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

Objective To investigate whether Heshouwuyin can delay the aging of Leydig cells in rat testis through DNA methyltransferase 1(DNMT1).Methods Totally 40 male Wistar rats were randomly divided into 4 groups,with 10 rats in each group.Immunohistochemistry was used to detect the expression levels of DNMT1 in testis tissue of rats.Testosterone content in serum of rats in each group was detected by ELISA test.A rat Leydig cell aging model was established by free radical oxidative damage.DNMT1 was knocked down by lentivirus in Leydig cells,and the cell senescence status and the testosterone content and testosterone synthesis key enzyme 3β-hydroxysteroid dehydrogenase(3β-HSD),cytochrome P450 family member 11A1(CYP11A1)content secreted by cells were detected by β-galactosidase(β-GAL)staining,immunofluorescenct staining and ELISA.Results Compared with the young control group(YCG),the expression of P16 protein and the positive rate of β-GAL in the testis tissue of rats in the natural aging group(NAG)increased significantly,and the expression of DNMT1 and serum testosterone levels decreased(P＜0.05).However,after Heshouwuyin intervention,the expression of P16 protein and the positive rate of β-GAL in the testis of aging rats were reduced,and DNMT1 expression and the serum testosterone levels increased(P＜0.05).The same trend was observed in Leydig cells.Knockdown of DNMT1 in Leydig cells,β-GAL positivity and P16 protein expression increased significantly,and testosterone secretion and testosterone synthesis key enzymes 3β-HSD,CYP11A1 content from Leydig cells decreased significantly,compared with the normal control group(NCG)(P＜0.05).When Heshouwuyin was added,the above phenomenon was improved.Conclusion Heshouwuyin can delay the aging of rat Leydig cells through DNMT1.

Objective The purpose of this study was to investigate the bacterial communities of biting midges and ticks collected from three sites in the Poyang Lake area,namely,Qunlu Practice Base,Peach Blossom Garden,and Huangtong Animal Husbandry,and whether vectors carry any bacterial pathogens that may cause diseases to humans,to provide scientific basis for prospective pathogen discovery and disease prevention and control. Methods Using a metataxonomics approach in concert with full-length 16S rRNA gene sequencing and operational phylogenetic unit(OPU)analysis,we characterized the species-level microbial community structure of two important vector species,biting midges and ticks,including 33 arthropod samples comprising 3,885 individuals,collected around Poyang Lake. Results A total of 662 OPUs were classified in biting midges,including 195 known species and 373 potentially new species,and 618 OPUs were classified in ticks,including 217 known species and 326 potentially new species.Surprisingly,OPUs with potentially pathogenicity were detected in both arthropod vectors,with 66 known species of biting midges reported to carry potential pathogens,including Asaia lannensis and Rickettsia bellii,compared to 50 in ticks,such as Acinetobacter lwoffii and Staphylococcus sciuri.We found that Proteobacteria was the most dominant group in both midges and ticks.Furthermore,the outcomes demonstrated that the microbiota of midges and ticks tend to be governed by a few highly abundant bacteria.Pantoea sp7 was predominant in biting midges,while Coxiella sp1 was enriched in ticks.Meanwhile,Coxiella spp.,which may be essential for the survival of Haemaphysalis longicornis Neumann,were detected in all tick samples.The identification of dominant species and pathogens of biting midges and ticks in this study serves to broaden our knowledge associated to microbes of arthropod vectors. Conclusion Biting midges and ticks carry large numbers of known and potentially novel bacteria,and carry a wide range of potentially pathogenic bacteria,which may pose a risk of infection to humans and animals.The microbial communities of midges and ticks tend to be dominated by a few highly abundant bacteria.

Objective To propose a sperm classification model based on convolutional neural network to enhance the accuracy of sperm morphological classification.Methods A FT-EfficientNet model was constructed using EfficientNetB0 as the base model,which was fine-tuned by data preprocessing enhancement,transfer learning and cosine decay.Classification experi-ments were performed on the sperm public datasets SCIAN-Morpho and HuSHeM,and the datasets were segmented and vali-dated using 5-fold cross-validation.The classification results by the FT-EfficientNet model were compared with those by the cascade ensemble of support vector machines(CE-SVM)model,the adaptive patch-based dictionary learning(APDL)model,fine tuning of visual geometry group(FT-VGG)model,morphological classification of human sperm heads(MH-HSH)model and transfer learning(TL)model.Ablation experiments were performed in the SCIAN-Morpho dataset to verify the effect of different fine-tuning methods on the model.Results The FT-EfficientNet model proposed had the accuracy,precision and F1 score on the SCIAN-Morpho validation set being 64.1％,63.8％and 64.8％,respectively,which were better than CE-SVM,APDL,FT-VGG and MC-HSH models.The recall rate of the model proposed(65.2％)was slightly lower than that of MC-HSH model(68.0％).The accuracy,precision,F1 score and recall rate on the HuSHeM validation set was 95.4％,95.8％,95.4％and 96.0％,respectively,which were slightly lower than those of TL model while better than those of CE-SVM,APDL,FT-VGG and MC-HSH models.Ablation experiments showed the FT-EfficientNet model behaved the best in fine-tuning.Conclusion The sperm classification model based on convolutional neural network facilitates sperm morphology classification with high accuracy and performance.[Chinese Medical Equipment Journal,2024,45(10):7-13]

Heart failure with preserved ejection fraction (HFpEF) accounts for about half of the number of patients with heart failure. In addition to the typical features of heart failure such as myocardial stiffness and diastolic function impairment, the key characteristic of HFpEF is the normal left ventricular ejection fraction, which increases the difficulty of clinical diagnosis. QiShenYiQi Dripping Pills (QSYQ) is a standardized traditional Chinese medicine approved by the China Food and Drug Administration (CFDA), and many clinical studies have demonstrated the efficacy and safety of QSYQ in the treatment of heart failure with reduced ejection fraction, but the role of QSYQ in HFpEF has not been clarified. In this paper, high fat diet (HFD) and drinking water containing N-nitro-L-arginine methyl ester (L-NAME, 0.5 g·L^-1, pH=7.4) were used in C57BL/6N male mice to construct the classical HFpEF model (the experiment was approved by the Animal Ethics Committee of Hefei University of Technology, the approval number is HFUT20220921002), and at the 8^th week, the mice were dosed with ① empagliflozin, ② low-dose QSYQ (LQ), ③ high-dose QSYQ (HQ), ④ empagliflozin plus low-dose QSYQ (ELQ) for 4 weeks, the body weight of the mice was recorded during the experiment, echocardiography, blood pressure and glucose tolerance were detected and the exercise capacity of mice was evaluated, and pathological and biochemical experiments were used to detect cardiac fibrosis, liver fibrosis and serum biochemical indexes in mice, RNAseq assay was performed with mice heart tissues. The results showed that QSYQ as well as QSYQ+empagliflozin could significantly reduce the body weight, improve diastolic function and hypertension, improve glucose tolerance and enhance exercise ability of HFpEF mice. At the biochemical and molecular levels, QSYQ and QSYQ+empagliflozin can reduce the cross-sectional area of cardiomyocytes and reduce cardiac collagen contents, thereby alleviating myocardial hypertrophy and fibrotic phenotypes, and improve metabolic disorders. The RNAseq results suggest that the function of QSYQ in improving HFpEF may be related to calsequestrin 1. In conclusion, this study shows that QSYQ and QSYQ+empagliflozin can significantly improve HFpEF-related cardiac dysfunction and metabolic disorders, which provides a theoretical basis for the clinical treatment of HFpEF.

OBJECTIVE: To evaluate the effectiveness and safety of Chinese medicine (CM) in the treatment of coronavirus disease 2019 (COVID-19) in China. METHODS: A multi-center retrospective cohort study was carried out, with cumulative CM treatment period of ⩾3 days during hospitalization as exposure. Data came from consecutive inpatients from December 19, 2019 to May 16, 2020 in 4 medical centers in Wuhan, China. After data extraction, verification and cleaning, confounding factors were adjusted by inverse probability of treatment weighting (IPTW), and the Cox proportional hazards regression model was used for statistical analysis. RESULTS: A total of 2,272 COVID-19 patients were included. There were 1,684 patients in the CM group and 588 patients in the control group. Compared with the control group, the hazard ratio (HR) for the deterioration rate in the CM group was 0.52 [95% confidence interval (CI): 0.41 to 0.64, P<0.001]. The results were consistent across patients of varying severity at admission, and the robustness of the results were confirmed by 3 sensitivity analyses. In addition, the HR for all-cause mortality in the CM group was 0.29 (95% CI: 0.19 to 0.44, P<0.001). Regarding of safety, the proportion of patients with abnormal liver function or renal function in the CM group was smaller. CONCLUSION This real-world study indicates that the combination of a full-course CM therapy on the basic conventional treatment, may safely reduce the deterioration rate and all-cause mortality of COVID-19 patients. This result can provide the new evidence to support the current treatment of COVID-19. Additional prospective clinical trial is needed to evaluate the efficacy and safety of specific CM interventions. (Registration No. ChiCTR2200062917).
Humans ; Retrospective Studies ; Male ; Female ; Middle Aged ; COVID-19/epidemiology* ; COVID-19 Drug Treatment ; Aged ; Medicine, Chinese Traditional/methods* ; Drugs, Chinese Herbal/adverse effects* ; SARS-CoV-2 ; Treatment Outcome ; China/epidemiology* ; Adult

Aim To explore he preventive and therapeutic effects of Tongbu-fangchan prescription on aceylcholine-calcium chloride (Ach-CaCl

A moxibustion device with the functions of auricular fumigation moxibustion and heat-sensitive moxibustion is designed. The smoke of the ignited moxa stick is used for the fumigation moxibustion at the external auditory canal, while the heat generated works on Dazhui (GV 14) for heat-sensitive moxibustion. The device consists of five parts, i.e. combustion chamber, smoke pipe, smoke processing chamber, power module and connector. It solves the limitations such as unpleasant experience in treatment, unfavorable temperature control, easy scalding and excessive manual dependence induced by usual fumigation moxibustion and during heat-sensitive moxibustion. This moxibustion device may improve the safety and convenience when delivering the treatment with fumigation moxibustion and heat-sensitive moxibustion, as well as the work efficiency of medical staff.
Humans ; Moxibustion ; Hot Temperature ; Fumigation ; Smoke ; Temperature

To explore the situation of 8 common respiratory pathogens in children with acute respiratory infection (ARI) from 2021 to 2022.The retrospective study selected 8 710 ARI patients from September 2021 to August 2022 in the Maternal and Child Health Hospital of Gansu Province as the study object, patients aged 0 to 17 years old, including 5 048 male children and 3 662 female children. Indirect immunofluorescence was used to detect 8 common respiratory pathogens, including influenza virus A (FluA), influenza virus B (FluB), parainfluenza virus (PIV), respiratory syncytial virus (RSV), adenovirus (ADV), Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Coxsackie virus group B (CoxB) IgM antibodies. χ² test was used to analyze the results. The results showed that 1 497 of 8 710 children with ARI were positive, with a positive rate of 17.19%. The detection rate of MP among 8 common respiratory pathogens was 11.34%, accounting for 66.0%, followed by FluB, CoxB, PIV, RSV, ADV, FluA and CP, accounting for 13.83%, 9.55%, 6.01%, 2.61%, 1.47%, 0.40% and 0.13%, respectively. Respiratory tract viruses (FluA, FluB, RSV, ADV, PIV, CoxB) accounted for 33.86%.There were significant differences in the detection rates of PIV, ADV and MP among children of different genders (χ²=6.814, 5.154 and 17.784, P<0.05). The detection rate of school-age children (6-17 years old) was the highest, accounting for 33.27% (184/553). The detection rates of 8 common respiratory pathogens in patients with ARI were higher in spring and winter and lower in summer and autumn. To sum up, from 2021 to 2022, MP and FluB infection were dominant in ARI patients in our hospital. The peak period of 8 common respiratory pathogens was in spring and winter. The physical examination rate of 8 common respiratory pathogens in ARI patients aged 6-17 years old was the highest.
Child ; Humans ; Male ; Female ; Infant ; Infant, Newborn ; Child, Preschool ; Adolescent ; Retrospective Studies ; Respiratory Tract Infections/epidemiology* ; Respiratory Syncytial Virus, Human ; Seasons ; Mycoplasma pneumoniae ; Adenoviridae ; Influenza B virus

Objective:To study influence of Status-Background-Assessment-Recommendation(SBAR)communi-cation mode intervention on patients with heart failure(HF).Methods:According to intervention method,a total of 187 HF patients treated in our hospital were divided into routine intervention group(n=96,received routine in-tervention based on routine treatment)and SBAR group(n=91,received SBAR communication mode intervention based on routine treatment).Both groups were intervened for six months.General clinical data,LVEF,left ven-tricular end-diastolic volume(LVEDV),left ventricular end-systolic volume(LVESV),scores of Morisky medi-cation compliance scale and the medical outcomes study 36-item short-form heath survey(SF-36),patients'satis-faction and incidence rate of major adverse cardiovascular events(MACE)within six-month follow-up were com-pared between two groups.Results:Compared with routine intervention group,after intervention,there were sig-nificant rise in LVEF[(53.44±2.11)%vs.(57.72±1.80)%],scores of Morisky medication compliance scale[(2.00±0.77)scores vs.(3.63±1.07)scores]and SF-36[(68.83±7.62)scores vs.(76.81±9.68)scores],and significant reductions in LVEDV[(132.63±5.59)ml vs.(119.15±6.89)ml]and LVESV[(64.36±5.82)ml vs.(56.62±4.21)ml]in SBAR group(P=0.001 all).Patients'satisfaction of SBAR group was significantly higher than that of routine intervention group(89.01%vs.73.96%,χ2=6.958,P=0.008),and incidence rate MACE during follow-up was significantly lower than the latter(5.49%vs.15.63%,LogRankχ2=5.043,P= 0.025).Conclusion:SBAR communication mode intervention can significantly improve heart function,medication compliance and quality of life in patients with heart failure,with high patients'satisfaction.