生态安全格局:设计原则与方法

区域生态安全格局概念的提出为区域生态恢复和生物保护提供了整体性对策。基于这一概念及其理论基础 ,提出了区域生态安全格局设计的初步原则和方法。通过对景观生态规划原则的增补 ,确定了区域生态安全格局的设计原则。根据区域生态环境问题和人类干扰的特点 ,综合集成了基于格局优化、干扰分析 2种规划途径和地理信息系统、空间模拟和预案研究等多种方法 ,形成了区域生态安全格局设计的方法框架。该方法突出体现了区域生态安全格局注重针对性、区域性、系统性和主动性的特点 ,满足了适应性生态系统管理的需求 ,为实现区域生态安全提供技术支持     

Estimating the size-selection curves of towed gears,traps, nets and hooks

A general statistical methodology for analysis of selectivity data from any type of fishing gear is presented. This formal statistical modelling of selectivity is built on explicit definitions of the selection process and specification of underlying assumptions and limitations, and this gives the resulting estimates of gear selectivity (and possibly fishing power) a clear interpretation. Application of the methodology to studies using subsampled catch data and to towed gears having windows or grids is outlined, and examples applied to passive nets and towed gears are presented. The analysis of data from replicate deployments is covered in detail, with particular regard to modelling the fixed and random effects of between-haul variation. Recent developments on the design of selectivity experiments are introduced and demonstrated.     

农地整理中路沟渠生态化设计研究进展

农地整理是人类对农田生态系统的强烈干扰.传统农地整理中路沟渠设计的主要技术经济指标是方便生产通行和调配水资源以及提高水的利用率,尚缺少系统化、生态化考量,已经影响了农田物种扩散,导致生物栖息地环境的退化,并对农田生态系统的生物竞争机制、缓冲和补偿能力、抵御虫害能力产生了显著的负效应.本文系统总结了路沟渠生态化的功能,着重介绍了近年来生态化路沟渠的形式与结构设计、路沟渠生态工法参数的试验以及路沟渠生态材料选择的技术和方法.未来的相关研究应根据各级道路、灌排沟渠的不同功能要求和规格,按照田间不同生物的习性和栖息要求,精确设计能够满足大多数生物需求的农田路沟渠形态、结构和控制性尺寸,切实解决农地整理带来的上述问题.     

葡激酶N端突变体的分子设计,高效表达与分离纯化

同源模建人微小纤溶酶原（ｍｉｃｒｏｐｌａｓｍｉｎｏｇｅｎ ,ｍｐｌｇ）的三维结构,建立葡激酶（ｓｔａｐｈｙｌｏｋｉｎａｓｅ,Ｓａｋ）,ｍｐｌｇ计算机分子对接的结构模型,模型与已有的实验结果基本相符。为研究葡激酶Ｎ端结构与功能的关系,进一步改造葡激酶分子,根据复合物结构模型设计了Ｎ端缺失１５个氨基酸的葡激酶突变体。     

Evaluation of AlphaFold2 structures as docking targets

AlphaFold2 is a promising new tool for researchers to predict protein structures and generate high-quality models, with low backbone and global root-mean-square deviation (RMSD) when compared with experimental structures. However, it is unclear if the structures predicted by AlphaFold2 will be valuable targets of docking. To address this question, we redocked ligands in the PDBbind datasets against the experimental co-crystallized receptor structures and against the AlphaFold2 structures using AutoDock-GPU. We find that the quality measure provided during structure prediction is not a good predictor of docking performance, despite accurately reflecting the quality of the alpha carbon alignment with experimental structures. Removing low-confidence regions of the predicted structure and making side chains flexible improves the docking outcomes. Overall, despite high-quality prediction of backbone conformation, fine structural details limit the naive application of AlphaFold2 models as docking targets.