Cattle Mammary Bioreactor Generated by a Novel Procedure of Transgenic Cloning for Large-Scale Production of Functional Human Lactoferrin

Large-scale production of biopharmaceuticals by current bioreactor techniques is limited by low transgenic efficiency and low expression of foreign proteins. In general, a bacterial artificial chromosome (BAC) harboring most regulatory elements is capable of overcoming the limitations, but transferring BAC into donor cells is difficult. We describe here the use of cattle mammary bioreactor to produce functional recombinant human lactoferrin (rhLF) by a novel procedure of transgenic cloning, which employs microinjection to generate transgenic somatic cells as donor cells. Bovine fibroblast cells were co-microinjected for the first time with a 150-kb BAC carrying the human lactoferrin gene and a marker gene. The resulting transfection efficiency of up to 15.79×10−2 percent was notably higher than that of electroporation and lipofection. Following somatic cell nuclear transfer, we obtained two transgenic cows that secreted rhLF at high levels, 2.5 g/l and 3.4 g/l, respectively. The rhLF had a similar pattern of glycosylation and proteolytic susceptibility as the natural human counterpart. Biochemical analysis revealed that the iron-binding and releasing properties of rhLF were identical to that of native hLF. Importantly, an antibacterial experiment further demonstrated that rhLF was functional. Our results indicate that co-microinjection with a BAC and a marker gene into donor cells for somatic cell cloning indeed improves transgenic efficiency. Moreover, the cattle mammary bioreactors generated with this novel procedure produce functional rhLF on an industrial scale

Current prescription status of antihypertensive drugs in Chinese patients with hypertension: analysis by type of comorbidities

In 2020, the National Center for Cardiovascular Diseases with National Committee on Hypertension Management in Primary Health Care in China issued revised national clinical practice guidelines on the management of hypertension in primary health care based on the 2018 Chinese guidelines for the management of hypertension. To evaluate adherence to the guidelines, this retrospective study assessed the real-world status of antihypertensive drug prescribing for Chinese patients with hypertension, classified by comorbidity: coronary heart disease, diabetes mellitus, heart failure, stroke, and renal disease. About 1088212 hypertensive patients who received their first prescription for antihypertensive therapy between January 2021 to June 2021, were obtained from a database of Hangzhou Kang Sheng Health Consulting CO., Ltd. Calcium channel blockers (CCBs) and angiotensin II receptor blockers (ARBs) were the most common drugs prescribed for each comorbidity subgroup. Whereas diabetes mellitus or renal disease is a compelling indication for use of renin–angiotensin system inhibitors, CCBs were often administered in these subgroups. The treatment pattern for patients with coronary heart disease was closely similar to that for the overall patient population. Beta-blockers (BBs) were prescribed more frequently for patients with heart failure than for those with other comorbidities. Although antihypertensive drug prescription varied by comorbidity, pharmacological decisions were largely made under Chinese recommendations while physicians could select antihypertensive drugs based on the patients’ comorbidities. However, educational initiatives are still necessary to inspire clinicians to better familiarize themselves with the guidelines and manage hypertension comorbid diseases