Diterpenoid ferruginol is a key intermediate in biosynthesis of active ingredients such as tanshinone and carnosic acid.However, the traditional process of obtaining ferruginol from plants is often cumbersome and inefficient. In recent years, the increasingly developing gene editing technology has been gradually applied to the heterologous production of natural products, but the production of ferruginol in microbe is still very low, which has become an obstacle to the efficient biosynthesis of downstream chemicals, such as tanshinone. In this study, miltiradiene was produced by integrating the shortened diterpene synthase fusion protein,and the key genes in the MVA pathway were overexpressed to improve the yield of miltiradiene. Under the shake flask fermentation condition, the yield of miltiradiene reached about(113. 12±17. 4)mg·L~(-1). Subsequently, this study integrated the ferruginol synthase Sm CYP76AH1 and Sm CPR1 to reconstruct the ferruginol pathway and thereby realized the heterologous synthesis of ferruginol in Saccharomyces cerevisiae. The study selected the best ferruginol synthase(Il CYP76AH46) from different plants and optimized the expression of pathway genes through redox partner engineering to increase the yield of ferruginol. By increasing the copy number of diterpene synthase, CYP450, and CPR, the yield of ferruginol reached(370. 39± 21. 65) mg·L~(-1) in the shake flask, which was increased by 21. 57-fold compared with that when the initial ferruginol strain JMLT05 was used. Finally, 1 083. 51 mg·L~(-1) ferruginol was obtained by fed-batch fermentation, which is the highest yield of ferruginol from biosynthesis so far. This study provides not only research ideas for other metabolic engineering but also a platform for the construction of cell factories for downstream products.
Saccharomyces cerevisiae/genetics* ; Diterpenes/metabolism* ; Metabolic Engineering ; Fermentation ; Abietanes

The phase changes and quantity-quality transfer of 17 batches of Ostreae Concha(Ostrea rivularis) during the raw material-calcined decoction pieces-standard decoction process were analyzed. The content of calcium carbonate(CaCO_3), the main component, was determined by chemical titration, and the extract yield and transfer rate were calculated. The CaCO_3 content in the raw material, calcined decoction pieces, and standard decoction was 94.39%-98.80%, 95.03%-99.22%, and 84.58%-90.47%, respectively. The process of raw material to calcined decoction pieces showed the yield range of 96.85% to 98.55% and the CaCO_3 transfer rate range of 96.92% to 99.27%. The process of calcined decoction pieces to standard decoction showed the extract yield range of 2.86% to 5.48% and the CaCO_3 transfer rate range of 2.59% to 5.13%. The results of X-ray fluorescence(XRF) assay showed that the raw material, calcined decoction pieces, and standard decoction mainly contained Ca, Na, Mg, Si, Br, Cl, Al, Fe, Cr, Mn, and K. The chemometric results showed an increase in the relative content of Cr, Fe, and Si from raw material to calcined decoction pieces and an increase in the relative content of Mg, Al, Br, K, Cl, and Na from calcined decoction pieces to standard decoction. X-ray diffraction(XRD) was employed to establish XRD characteristic patterns of the raw material, calcined decoction pieces, and standard decoction. The XRD results showed that the main phase of all three was calcite, and no transformation of crystalline form or generation of new phase was observed. Fourier transform infrared spectroscopy(FTIR) was employed to establish the FTIR characteristic spectra of the raw material, calcined decoction pieces, and standard decoction. The FTIR results showed that the raw material had internal vibrations of O-H, C-H, C=O, C-O, and CO■ groups. Due to the loss of organic matter components after calcination, no information about the vibrations of C-H, C=O, and C-O groups was observed in the spectra of calcined decoction pieces and standard decoction. In summary, this study elucidated the quantity-quality transfer and phase changes in the raw material-calcined decoction pieces-standard decoction process by determining the CaCO_3 content, calculating the extract yield and transfer rate, and comparing the element changes, FTIR characteristic spectra, and XRD characteristic pattern. The results were reasonable and reliable, laying a foundation for the subsequent process research and quality control of the formula granules of calcined Ostreae Concha(O. rivularis Gould), and providing ideas and methods for the quality control of the whole process of raw material-decoction pieces-standard decoction-formula granules of Ostreae Concha and other testacean traditional Chinese medicine.
Drugs, Chinese Herbal/isolation & purification* ; Calcium Carbonate/analysis* ; Quality Control

OBJECTIVES: To evaluate the safety and efficacy of thiotepa (TT)-containing conditioning regimens for allogeneic hematopoietic stem cell transplantation (HSCT) in children with inborn errors of immunity (IEI). METHODS: Clinical data of 22 children with IEI who underwent HSCT were retrospectively reviewed. Survival after HSCT was estimated using the Kaplan-Meier method. RESULTS: Nine patients received a traditional conditioning regimen (fludarabine + busulfan + cyclophosphamide/etoposide) and underwent peripheral blood stem cell transplantation (PBSCT). Thirteen patients received a TT-containing modified conditioning regimen (TT + fludarabine + busulfan + cyclophosphamide), including seven PBSCT and six umbilical cord blood transplantation (UCBT) cases. Successful engraftment with complete donor chimerism was achieved in all patients. Acute graft-versus-host disease occurred in 12 patients (one with grade III and the remaining with grade I-II). Chronic graft-versus-host disease occurred in one patient. The incidence of EB viremia in UCBT patients was lower than that in PBSCT patients (P<0.05). Over a median follow-up of 36.0 months, one death occurred. The 3-year overall survival (OS) rate was 100% for the modified regimen and 88.9% ± 10.5% for the traditional regimen (P=0.229). When comparing transplantation types, the 3-year OS rates were 100% for UCBT and 93.8% ± 6.1% for PBSCT (P>0.05), and the 3-year event-free survival rates were 100% and 87.1% ± 8.6%, respectively (P>0.05). CONCLUSIONS TT-containing conditioning for allogeneic HSCT in children with IEI is safe and effective. Both UCBT and PBSCT may achieve high success rates.
Humans ; Retrospective Studies ; Transplantation Conditioning/methods* ; Thiotepa/therapeutic use* ; Hematopoietic Stem Cell Transplantation/adverse effects* ; Male ; Female ; Child, Preschool ; Infant ; Child ; Transplantation, Homologous ; Graft vs Host Disease ; Adolescent

OBJECTIVE: To evaluate the safety and effectiveness of robot-assisted percutaneous coronary intervention (R-PCI) compared to traditional manual percutaneous coronary intervention (M-PCI). METHODS: This prospective, multicenter, randomized controlled, non-inferior clinical trial enrolled patients with coronary heart disease who met the inclusion criteria and had indications for elective percutaneous coronary intervention. Participants were randomly assigned to either the R-PCI group or the M-PCI group. Primary endpoints were clinical and technical success rates. Clinical success was defined as visually estimated residual post-percutaneous coronary intervention stenosis < 30% with no 30-day major adverse cardiac events. Technical success in the R-PCI group was defined as successful completion of percutaneous coronary intervention using the ETcath200 robot-assisted system, without conversion to M-PCI in the event of a guidewire or balloon/stent catheter that was unable to cross the vessel or was poorly supported by the catheter. Secondary endpoints included total procedure time, percutaneous coronary intervention procedure time, fluoroscopy time, contrast volume, operator radiation exposure, air kerma, and dose-area product. RESULTS: The trial enrolled 152 patients (R-PCI: 73 patients, M-PCI: 79 patients). Lesions were predominantly B2/C type (73.6%). Both groups achieved 100% clinical success rate. No major adverse cardiac events occurred during the 30-day follow-up. The R-PCI group had a technical success rate of 100%. The R-PCI group had longer total procedure and fluoroscopy times, but lower operator radiation exposure. The percutaneous coronary intervention procedure time, contrast volume, air kerma, and dose-area product were similar between the two groups. CONCLUSIONS For certain complex lesions, performing percutaneous coronary intervention using the ETcath200 robot-assisted system is safe and effective and does not result in conversion to M-PCI.

Objective To construct a quality evaluation index system for emergency triage and form a quality evalua-tion table for application,it provides a reference basis for the supervision of the quality of emergency triage.Methods Based on the Donabedian structural model and referring to the latest released relevant policies and stan-dards,a Delphi method were used to determine the quality evaluation indicators of emergency triage,as well as the scores and evaluation methods of each level of indicators.The convenience sampling method was applied to seven tertiary hospitals in Jiangsu Province to test the reliability and validity of the emergency triage quality evaluation scale and to analyze the application.Results The effective recovery rate of the two rounds of expert correspondence ques-tionnaires was 100％,and the expert authority coefficients were 0.869 and 0.927,respectively,and the resulting emergency triage quality evaluation index system included 3 first-level indexes,12 second-level indexes,and 43 third-level indexes.The Cronbach coefficient of the emergency triage quality evaluation table was 0.719,and the average score of emergency triage quality of the seven hospitals was 93.36 points,which was at a good level.Conclusions The constructed emergency triage quality evaluation index system has good expert recognition,specialty and good reliability and validity,and the contents included are reasonable and clear with feasibility.

Objective To construct a quality evaluation index system for emergency triage and form a quality evalua-tion table for application,it provides a reference basis for the supervision of the quality of emergency triage.Methods Based on the Donabedian structural model and referring to the latest released relevant policies and stan-dards,a Delphi method were used to determine the quality evaluation indicators of emergency triage,as well as the scores and evaluation methods of each level of indicators.The convenience sampling method was applied to seven tertiary hospitals in Jiangsu Province to test the reliability and validity of the emergency triage quality evaluation scale and to analyze the application.Results The effective recovery rate of the two rounds of expert correspondence ques-tionnaires was 100％,and the expert authority coefficients were 0.869 and 0.927,respectively,and the resulting emergency triage quality evaluation index system included 3 first-level indexes,12 second-level indexes,and 43 third-level indexes.The Cronbach coefficient of the emergency triage quality evaluation table was 0.719,and the average score of emergency triage quality of the seven hospitals was 93.36 points,which was at a good level.Conclusions The constructed emergency triage quality evaluation index system has good expert recognition,specialty and good reliability and validity,and the contents included are reasonable and clear with feasibility.

Objective To construct a quality evaluation index system for emergency triage and form a quality evalua-tion table for application,it provides a reference basis for the supervision of the quality of emergency triage.Methods Based on the Donabedian structural model and referring to the latest released relevant policies and stan-dards,a Delphi method were used to determine the quality evaluation indicators of emergency triage,as well as the scores and evaluation methods of each level of indicators.The convenience sampling method was applied to seven tertiary hospitals in Jiangsu Province to test the reliability and validity of the emergency triage quality evaluation scale and to analyze the application.Results The effective recovery rate of the two rounds of expert correspondence ques-tionnaires was 100％,and the expert authority coefficients were 0.869 and 0.927,respectively,and the resulting emergency triage quality evaluation index system included 3 first-level indexes,12 second-level indexes,and 43 third-level indexes.The Cronbach coefficient of the emergency triage quality evaluation table was 0.719,and the average score of emergency triage quality of the seven hospitals was 93.36 points,which was at a good level.Conclusions The constructed emergency triage quality evaluation index system has good expert recognition,specialty and good reliability and validity,and the contents included are reasonable and clear with feasibility.

Objective To construct a quality evaluation index system for emergency triage and form a quality evalua-tion table for application,it provides a reference basis for the supervision of the quality of emergency triage.Methods Based on the Donabedian structural model and referring to the latest released relevant policies and stan-dards,a Delphi method were used to determine the quality evaluation indicators of emergency triage,as well as the scores and evaluation methods of each level of indicators.The convenience sampling method was applied to seven tertiary hospitals in Jiangsu Province to test the reliability and validity of the emergency triage quality evaluation scale and to analyze the application.Results The effective recovery rate of the two rounds of expert correspondence ques-tionnaires was 100％,and the expert authority coefficients were 0.869 and 0.927,respectively,and the resulting emergency triage quality evaluation index system included 3 first-level indexes,12 second-level indexes,and 43 third-level indexes.The Cronbach coefficient of the emergency triage quality evaluation table was 0.719,and the average score of emergency triage quality of the seven hospitals was 93.36 points,which was at a good level.Conclusions The constructed emergency triage quality evaluation index system has good expert recognition,specialty and good reliability and validity,and the contents included are reasonable and clear with feasibility.

Objective To construct a quality evaluation index system for emergency triage and form a quality evalua-tion table for application,it provides a reference basis for the supervision of the quality of emergency triage.Methods Based on the Donabedian structural model and referring to the latest released relevant policies and stan-dards,a Delphi method were used to determine the quality evaluation indicators of emergency triage,as well as the scores and evaluation methods of each level of indicators.The convenience sampling method was applied to seven tertiary hospitals in Jiangsu Province to test the reliability and validity of the emergency triage quality evaluation scale and to analyze the application.Results The effective recovery rate of the two rounds of expert correspondence ques-tionnaires was 100％,and the expert authority coefficients were 0.869 and 0.927,respectively,and the resulting emergency triage quality evaluation index system included 3 first-level indexes,12 second-level indexes,and 43 third-level indexes.The Cronbach coefficient of the emergency triage quality evaluation table was 0.719,and the average score of emergency triage quality of the seven hospitals was 93.36 points,which was at a good level.Conclusions The constructed emergency triage quality evaluation index system has good expert recognition,specialty and good reliability and validity,and the contents included are reasonable and clear with feasibility.