Objective To extract hyperoside from the leaves of Acanthopanax sessiliflorus by complex enzyme method, and optimize the extraction process by orthogonal experiment. Methods Hyperoside was determined by HPLC. Effects of temperature,α-amylase, neutral protease and cellulase on extraction rate were detected by the orthogonal tests, and the optimum extraction condition of hyperoside from the leaves of Acanthopanax sessiliflorus was determined by complex enzyme method. Results The main influence factor was temperature,follows byα-amylase, neutral protease and cellulase according to orthogonal analysis.The best condition was as follows: dose of cellulase, neutral protease and α-amylase was 2%, 0. 5% and 3%, respectively, extract at temperature of 30 ℃for 10 min. Under this condition, the extraction rate of hyperoside in the leaves of Acanthopanax senticosus was 0.52%. Conclusion As compared with the traditional technics, compound enzyme increases the productivity of hyperoside.

This study was aimed to optimize the extraction process of double-marker components for Arctium lappa L. The central composite design and response surface methodology was used. According to 3 main factors, the extraction rates of arctiin and arctigenin was used as evaluation indexes. Multiple linear regression and two-order polynomial equation were used. The binomial fitting model was performed in the optimization of arctiin and arctigenin extraction technology. The results showed that the indentified optimized extraction technology of arctiin and arctigenin was 70% ethanol, 24-fold, ultrasonic solvent extraction for 15 minutes. It was concluded that this technology was able to extract large amount of arctiin and arctigenin, which provided experiment evidences for arctiin and arctigenin preparation. It also provided references for the development and utilization of arctiin and arctigenin.

AIM: Terminalia chebula contained hydrolysable tannins up to about 35%. It was necessary to establish a chromatographic fingerprint to meet the quality control need effectively. METHODS: HPLC method was carried out with 3 kinds of the mobile phase , namely, A∶ 0.05mol?L -1 Phosphoric acid/ 0.05mol?L -1 Potassium dihydrogen phosphorate aqueous solution, B: methanol and C: Ethyl acetate, running in gradient mode based on the previous experiment. RESULTS: A marked peaks of HPLC fingerprint of the raw material, the extracts and its final product consisted of gallic acid, terchebulin, chebulamin, chebulagic acid and chebulinic acid. CONCLUSION: The fact has depicted that chromatographic fingerprint is a powerful tool for in-process-quality supervisory control and dynamic analysis of the active constituents during manufacture procedure of immature fruit products of Terminalia chebula.

Rhizopus oryzae lipase (ROL) is not only a biocatalyst used in a broad range of biotechnological fields, but also a model to investigate the function of intramolecular chaperone in the post-translational processing of lipase. In this study, we cloned and expressed the mature lipase gene (m-ROL) containing the pre-sequence (pro-ROL) of R. oryzae HU3005 in Pichia pastoris GS115 and characterized their enzymatic activities. m-ROL exhibited higher hydrolysis activity towards middle-chain substrates (C10 and C12) at pH 9.0, whereas pro-ROL preferred short-chain substrates (C4) and displayed maximal activity at pH 8.0. Moreover, pro-ROL possessed better thermal stability than m-ROL. This enzymatic discrepancy between m-ROL and p-ROL may be due to the pre-sequence that affects the folding and conformation of the mature lipase domain. To improve the expression level of m-ROL in P. pastoris, overlap extension PCR was conducted to substitute eight less-frequently used codons of m-ROL with frequently used codons. After methanol-induced expression for 72 h, the activity and protein content of the codon optimized m-ROL reached 132.7 U/mL and 50.4 mg/L, while the activity of the parental m-ROL and pro-ROL are 28.7 U/mL and 14.4 mg/L, 29.6 U/mL and 14.1 mg/L, respectively.
Codon ; Enzyme Precursors ; biosynthesis ; chemistry ; genetics ; Enzyme Stability ; Lipase ; biosynthesis ; genetics ; metabolism ; Pichia ; enzymology ; genetics ; Protein Engineering ; methods ; Protein Folding ; Recombinant Proteins ; biosynthesis ; genetics ; Rhizopus ; enzymology ; genetics ; Substrate Specificity

Aspergillus niger lipases are important biocatalysis widely used in industries for food processing and pharmaceutical preparation. High-level expression recombinants can lead to cost effective lipase large scale production. Full length gene synthesis is an efficient measure to enhance the expression level of the gene. In order to reduce the non-specific binding between oligonucleotides and bases mutation caused by the complicate secondary structure of DNA and excessive PCR amplification, a frequently phenomenon in one-step gene synthesis, we used a two-step method including assembly PCR (A-PCR) and digestion-ligation step to synthesis Aspergillus niger lipase gene lipA. Assisted by DNA2.0 and Gene2Oliga software, we optimized the codon usage and secondary structure of RNA and induced enzyme sites Cla I (237 site) and Pst I (475 site) into the gene. In the first step, fragments F1 (237 bp), F2 (238 bp) and F3 (422 bp) were separately synthesized by assembly PCR. In the second step, fragments F1, F2 and F3 were separately digested by Cla I and Pst I, and then ligated into a full length lipA gene. Two-step method efficiently enhanced successful ratio for full-length gene synthesis and dispersed the risk for gene redesign. The synthesized gene was cloned into pPIC9K vector and transferred into Pichia pastoris. After methanol inducement, the expression level of the codon optimized lipA-syn gene reached 176.0 U/mL, 10.8-fold of the original lipA gene (16.3 U/mL) in Pichia pastoris GS1115. The recombinant offers the possibility for lipase large-scale production.
Base Sequence ; Carboxylic Ester Hydrolases ; biosynthesis ; genetics ; Cloning, Molecular ; Genes, Synthetic ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

At present, interdisciplinary integration has become a major feature of the development of science and technology, and multi-disciplinary integration will gradually become the norm. Professional and technological multi-disciplinary integration has unpredictable potential, which will produce new disciplinary frontiers, new fields of science and technology and new patterns of innovation. Organ donation is a new discipline in China's new era. Constructing and promoting an organ donation disciplinary system with the overall goal of safeguarding legal and reasonable rights and interests of donors and their families and the health rights and interests of the recipients are in line with the fundamental requirements of maintaining high-quality development of organ donation and transplantation in China. Meantime, organ donation is a complex medical and social behavior, and organs donated by citizens belong to national resources, which also endows organ donation with a social welfare attribute and relevance with all parties in society. In this article, the essence of current problems encountered during organ donation in China, the whole process of organ donation and theoretical knowledge, professional skills and personnel support required by donors in different clinical stages were analyzed to illustrate the necessity and feasibility of establishing an organ donation disciplinary system based on multi-disciplinary integration. Besides, how to integrate organ donation disciplinary construction into the national policy was also investigated. Taking safeguarding the rights and interests of donors, family members and recipients as the core and taking organ donation and transplantation as the main line, cooperative principles of co-creation, co-construction, mutual promotion, sharing and win-win should be upheld, aiming to promote multi-disciplinary integration and comprehensive talent cultivation of organ donation, jointly enhance the recognition rate and donation rate of organ donation, and make organ donation widely recognized by citizens from all walks of life.

Objective:To construct a sensitive index system of nursing quality for patients with acute ischemic stroke patients undergoing interventional thrombectomy, so as to provide a scientific basis for interventional thrombectomy care for acute ischemic stroke.Methods:Taking the "structure-process-result" three-dimensional quality model as the theoretical framework, through evidence-based literature search, the item pool was drawn up, and finally the indicators and their weights at each level were determined by two rounds of Delphi method in December 2021 and January 2022 and analytic hierarchy process.Results:The positive coefficients of the two rounds of inquiries to experts were 0.92 and 1.00, respectively, the authority coefficients were 0.913 and 0.917, and the Kendall coefficients were 0.141 and 0.202, respectively. The final index system consisted of 3 primary indicators, 9 secondary indicators and 50 three-level indicators.Conclusions:The nursing sensitive quality index system for patients with acute stroke patients undergoing interventional thrombectomy constructed in this study has high reliability and scientificity, which can provide target incentives for clinical nurses and provide reference for the development of specialized nursing sensitive quality indicators.

Objective At present, the novel coronavirus pneumonia (COVID-19) pandemic is still raging in certain regions around the globe, and the prevention and control of the pandemic should be strengthened. Under the challenges of respective social environment and allocation of medical resources, and support from the inertia and inherent productivity of the system on which the industry depends, extensive attempts are being delivered to push forward the work of organ donation and transplantation in each country. Under the guidance of national experts and committee members, Shanxi Provincial Human Organ Procurement and Allocation Service Center was established on August 28, 2018 approved by the former Shanxi Provincial Health and Family Planning Commission. It is the only independent non-profit medical institution in Shanxi Province. In this article, the system construction of citizen's organ donation and transplantation fitting national and provincial conditions was further explored according to the data analysis of organ donation and transplantation in the United States and Spain during the COVID-19 pandemic combined with the implementation of organ donation work in Shanxi Provincial Human Organ Procurement and Allocation Service Center.

Objective To investigate the application of Rotarex mechanical thrombectomy system in lower extremity arterial thrombosis and to evaluate its clinical efficacy.Methods The clinical data of 61 patients(71 limbs,35 cases in acute phase,21 cases in subacute phase and 5 cases in chronic phase)with lower extremity arterial thrombosis treated with Rotarex were analyzed retro-spectively.Distal protective device was used in patients with poor distal artery outflow.High pressure saline was used during the pro-cedure and stent was used in patients with flow-limiting dissection.Catheter aspiration was performed in patients with distal artery embo-lization.Anticoagulant therapy was used in patients with thromboembolism and dual antiplatelet therapy was used in patients with in-situ thrombosis.Postoperative follow-up was performed with color Doppler ultrasound or computed tomography angiography(CTA)at 1 month,3 months and 6 months.Results Fifty-nine cases were treated with 6F Rotarex catheters and 2 cases were treated with 8F Rotarex catheters.Distal protective device was used in 10 cases,balloon dilation was performed in 49 cases and stent was used in 5 cases.Catheter aspiration was performed in 10 cases.Vessel rupture occurred in 4 cases,among whom 3 cases were successfully treated with the method of balloon compression and 1 case was treated with covered stent.Severe adverse events occurred in 4 cases and perioperative toe amputation was performed in 7 cases.Follow-up time was 3 to 6 months(mean 4.9 months).Lower extremity ischemic event occurred in 1 case at 6th month follow-up and was treated with stent.No other lower extremity ischemic events occurred in the course of follow-up.Conclusion For the treatment of lower extremity arterial thrombosis,Rotarex mechanical thrombectomy sys-tem has the advantages of minimally invasion,rapid and high efficiency.Combined with the therapy of catheter aspiration and stent place-ment,vascular patency can be further maintained and the lower extremity ischemic symptoms can be quickly relieved.

Objective:To discuss the method of modified balloon-occluded retrograde transvenous obliteration (M-BRTO) combined with antegrade transvenous obliteration (ATO) using tissue adhesive and (or) coils in the treatment of portal hypertension with spontaneous portosystemic shunt (SPSS), and to evaluate its clinical efficacy.Methods:From February 2018 to October 2022,clinical data of patients with portal hypertension with SPSS treatment in Henan Anyang District Hospital were retrospectively analyzed. A total of 21 patients were enrolled. Under the blood flow limit of SPSS outflow tract, ATO was firstly performed, then followed by M-BRTO. The ATO route could be performed from percutaneous transhepatic portal vein, percutaneous transumbilical vein or transjugular intrahepatic portal vein shunt (TIPS) approach and the M-BRTO route could be performed from femoral vein (FV), jugular vein (JV) or anterior cubical vein (ACV). The operation of M-BRTO+ATO was performed under local anesthesia and was suitable for patients with isolated gastric varicose bleeding, hepatic encephalopathy or cardiac insufficiency. TIPS combination with M-BRTO+ATO was performed under general anesthesia and was suitable for patients with gastrointestinal hemorrhage complicated with severe gastrorenal or splenorenal shunt, or with portal thrombosis. Abdominal plain CT scan was performed 1 week later to show the deposition of embolic agent. Abdominal color ultrasound was done 1 month later, contrast-enhanced CT scan was performed 3 months and 6 months later, and then color ultrasound or contrast-enhanced CT was performed every 6 months to show the portal vein blood flow or the patency of TIPS stent. Hepatic artery chemoembolization was performed 1 month later for patients with liver cancer.Results:A total of 23 times of operation were performed in 21 patients, including 1 case with 3 times of operation. The approach of percutaneous transhepatic route was used in 11 cases (7 cases combined with FV, 3 cases combined with JV and 1 case combined with ACV), the approach of TIPS route combination with FV was used in 9 cases, paraumbilical vein combination with FV was used in 2 cases and paraumbilical vein combination with ACV was used in 1 case. Ectopic embolization occurred in 3 cases (1 case to the spleen vein, 2 cases to the liver). Perioperative fever occurred in 5 cases, bleeding of hepatic puncture tract occurred in 1 case, and death happened in 2 cases (1 case of acute liver failure induced by bile duct stone, 1 case of acute heart failure combined with acute gastrointestinal massive hemorrhage). During the follow-up, 4 cases died (3 cases of liver cancer and 1 case of infection). The remaining 15 patients were followed up for 2 to 47 (22±13) months and there was no recurrence of hepatic encephalopathy and gastrointestinal hemorrhage during follow-up.Conclusions:The operation of M-BRTO+ATO using tissue adhesive or combining with coils as embolic agent can quickly obliterate outflow tract of SPSS and completely block the whole tract of SPSS, so it is a fast, safety and effective method for the treatment of PH with SPSS.