Purpose To indicate the expression of nattokinase (NK) in Pichia pastoris , an emulsion was prepared with the purified NK to prepare polyclonal antibody. In order to establish sandwich enzyme-linked immunosorbent assay (ELISA) to assay NK in organism, furthermore to lay the foundation for researching in vivo metabolism and function of NK. Methods The NK gene was cloned into a Pichia pastoris expression vector pHBM905A to construct the recombinant plasmid pPRONK.The recipient cell of Pichia pastoris GS115 was transformed with pPRONK which had been cut by restriction enzyme Sal I , under the induction of methanol. The expressed production is purified by salting out and ultrafiltration membrane. An emulsion was prepared with the purified NK and injected into rabbits to prepare polyclonal antibody. Results NK was expressed and identified by SDS-PAGE.The molecular mass of expressed production is about 27 kD.The fibrin plate assay indicated that the NK protein can cleavage fibrin effectively. ELISA analysis indicated that the polyclonal antibody titer is about 1:8 000. Western blot demonstrated that there was a special strap nearby 27 kD. Conclusion NK was successfully expressed in Pichia pastoris , the production can cleavage fibrin effectively and it had great immunogenicity.

Objecfive To investigate possibihty of monitoring myocardial isehemia by eontinuous EASI 12-lead ST-segment trend analysis during orthopaedic surgery and find its rules.Methods Three hundred and eleven consecutive patients who underwent orthopaedic surgery were monitored bv continuous EASI 12-lead ST-segment analysis during surgery.Results Among 311 patients 81 patients(26.0%) had myocardial ischemic events.The ischemie incidence of general anesthetics WaS signifieantly higher than local anesthetics(the spinal or epidural anesthetics and nerve-block anesthetics)(38.1%vs 23.0%.P＝ 0.01 5).During ischemie events the patients whose heart rate exceeded 90 beat per minute was 71.6% (58/81).Among the 177 isehemie events,the duration ofless than 5 minutes was 79.7%(141/177).ST-seg- ment deviation ofless than 0.15 mV was 71.2%(126/177).Conclusions During orthopaedic surgery EASt 12-lead electrocardiogram is the optimal approach for monitoring myocardial isehemia.The ischemic incidence of general anesthetics is significantly higher than local anesthetics.The myocardial ischemia events correlates with fast heart rate.The intraoperative myocardial ischemia events are more often transient.In most of these ischemic events ST-segment deviation is not very serious.

Corneal confocal microscopy (CCM) is an in vivo corneal imaging technique, which can directly quantify corneal nerve fibers in real time. It has the characteristics of non-invasive, objective and high sensitivity. CCM can not only be used for the diagnosis and treatment evaluation of corneal diseases, but also plays an important role in the diagnosis and prognosis evaluation of some peripheral and central nervous system diseases, such as diabetes peripheral neuropathy and Parkinson's disease. In addition, the changes of corneal nerve fibers can indirectly reflect the severity of ischemic cerebrovascular disease, and it is expected to become a noninvasive bioimaging marker of ischemic cerebrovascular disease. This article reviews CCM and its application in ischemic cerebrovascular disease, in order to provide better means for early diagnosis and prognosis evaluation of ischemic cerebrovascular disease.

Corneal confocal microscopy (CCM) is a non-invasive, simple and rapid visual corneal imaging technique, which can directly conduct real-time collection and quantitative analysis of corneal nerve fibers. Studies have shown that CCM can be used in the diagnosis and prognosis evaluation of degenerative diseases, demyelinating diseases, degenerative diseases and other types of diseases of the central nervous system. In this paper, the recent advance in CCM in neurological diseases is summarized to provide new ideas for their diagnosis and prognosis evaluation.

Objective:To explore the current status of deep vein thrombosis (DVT) prevention and nursing in Department of Orthopedics of China hospitals, and provide a basis for hospitals and related departments to promote systematic DVT prevention and nursing management.Methods:This study was a cross-sectional study, and convenience sampling method was used. In September 2019, the self-designed Domestic Orthopedic Deep Vein Thrombosis Prevention and Nursing Questionnaire was used to investigate the members of the Orthopedic Nursing Professional Committee who signed up for the China Health Promotion Foundation Orthopedic Venous Thromboembolism (VTE) Prevention and Nursing Management Forum. The survey subject came from 125 hospitals in 29 provinces, municipalities and autonomous regions across the country. Respondents completed the electronic questionnaire survey by scanning the quick response code, and 118 valid questionnaires were finally recovered.Results:The proportion of 118 hospitals carrying out in-hospital DVT special training was 82.2% (97/118) . Among the ways for nurses to acquire knowledge, the exchanges outside the hospital, literature review, and international exchange accounted for 70.3% (83/118) , 60.2% (71/118) and 14.4% (17/118) respectively. The proportion of establishing hospital DVT risk early warning information system was 57.6% (68/118) . The hospital-level professional group formulated a complete DVT prevention and nursing system process accounting for 44.1% (52/118) , and the department-level professional group formulated a complete DVT prevention and nursing system process accounting for 36.4% (43/118) . In the comprehensive evaluation of DVT prevention and nursing management, 65.3% (77/118) of hospitals rated it as excellent, and 34.7% (41/118) of hospitals rated it as fair and lacking.Conclusions:The Department of Orthopedics of China hospitals attaches great importance to DVT prevention, but in the construction of DVT prevention and nursing management system for orthopedic patients, the popularization of DVT risk early warning information system, the application of DVT preventive equipment measures, the improvement of DVT prevention and nursing related systems, and the implementation of professional management specifications needs to be further strengthened.