Anthoceros angustus Steph. is rich in phenolic acids such as rosmarinic acid (RA). Phenylalanine ammonia-lyase (PAL) is an entry enzyme in the phenylpropanoid pathway of plants, playing an important role in the biosynthesis of RA. To investigate the important role of PAL in rosmarinic acid synthesis, two PAL genes (designated as AanPAL1 and AanPAL2) were cloned from A. angustus, encoding 755 and 753 amino acid residues, respectively. The AanPAL deduced amino acid sequences contain the conserved domains of PAL and the core active amino acid residues Ala-Ser-Gly. The phylogenetic analysis indicated that AanPAL1 and AanPAL2 were clustered with PALs from bryophytes and ferns and had the shortest evolutionary distance with the PALs from Physcomitrella patens. Quantitative real-time PCR results showed that the expression of AanPAL1 and AanPAL2 was induced by exogenous methyl jasmonate (MeJA). HPLC results showed that the MeJA treatment significantly increased the accumulation of RA. AanPAL1 and AanPAL2 were expressed in Escherichia coli and purified by histidine-tag affinity chromatography. The recombinant proteins catalyzed the conversion of L-phenylalanine to generate trans-cinnamic acid with high efficiency, with the best performance at 50 ℃ and pH 8.0. The K_m and k_cat of AanPAL1 were 0.062 mmol/L and 4.35 s^-1, and those of AanPAL2 were 0.198 mmol/L and 14.48 s^-1, respectively. The specific activities of AanPAL1 and AanPAL2 were 2.61 U/mg and 8.76 U/mg, respectively. The two enzymes had relatively poor thermostability but good pH stability. The high activity of AanPAL2 was further confirmed via whole-cell catalysis with recombinant E. coli, which could convert 1 g/L L-phenylalanine into trans-cinnamic acid with a yield of 100% within 10 h. These results give insights into the regulatory role of AanPAL in the biosynthesis of RA in A. angustus and provide candidate enzymes for the biosynthesis of cinnamic acid.
Phenylalanine Ammonia-Lyase/metabolism* ; Cloning, Molecular ; Cinnamates/metabolism* ; Recombinant Proteins/metabolism* ; Rosmarinic Acid ; Depsides/metabolism* ; Escherichia coli/metabolism* ; Amino Acid Sequence ; Plant Proteins/metabolism* ; Phylogeny ; Acetates/pharmacology* ; Cyclopentanes ; Oxylipins

Objective:To construct an evidence-based practice program for dietary management of patients with non-dialysis chronic kidney disease (CKD) based on best evidence and to evaluate the effectiveness of its application.Methods:The best evidence for dietary management of non-dialysis CKD patients was summarized. From September to October 2022, following the evidence clinical transformation model of the Fudan University Centre for Evidence-based Nursing, the best evidence was screened and evidence-based practice program were developed, taking into account patients' wishes, expert opinions, and clinical contexts. From November 2022 through March 2023, baseline reviews, analysis of barriers and facilitators were implemented. Between April 2023 and April 2024, evidence-based practice was carried out in the Department of Nephrology of the Second Affiliated Hospital of Guangzhou Medical University to compare the implementation rate of review indicators at the system, practitioner, and patient levels, and practitioners' knowledge before and after the application of evidence.Results:A total of 14 review indicators were developed. The implementation rate of the 12 review indicators and the practitioners' knowledge of the CKD diet were elevated after the evidence-based practice ( P<0.05) . Conclusions:Evidence-based practice program for dietary management of patients with non-dialysis CKD has a positive effect on improving practitioners' knowledge of non-dialysis CKD diets, implementation rate of dietary management behaviors, and patients' dietary behaviors.

Objective:To construct an evidence-based practice program for dietary management of patients with non-dialysis chronic kidney disease (CKD) based on best evidence and to evaluate the effectiveness of its application.Methods:The best evidence for dietary management of non-dialysis CKD patients was summarized. From September to October 2022, following the evidence clinical transformation model of the Fudan University Centre for Evidence-based Nursing, the best evidence was screened and evidence-based practice program were developed, taking into account patients' wishes, expert opinions, and clinical contexts. From November 2022 through March 2023, baseline reviews, analysis of barriers and facilitators were implemented. Between April 2023 and April 2024, evidence-based practice was carried out in the Department of Nephrology of the Second Affiliated Hospital of Guangzhou Medical University to compare the implementation rate of review indicators at the system, practitioner, and patient levels, and practitioners' knowledge before and after the application of evidence.Results:A total of 14 review indicators were developed. The implementation rate of the 12 review indicators and the practitioners' knowledge of the CKD diet were elevated after the evidence-based practice ( P<0.05) . Conclusions:Evidence-based practice program for dietary management of patients with non-dialysis CKD has a positive effect on improving practitioners' knowledge of non-dialysis CKD diets, implementation rate of dietary management behaviors, and patients' dietary behaviors.

Objective To explore the efficacy and safety of lung recruitment maneuver(LRM)on postoperative hypoxemia in patients with acute type A aortic dissection(ATAAD).Methods A total of 56 ATAAD patients with postoperative hypoxemia in the First Affiliated Hospital of Guangxi Medical University from November 2019 to May 2022 were selected and randomly divided into LRM group(n=36)and conventional treatment group(n=20).Patients in conventional treatment group received routine mechanical ventilation on the basis of lung protective ventilation.The patients in LRM group were treated with incremental positive end expiratory pressure(PEEP).Arterial blood gas analysis,respiratory parameters,hemodynamics parameters and serum interleukin(IL)-6 and IL-10 levels were compared between two groups before and after treatment.Results At 12h and 24h after treatment,arterial partial pressure of oxygen(PaO2),oxygenation index(OI),static compliance(Cstat)and dynamic compliance(Cdyn)in two groups were significantly higher than before treatment,the alveolar-arterial gradient of oxygen[PO2(A-a)],respiratory index(RI),peak inspiratory pressure(Ppeak)and plateau pressure(Pplat)were significantly lower than before treatment(P<0.05).PaO2,OI,Cstat and Cdyn in LRM group were significantly higher than those in conventional treatment group,PO2(A-a),RI,Ppeak and Pplat were significantly lower than those in conventional treatment group(P<0.05).Systolic blood pressure and mean arterial pressure decreased and central venous pressure increased during pulmonary reexpansion in LRM group(P<0.05),and all patients returned to baseline level after pulmonary reexpansion.At 12h after treatment,serum IL-6 and IL-10 levels in both groups were significantly lower than before treatment(P<0.05).Conclusion Incremental PEEP can improve oxygenation and lung compliance in patients with hypoxemia after ATAAD surgery,but it has transient effects on hemodynamics,and should be closely monitored during treatment.

OBJECTIVE To investigate the effect of polymorphisms of solute carrier organic anion transporter family, member 1B3(SLCO1B3) gene on the pharmacodynamics of mycophenolate mofetil(MMF) in patients with lupus nephritis. METHODS Patients with lupus nephritis who were treated in Jieyang People’s Hospital from September 2019 to April 2021 were selected. All subjects were treated with MMF for at least 12 months, or discontinued due to poor efficacy. The efficacy of MMF was evaluated. The SLCO1B3 334T>G/699G>A(rs4149117/rs7311358) genotype was detected using Agena MassARRAY®, and the correlation between gene polymorphisms and MMF pharmacodynamics was analyzed using SPSS 25.0 software. RESULTS The genotype frequencies of SLCO1B3 334T>G/699G>A were in Hardy-Weinberg equilibrium. The probability of poor MMF treatment effect of 334GG/699AA carriers was significantly higher than that of 334TT/699AA and 334TG/699GA carriers(P<0.001); Logistic regression showed that both 334GG/699AA and urine protein>2.5 g·(24 h)−1 were the risk factors for poor MMF treatment[OR=4.038(1.731, 9.420), P<0.001; OR=4.157(1.705, 10.137), P=0.002]. Combined analysis showed that patients with both 334GG/699AA genotype and urine protein>2.5 g·(24 h)−1 were at higher risk for poor efficacy[OR=8.563(3.301, 22.216), P<0.001]. CONCLUSION SLCO1B3 334T>G/699G>A is related to the efficacy of MMF treating lupus nephritis, and 334GG/699AA carriers are more likely to result in poor efficacy.

Objective:To summarize the evidence of diet management in patients with chronic kidney disease (CKD) in stage 3-5 D, so as to provide a reference for clinical nursing.Methods:After establishing evidence-based nursing questions, according to the "6S evidence model", the evidence on diet management of patients with CKD in stage 3-5 D was searched by computer, including computer decision support system evidence, guidelines, best evidence summary and systematic review. The search time limit was from the establishment of the database to September 2020. The guideline quality assessment was independently completed by two research nurses and a doctor of nephrology. The quality assessment of non-guideline article was independently completed by two research nurses, and an evidence-based instructor participated in decision-making. The evidence was extracted, evaluated and graded using the Joanna Briggs Institute (JBI) 2014 version of the intervention research evidence pre-grading system.Results:A total of 19 articles that met the requirements were included, including 5 guidelines, 8 systematic reviews, 4 randomized controlled trials, 1 clinical decision, and 1 national standard. A total of 22 pieces of the best evidence on diet management of patients with CKD in stage 3-5 D were summarized from three aspects, namely, diet/nutrition assessment, diet management, teamwork and education.Conclusions:The best evidence of diet management in patients with CKD in stage 3-5 D provides a certain reference for clinical practice, so as to improve the quality of diet management in patients with CKD and the clinical outcomes of patients.

Objective: To investigate hand hygiene of children in kindergartens in Huzhou City, so as to provide basis for improving hand hygiene and conduct health education on hand hygiene related diseases. Methods: A total of 343 children in 6 kindergartens in two districts of Huzhou City were observed by stratified cluster random sampling and observation. Results: A total of 1 042 hand hygiene indications and 886 hand washing (85.03%). The overall hand-washing qualification rate was 53.35%. Within different kindergartens, children in the lowest level kindergartens had poor hand washing habits. There was a positive correlation between levels of kindergarten and children’s hand washing habits. Boys’ hand washing habits were relatively poor, 45.35 percent of boys’ had substandard hand washing habits, which was only 19.88 percent of girl. The hand hygiene behavior of children in primary class was better than that of middle class. The proportion of substandard hand washing of children in primary class and middle class was 23.13% and 39.80% respectively. Conclusion There is a big promotion space of hand hygiene habits of children in kindergartens, so it is necessary to strengthen compliance with hand hygiene and cultivate correct hand hygiene habits.

Objective: To establish an evaluation index system for health education of hand,foot and mouth disease（HFMD）in nursery children by Delphi method. Methods: After referring to the relevant literature,an initial health education index system for HFMD of nursery children was established,including four first-level indicators,twelve second-level indicators and forty-six third-level indicators. Two rounds of expert consultation were conducted according to Delphi method. The enthusiasm of experts was evaluated by response rate,and the authority of experts was evaluated by authority and variation coefficient. The consultation questionnaire in the second round was based on the results of the first round and was scored again in the same way. Then the evaluation index system of HFMD health education for nursery children was finally determined. Results: Thirteen experts participated in two rounds of consultation,including four aged 40-49 years and nine aged 50-59 years;two of deputy senior title and eleven of senior title;one worked in the health administration department,six in the CDC,five in health education institutions and one in kindergarten. The two rounds of expert consultation were carried out effectively within the time set and the response rate reached 100%. In the first round,the judgment coefficient,familiarity coefficient and authority coefficient were 0.92±0.06,0.85±0.12 and 0.88±0.06,respectively. One first-level indicator,two second-level indicators and twelve third-level indicators were added,seven third-level indicators were deleted,and the contents of one second-level indicator and ten third-level indicators were improved in the first round. In the second round,the judgment coefficient,familiarity coefficient and authority coefficient were 0.95±0.07,0.88±0.10 and 0.91±0.06,respectively. The coordination coefficients of the first-,second- and third-level indicators in the second round were 0.170,0.166 and 0.283,respectively,and the coefficients of variation were all less than 0.25. After two rounds of discussion,five first-level indicators,fourteen second-level indicators and fifty-two third-level indicators were finally established as the evaluation index system of HFMD health education for nursery children. Conclusion The evaluation index system of HFMD health education for nursery children established by Delphi method has high authority and practicability,and it can be used to comprehensively evaluate the effects of HFMD health education on nursery children.

Objective: To understand knowledge and awareness of hand-foot-mouth disease (HFMD) and associated factors among parents of kindergarten children in urban district of Huzhou City and to provide a veference for making effective measure of health education of HFMD. Methods: Self-designed questionnaire was used to investigate 851 parents from 6 kindergartens by stratified cluster random sampling. Results: The overall recognition of HFMD was 8.15±3.43, and the qualified rate was 5.17%. Awareness rates of sources, transmission routes, symptoms and signs, as well as preventive measures were 34.08%, 20.80%, 3.41% and 30.32%, respectively. Multiple linear regression analysis showed that kindergarten type(B=-1.07), gender(B=0.70), age(B=-0.41), education level(B=1.60), occupation (B=-1.37) associated with awareness of HFMD (P<0.05).The top three sources of HFMD prevention and treatment were mobile messages(45.24%), kindergarten lectures (43.24%) and brochure/propaganda column(40.19%) . Conclusion The general knowledge and awareness of HFMD among parents of kindergartens children’s parents is low in urban district of Huzhou city. In order to improve the awareness and health-related behaviors among parents of kindergarten children to prevent HFMD, child care institutions and basic public health service health education programs should be relied on, to carry out appropriate health communication and intervention.

In order to construct recombinant baculovirus carrying Schistosoma japonicum 26 ku glutathione S-transferase gene (Sj26), and observe the expression of Sj26 in mammalian cells, the Sj26 gene was amplified with plasmid pGEX-3X as template by PCR, and then recombined into T vector for sequencing. Sj26 gene was inserted into the downstream of CMV promoter of donor plasmid pFBDGC, and the recombinant donor plasmid pFBDGC-Sj26 transformed into DH10Bac, then the recombinant bacmid AcCMVSj26 was isolated and transfected into Sf9 cells. The recombinant baculovirus was harvested and final titer of vAcCMVSj26 was measured. BHK cells were transducted with recombinant baculovirus in vitro. By using Western blot, the expression of 26 ku glutathione S-transferase (GST) was detected. The results showed that after enzyme digestion and sequencing, the donor plasmid was successfully constructed. PCR confirmed that pFBDGC-Sj26 and Bacmid homologous recombination occurred in E. coli. After transfection of Sf9 cells with recombinant Bacmid, recombinant baculovirus was replicated in Sf9 cells and expressed green fluorescent protein. PCR further revealed recombinant baculovirus contained Sj26. The titer of the harvested baculovirus was 1.24 x 10(8). Western blot demonstrated that recombinant baculovirus could express 26 ku GST in BHK cells. It was concluded that Sj26 recombinant baculovirus was successfully constructed, and the 26 ku GST was expressed in mammalian cells.