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*

Autism spectrum disorder（ASD）is a multifactorial，pervasive neurodevelopmental disorder.As the morbidity rate of ASD in children increases year by year，feeding and eating behaviors，as an important and common clinical problem in children with ASD，are gaining more and more attention.Many children with ASD often have food selection issues，chewing problems，food allergy and related gastrointestinal symptoms，and even serious diseases such as eating disorders，which negatively impact on their growth and development.There are many factors affecting feeding and eating behavior problems in children with ASD，such as sensory processing，ritualistic eating behavior，gastrointestinal symptoms，age，and parenting pressure.There are also a variety of interventions that can help to improve feeding and eating behavior problems in children with ASD.Strengthening the understanding of these influencing factors and intervention treatment methods is beneficial for improving the quality of life in children with ASD.

Objective To investigate the molecular targets and signaling pathways involved in the therapeutic effects of Lishukang Capsule(LSK)in a rat model of high-altitude pulmonary edema(HAPE)using a proteomics-based approach.Methods A total of 60 male Wistar rats were randomly assigned to a control group,a HAPE model group,3 LSK treatment groups receiving low-,medium-,and high-dose LSK,respectively,and a Rhodiola rosea(also known as Hongjitian[HJT]in pinyin,a Chinese Romanization system)control group.After HAPE modeling,the pharmacodynamic effects were assessed and the optimal LSK dose was determined using HE stains,inflammatory cytokine quantification,lung tissue water content,and the protein concentration in bronchoalveolar lavage.Label free quantitative proteomic profiling was then applied to identify differentially expressed proteins(DEPs)in the optimal dose group,using a screening threshold of over 1.5-fold change and P<0.05.The selected DEPs were validated with Western blotting,followed by Gene Ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Results The medium-dose LSK group exhibited significant anti-HAPE effects.Findings from the proteomic analysis revealed,in the comparison with the control group,267 DEPs were identified in the HAPE group.In the comparison with the HAPE group,225 DEPs were identified in the medium-dose LSK group.A total of 112 DEPs in the control group were normalized following LSK treatment in the medium-dose LSK group.In addition,GO enrichment analysis of proteins differentially expressed between the HAPE and LSK group showed that these DEPs were mainly enriched in 12 biological processes,2 cellular components,and 5 molecular functions.KEGG pathway analysis showed that LSK activated pathways associated with cell adhesion molecules,glycosaminoglycan biosynthesis,DNA replication/nucleotide excision repair,transcriptional dysregulation in cancer,and Herpes simplex virus type 1(HSV-1)infection,while inhibiting pathways associated with glycerophospholipid metabolism.Some differentially expressed proteins with potential functions were verified by Western blotting,including AGPAT5,NCAM1,SRSF3,and PLA2.These differentially expressed proteins were significantly expressed in the normal group,HAPE group,and LSK group,and the validation results were consistent with proteomic findings,indicating the high reliability of the proteomic results.Conclusion LSK exerts a significant protective effect against HAPE.Proteomic analysis suggests that its therapeutic action may be mediated through activating pathways involved in cell adhesion molecules,glycosaminoglycan biosynthesis,DNA replication/nucleotide excision repair,transcriptional dysregulation in cancer,and HSV-1 infection,alongside inhibition of pathways associated with glycerophospholipid metabolism.The key DEPs identified in these pathways may play crucial roles in the preventive and therapeutic effects of LSK on HAPE.

Objective·To explore the predictive capability of retinal vascular features in preeclampsia(PE)based on artificial intelligence(AI)models.Methods·This retrospective study enrolled 789 pregnant women who registered from June 2020 to January 2021 at Shanghai First Maternity and Infant Hospital of Tongji University in the first 16 weeks of gestation.These women underwent regular prenatal examinations,had retinal fundus images captured,and delivered singleton live births at the hospital.According to whether they developed hypertensive disorders of pregnancy(HDP),they were divided into unaffected group(n=685)and HDP group(n=104).Within the HDP group,pregnancies were further categorized into gestational hypertension(GH)group(n=36)and PE group(n=68)based on the occurrence of PE.Based on the gestational age at onset,the PE group was further divided into early-onset PE group(gestational age<34 weeks)and late-onset PE group(gestational age≥34 weeks).Fundus images of the pregnant women were obtained,and an AI algorithm was utilized to diagnose retinal lesions and quantify retinal vascular features.Comparative analyses were conducted on fundus features and retinal vascular features.Univariate Logistic regression model was employed to analyze the influencing factors of PE occurrence,and multivariate Logistic regression model was further utilized to assess the correlation between retinal vascular features and the occurrence of PE.The predictive capability of retinal vascular features for PE(both early-and late-onset PE)was analyzed by using area under the curve(AUC)of receiver operator characteristic curve(ROC curve).Results·The comparative analysis of fundus features and retinal vascular features demonstrated statistically significant differences between the unaffected group and PE group in terms of central retinal artery equivalent(CRAE),central retinal vein equivalent(CRVE),arteriole-to-venular ratio(AVR),retinal artery tortuosity and retinal artery fractal dimension(all P<0.05).Univariate Logistic regression analysis indicated that second-trimester mean arterial pressure(MAP),second-trimester estimated fetal weight(EFW),CRAE,CRVE,AVR,retinal artery tortuosity and retinal artery fractal dimension were the influencing factors for PE occurrence(all P<0.05).Multivariate Logistic regression analysis revealed that second-trimester EFW,CRAE,CRVE,AVR,retinal artery tortuosity and retinal artery fractal dimension were the protective factors for the occurrence of PE,while second-trimester MAP was the risk factor for PE(all P<0.05).The analysis of ROC curves revealed that maternal risk factors along with second-trimester prenatal examination data(including MAP and EFW)and retinal vascular features model had good predictive ability for PE[AUC(95%CI)=0.784(0.725-0.843),and this model exhibited better predictive capability for early-onset PE,with an AUC(95%CI)of 0.840(0.756-0.924).Conclusion·The integration of quantified retinal vascular features based on AI models with maternal risk factors and second-trimester prenatal examination data(including MAP and EFW)enables a more effective prediction of PE occurrence,particularly early-onset PE.

Objective:To explore the teaching effect of 12-lead electrocardiogram (ECG) interpretation based on self-drawing method.Methods:This study recruited 489 students from 10 classes of clinical medicine major of Shandong Medical College as research objects. They were divided into control group and experimental group. Theoretical teaching and traditional practical training were adopted to the control group while theoretical teaching and self-drawing method were adopted to the experimental group. The ECG test scores, the number of students with excellent performance and the ECG scores of the final exam were compared between the two groups. Questionnaires were issued to assess the effect of different teaching methods on students. SPSS 25.0 was used for t test and chi-square test. Results:Compared with the experimental group, the results of the ECG special test, the number of excellent students and the ECG scores of the final exam in the control group all decreased, and the difference was statistically significant ( P<0.01). The results of the questionnaire showed that the experimental group had higher mastery of ECG abnormal patterns, learning initiative, self-confidence and better satisfaction with the overall teaching of ECG ( P<0.01). Conclusion:The practical training based on self-drawing method can improve ECG interpretation skills, learning initiative and self-confidence of students, which has achieved good teaching effect.

Objective:To investigate the 2 years’ efficacy of intravesical instillation of domestic BCG versus epirubicin in the prevention of recurrence of intermediate-risk or high-risk non-muscular invasive bladder cancer and predictive factors of BCG instillation.Methods:From July 2015 to June 2020, 18-75 years old patients with moderate to high-risk non muscle invasive bladder cancer (NMIBC) confirmed by pathological examination were involved. The ECOG score was 0-2. Exclusion criteria included ①immune deficiency or impairment (such as AIDS), using immunosuppressive drugs or radiotherapy, suspected allergic to BCG or epirubicin or excipients of the two drugs, fever or acute infectious diseases including active tuberculosis or receiving anti tuberculosis treatment, with severe chronic cardiovascular and cerebrovascular diseases or chronic kidney disease; ②combined with other urogenital system tumors or other organ tumors; ③combined with muscle invasive bladder urothelial carcinoma (≥T 2); ④undergoing chemotherapy, radiotherapy or immunotherapy within 4 weeks (immediate instillation after surgery not included); ⑤ pregnant or lactating women; ⑥ comfirmed or suspected bladder perforation; ⑦gross hematuria; ⑧cystitis with severe bladder irritation that may affect the evaluation; ⑨participat in other clinical trials within 3 months; ⑩alcohol or drug addiction; ?any risk factors that may increasing the risk of patients. Epirubicin 50 mg was irrigated immediately after the operation(TURBT or laser resection). The patients were randomly divided into BCG15 group, BCG19 group and epirubicin group by the ratio of 2∶2∶1, and the patients were maintained intravescical instillation for 1 year. The recurrence and adverse events of the three groups were compared. Univariate and multivariate analysis was performed to predict the risk factors of BCG irrigated therapy failure. Result:By June 15, 2020, the median follow-up duration was 22.1 months(12.1, 32.3), and there was no statistical difference between the groups ( P=0.9024). There were 274 patients enrolled in BCG19 group, 277 patients enrolled in BCG15 group and 130 patients enrolled in the epirubicin group. The drop-off rate was 16.6%(113 cases)and made no difference between groups( P=0.6222). There were no significant difference in age, gender, BMI, or ECOG score( P>0.05). During the follow-up, 116 cases was detected recurrence or progression. The recurrence rate of the three groups was 14.2% and 14.8% in BCG19 group and BCG15 group, and 27.7% in the epirubicin group. There was no difference in recurrence rate between BCG19 and BCG15 group( P=0.9464). The recurrence rate of BCG19 group was lower than that of the epirubicin group ( P=0.0017). The recurrence rate of BCG15 group was lower than that of the epirubicin group ( P=0.0020). There was no difference in the cumulative recurrence free survival rate between BCG19 and BCG15 group (95% CI0.57-1.46, P=0.7173). The cumulative recurrence free survival rate of BCG 19 group was better than that of the epirubicin group( HR=0.439, 95% CI0.26-0.74, P=0.0006), and the cumulative recurrence free survival rate of BCG15 group was better than that of the epirubicin group ( HR=0.448, 95% CI0.29-0.80, P=0.0021). The total incidence of adverse events in 19 BCG19, BCG15 and epirubicin group were 74.5%, 72.6% and 69.8% respectively. There was no difference in the incidence of adverse events between BCG19 and BCG15 group( P=0.6153). The incidence of adverse events in epirubicin group was lower than that of BCG19( P=0.0051) and BCG15( P=0.0167) groups.There was no significant difference in the incidence of serious adverse events (SAE) among the three groups ( P=0.5064). Log rank test univariate analysis and Cox risk regression model multivariate analysis showed that the history of bladder cancer recurrence( HR=6.397, 95% CI1.95-20.94, P=0.0001)was independent risk factor for BCG irrigation failure. Conclusions:The 2 years’ efficacy of intravesical instillation of domestic BCG is better than than of epirubicin with good tolerance and safety. There is no difference between BCG19 and BCG15 group. BCG doesn’t increase SAE compared with epirubicin. Recurrence status was an independent prognostic factor regarding recurrence-free survival.

Objective To investigate the efficacy and safety of intravesical instillation of BCG vaccine in the prevention of early recurrence of middle and high risk non-muscle invasive bladder cancer.Methods From July 2015,patients with non-muscle invasive bladder cancer aged 18-75 years with informed consent were screened and underwent transurethral resection of bladder tumor (TURBT).Immediately intravesical instillation of epirubicin 50 mg was given postoperatively.After pathology was comfirmed,patients was enrolled in group 1 (BCG15) or group 2 (BCG 19) or the control group (epirubicin 18) randomly with SAS 9.3 software.Data of follow-up and Adverse event was collected and analyzed.Results By May 31,2019,531 patients were enrolled in the study.The drop-off rate was 20.1％.167 patients (143 males and 24 females)in group 1,172 patients (141 males and 31 females)in group2 and 84(75 males and 9 females) in the control group with follow-up data were analyzed.There were no significant differences in age,gender,BMI,ECOG score,risk stratification between the three groups (P =0.8641,P =0.2906,P =0.9384,P =0.6126).The median follow-up time makes no statistical difference between the groups (P =0.9251),12.0 (6.0,22.5) months,13.0 (6.0,22.3) months,and 13.0 (7.0,22.3) months.The median recurrence time of the three groups was 4.0 (3.0,6.0) months,4.5 (3.0,9.8) months,4.5 (3.0,8.8) months.There was no statistical difference between the three groups (P =0.2852).Risk stratification in the patients got no significant difference between the three groups (P ＞ 0.05).The 1-year recurrence-free survival rates were 80.0％ in the group 1 and 88.3％ in the group 2 and 73.7％ in the control group.The group 2 was superior to the group 1 and the control group (P =0.0281,P =0.0031).There was no significant difference between group 1 and control group (P =0.2951).There was no significant difference in the cumulative recurrence-free survival between the experimental group 1 and the experimental group 2,(95％ CI 0.80-2.43,P =0.2433).The cumulative recurrence-free survival in the group 1 and the group 2 was better than the control group (95 ％ CI 0.31-0.92,P =0.0266;95 ％ CI 0.20-0.65,P =0.0008).All the cases underwent instillation were analyzed for adverse events.The incidence of overall AE(adverse events) in group 1 was 68.5％ (152/222),the incidence of grade Ⅰ-Ⅱ AE was 53.2％ (118/222),the incidence of grade Ⅲ-Ⅳ AE was 15.3％ (32/222).The incidence of overall AE in the group 2 was 71.8％ (160/223),the incidence of grade Ⅰ-Ⅱ AE was 60.1％ (134/223),and the incidence of grade Ⅲ-Ⅳ AE was 11.7％ (26/223).The overall AE rate in the control group was 53.2％ (59/111),of which the incidence of grade Ⅰ-Ⅱ AE was 42.4％ (47/111),and the incidence of grade Ⅲ-Ⅳ AE was 10.8％ (12/111).There was no difference in the incidence of overall AE between the group 1 and the group 2 (P =0.4497).The incidence of AE in the two experimental groups was higher than that in the control group (P =0.0062,P =0.0008).There was no difference in the incidence of grade Ⅲ-Ⅳ AE between the three groups (P =0.3902).Conclusions BCG(19 instillation schedule) has a better effect on preventing recurrence after 1 year of bladder surgery,which is superior to epirubicin group.The long-term efficacy of BCG in preventing recurrence and the efficacy of different schedules need to be further followed up.The lower urinary tract symptoms,which are mainly urinary frequency,are one of the causes of case fallout and should be fouced in future.Compared with epirubicin,BCG perfusion does not increase the incidence of grade Ⅲ-Ⅳ adverse reactions,and is safe to use.

Background:Previous study has found that harmine inhibited the proliferation of gastric cancer cells by down-regulating cyclooxygenase-2(COX-2)expression. However,its molecular mechanism is not fully clear. Aims:To investigate the effect of harmine on proliferation and apoptosis of gastric cancer cells,and explore the role of PTEN/Akt/MDM2 signaling pathway in this process. Methods:Human gastric adenocarcinoma cell line SGC-7901 and MKN-45 were treated with harmine at different concentrations(2,4,8,16,32 μg/mL)for 24,48,and 72 hours. The cell proliferation and apoptosis were detected by MTT assay and Hoechst staining,respectively. The expressions of PTEN,COX-2, phosphorylated Akt(p-Akt)and p-MDM2 were measured by Western blotting. Results:Harmine dose- and time-dependently inhibited proliferation and induced apoptosis of SGC-7901 and MKN-45 cells. Also,harmine dose-dependently increased PTEN expression,and inhibited p-Akt,p-MDM2 and COX-2 expressions in SGC-7901 and MKN-45 cells. Conclusions:Harmine may inhibit proliferation and induce apoptosis of gastric cancer cells via down-regulating COX-2 expression through PTEN/Akt/MDM2 signaling pathway.

Objective The expression of TNF-α was detected in sera of rabbits treated by ulinastatin with acute lung injury in duced by underwater explosion.Methods Rabbits were randomly divided into two groups such as the injured group and ulinastatin therapy group.Established underwater explosion device was used to cause acute lung injury in rabbits.TNF-α in sera of the rabbits were measured by ELISA at 4,12 and 24 hours after the explosion.The SPSS17.0 software was used to analyze the data and P＜0.05 was considered to be significant.Results There was no significant difference between the concentrations of TNF-α in the sera of rabbits in the injure group (538.20±201.43 ng/L) and that of in the ulinastatin group (386.90± 109.22 ng/L,t=2.088,P=0.051) at 4 hours after burst.However,there was evidently decreased in the level of TNF-α in the ulinastatin group (400.60 ± 78.98 ng/L) compared with the injury group (573.80 ± 178.24 ng/L,t =2.809,P =0.012) at 12 hours after burst.Moreover and TNF-α in the ulinastatin group (356.10 ± 130.99 ng/L) was also decreased compared to the injure group (552.30± 169.64 ng/L,t=2.895,P=0.010) at 24 hours after burst.Conclusion The TNF-α expression in sera of the rabbits in ulinastatin group were dramatically decreased than thai of in injury group at 12 hours after burst,which may be benefit to rabbits with acute lung injury induced by underwater explosion.

Objective To evaluate the relationship of preoperative neutrophil-lymphocyte ratio (NLR) with clinicopathological features and prognosis of colorectal cancer in middle-aged and elderly patients.Methods A retrospective analysis was performed in 212 patients with colorectal cancer in the First Affiliated Hospital of Nanjing Medical University from January 2011 to June 2013.All patients were divided into middle-aged group (46-65 year old,n=130) and old-aged group (66-89 year old,n=82),The optimal cut-off point of NLR was identified by the area under receiver operating characteristic curve,while NLR ＞ 3.13 and NLR≤3.13 were classified as high and low NLR group.The clinicopathological features and prognosis between the two groups were compared.Results There was no difference in gender,tumor growth site,depth of invasion,tumor embolus,lymphatic metastasis,distant metastasis,TNM stage between low and high NLR group (allP＞ 0.05).However,the difference between high NLR group and low NLR group in old-aged group with diabetes mellitus was statistically significant (P＜0.05).The 1-,2-,and 3-year survival rate of the overall 212 patients were 96.2％ (204/212),87.7％ (186/212) and 74.5％ (158/212) In middle-aged group,the 1-,2-,and 3-year survival rates were 98.8％ (85/86),90.7％ (78/86) and 84.9％ (73/86) respectively in low NLR group,but 95.5％ (42/44),84.1％ (37/44) and 72.7％ (32/44) respectively in high NLR group,(allP＜0.05).In old-aged group,the 1-,2-,and 3-year survival rates were 95.7％ (44/46),89.1％ (41/46) and 73.9％ (34/46) respectively in low NLR group,but 91.7％ (33/ 36),83.3％ (30/36) and 52.8％ (19/36) respectively in high NLR group (all P＜0.05).Cox regression showed that TNM stage and NLR were independent risk factors for the prognosis of the middle-aged and elderly patients with colorectal cancer (P＜0.05 or P＜0.01).Conclusions Preoperative NLR ＞ 3.13 suggest that the prognosis is poor in middle-aged and elderly patients with colorectal cancer.