ObjectiveThis study aims to enhance of the farnesyl pyrophosphate（FPP） pool in Saccharomyces cerevisiae by heterologously expressing different farnesyl diphosphate synthases（FPSs） from various plants， thereby increasing the production of terpenoid compounds by the engineered yeast. MethodsRNA from mixed samples of roots， stems， and leaves of seven plants including Arabidopsis thaliana， Rosa rugosa， Artemisia annua， Centella asiatica， Humulus lupulus， Medicago sativa， and Panax ginseng was extracted by column chromatography and reverse transcribed into the first strand of complementary DNA（cDNA）， and based on the transcriptome data of the seven species of plants， sequence-specific primers were designed for CaFPS， RrFPS， MsFPS， HiFPS， PgFPS， AtFPS， and AaFPS， the full-length of the genes was cloned， and the genes were analyzed for bioinformatics in order to construct a pESC yeast shuttle vector. These seven plant-derived FPSs were further heterologously expressed in the previous constructed β-elemene-producing yeast， and the yield of β-elemene was indicated for their catalytic acivities. ResultsThe coding sequences of CaFPS， RrFPS， MsFPS， HiFPS， PgFPS， AtFPS， and AaFPS were all of 1 021 bp in length and encoding 301 amino acids， all of which were similarly related to the endogenous FPS-encoding gene（ERG20） in S. cerevisiae. After heterologous expression， RrFPS was identified as the most effective in catalyzing the synthesis of FPP from isopentenyl pyrophosphate（IPP） and dimethylallyl pyrophosphate（DMAPP）. Compared to the control strains， the RrFPS overexpressed yeast strains YB-1-Rr and YB-3-Rr increased the production of β-elemene by 231.25% and 189.3%， respectively. ConclusionBy comparing the functions of FPS-encoding genes from seven different plant sources， it is determined that the protein encoded by the RrFPS from R. rugosa has the best catalytic ability， which can provide key genetic elements for the construction of engineered yeast strain constructs with high terpenoid production.

ObjectiveTo analyze the "compatibility" relationship of sugars and glycosides and the heat-clearing and blood-cooling effect of the roots of four varieties of Rehmannia glutinosa and provide a basis for research on the pharmacodynamic material basis and quality control of R. glutinosa. MethodsThe content of sugars and glycosides in the roots of four varieties of R. glutinosa was determined during the growth period. The principal component analysis （PCA）， orthogonal partial least squares-discriminant analysis （OPLS-DA）， and the "compatibility" relationship of active components were employed to screen out the differential samples. A rat model of bleeding due to blood heat was used to verify the pharmacodynamic differences and the potential active components of differential samples. ResultsThe content and proportion characteristics of various components in roots of the four varieties of R. glutinosa during the expansion stage and the maturity stage had obvious differences. The proportion of phenylethanoid glycosides at the maturity stage was higher than that at the expansion stage. The R. glutinosa variety 85-5 had special quality characteristics among the tested varieties. All the samples alleviated the symptoms in the rat model. The effect of clearing heat and cooling blood was different between the maturity stage and the expansion stage， as well as between 85-5 samples at the maturity stage and other samples. The effect of clearing heat and cooling blood of R. glutinosa roots was the result of the combined action of multiple components in R. glutinosa roots and might be related to the high proportions of polysaccharides， iridoid glycosides， and phenylethanoid glycosides. ConclusionThe growth stage and variety affect the quality of R. glutinosa roots. The effect of clearing heat and cooling blood of R. glutinosa roots was related to the content and proportions of various components. The study can provide a basis for the basic research on the active components and quality control of R. glutinosa.

ObjectiveTo analyze the "compatibility" relationship of sugars and glycosides and the heat-clearing and blood-cooling effect of the roots of four varieties of Rehmannia glutinosa and provide a basis for research on the pharmacodynamic material basis and quality control of R. glutinosa. MethodsThe content of sugars and glycosides in the roots of four varieties of R. glutinosa was determined during the growth period. The principal component analysis （PCA）， orthogonal partial least squares-discriminant analysis （OPLS-DA）， and the "compatibility" relationship of active components were employed to screen out the differential samples. A rat model of bleeding due to blood heat was used to verify the pharmacodynamic differences and the potential active components of differential samples. ResultsThe content and proportion characteristics of various components in roots of the four varieties of R. glutinosa during the expansion stage and the maturity stage had obvious differences. The proportion of phenylethanoid glycosides at the maturity stage was higher than that at the expansion stage. The R. glutinosa variety 85-5 had special quality characteristics among the tested varieties. All the samples alleviated the symptoms in the rat model. The effect of clearing heat and cooling blood was different between the maturity stage and the expansion stage， as well as between 85-5 samples at the maturity stage and other samples. The effect of clearing heat and cooling blood of R. glutinosa roots was the result of the combined action of multiple components in R. glutinosa roots and might be related to the high proportions of polysaccharides， iridoid glycosides， and phenylethanoid glycosides. ConclusionThe growth stage and variety affect the quality of R. glutinosa roots. The effect of clearing heat and cooling blood of R. glutinosa roots was related to the content and proportions of various components. The study can provide a basis for the basic research on the active components and quality control of R. glutinosa.

Objective To investigate and analyze the risk factors of cardiovascular events by low density lipoprotein cholesterol (LDL-C). Methods A total of 430 patients with stable angina pectoris (SAP) in the hospital were included from June 2021 to June 2024 for retrospective analysis. According to whether acute myocardial infarction (AMI) occurred, the enrolled patients were divided into stable group (n=257) and deteriorating group (n=173). The general data were compared between groups, and the risk factors affecting AMI in SAP patients were analyzed. The predictive value of the above risk factors on predicting AMI in SAP patients was analyzed. Results Compared with the stable group, the levels of LDL-C, TG, LP-a and Hcy in the deteriorating group were higher (t=4.033, P<0.001; t=4.104, P<0.001; t=6.342, P<0.001; t=4.883, P<0.001) while the HDL-C level was lower (t=5.129, P<0.001). Multivariate logistic regression analysis suggested that the elevated levels of LDL-C, TG, LP-a and Hcy were the risk factors of AMI in SAP patients (P<0.05), and the elevated level of HDL-C was a protective factor (P<0.05). In the ROC curve, the area under the curve (AUC), sensitivity and specificity of combination of LDL-C, HDL-C, TG, LP-a and Hcy in predicting AMI in SAP patients were 0.777, 63.01% and 81.71%. Conclusion LDL-C is a risk factor of AMI in SAP patients. Combination of HDL-C, TG, LP-a and Hcy has certain value on predicting AMI in SAP patients.

ObjectiveTo investigate the alleviating effect of calcium cyanamide （CaCN₂） soil fumigation on replanting problems of Rehmannia glutinosa. MethodsNewly soil （NP） was used as the control group， while three treatment groups were established： replanted soil （RP）， newly soil treated with CaCN₂ （120 g·m²， tillage depth 25 cm） （NPCC）， and replanted soil treated with CaCN₂ （RPCC）. R. glutinosa was cultivated in all groups. At harvest， the tuber agronomic traits （number of enlarged roots， maximum root diameter， fresh weight， dry weight） were measured. The content of catalpol and rehmannioside D was quantified by ultra-high-performance liquid chromatography （UPLC） to evaluate medicinal quality. Rhizosphere soil available nutrients and enzyme activities were analyzed by assay kits. The community structure and composition of fungi and bacteria in rhizosphere soil were assessed via internal transcribed spacer 2 （ITS2） sequencing and 16S rDNA sequencing， respectively. ResultsCompared with NP， the RP group showed obviously reduced in tuber agronomic traits and quality indicators （P0.05）. However， the RPCC group showed significant improvement in agronomic traits and a notable increase in rehmannioside D content compared to RP （P0.05）. The contents of available phosphorus and potassium in RPCC and NP groups were obviously lower than those in RP （P0.05）. The polyphenol oxidase soil （S-PPO） activity in RP was obviously lower than in NP （P0.05）， while sucrose soil （S-SC）， acid phosphatase soil （S-ACP）， and S-PPO activities in RPCC were obviously higher than in RP （P0.05）. Microbial richness and diversity in RP were obviously higher than in NP （P0.05）， whereas no significant differences were observed between the RPCC and NP. The relative abundances of fungal genera Nectria， Myrothecium， Tomentella， and bacterial genus Skermanella were obviousl lower in RPCC and NP than in RP （P0.05）. Correlation analysis that S-ACP activity was positively correlated with the content of rehmannioside D （P0.05）. Fungal genera Engyodontium and Alternaria， and bacterial genera Pir4 lineage， Pirellula， Methyloversatilis， Brevundimonas， Ralstonia， and Acidibacter were obviously positively correlated with tuber dry weight （P0.05）. Conversely， fungal genera Pseudaleuria， Nectria， Haematonectria， Ceratobasidium， and bacterial genera Streptomyces， Skermanella， RB41， Gemmatimonas， and Bacillus were obviously negatively correlated with dry weight （P0.05）. The fungal genus Alternaria and bacterial genera Brevundimonas， Ralstonia， Acidibacter， and Dongia showed positive correlations with medicinal quality of R.glutinosa tuber， while fungal genera Pseudaleuria， Nectria， Stachybotrys， Fusarium， Gibberella， Ceratobasidium， and bacterial genera Sphingomonas， Skermanella， RB41， Gemmatimonas， and Bacillus were obviously negatively correlated （P0.05）. ConclusionCaCN₂ soil fumigation can significantly improve enzyme activities in replanted Rehmannia rhizosphere soil， enhance the utilization of available nutrients， reshape microbial community structure of replanted R.glutinosa at the family and genus level， and notably improve tuber agronomic traits and medicinal quality. This study provides a novel approach to alleviating replanting problems and offers insights for the integrated development of standardized cultivation techniques， including soil disinfection， nutrient-targeted regulation， and microbial inoculant application.

BACKGROUND:Clinical studies have found good analgesic effects of silver needle-thermal conduction therapy in patients with myofascial pain syndrome,but the exact mechanism remains unclear. OBJECTIVE:To observe the effect of silver needle-thermal conduction therapy on silent information regulator homolog 3(SIRT3)changes and mitochondrial ultrastructure in a rat model of myofascial pain syndrome. METHODS:Twenty rats were randomly selected from 26 Sprague-Dawley rats and were subjected to percussion combined with motor fatigue for replicating the rat model of myofascial pain syndrome.Sixteen rats that were successfully modeled were randomly divided into model group and silver needle-thermal conduction therapy group(treatment group),with eight rats in each group.The remaining rats were used as controls(normal group).The treatment group was treated with silver needle-thermal conduction therapy.Mechanical withdrawal threshold and thermal withdrawal latency of rats were measured at 1 day before modeling,1 day after modeling and 14 days after treatment.Electromyographic activities of the right medial femoral muscle were measured at 14 days after treatment.The right medial femoral muscle tissue was taken for hematoxylin-eosin staining to observe the local morphology and for transmission electron microscopy to observe the mitochondrial ultrastructure.Western blot assay was performed to detect SIRT3 expression. RESULTS AND CONCLUSION:Pain threshold:The mechanical withdrawal threshold and thermal withdrawal latency of the model and treatment groups were significantly decreased compared with those in the normal group and before modeling(P＜0.01).After treatment,the mechanical withdrawal threshold and thermal withdrawal latency of rats were significantly higher in the treatment group compared with the model group(P＜0.01).Electromyography:The rats in the model group showed spontaneous electrical activity in the right medial femur,while the rats in the treatment group showed reduced spontaneous electrical activity,longer time frame(P＜0.01)and lower wave amplitude(P＜0.05)compared with the model group.Hematoxylin-eosin staining:In the normal group,rat muscle fibers arranged closely and regularly.In the model group,the muscle fibers of rats were atrophied,degenerated,and disordered in arrangement.In the treatment group,rat muscle structure disorder improved.Mitochondrial microstructure:Under the transmission electron microscope,mitochondrial structure in the normal group was normal;mitochondrial swelling with broken or disappeared cristae appeared in the model group;mitochondrial swelling in the treatment group was obviously relieved or tended to be normal.SIRT3 expression:SIRT3 expression was significantly downregulated in the model group compared with the normal group,but was significantly upregulated in the treatment group compared with the model group(P＜0.05).To conclude,abnormalities in local muscle mitochondria and downregulation of SIRT3 expression suggest the presence of impaired energy metabolism in the rat model of myofascial pain syndrome.Mitochondrial changes recover and are close to normal after the silver needle-thermal conduction therapy,and the expression of SIRT3 is also upregulated close to the normal group,indicating the silver needle-thermal conduction therapy may play a therapeutic role by promoting mitochondrial repair and improving energy metabolism disorder.

Objective:To evaluate the value of the ROX index [blood oxygen saturation (SpO 2)/fraction of inspiration O 2 (FiO 2)/respiratory rate (RR)], ROX-heart rate (HR) index (ROX index/HR × 100), modified ROX (mROX) index [partial pressure of oxygen in the blood (PaO 2)/FiO 2/RR], and mROX-HR index (mROX index/HR × 100) in predicting prognosis for patients with acute respiratory distress syndrome (ARDS) treated with high-flow nasal cannula oxygen therapy (HFNC). Methods:The clinical data of 100 patients with ARDS who received HFNC between January 2018 and December 2022 at The Third People's Hospital of Hubei Province, Jianghan University, were retrospectively analyzed. These patients were divided into two groups based on whether HFNC treatment was successful or not: a success group with 65 patients and a failure group with 35 patients. The differences in the ROX index, ROX-HR index, mROX index, and mROX-HR index in the observation group were observed at the designated time points: 2, 12, and 24 hours after HFNC treatment. Receiver operating characteristic (ROC) curves were utilized to evaluate the value of ROX index, ROX-HR index, mROX index, and mROX-HR index in predicting the success or failure of HFNC treatment at 2, 12, and 24 hours. Cutoff values were determined.Results:There were no significant differences in age, gender, body mass index, Acute Physiology and Chronic Health Evaluation (APACHE II) score, Sequential Organ Failure Assessment score, or the proportions of underlying diseases and pulmonary causes between the success and failure groups (all P > 0.05). Furthermore, there were no significant differences in baseline HR, RR, FiO 2, SpO 2, partial pressure of carbon dioxide (PaCO 2), PaO 2, pH, lactate, oxygenation index, ROX index, mROX index, ROX-HR index, or mROX-HR index between the two groups (all P > 0.05). The ROX index in the success group at 2, 12, and 24 hours after HFNC treatment was 6.86 ± 1.09, 6.31 ± 1.61, and 8.24 ± 2.29, respectively. These values were significantly higher than those in the failure group (6.36 ± 0.67, 5.65 ± 1.44, and 5.41 ± 0.84) at the corresponding time points ( F = 5.97, 4.04, 49.40, all P < 0.05). At 2, 12, and 24 hours after HFNC treatment, the mROX index in the success group was 5.94 ± 1.28, 5.74 ± 1.23, and 8.51 ± 2.64, respectively. These values were significantly higher than those in the failure group (5.26 ± 0.74, 4.80 ± 0.97, 4.81 ± 1.17) at the corresponding time points ( F = 8.23, 15.38, 61.79, all P < 0.05). At 2, 12, and 24 hours after HFNC treatment, the ROX-HR index in the success group was 6.53 ± 1.32, 6.85 ± 1.44, and 7.57 ± 1.47, respectively. These values were significantly higher than those in the failure group (5.79 ± 1.04, 5.87 ± 1.03, 5.57 ± 0.63) at the corresponding time points ( F = 8.28, 12.61, 58.34, all P < 0.05). At 2, 12, and 24 hours after HFNC treatment, the mROX-HR index in the success group was 6.11 ± 1.30, 6.86 ± 1.13, and 7.79 ± 1.79, respectively. These values were significantly higher than those in the failure group (5.20 ± 1.06, 5.66 ± 1.46, 4.92 ± 0.90) at the corresponding time points ( F = 12.60, 20.87, 78.56, all P < 0.05). The receiver operating characteristic curve analysis revealed that the optimal thresholds were 6.56, 6.02, 6.24, and 5.25 for the ROX index, mROX index, ROX-HR index, and mROX-HR index, respectively. The area under the curve (AUC) values were 0.63, 0.66, 0.68, and 0.72, with sensitivity of 55.4%, 47.7%, 56.9%, and 76.9%, and specificity of 71.4%, 91.4%, 77.1%, and 62.9%, respectively. At 12 hours after treatment, the optimal thresholds were 6.09, 5.53, 6.52, and 5.99, with AUC values of 0.62, 0.70, 0.67, and 0.80, sensitivity of 55.4%, 53.8%, 61.5%, and 80.0%, and specificity of 74.3%, 77.1%, 71.4%, and 74.3%, respectively. At 24 hours after treatment, the optimal thresholds were 6.23, 6.4, 5.99, and 6.22, with AUC values of 0.88, 0.90, 0.91, and 0.93, sensitivity of 81.5%, 80.0%, 87.7%, and 83.1%, and specificity of 91.4%, 94.3%, 80.0%, and 91.4%, respectively. Conclusion:The use of the ROX index, mROX index, ROX-HR index, and mROX-HR index can aid in predicting the prognosis of ARDS patients. The predictive value of these indices increases as treatment time progresses. The mROX-HR index offers marked advantages during the initial stages of treatment and could serve as a reliable early predictor.

Objective:To confirm the transportation and evaporation efficiency of micro droplets sprayed by high-pressure nozzles from purified tritium-containing wastewater with carrier gas in the horizontal evaporation chamber.Methods:A scale test bench with a single high-pressure nozzle had been built based on design conditions and similarity criteria to explore the designed evaporation characteristics of generated micro droplets in the horizontal evaporation chamber. At the same time, combined with experimental data, a suitable evaporation model was constructed using the Discrete Phase Model (DPM) in Ansys Fluent. On this basis, further simulation and analysis were conducted on the evaporation process and efficiency under the condition of multiple nozzles in the desighed horizontal evaporation chamber.Results:For single nozzle tests, the particle size of micro droplets along the nozzle centerline was between 12-50 μm and the particle size distribution was similar to the Rosin-Rammler distribution. Besides, the relative light intensity decreased exponentially with distance, indicating that the particle size and concentration of micro droplets both decreased rapidly, which means the evaporation rate of micro droplets was rapid. For the simulation of multiple nozzles injection in the desighed horizontal evaporation chamber, even for the hardest design condition, the evaporation percentage reached up to 99%, and small amount of the escaping micro droplets continued to evaporate during the process of mixing with other process exhaust until complete evaporation in the vertical chimney section.Conclusions:Under the conditions of desighed sprayed particle size distribution, typical operating conditions and the incoming flow similar to the supposed area, the complete evaporation of the micro droplets can be basically achieved in the horizontal evaporation chamber, so as to ensure the complete evaporation of purified tritium-containing wastewater from the outlet of the chimney.

Objective:To evaluate clinical prognosis and prognostic factors of patients with early stage (Ⅰ stage) and locally advanced (Ⅱ/Ⅲ stage) lung cancer treated with carbon ion radiotherapy (CIRT).Methods:Clinical data, treatment, adverse reactions, survival and so on of 54 lung cancer patients who received CIRT and follow-up in the Heavy Ion Center of Wuwei Cancer Hospital of Gansu Province from March 2020 to September 2022 were retrospectively analyzed. The survival curve was plotted using Kaplan-Meier method. Difference tests were performed using log-rank test. Logistic regression analysis was used to identify prognostic factors.Results:According to inclusion and exclusion criteria, 54 patients were enrolled in the study, including 10 patients with early stage lung cancer and 44 patients with locally advanced lung cancer. The median follow-up time for 10 patients with early stage lung cancer was 11.0 (6.75, 17.25) months, and the median dose of irradiation was 60 Gy [relative biological effect (RBE)]. Upon the last follow-up, 3 patients had complete response (CR) and 3 patients had partial response (PR). Four patients had stable disease (SD) and no progressive disease (PD). The 1-year and 2-year local control rates (LCR), progression-free survival (PFS) rates and overall survival (OS) rates were 100%. During treatment and follow-up, 2 patients developed grade 1 radiation pneumonia, 1 case of grade 2 radiation pneumonia, 1 case of chest wall injury (chest wall pain), and there were no adverse reactions greater than grade 2. The median follow-up time of 44 patients with locally advanced stage was 12.5 (4.25, 21.75) months, and the median irradiation dose was 72 Gy (RBE). Thirty-two (73%) patients received concurrent chemotherapy during treatment, 20 (45%) patients received sequential chemotherapy after treatment, 14 (32%) patients received immune maintenance therapy and 3 (7%) patients obtained PD and received targeted drugs. Upon the last follow-up, 3 (7%) patients had CR, 17 (39%) patients had PR, 19 (43%) patients obtained SD, and 5 (11%) patients had PD. The 1-year and 2-year LCR were 96.0% and 87.3%, 90.9% and 84.1% for the 1-year and 2-year PFS rates, and 93.2% and 86.4% for the 1-year and 2-year OS rates, respectively. The median OS and PFS of patients were not reached. Multivariate logistic regression analysis showed that maintenance therapy after radiotherapy ( P=0.027) and clinical target volume (CTV) irradiation volume ( P=0.028) were the factors affecting PFS. Simultaneous chemoradiotherapy ( P=0.042) and maintenance therapy after radiotherapy ( P=0.020) were the factors affecting OS. And gross tumor volume (GTV) ≥215 ml ( P=0.068) might be an independent risk factor for grade 2 and above radiation pneumonia. Conclusions:The domestic carbon ion system has definite clinical effect and controllable toxic and side effects in the treatment of early stage and locally advanced lung cancer. The combination of synchronous chemotherapy and further maintenance treatment can significantly improve clinical prognosis of patients without significantly increasing the risk of toxic and side effects.

Objective To explore the influencing factors of over active bladder(OAB)in patients with cerebral small vessel disease(CSVD)and its correlation with CSVD imaging markers.Meth-ods A total of 163 elderly CSVD patients admitted in our hospital from January 2021 to Decem-ber 2022 were enrolled and divided into OAB group(37 cases)and non-OAB group(126 cases)based on the results of OAB rating scale.Mini-mental State Examination(MMSE)score,Fazekas scale score,and total CSVD burden score were recorded and compared between the two groups.Results The OAB group had older age,higher urinary frequency,larger proportions of nocturia,urgency,and urge incontinence ratio,increased Fazekas score,periventricular white matter hyper-intensity(PWMH)score and deep white matter hyperintensity(DWMH)score,and elevated total CSVD burden score and lower MMSE score than the non-OAB group(P＜0.05,P＜0.01).PWMH score and DWMH score were risk factors for the occurrence of OAB(P＜0.01).The OAB score was positively correlated with Fazekas score,PWMH score,and DWMH score in the CSVD patients(r=0.533,P=0.001;r=0.462,P=0.004;r=0.398,P=0.015).The occurrence of urgency urinary incontinence was positively correlated with Fazekas score and PWMH score in the CSVD patients(r=0.352,P=0.033;r=0.346,P=0.036).Conclusion PWMH and DWMH are risk factors for OAB occurrence in CSVD patients.