Genetic and Epigenetic Variation across Genes Involved in Energy Metabolism and Mitochondria of Chinese Hamster Ovary Cell Lines

The increasingdemandfor biopharmaceutical products drives the search for efficient cell factories that are able to sustainably support rapid growth, high productivity, and product quality. As these depend on energy generation, here the genomic variation in nuclear genes associated with mitochondria and energy metabolism and the mitochondrial genome of 14 cell lines is investigated. The variants called enable reliable tracing of lineages. Unique sequence variations are observed in cell lines adapted to grow in protein‐free media, enriched in signaling pathways or mitogen‐activated protein kinase 3. High‐producing cell lines bear unique mutations in nicotinamide adenine dinucleotide (NADH) dehydrogenase (ND2 and ND4) and in peroxisomal acyl‐CoA synthetase (ACSL4), involved in lipid metabolism. As phenotypes are determined not only by functional mutations, but also by the exquisite regulation of expression patterns, it is not surprising that ≈50% of the genes investigated here are found to be differentially methylated and thus epigenetically controlled, enabling a clear distinction of high producers, and cells adapted to a minimal, glutamine (Gln)‐free medium. Similar pathways are enriched as those identified by genome variation. This strengthens the hypothesis that these phenomena act together to define cell behavior.     

PMab-219: A monoclonal antibody for the immunohistochemical analysis of horse podoplanin

Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, and bovine podoplanin (PDPN), a lymphatic endothelial cell marker, have been established in our previous studies. However, mAbs against horse PDPN (horPDPN), which are useful for immunohistochemical analysis, remain to be developed. In the present study, mice were immunized with horPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/horPDPN), and hybridomas producing mAbs against horPDPN were screened using flow cytometry. One of the mAbs, PMab-219 (IgG_2a, kappa), specifically detected CHO/horPDPN cells via flow cytometry and recognized horPDPN protein using Western blotting. Furthermore, PMab-219 strongly stained CHO/horPDPN via immunohistochemistry. These findings suggest that PMab-219 is useful for investigating the function of horPDPN.     

Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses

Podoplanin (PDPN) is known as a lymphatic endothelial cell marker. Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, bovine, pig, and horse PDPN have been established in our previous studies. However, mAbs against alpaca PDPN (aPDPN), required for immunohistochemical analysis, remain to be developed. In the present study, we employed the Cell-Based Immunization and Screening (CBIS) method for producing anti-aPDPN mAbs. We immunized mice with aPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/aPDPN), and hybridomas producing mAbs against aPDPN were screened using flow cytometry. One of the mAbs, PMab-225 (IgG_2b, kappa), specifically detected CHO/aPDPN cells via flow cytometry and recognized the aPDPN protein on Western blotting. Further, PMab-225 strongly stained lung type I alveolar cells, colon lymphatic endothelial cells, and kidney podocytes via immunohistochemistry. These findings demonstrate that PMab-225 antibody is useful to investigate the function of aPDPN via different techniques.     

Tiankengomelania guangxiense,gen. et sp. nov., a dark septate endophytic fungus,promotes the growth of the medicinal orchid Dendrobium officinale

Dendrobium officinale (Orchidaceae) is a traditional Chinese medicinal plant. Its growth is slow, however many dark septate endophytic fungi (DSEs) are considered useful to plant growth and as biocontrol agents against plant pathogens. The goals of this study were to identify a new DSE and evaluate its plant-growth promotion characteristics. Based on morphological and molecular phylogenetic evidence, a DSE fungal strain TK815 isolated from Dashiwei Tiankengs in Leye county Guangxi Province, China, was classified as a novel genus in the order Cheatothyriales, namely Tiankengomelania gen. nov. typified with T. guangxiense sp. nov. Tiankengomelania guangxiense TK815 can significantly promote the growth of D. officinale in stem length (11.25%), seedling height (16.97%), root length (10.34%), and dry weight (41.05%). This study discovered, described, and illustrated a new DSE fungus, and evaluated its biological function in contributing to the growth and production of the Chinese medicinal plant D. officinale.     

Higher insulin, triglycerides, and blood pressure with greater trunk fat in Tanner 1 Chinese

Objective: The aims of this study were to investigate the body fat distribution pattern in prepubertal Chinese children and to investigate the relationship between central fat distribution and specific biomarkers of cardiovascular disease. Research Methods and Procedures: The study was conducted in an urban Mainland Chinese (Jinan, Shandong) sample of children using a cross‐sectional design. Pubertal status was determined by Tanner criteria. Measurements included weight, height, waist circumference, DXA measures of total body fat and trunk fat; fasting serum measures of glucose, insulin, triglyceride, cholesterol, high‐density lipoprotein‐cholesterol; and systolic and diastolic blood pressure. Multiple regression models were developed with the biomarkers of cardiovascular risk factor as the dependent variables, and adjustments were made for significant covariates, including sex, age, height, weight, waist circumference, total body fat, trunk fat, and interactions. Results: A total of 247 healthy prepubertal subjects were studied. After co‐varying for age, weight, height, and extremity fat (the sum of arm fat and leg fat), girls had greater trunk fat than boys (p < 0.0001, R² for model = 0.95). Insulin and triglyceride were positively related to central fat measured by DXA‐trunk fat (p < 0.05) but not related to the waist circumference. In the blood pressure model, waist circumference was a significant predictor of both systolic blood pressure and diastolic blood pressure, while DXA‐trunk fat was associated with diastolic blood pressure only. Significant interactions between sex and trunk fat, and sex and total fat, were found in relation to diastolic blood pressure. Discussion: In prepubertal Chinese children, greater trunk fat was significantly associated with higher insulin and triglyceride in boys and girls and was associated with higher diastolic blood pressure in boys only.     

Waist circumference and BMI cut-off based on 10-year cardiovascular risk: evidence for "central pre-obesity"

Objective: The objective was to assess the waist circumference (WC) cut‐off point that best identifies a level of 10‐year cardiovascular disease (CVD) risk with optimal balance of sensitivity and specificity in Chinese subjects according to their predicted 10‐year CVD risk. Research Methods and Procedures: A community‐based cross‐sectional observational study involving 14,919 Hong Kong Chinese subjects. The 10‐year CVD risk based on various prediction models was calculated. The projected WC cut‐off points were then determined. Results: There were 4837 (32.4%) men and 10,082 (67.6%) women (mean age ± standard deviation, 47.3 ± 13.5 years; age range, 18 to 93 years; median age, 45.0 years). The mean optimal WC or BMI predicting a 15% to 30% 10‐year CVD risk were 83 to 88 cm and 25 kg/m² for men, and 76 cm and 23 kg/m² for women, respectively. With WC ≥90 cm in men and ≥80 cm in women, the likelihood ratio at various WC cut‐off points to develop a ≥20% 10‐year CVD risk is 1.5 to 2.0 in men and 3.0 in women. The likelihood ratio was 1.5 in men with WC at 84 cm and in women at 70 cm. Discussion: Our results agree with the present guidelines on the definition of general and central obesity in Asia‐Pacific regions. We propose the creation of an intermediate state of high WC, the “central pre‐obesity” for Chinese men with WC ≥84 to 90 cm (≥33 to 36 inches) and women with WC ≥74 to 80 cm (≥29 to 32 inches). People with central pre‐obesity, similar to those with overweight (BMI ≥23 to 25 kg/m²), already have an increased risk of co‐morbidities.     

难治性急性白血病研究现状与进展

难治性急性白血病治疗反应差,诱导缓解率低,复发率高,生存期短,因而是白血病治疗中的难题。本文从难治性急性白血病诊断标准与免疫学、细胞遗传学及分子生物学特征,多药耐药与难治性白血病,难治性急性白血病治疗现状以及治疗展望五大方面阐述了难治性急性白血病研究现状与进展。基础与初步临床研究显示中药配合化疗能够提高难治性急性白血病围化疗期临床缓解率。认为中药在提高难治性白血病临床疗效方面具有潜在临床应用前景和开发的商业价值,值得进行深入研究。     

骨折电流疗法的改良

目的:针对电流治疗骨折等疾病时存在的问题,提出相应的改进方法。方法:分析以往使用电流对骨折等疾病进行治疗时存在的电化学副作用、创口易于感染、重复施术不便等问题,提出相应的诸如使用溶液导电、隔离电解物、使电极与创口闭合生长等改进方法;并提出将电流治疗与药物治疗相结合的骨科疾病电流疗法改良方案。结果:将电流与药物相结合治疗骨折等疾病是很有可行性的治疗方法,本法不仅克服了已往疗法存在的一些问题,还为骨伤药物直接进入病患部位提供了有效的平台,使骨折、骨不连等疾病能够快速愈合。今后应加强研究骨折愈合各阶段所需的直接注射药物及其配比。完善电流—药物复合电极使用方法,以便与本法尽快进入临床实践。结论:将电流与药物相结合治疗骨折等疾病是很有可行性的治疗方法。     

福建省安溪县药用植物资源调查与分析

依托第四次全国中药资源普查,采用代表区域-样地-样方-样方套,结合样线调查方法,通过野外调查、走访调查及查阅相关资料等,对福建安溪药用植物资源种类和分布进行分析。结果显示,安溪县药用植物有1515种,隶属215科844属,草本为主要生活类型,药用部位主要以根及根茎类和全草类为主;其中福建重点药材115种、特色药材46种。安溪县药用植物资源丰富,物种多样性高,今后应加强资源保护与规划,合理开发利用该地药用植物资源。