元素衡算和代谢衡算在类人胶原蛋白表达期的应用研究

应用元素衡算和代谢衡算方法,建立用重组大肠杆菌发酵生产类人胶原蛋白表达期的数学模型,并利用非线性优化法对模型中的未知参数进行估算。结果表明该模型与重组大肠杆菌的生长、代谢动力学模型一致,且模型的关键计算参数αh、αp和mx分别为1.173 molo C-mol-1、293.814 molo C-mol-1和17.878 molo C-mol-1oh-1。该模型能较好地预测重组类人胶原蛋白表达期中的宏观反应速率,且在类人胶原蛋白发酵表达期,必须通过控制葡萄糖的补料速率来控制菌体的生长,当?=0.04 h-1时,类人胶原蛋白的比生成速率达最大值。     

玉米秸秆直接液化产物分析与热量衡算

由于反应条件相对温和,产物具有良好的环保性和可再生性,生物质直接液化是当前生物质能源化技术领域研究热点之一.本文进行了不同反应温度下玉米秸秆在水中直接液化制取生物燃油及相关产物的小型试验,并对其各组试验进行热量衡算.通过试验和衡算,获得了不同反应温度产物得率和直接液化反应热,表明直接液化反应终点温度为340℃.试验和衡算结果对反应过程研究有一定的意义,也可为该工艺的中试放大提供设计参考.     

年产一万吨香肠工厂物料衡算与主要设备选型优化设计

针对一万吨香肠工厂中的物料衡算与主要设备选型进行了研究和设计,主要研究了以下几个内容:物料、水、电、汽衡算,以及相关主要设备的选型。根据本工厂的生产需要,由物料衡算、热量衡算及日产量对水电气的使用量进行估算并确定主要设备规格类型,同时对水、电、汽的使用量进行了估算最大量为依次为200吨、5 000千瓦、50吨。     

如何正确使用杜南平衡公式计算离子的积累量

社南平衡是杜南河（F·G·Donnan）提出的一种说明离子积累现象的特殊平衡,即细胞内可扩散正离子和负离子浓度的乘积,等于细胞外正负离子浓度的乘积时的平衡。如何应用这个平衡公式计算离子积累量,在曹宗类和吴相任[1]合编的《植物生理学》一书中是这样演示解释权南平衡和说明细胞内离子积累量的：将适当pH的蛋白质溶液（蛋白质带负电荷）放在火棉胶袋中,呈阴离子状态的蛋白质可以与一定量的K 结合。当火棉胶袋浸在一KCl溶液中,开始时,袋内的K 浓度为Ci,CI-浓度为零,蛋白质浓度亦为Ci;外界溶液中,开始时K”的浓度为C。,CI…     

重组人血清白蛋白发酵过程生长期的代谢计算

结合元素平衡和代谢平衡,建立了重组人血清白蛋白发酵过程生长期的数学模型,并按单纯形多变量最优化方法估算了模型的未知参数。该模型能较好地描述重组人血清白蛋白发酵过程生长期中各宏观反应速率之间的关系,为重组人血清白蛋白宿主菌—Pichia pastoris的高密度培养提供了依据。     

基于参数优化的人工灌丛生态系统碳水通量模拟

荒漠草原是陆地生态系统中最为脆弱且受人类干扰较为严重的生态类型之一,精准模拟其碳水通量及对人为干扰的响应,不仅能够揭示其复杂的生态学过程,而且还可为人为生态修复和保护提供决策依据。生态模型能够有效地模拟陆地生态系统的碳水循环过程,但模型众多的参数及其取值的合理性限制了其普遍应用,故探索参数优化是提升生态模型应用的有效途径。利用PEST参数优化方法和涡度相关观测数据对Biome-BGC模型的生理生态参数进行优化,在评估参数优化效果的基础上模拟了1986-2018年宁夏盐池荒漠草原区人工灌丛生态系统的总初级生产力（Gross primary productivity,GPP）和蒸散（Evapotranspiration,ET）。结果表明：（1）参数优化可以改善Biome-BGC模型对荒漠草原区人工灌丛生态系统GPP和ET的模拟效果,参数优化后模拟的GPP和ET均更接近于观测值,其中月尺度的模拟效果更佳;（2）基于PEST的Biome-BGC模型参数优化方法具有较强的普适性,优化后的参数可推广应用于荒漠草原区人工灌丛生态系统长时间序列的GPP和ET模拟;（3）宁夏盐池荒漠草原区人工灌丛生态系统的GPP在1986-2018年呈缓慢上升趋势,增幅为1.47 g C m^-2 a^-1,但ET的年际变化率较大,且无显著变化趋势。     

运用公式求算两种遗传病的概率

遗传病概率计算是高中生物学教学的一个重点和难点。学生一般能熟练地进行一种遗传病概率的求算 ,而对两种遗传病概率的求算则感到十分困难。笔者在实际教学中 ,首先帮助学生总结归纳出求算两种遗传病概率的一般公式 ,然后应用公式解题 ,从而使计算简易准确 ,极大地调动了学生的学习积极性 ,收到了较好的教学效果。下面简述公式的归纳过程及其应用。1　公式的归纳1.1　首先研究一种遗传病的有关情况 ,下面以白化病遗传为例 :表 1亲本子代基因型及比例 子代表现型及比例肤色正常 ( A )白化 ( aa)AA×aa 1Aa 10Aa×Aa 1/ 4AA· 2 / 4Aa· …     

乙醇预处理麦秆的酶解性能研究简

采用H2 SO4催化和自催化乙醇法对麦秆进行预处理,比较预处理后麦秆的主要化学组成、纤维素酶解性能和半同步糖化发酵生产乙醇特性,并进行物料衡算。结果表明：H2 SO4催化和自催化乙醇预处理过程中纤维素固体回收率大于90％。添加非离子表面活性剂吐温20和吐温80没有显著提高H2 SO4催化乙醇预处理后纤维素的酶解葡萄糖得率及半同步糖化发酵过程中乙醇的产量,而对自催化乙醇处理后麦秆的酶解和半同步糖化发酵过程有一定程度的促进作用,相应的酶解葡聚糖转化率由72.7％提高到85.0％,而半同步糖化发酵过程中乙醇质量浓度提高了11.4％。物料衡算结果表明：酸催化和自催化乙醇预处理后葡聚糖回收率分别为91.0％和95.4％;半同步糖化发酵生产乙醇的得率分别为10.4和11.6 g（按100 g原料计）。     

代谢调衡饮治疗代谢综合征的临床疗效观察

目的:考察代谢调衡饮治疗代谢综合征的临床疗效。方法:将60例代谢综合征患者随机分为试验组和对照组,对照组口服卡托普利和二甲双胍,试验组在此基础上加服代谢调衡饮,疗程3个月,观察两组治疗前后体重指数、血压、血糖、血脂等指标变化。结果:试验组有效率明显高于对照组(P<0.05)。与对照组相比,试验组患者治疗后体重指数、血压、血糖、血脂指标改善更明显(P<0.05 or P<0.01)。结论:代谢调衡饮对代谢综合征患者具有减重降压,降糖调脂功效,是干预代谢调衡饮的有效方剂。     

利用城市垃圾发酵生产绿色木霉孢子*

研究利用城市生活垃圾发酵生产绿色木霉孢子的可行性,培养条件和发酵过程中的种群密度变化规律。结果表明,城市生活垃圾适宜作为生产木霉孢子的培养基质;合适的培养条件为垃圾培养基质中添加 ３０％（Ｗ／Ｗ）的麸皮,接种量 ３０％（ｖ／ｗ）,发酵 ８—１０ｄ,孢子密度可达 １０^９／ｇ以上。     

重组人血清白蛋白发酵过程表达期的定量研究

结合元素平衡和代谢平衡,建立了重组人血清白蛋白发酵过程表达期的数学模型,按单纯形多变量最优化方法估算了模型的未知参数,并探讨了导致表达期不同阶段产物形成速率存在差异的可能原因。该模型能较好地描述重组人血清白蛋白发酵过程表达期中各宏观反应速率之间的关系,为重组人血清白蛋白的高效表达提供了线索。     

Application of dynamic metabolic flux analysis for process modeling: Robust flux estimation with regularization,confidence bounds,and selection of elementary modes

In macroscopic dynamic models of fermentation processes, elementary modes (EM) derived from metabolic networks are often used to describe the reaction stoichiometry in a simplified manner and to build predictive models by parameterizing kinetic rate equations for the EM. In this procedure, the selection of a set of EM is a key step which is followed by an estimation of their reaction rates and of the associated confidence bounds. In this paper, we present a method for the computation of reaction rates of cellular reactions and EM as well as an algorithm for the selection of EM for process modeling. The method is based on the dynamic metabolic flux analysis (DMFA) proposed by Leighty and Antoniewicz (2011, Metab Eng, 13(6), 745–755) with additional constraints, regularization and analysis of uncertainty. Instead of using estimated uptake or secretion rates, concentration measurements are used directly to avoid an amplification of measurement errors by numerical differentiation. It is shown that the regularized DMFA for EM method is significantly more robust against measurement noise than methods using estimated rates. The confidence intervals for the estimated reaction rates are obtained by bootstrapping. For the selection of a set of EM for a given st oichiometric model, the DMFA for EM method is combined with a multiobjective genetic algorithm. The method is applied to real data from a CHO fed-batch process. From measurements of six fed-batch experiments, 10 EM were identified as the smallest subset of EM based upon which the data can be described sufficiently accurately by a dynamic model. The estimated EM reaction rates and their confidence intervals at different process conditions provide useful information for the kinetic modeling and subsequent process optimization.     

Development of a structured dynamic model for the production of polyhydroxybutyrate (PHB) in Azohydromonas lata cultures

The development of an accurate model representation of the fermentative production of polyhydroxybutyrate (PHB) is a prerequisite for the optimal operation and control of the microbial process. In the present work, a macroscopic model is developed to quantify the intracellular production of PHB in Azohydromonas lata bacteria. The proposed two-compartment structured model provides an accurate prediction of the metabolic and macroscopic phenomena occurring in A. lata bacterial cultures. Precisely, the proposed dynamic model accounts for biomass growth, polymer accumulation, carbon and nitrogen sources utilization and oxygen mass transfer rates. It is shown that the model can closely describe the time evolution of the bacterial culture via its direct comparison with experiments performed in flasks or/and a bioreactor. Moreover, it is shown that the model can be used as a simulation tool for process optimization and scaling-up studies of the PHB fermentative production in A. lata cultures.     

Optimization of an enrichment process for circulating tumor cells from the blood of head and neck cancer patients through depletion of normal cells

The optimization of a purely negative depletion, enrichment process for circulating tumor cells (CTCs) in the peripheral blood of head and neck cancer patients is presented. The enrichment process uses a red cell lysis step followed by immunomagnetic labeling, and subsequent depletion, of CD45 positive cells. A number of relevant variables are quantified, or attempted to be quantified, which control the performance of the enrichment process. Six different immunomagnetic labeling combinations were evaluated as well as the significant difference in performance with respect to the blood source: buffy coats purchased from the Red Cross, fresh, peripheral blood from normal donors, and fresh peripheral blood from human cancer patients. After optimization, the process is able to reduce the number of normal blood cells in a cancer patient's blood from 4.05 × 10⁹ to 8.04 × 10³ cells/mL and still recover, on average, 2.32 CTC per mL of blood. For all of the cancer patient blood samples tested in which CTC were detected (20 out of 26 patients) the average recovery of CTCs was 21.7 per mL of blood, with a range of 282 to 0.53 CTC. Since the initial number of CTC in a patient's blood is unknown, and most probably varies from patient to patient, the recovery of the CTC is unknown. However, spiking studies of a cancer cell line into normal blood, and subsequent enrichment using the optimized protocol indicated an average recovery of approximately 83%. Unlike a majority of other published studies, this study focused on quantifying as many factors as possible to facilitate both the optimization of the process as well as provide information for current and future performance comparisons. The authors are not aware any other reported study which has achieved the performance reported here (a 5.66 log₁₀) in a purely negative enrichment mode of operation. Such a mode of operation of an enrichment process provides significant flexibility in that it has no bias with respect to what attributes define a CTC; thereby allowing the researcher or clinician to use any maker they choose to define whether the final, enrich product contains CTCs or other cell type relevant to the specific question (i.e., does the CTC have predominately epithelial or mesenchymal characteristics?). Biotechnol. Bioeng. 2009;102: 521–534. © 2008 Wiley Periodicals, Inc.     

Constitutive basal and stimulated human small bowel contractility is enhanced in obesity

Background

Between 5 and 10% of the patients undergoing a colonoscopy cannot have a complete procedure mainly due to stenosing neoplastic lesion of rectum or distal colon. Nevertheless the elective surgical treatment concerning the stenosis is to be performed after the pre-operative assessment of the colonic segments upstream the cancer. The aim of this study is to illustrate our experience with the Computed Tomographic Colonography (CTC) for the pre-operative assessment of the entire colon in the patients with stenosing colorectal cancers.

Methods

From January 2005 till March 2009, we observed and treated surgically 43 patients with stenosing colorectal neoplastic lesions. All patients did not tolerate the pre-operative colonoscopy. For this reason they underwent a pre-operative CTC in order to have a complete assessment of the entire colon. All patients underwent a follow-up colonoscopy 3 months after the surgical treatment. The CTC results were compared with both macroscopic examination of the specimen and the follow-up coloscopy.

Results

The pre-operative CTC showed four synchronous lesions in four patients (9.3% of the cases). The macroscopic examination of the specimen revealed three small sessile polyps (3 - 4 mm in diameter) missed in the pre-operative assessment near the stenosing colorectal cancer. The follow-up colonoscopy showed four additional sessile polyps with a diameter between 3 - 11 mm in three patients. Our experience shows that CTC has a sensitivity of 83,7%.

Conclusion

In patients with stenosing colonic lesions, CTC allows to assess the entire colon pre-operatively avoiding the need of an intraoperative colonoscopy. More synchronous lesions are detected and treated at the time of the elective surgery for the stenosing cancer avoiding further surgery later on.     

Development of a Highly Sensitive and Specific Method for Detection of Circulating Tumor Cells Harboring Somatic Mutations in Non-Small-Cell Lung Cancer Patients

Background

Oncogenic mutations are powerful predictive biomarkers for molecularly targeted cancer therapies. For mutation detection patients have to undergo invasive tumor biopsies. Alternatively, archival samples are used which may no longer reflect the actual tumor status. Circulating tumor cells (CTC) could serve as an alternative platform to detect somatic mutations in cancer patients. We sought to develop a sensitive and specific assay to detect mutations in the EGFR gene in CTC from lung cancer patients.

Methods

We developed a novel assay based on real-time polymerase chain reaction (PCR) and melting curve analysis to detect activating EGFR mutations in blood cell fractions enriched in CTC. Non-small-cell lung cancer (NSCLC) was chosen as disease model with reportedly very low CTC counts. The assay was prospectively validated in samples from patients with EGFR-mutant and EGFR-wild type NSCLC treated within a randomized clinical trial. Sequential analyses were conducted to monitor CTC signals during therapy and correlate mutation detection in CTC with treatment outcome.

Results

Assay sensitivity was optimized to enable detection of a single EGFR-mutant CTC/mL peripheral blood. CTC were detected in pretreatment blood samples from all 8 EGFR-mutant lung cancer patients studied. Loss of EGFR-mutant CTC signals correlated with treatment response, and its reoccurrence preceded relapse.

Conclusions

Despite low abundance of CTC in NSCLC oncogenic mutations can be reproducibly detected by applying an unbiased CTC enrichment strategy and highly sensitive PCR and melting curve analysis. This strategy may enable non-invasive, specific biomarker diagnostics and monitoring in patients undergoing targeted cancer therapies.