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

AIM To investigate the protective effects and the mechanism of the Liuwei Dihuang Pills on mouse brain microvascular endothelial(bEnd.3)cells damaged by β-Amyloid protein1-40(Aβ1-40).METHODS CCK8 method was used to detect the effects of Aβ1-40 and medicated serum of Liuwei Dihuang Pills(MSLDP)on cell activity,and to screen the appropriate concentration.bEnd.3 cells of the control group,the Aβ1-40 group,the MSLDP+Aβ1-40 group and the MSLDP group had their low density lipoprotein-associated protein 1(LRP1),receptor for advanced glycation end products(RAGE),matrix metalloproteinase-2(MMP-2),MMP-9,scaffold protein zonule protein-1(ZO-1)detected by Western blot.bEnd.3 cells assigned into the control group,the Aβ1-40 group,the FPS-ZM1(RAGE inhibitor)+Aβ1-40 group and the FPS-ZM1+Aβ1-40+MSLDP group had their expressions of RAGE,MMP-9,MMP-2 and ZO-1 detected by Western blot as well.RESULTS The cell activity of bEnd.3,was dose-dependently decreased by Aβ1-40(P<0.01),but was protected by MSLDP(P<0.05,P<0.01).And 10 μmol/L Aβ1-40 and 10%MSLDP were selected for subsequent experiments.Compared with the control group,the Aβ1-40 group displayed increased protein expressions of RAGE,MMP-2 and MMP-9(P<0.01),decreased protein expressions of LRP1,ZO-1 and BDNF(P<0.05,P<0.01),and decreased fluorescence intensities of LRP1 and ZO-1(P<0.01).Compared with the Aβ1-40 group,the MSLDP group shared decreased expressions of RAGE,MMP-2,MMP-9 proteins and RAGE fluorescence intensity(P<0.05,P<0.01),and increased expressions of LRP1,ZO-1 and BDNF proteins,and the fluorescence intensity of LRP1 and ZO-1(P<0.05,P<0.01);the Aβ1-40+FPS-ZM1 group displayed decreased protein expressions of MMP-2,MMP9 and RAGE(P<0.05,P<0.01),and increased ZO-1 protein expression(P<0.05);and the Aβ1-40+FPS-ZM1+ MSLDP group displayed an even more decreased protein expressions of MMP-2,MMP9 and RAGE(P<0.01),increased ZO-1 protein expression(P<0.01)due to the the combination use of FPS-ZM1 and MSLDP.CONCLUSION Liuwei Dihuang Pills can protect the tight junction of bEnd.3 injured by Aβ1-40 and neurovascular units from Alzheimer's disease by alleviating the dysfunction of the blood-brain barrier via RAGE-mediated MMP-2/MMP-9 pathway inhibition.

AIM: To observe the imaging characteristics of the affected eyes of patients with central serous chorioretinopathy(CSC)of different ages and their asymptomatic fellow eyes.METHODS: Retrospective study. A total of 76 cases(88 eyes)of CSC patients diagnosed in the ophthalmology department of our hospital from April to September, 2023 and 35 cases(35 eyes of asymptomatic fellow eyes of patients with unilateral CSC)were selected for the study. According to age, they were divided into young and middle-aged groups(<40 years old), middle-aged groups(40-50 years old)and middle-aged and elderly groups(>50 years old). The imaging features of the affected eyes of CSC patients of different ages and their asymptomatic fellow eyes were observed.RESULTS: The subfoveal choroidal thickness(SFCT)of CSC eyes in the young and middle-aged patients(487.30±83.33 μm)was significantly greater than that of the middle-aged group(414.17±96.02 μm, P<0.05)and the middle-aged and elderly group(409.4±107.42 μm, P<0.05). The incidence of choroidal neovascularization(CNV)in CSC patients of the middle-aged and elderly group was significantly higher than that in the young and middle-aged group(P<0.0167). The SFCT of the asymptomatic fellow eye of the unilateral CSC patient in the young and middle-aged group(511.29±40.89 μm)was significantly larger than that of the middle-aged and elderly group(364.76±82.26 μm, P<0.05). Among them, the vortex vein anastomosis rate in eyes with CSC is higher than 90%, and vortex vein anastomosis or dilatation is present in all asymptomatic fellow eyes of CSC patients.CONCLUSION: There are differences in the imaging manifestations of CSC-affected eyes and their asymptomatic fellow eyes of different age groups. SFCT is generally thickened and gradually becomes thinner with the growth of age. The incidence of CNV in CSC-affected eyes is the highest in the middle-aged and elderly group. In addition, vortex vein anastomosis and dilatation are common in CSC-affected eyes and asymptomatic fellow eyes.

Objective To observe the efficacy and safety of Morinda officinalis oligosaccharides in the continuation treatment of mild and moderate depression.Methods An open,single-arm,multi-center design was adopted in our study.Adult patients with mild and moderate depression who had received acute treatment of Morinda officinalis oligosaccharides were enrolled and continue to receive Morinda officinalis oligosaccharides capsules for 24 weeks,the dose remained unchanged during continuation treatment.The remission rate,recurrence rate,recurrence time,and the change from baseline to endpoint of Hamilton Depression Scale(HAMD),Hamilton Anxiety Scale(HAMA),Clinical Global Impression-Severity(CGI-S)and Arizona Sexual Experience Scale(ASEX)were evaluated.The incidence of treatment-related adverse events was reported.Results The scores of HAMD-17 at baseline and after treatment were 6.60±1.87 and 5.85±4.18,scores of HAMA were 6.36±3.02 and 4.93±3.09,scores of CGI-S were 1.49±0.56 and 1.29±0.81,scores of ASEX were 15.92±4.72 and 15.57±5.26,with significant difference(P＜0.05).After continuation treatment,the remission rate was 54.59％(202 cases/370 cases),and the recurrence rate was 6.49％(24 cases/370 cases),the recurrence time was(64.67±42.47)days.The incidence of treatment-related adverse events was 15.35％(64 cases/417 cases).Conclusion Morinda officinalis oligosaccharides capsules can be effectively used for the continuation treatment of mild and moderate depression,and are well tolerated and safe.

Through a compound induction method, combined with neurobehavioral, macroscopic characterization and objective pathological evaluation indicators, a murine depression model of liver depression transforming into fire syndrome was constructed and confirmed. The model was constructed using a combination of sleep deprivation, light exposure, and alternate-day food deprivation. Evaluation was conducted at three levels: face validity, constructs validity, and predictive validity. The establishment of the liver depression transforming into fire syndrome depression model was further validated through the counterproof of traditional Chinese medicine formulas. In terms of face validity, compared to the control group, mice in the model group exhibited typical depressive symptoms in neurobehavioral assessments; the general observation of the model group mice reveals disheveled and lackluster fur, along with delayed and easily agitated responses. Additionally, there is a substantial increase in water consumption. In the sleep phase detection of mouse, the model group showed a significant increase in the proportion of time spent in the wake phase during sleep, accompanied by a significant decrease in the proportions of time spent in both non-rapid eye movement (NREM) and rapid eye movement (REM) sleep phases. There are significant differences in physiological indicators such as average blood flow velocity, blood flow rate, tongue, urine, and claw color (r values) in the internal carotid artery. Structural validity demonstrated that levels of 5-hydroxytryptamine (5-HT), dopamine (DA), and γ-aminobutyric acid (GABA) in the hippocampus of model mice decreased significantly, while acetylcholine (ACh), tryptophan (Try), and glutamic acid (Glu) levels increased significantly. Treatment with Danzhi Xiaoyao San led to varying degrees of restoration in the aforementioned neurotransmitters. In terms of predictive validity, the antidepressant paroxetine effectively ameliorated depressive behaviors in the model mice, and the classic formula Danzhi Xiaoyao San demonstrated varying degrees of improvement in depression-related indicators. In conclusion, this study has established a novel animal model of liver-stagnation with the fire depression, thereby expanding the repertoire of traditional Chinese medicine depression models. This development contributes to the diversification of melancholic syndrome models in Chinese medicine, offering a broader spectrum of options for scientific exploration, efficacy evaluation, and drug screening in the future prevention and treatment of depression with traditional Chinese medicine. Animal experiments were conducted with the approval and supervision of the Animal Ethics Committee of Jiangxi University of Traditional Chinese Medicine (ethics number: 20230313040).

Central serous chorioretinopathy (CSC) is one of the main causes of impaired visual function in middle-aged men. CSC is characterized by a thickening of the choroid and hyperpermeability of the choroidal vessels, resulting in serous subretinal fluid. The pathogenesis of CSC is not fully understood. Since the introduction of indocyanine green angiography, the detection of the influence of the vortex veins in CSC, it has been established that the presence of a thick choroid may be caused by congestion of the vortex vein, resulting in new choroidal drainage through a vortex vein anastomosis. The study of vortex venous blood hemodynamics has elucidated the new concept of the pathogenesis of CSC, deepened our understanding of the disease, and provided a theoretical basis for new treatment methods. With a better understanding of the pathogenesis of CSC, we expect to be able to stratify patients at risk in the clinic and evaluate optimized treatment options for patients with CSC

Objective:To observe the changes of chorioidal thickness (ChT) in patients with central serous chorioretinopathy (CSC) in different mode of vortic venous dilation.Methods:A prospective cross-sectional observational study. A total of 80 patients with 89 eyes (CSC group) diagnosed in Department of Ophthalmology, General Hospital of Central Theater Command from April to October 2023 were included in the study. Among them, 64 males had 71 eyes and 17 females had 18 eyes. A total of 15 healthy volunteers matched in age and sex were selected as the control group. Among them, 14 men had 26 eyes and one woman had two eyes. The macular region was examined by ultra-wide-angle scanning frequency source optical coherence tomography (OCTA) with BM400K BMizar made by TowardPi (Beijing) Medical Technology Co., LTD. Scanning rate 1 536 A scanning×1 280 B scanning, scanning range 24 mm×20 mm. The accompanying software delineated nine subfields (superotemporal, upper, superonasal, temporal, central, nasal, inferotemporal, lower, inferonasal regions) to record ChT. En-face OCTA mode was utilized to observe the anatomy and functional anastomosis of the vortex veins above and below the choroidal blood layer. Eyes in the CSC group were further categorized into upper-dominant, symmetrical, and lower-dominant groups based on the difference in vortex vein expansion shown in the choroidal layer of the en-face image, with 36, 35, and 18 eyes respectively. Statistical analysis included the use of independent samples t-test or Mann-Whitney test for comparison between two groups, one-way analysis of variance or Kruskal-Wallis H test for comparison between multiple groups, and the χ test or Fisher test for categorical variables. Results:Compared with the control group, ChT in the CSC group was thickened in the foveal area and different areas of the macula, with the greatest difference in the fovea, and the differences were statistically significant ( t=3.345, 5.018, 2.902, 4.667, 7.276, 3.307, 3.868, 4.795, 2.583; P<0.05). Compared with the ChT of the control group, there was no statistically significant difference in the superotemporal, region of the upper-dominant group ( t=1.510, P>0.05); in other regions, the differences were statistically significant ( t=3.207, 5.163, 2.526, 4.310, 6.285, 2.656, 3.812, 2.173; P<0.05). The differences in the foveal area and other areas in the symmetrical group were statistically significant ( t=4.488, 5.554, 3.457, 5.314, 7.256, 3.507, 5.584, 6.019, 2.994; P<0.05). In the superotemporal, and superonasal, regions of the lower dominant group, the differences were not statistically significant ( t=1.150, 1.465; P<0.05); in other regions, the differences were statistically significant ( t=2.278, 4.168, 5.244, 2.783, 5.040, 3.432, 2.095; P<0.05). Conclusion:The dilated distribution of vortex veins on en-face ultra-wide-angle OCTA has a corresponding relationship with ChT. In eyes with CSC, the superior vortex vein drainage system may be the primary route for choroidal drainage.

Objective:To observe the long-term effects of anti-vascular endothelial growth factor (VEGF) drug initiation combined with dexamethasone intravitreal implant (DEX) on the structural integrity of the outer macular region of the eye in patients with macular edema (ME) secondary to central retinal vein occlusion (CRVO).Methods:A retrospective clinical study. From February 2018 to August 2022, 54 patients diagnosed with CRVO combined with ME (CRVO-ME) in Department of Ophthalmology of Central Theater Command General Hospital were included in the study. Among them, there were 30 males and 24 females, all with monocular disease. According to different treatment regiments, patients were divided into anti-VEGF and DEX combination therapy group (initial combination group), anti-VEGF drug monotherapy group (monotherapy group) with 21 eyes and 33 eyes, respectively. Best corrected visual acuity (BCVA), optical coherence tomography (OCT) examination were performed in all eyes. The thickness of foveal retina (CRT) and the deficiency length of outer membrane (ELM), ellipsoid band (EZ) and chimaera band (IZ) in the 1 mm macular area were measured by OCT. The initiating combination group was treated with anti-VEGF agents or DEX as assessed on demand (PRN) after the combination therapy, and the monotherapy group received 3+PRN regimen. Relevant examinations were performed at 1 (V1), 6 (V6), 12 (V12) months and observation cut-off or the last visit (Vf) after treatment using the same equipment before treatment. The deletion length of ELM, EZ and IZ in V1, V6, V12 and Vf after treatment were compared between the two groups. Repeated measurement ANOVA was used to compare BCVA, CRT and deletion length of ELM, EZ and IZ at different follow-up times. Spearman rank correlation test was used to analyze the correlation between the two groups of continuous variables.Results:The follow-up time of patients in the initial combination group and monotherapy group was (18.05±5 .66) and (21.90±10.80) months, respectively, with no statistical significance ( F=13.430, P=0.229). Compared with baseline, the deletion lengths of ELM, EZ and IZ were significantly improved ( F=11.848, 10.880, 29.236), BCVA was increased ( F=10.541) and CRT was decreased ( F=52.278) in the initial combination group and the monotherapy group at different follow-up times after treatment. The differences were statistically significant ( P<0.001). At V1, EZ and IZ deletion lengths were (344.10±413.03), (593.33±372.96) μm and (354.71±321.75), (604.85±385.77) μm in the initial combination group and monotherapy group, respectively. The improvement of EZ and IZ deletion lengths in the initial combination group was better than that in the single drug group, and the difference was statistically significant ( F=5.272, 6.106; P=0.026, 0.017). The CRT of the initial combination group and the monotherapy group were (248.86±59.99) and (314.72±214.91) μm, respectively, and the CRT of the initial combination group was significantly lower than that of the monotherapy group, with statistical significance ( F=6.102, P=0.017). At V6, V12 and Vf, the deletion length of ELM, EZ and IZ and BCVA and CRT showed no statistical significance ( P>0.05). Correlation analysis showed that ELM, EZ, IZ were positively correlated with BCVA and CRT in the initial combination group and monotherapy group ( P<0.001). In V6, V12 and Vf, the number of anti-VEGF drug injections in the initial combination group and monotherapy group was (2.67±1.32), (4.43±2.27), (6.05±3.51), (4.58±0.90), (7.33±1.93), (11.33±6.10) times, respectively. The number of injections in the initial combination group was significantly lower than that in the monotherapy group, and the difference was statistically significant ( F=5.150, 0.646, 3.433; P<0.001). Conclusions:The improvement of BCVA and CRT in the initial combination group is similar to that in the monotherapy group. Compared with the monotherapy group, EZ and IZ deletion are improved more significantly in the initial combination group, and CRT decreased more rapidly and significantly. The initial combination group receives fewer anti-VEGF injections than the monocular group.

Objective To investigate the causes of a cluster of suspected neonatal carbapenem-resistant Klebsiella pneumoniae(CRKP)bloodstream infection(BSI)in the neonatal department of a hospital,and provide references for the effective control of the occurrence of healthcare-associated infection(HAI).Methods Epidemiological in-vestigation on 3 neonates with CRKP BSI in the neonatal department from January 31 to February 6,2023 was per-formed.Specimens from environmental object surfaces were taken for environmental hygiene monitoring,and effec-tive control measures were taken according to the risk factors.Results From January 31 to February 6,2023,a to-tal of 60 neonates were admitted in the neonatal department,including 16 with peripherally inserted central venous catheter(PICC).Three neonates had CRKP BSI,with a incidence of 5.00％.There were 33 hospitalized neonates on the day(February 7)when the cluster of HAI was reported,with a prevalence rate of 9.09％(3/33).CRKP BSI rate in the neonatal department of this hospital from January 31 to February 6,2023 was higher than that in 2022(P＜0.001).The incubators of the 3 neonates with CRKP BSI were in the same ward and adjacent to each other.The first neonate with CRKP BSI(who developed BSI on January 31)underwent PICC maintenance on Feb-ruary 4,and the other 2 neonates with PICC maintenance immediately following the first one also developed CRKP BSI.CRKP were isolated from blood culture of all 3 neonates,and antimicrobial susceptibility testing results were consistent.Conclusion The occurrence of the cluster event of neonatal CRKP BSI may be related to the failure of strict implementation of aseptic procedures during PICC maintenance and cross contamination among items.

Objective To compare choroidal parameters and morphological changes in asymptomatic contralateral eyes of patients with pachychoroid neovasculopathy(PNV)and central serous chorioretinopathy(CSC)using ultra-wide-field swept-source optical coherence tomography angiography(UWF SS-OCTA).Methods A prospective,cross-section-al,observational study was conducted,including 23 asymptomatic contralateral eyes of 23 CSC patients and 19 asympto-matic contralateral eyes of 19 PNV patients treated in our Ophthalmology Department from April 2023 to September 2023,as well as 22 eyes of 22 age-and sex-matched healthy individuals.These eyes were divided into the CSC contralateral eye group,PNV contralateral eye group,and healthy eye group,respectively.There was no statistically significant difference in age,male ratio,and best corrected visual acuity among the three groups(all P＞0.05).All subjects underwent UWF SS-OCTA imaging examination and comparison,and their choroidal thickness(CT),vascular density(VD)in the major vas-cular layer of the choroid(ratio of blood flow pixel area to total area of the target area),and choroidal vascular index(CVI)(ratio of choroidal vascular lumen volume to total volume of choroidal vessels and stromata)in 9 zones(superior temporal,superior,superior nasal,temporal,foveal,nasal,inferior temporal,inferior,and inferior nasal)were measured and recorded,respectively.The macular subfoveal retinal thickness,subfoveal choroidal thickness(SFCT),and asymmet-ric dilation and anastomosis of the vortex veins were examined and compared in each group of patients.Results There was a statistically significant difference in macular SFCT among the three groups(all P＜0.01).There was a statistically significant difference in the proportion of asymmetric dilation or anastomosis of the vortex veins among the three groups(all P＜0.001).The CVI in the superior nasal and nasal zones in the CSC contralateral eye group were higher than those in the PNV contralateral eye group(both P＜0.05).The CVI and average CVI in the superior nasal,temporal,foveal,inferi-or,and inferior nasal zones in the CSC contralateral eye group were higher than those in the healthy eye group(all P＜0.05).The CVI in the temporal,foveal,and inferior zones in the PNV contralateral eye group were higher than those in the healthy eye group(all P＜0.05).The VD in the major vascular layer of the choroid in the superior temporal,superior,su-perior nasal,foveal,and inferior temporal zones in the CSC contralateral eye group were lower than those in the healthy eye group(all P＜0.05).The VD in the major vascular layer of the choroid in the PNV contralateral eye group were lower than that in the healthy eye group in all zones(all P＜0.001).The CT in the superior temporal and superior nasal zones in the CSC contralateral eye group were higher than those in the PNV contralateral eye group(both P＜0.05).The CT in the superior temporal,superior,superior nasal,temporal,foveal,inferior,and inferior nasal zones in the CSC contralateral eye group were higher than those in the healthy eye group(all P＜0.05).The CT in the superior,temporal,foveal,and in-ferior zones in the PNV contralateral eye group were higher than those in the healthy eye group(all P＜0.05).Conclu-sion This study provides validation from the perspective of the fellow eye that CSC and PNV may not be different stages of the same disease.CSC and PNV are not unilateral diseases,but bilateral choroida diseases that manifest in one eye.