Sesquiterpenes are natural terpenes containing 15 carbon atoms. They are widely used in the perfume, pharmaceutical, and biofuel industries due to their remarkable biological activities. The traditional production of sesquiterpenes relies on chemical synthesis or plant extraction, which has the disadvantages of low yields and waste of resources. The construction of microbial cell factories for the efficient synthesis of sesquiterpenes by means of synthetic biology provides a new option. In recent years, with the development of metabolic engineering and synthetic biology, the heterologous synthesis of a variety of sesquiterpenes has been successfully achieved by metabolic engineering of the oleaginous yeast, Yarrowia lipolytica. In this paper, we review the research progress in the heterologous synthesis of different sesquiterpenes by Y. lipolytica, discuss the synthetic biology strategies commonly used in this field, and make an outlook on the research directions and engineering approaches to further enhance the sesquiterpene yield in this host. This paper provides a reference for strategies such as synergistic optimization of synthetic biology and metabolic engineering, enhanced precursors, and opens up new directions for the application of synthetic biology in green chemistry and sustainable production.
Yarrowia/genetics* ; Sesquiterpenes/metabolism* ; Metabolic Engineering/methods* ; Synthetic Biology/methods*

Objective:To investigate the effect of different timing of arterial -venous extracorporeal membrane oxygenation (VA-ECMO) on the prognosis of patients with acute myocardial infarction complicated with cardiogenic shock (AMICS).Methods:This study was a prospective cohort study. AMICS patients received VA-ECMO support primary percutaneous coronary intervention in Henan Provincial People's Hospital from May 2017 to July 2023 were divided into early VA-ECMO group and late VA-ECMO group. 64 AMICS patients who met the indications for VA-ECMO implantation, but did not revive VA-ECMO were included as control group. Demographic characteristics, coronary interventional (PCI) information and complications after VA-ECMO implantation were collected. The primary end points was 1-year survival, minor end point were in-hospital and perioperative death. Multivariate Logistic and Cox regression models were used to evaluate the effect of timing of VA-ECMO on prognosis of AMICS patients. Kaplan-Meier survival curve was used to analyze the 1-year survival outcome of the 3 groups.Results:A total of 143 AMICS patients were included, and materials of 136 patients entered in the final analysis, including 42 in the early VA-ECMO group, 34 in the late VA-ECMO group, and 60 in the non-VA-ECMO group. Compared with the late VA-ECMO group, the early VA-ECMO group had a higher ratio of PPCI after VA-ECMO, a longer D-to-B time, a shorter VA-ECMO support time, a higher success rate of VA-ECMO withdrawal, and a lower complication rate (all P<0.05). Compared with the early VA-ECMO group, the perioperative, in-hospital and 1-year mortality were significantly higher in Non-ECMO support (all P<0.05). There was no difference in perioperative and in-hospital mortality between the early VA-ECMO group and the late VA-ECMO group, but the 1-year mortality in the late VA-ECMO group was significantly higher ( P<0.05). Perioperative, in-hospital and 1-year mortality rates were lower in the late VA-ECMO group than in the no-VA-ECMO group, but the differences were not statistically significant. Multivariate Logistic and Cox regression analysis showed that after adjusting interference factors, early VA-ECMO was still a protective factor for in-hospital ( OR=0.244, P=0.015) and one year ( HR=0.308, P=0.001)mortality. Kaplan-Merier survival curve showed that compared with the late VA-ECMO group and the group without VA-ECMO, the early VA-ECMO group had the highest 1-year survival rate. Conclusion:Patients with AMICS may benefit more from early VA-ECMO than from late VA-ECMO support for PPCI.

To determine the efficacy and safety of selinexor combined with venetoclax (VEN) and azactitidine (AZA) for patients with relapsed and/or refractory acute myeloid leukemia (R/R AML) . Twelve patients with R/R AML treated with selinexor plus VEN and AZA in the Affiliated Cancer Hospital of Zhengzhou University from May 2022 to May 2023 were included. Their clinical data were retrospectively analyzed. Among the 12 R/R AML patients, 5 (41.7%) achieved complete remission (CR) , 1 (8.3%) achieved CR with incomplete hematological recovery, and 5 (41.7%) achieved partial remission. The median time to reach CR was 28 (16-59) days. The median PFS was 61 (15-300) days. The main adverse event of the regimen was hematological toxicity. No chemotherapy-related deaths were observed. The combination of selinexor plus VEN and AZA is an effective treatment for R/R AML patients.

Podophyllotoxin is an aryl naphthalene lignan-like secondary metabolite, which is widely existed in plants of the Podophyllum. It exhibits a broad spectrum of antitumor activity and holds significant clinical value as a medicinal natural product of plant origin. This review discusses podophyllotoxin from three aspects: plant distribution, biosynthesis and pharmacological effects. Distribution of podophyllotoxin in nature: Podophyllotoxin and its analogues are mainly found in 12 genera of three families, which are Berberidaceae, Linaceae and Cupressaceae, respectively. They are mostly distributed in East Asia and North America, especially in China, which is not only the diversity center of the Podophyllum, but also the distribution center. Among all these plants, the genera Dysosma and Sinopodophyllum are the focus of the relevant research. Biosynthesis of podophyllotoxin: In order to take the alleviation of the pressure of podophyllotoxin that is in short supply of natural resources, and to protect its existing resources, the biosynthesis of podophyllotoxin, especially synthetic biology, has gradually become a new direction of current research. Compared with chemical synthesis which has the shortcoming of low synthesis efficiency, high production cost and cumbersome steps, biosynthesis has a variety of advantages. It’s not limited by resources, easy to cultivate on a large scale and has fast production speed, low production cost and simple genetic manipulation, so there is a broader prospect for the exact purpose of biosynthetic development. The biosynthesis of podophyllotoxin is divided into four steps in general, including the synthesis of coniferyl alcohol, the synthesis of matairesinol, the synthesis of deoxypodophyllotoxin and the synthesis of podophyllotoxin. It requires various enzymes catalyzed during the synthesis process, such as pinoresinol-lariciresinol reductase(PLR), secoisolariciresinol dehydrogenase(SDH) and others. At present, by means of synthetic biology, heterologous production of natural drugs in microbial cell factories, the synthesis of the intermediate coniferyl alcohol has been achieved in Escherichia coli with an increased yield of (124.9±5.1)mg·L−1, and the concentration of (-)-pluviatolide was elevated to 137 mg·L−1. In addition, p-coumaric acid in Saccharomyces cerevisiae was obtained to 71.71 mg·L−1, and in a tobacco plant production chassis, highly purified (-)-deoxypodophyllotoxin was increased to 0.71 mg·g−1 (dry weight) after chromatographic separation. Overall, the utilization of biosynthesis can significantly increase the yield of podophyllotoxin and improve the research and production efficiency of anti-tumor drugs greatly, which brings a more effective strategy for the treatment of related diseases. The pharmacological activity of podophyllotoxin: Podophyllotoxin and its analogues have strong cytotoxicity and they exert good inhibitory effects on the proliferation of multifarious tumor cells, viruses and bacteria. So they are widely applied to the clinical treatment of various types of tumors, such as colorectal cancer, lung cancer, cervical cancer, hemangioma, breast cancer, esophageal cancer, etc. The molecular mechanism of its cancer treatment is mainly in the G1, G2/M to block the cell cycle to make drugs work. Meanwhile, podophyllotoxin has relatively significant toxic side effects and drawbacks that can’t be ignored as a drug with superior therapeutic effect on specific diseases, which tends to increase the risk of bone marrow suppression, chromosomal aberrations and so forth. In conclusion, this literature sums up the distribution of podophyll plants in China, making review of the biosynthetic steps, applications, and biological research of podophyllotoxin, and outlines the pharmacological activity of podophyllotoxin. It aims to set the foundation for the reasonable application of podophyllotoxin resources, the efficient synthesis of podophyllotoxin, and the development of Podophyllum drugs which are highly effective and low toxic. At the same time, this paper gives full play to the clinical medicinal value of podophyllotoxin, which has important influence and far-reaching significance in the treatment of malignant diseases, and promotes the modernization of traditional Chinese medicine.

Lactic acid bacteria(LAB)were isolated and cultivated from the intestinal contents of rabbits by MRS-CaCO3 solid medium.Identification was achieved through morphological observa-tion,Gram staining,physiological and biochemical characterisation,16S rDNA sequence analysis,and ERIC-PCR analysis.Strains displaying typical Lactobacilli characteristics were exanimated for their biological characteristics,resistance properties,adherence capacity in vitro,colonization abili-ty in vivo,and safety profile.In this study,a total of four strains of Lactobacillus reuteri were iso-lated from rabbits,all of which exhibited typical biological characteristics of LAB.These strains demonstrated inhibitory effects on common pathogenic bacteria in the gastrointestinal tract,with the primary inhibitory substance being bacteriocin.Furthermore,they showed sensitivity to chlor-amphenicol,rifampicin,and erythromycin,and displayed a degree of tolerance to gastrointestinal conditions and high temperature.These stains were capable of successful colonization in rabbits with a higher degree of safety.This study lays a foundation for the development of LAB prepara-tions for the prevention and treatment of rabbit intestinal diseases.

Objective:To observe the clinical efficacy of Dushen Decoction and Zhenwu Decoction in the treatment of chronic congestive heart failure (CCHF) of Yang deficiency and blood stasis type. Methods:From March 2017 to December 2019, 120 patients with CCHF of Yang-deficiency and blood stasis type admitted to Shanghai Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine were selected, and they were divided into study group and control group according to the randomized number table method, with 60 in each group. The control group was given western medicine of conventional treatment, and the study group was combined with Dushen Decoction and Zhenwu Decoction on the basis of the control group. Both groups were treated for 1 month. Before and after treatment, TCM syndrome scores were performed, serum TNF-α, IL-17 and CRP were detected by ELISA, left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVEDD) and left ventricular end systolic diameter (LVESD) were detected by color doppler echocardiography the adverse reactions during the treatment were recorded and the clinical efficacy was evaluated. Results:The total effective rate was 91.7% (55/60) in the study group and 71.7% (43/60) in the control group, with significant difference between the two groups ( χ2=8.015, P=0.005). After treatment, the scores of palpitation, dyspnea, chilly limbs, dull tongue and edema in the study group were significantly lower than those in the control group ( t=13.953, 13.915, 30.945, 32.339, 20.403, P<0.001). After treatment, LVEF [(56.28 ± 4.34)% vs. (42.47 ± 4.56)%, t=16.993] in the study group was significantly higher than that of the control group ( P<0.01); LVEDD [(44.32 ± 6.23) mm vs. (53.81 ± 5.19) mm, t=9.066] and LVESD [(31.28 ± 4.62) mm vs. (37.51 ± 4.73) mm, t=7.299] in the study group were significantly lower than those in the control group ( P<0.01). After treatment, the serum levels of TNF-α, IL-17 and CRP in the study group were significantly lower than those in the control group ( t=12.644, 15.975 and 14.379, P<0.001). The incidence of adverse reactions was 6.7% (4/60) in the control group and 8.3% (5/60) in the study group, and there was no significant difference between the two groups ( χ2=0.120, P=0.729). Conclusion:Dushen Decoction and Zhenwu Decoction in the treatment of CCHF can improve the clinical symptoms, improve the cardiac function, reduce inflammatory factors, improve the treatment efficiency with good safety.

Objective:To investigate the difference of metabolomics between acute heart failure (AHF) patients and control. To find and validate new metabolic biomarkers.Methods:This was a single-center case-control study which included 89 acute heart failure patients admitted to the emergency department of Henan Provincial People's Hospital from January 2018 to June 2019. Eighty people without heart failure and diastolic dysfunction were enrolled as control group whose age and sex were matched to the study group. The fasting blood samples were collected from femoral arterial. Qualitative and quantitative analyses of plasma metabolites were performed in 2 groups by high performance liquid chromatography tandem mass spectrometry (UHPLC-MS), Orthogonal partial least squares-discriminant analysis (OPLS-DA) model and ROC curve method were applied.Results:Compared with the control group, we found that AHF group had higher likelihood to groups with coronary heart disease (37% vs. 7%, P<0.001), hypertension (58% vs. 28%, P<0.001), diabetes (33% vs. 18%, P=0.033), atrial fibrillation (24% vs. 4%, P<0.001), smoking history (42% vs. 18%, P=0.001), and that AHF group had higher creatinine level [(121.6 ± 78.4) vs. (69.0 ± 21.0), P<0.001], higher urea level [(11.5 ± 7.6) vs. (6.2 ± 2.0), P<0.001], higher heart rate [(92 ± 23) vs. (78 ± 14), P<0.001], hypoproteinemia [(32.4 ± 5 .2) vs. (40.4 ± 2.2), P<0.001], and significantly increased BNP level [(4 200 ± 5 229) vs. (100 ± 68), P<0.001], lower left ventricular ejection fraction[(45 ± 8) vs. (57 ± 6), P<0.001], low serum sodium level ( P<0.001). The metabolites of AHF group were significantly different from those of the control group. The metabolites involved amino acids, fatty acids, lipids, nucleosides and their derivatives. Adenine, N-acetyl-L-glutamic, pseudouridine and Gamma-Glutamylcysteine had certain diagnostic value for AHF comparing to control. The AUC were 0.995, 0.932, 0.920 and 0.900. And the AUC value for BNP diagnosis of AHF is 0.978. Conclusions:There were significant differences in metabolism between AHF group and control group including multiple substances. Adenine, N-acetyl-L-glutamic, pseudouridine and Gamma-Glutamylcysteine has similar diagnostic value compared with BNP for diagnosing AHF.

Objective:To investigate the effect of hourly ambient temperature exposure on emergency calls in Xuchang city and Zhengzhou city.Methods:The hourly meteorological data, air pollution data and emergency calls of Xuchang city and Zhengzhou city were collected from January 1, 2017 to October 31, 2019. A distributed lag non-linear model was used to calculate the excess relative risk ( ERR). The lag effect and cumulative effect of extreme temperature exposure on emergency calls were evaluated. Results:The relationship between hourly temperature exposure and emergency calls was a U-shaped curve. In Xuchang city and Zhengzhou city, both low and high temperatures would increase the number of hourly emergency calls. The earliest effect of low temperature occurred at a lag of 22 h and 52 h, with ERR values (95% CI) about 0.20% (0.00%, 0.39%) and 0.11% (0.00%, 0.22%), respectively. The earliest effect of high temperature occurred at a lag of 0 h with ERR values about 1.59% (1.09%, 2.09%) and 1.45% (1.22%, 1.69%), respectively. High temperature had the greatest impact on the number of emergency calls of cardiovascular disease at a lag of 4-8 h. The cumulative ERR values (95% CI) of the two cities were 8.70% (4.98%, 12.75%) and 3.89% (2.61%, 5.22%), respectively. Conclusion:High temperature exposure could increase the number of emergency calls within a few hours, while the effect of low temperature would not occur until 22 hours later.

Objective:To investigate the effect of hourly ambient temperature exposure on emergency calls in Xuchang city and Zhengzhou city.Methods:The hourly meteorological data, air pollution data and emergency calls of Xuchang city and Zhengzhou city were collected from January 1, 2017 to October 31, 2019. A distributed lag non-linear model was used to calculate the excess relative risk ( ERR). The lag effect and cumulative effect of extreme temperature exposure on emergency calls were evaluated. Results:The relationship between hourly temperature exposure and emergency calls was a U-shaped curve. In Xuchang city and Zhengzhou city, both low and high temperatures would increase the number of hourly emergency calls. The earliest effect of low temperature occurred at a lag of 22 h and 52 h, with ERR values (95% CI) about 0.20% (0.00%, 0.39%) and 0.11% (0.00%, 0.22%), respectively. The earliest effect of high temperature occurred at a lag of 0 h with ERR values about 1.59% (1.09%, 2.09%) and 1.45% (1.22%, 1.69%), respectively. High temperature had the greatest impact on the number of emergency calls of cardiovascular disease at a lag of 4-8 h. The cumulative ERR values (95% CI) of the two cities were 8.70% (4.98%, 12.75%) and 3.89% (2.61%, 5.22%), respectively. Conclusion:High temperature exposure could increase the number of emergency calls within a few hours, while the effect of low temperature would not occur until 22 hours later.

Objective:To observe the changes of follistatin-like protein 1 (FSTL1) in serum of patients with proliferative diabetic retinopathy (PDR).Methods:Twenty PDR patients confirmed by clinical examination and 20 normal people were included in the study. Human retinal vascular endothelial cells (HRCEC) were divided into HRCEC blank control group, 3 h hypoxia group, 6 h hypoxia group. Human umbilical vein endothelial cell (HUVEC) were divided into HUVEC blank control group, 3h hypoxia group, 6h hypoxia group. Real-time quantitative PCR (RT-PCR) and ELISA were used to determine the expression of FSTL1, TGF-β, VEGF, connective tissue growth factor (CTGF) mRNA and protein in peripheral blood and cells of all groups from all subjects.Results:The expressions of FSTL1, TGF-β1, CTGF, VEGF mRNA in blood samples of patients with PDR were 1.79±0.58, 0.97±0.21, 1.85±0.69 and 1.38±0.44. The expressions of FSTL1, TGF-β1 protein were 1.19±0.50, 0.71±0.24 ng/ml and 734.03±116.45, 649.36±44.23 ng/L. Compared with normal people, the differences were statistically significant ( tmRNA=0.90, 0.21, 2.85, 1.77; P=0.00, 0.00, 0.04, 0.02. tprotein=1.88, 7.68; P=0.00, 0.02). The cell viability of HRCEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.66±0.05 and 0.64±0.04, respectively. Compared with the blank control group, the difference was statistically significant ( F=13.02, P=0.00). The cell viability of HUVEC cells in the 3 h hypoxia group and the 6 h hypoxia group were 0.63±0.06 and 0.68±0.06, respectively. Compared with the blank control group, the difference was statistically significant ( F=26.52, P=0.00). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HRCEC blank control group and 3 h hypoxia group, the differences were statistically significant ( F=14.75, 44.93, 85.54, 6.23; P=0.01, 0.00, 0.00, 0.03). Compared with the HRCEC blank control and 3 h hypoxia group, the expressions of FSTL1 and TGF-β1 protein were statistically significant ( P<0.05). There was a statistically significant difference in TGF-β1 protein expression in the hypoxic 6 h group ( P=0.03) and no significant difference in FSTL1 protein expression ( P=0.68). Comparison of FSTL1, TGF-β1, CTGF, and VEGF mRNA expression in HUVEC blank control group and 3h hypoxia group, the differences were statistically significant ( F=19.08, 25.12, 22.89, 13.07; P=0.00, 0.00, 0.00, 0.01). Immunofluorescence staining results showed that FSTL1, TGF-β1, CTGF, and VEGF proteins were positively expressed in cells in the 3h hypoxia and 6h hypoxia groups. Conclusion:The expression of FSTL1 gene and protein in serum of PDR patients was significantly higher than that of normal people.