ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a serious impact on global public health and the economy. SARS-CoV-2 infiltrates host cells via its surface spike protein, which binds to angiotensin-converting enzyme 2 on the host cell membrane. As a result, small molecules targeting spike protein have emerged as a hotspot in anti-SARS-CoV-2 drug research. Activity screening is an important step in seeking small molecule drugs. Therefore, this article aims to review the biological activity evaluation methods of small molecule inhibitors targeting SARS-CoV-2 spike protein, with the goal of laying the foundation for the discovery of new anti-SARS-CoV-2 drugs.

Objective To study the effect of changes in estradiol(E2)levels on the reproductive outcomes of pa-tients with polycystic ovary syndrome(PCOS)undergoing controlled ovarian stimulation(COS)with gonadotropin-releasing hormone antagonist(GnRH-ant)protocol.Methods A retrospective study was conducted involving 338 patients with polycystic ovary syndrome(PCOS)infertility who underwent GnRH-ant protocol for controlled ovarian hyperstimulation followed by in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI),and subse-quently underwent their first frozen-thawed embryo transfer(FET).The clinical data of these patients were ana-lyzed.Patients were grouped based on the changes in serum E2 levels on the first and fourth day of GnRH-ant ad-ministration(blood samples collected before GnRH-ant injection):the E2 elevation group(Group A,E2 value in-creased more than 30％,165 cases),the E2 stable group(Group B,E2 rate of change was within-30％～30％,162 cases),and the E2 decline group(E2 value declined more than 30％,11 cases,not included in statistical a-nalysis due to small sample size).The differences in demographic characteristics,ovulation induction outcomes,embryo outcomes and clinical pregnancy-related indicators were analyzed between Groups A and B.Results There were no statistically significant differences in basic information such as age,duration of infertility,body mass index(BMI),basal endocrine levels,and anti-Müllerian hormone(AMH)levels between groups A and B.Regarding embryo characteristics,there were no statistically significant differences in fertilization method and number of trans-ferred embryos between groups A and B.However,group A had a higher total number of retrieved oocytes,normal fertilization rate,number of high-quality embryos,and rate of high-quality embryos compared to group B,with sta-tistically significant differences(P＜0.05).In terms of clinical pregnancy outcomes,there were no statistically significant differences in the incidence of severe ovarian hyperstimulation syndrome(OHSS)between groups A and B.However,group A had higher rates of clinical pregnancy,implantation,and live birth compared to group B,with statistically significant differences.Group A also had a lower rate of early miscarriage compared to group B,with statistically significant differences.Conclusion Choosing the GnRH-ant protocol for IVF/ICSI-FET in PCOS patients,if the blood E2 level increases by more than 30％after 4 days of adding the antagonist(blood sample col-lected before administering GnRH-ant),the clinical pregnancy outcome will be better.

WRKY transcription factor is a type of transcription factor unique to plants and plays an important role in various physiological processes of plants. This study is based on the transcriptome data of Atractylodes lancea, the correlation between the FPKM values of the AlWRKY gene and the AlTPS gene of Atractylodes lancea was analyzed. Combined with the analysis results, the candidate gene AlWRKY65 with a significant positive correlation with the expression levels of AlTPS1 and AlTPS6 genes and a relatively high FPKM value was screened. The candidate gene was cloned to obtain the open reading frame ORF of the AlWRKY65 gene, and the related information of its encoded protein was analyzed and the gene expression was studied. The results showed that AlWRKY65 contains a 681 bp open reading frame and encoding 226 amino acids. Through amino acid sequence homology analysis, it was found that AlWRKY65 amino acid sequence had high homology with several plants such as HaWRKY65 and LsWRKY65; AlWRKY65 protein had a typical WRKYGQK domain, belonging to IIe subgroup of WRKY transcription factor family; phylogenetic analysis indicated that AlWRKY65 protein had the higher homology with CcWRKY65 protein; the expression of AlWRKY65 gene in different tissues of two producing areas of Atractylodes lancea were assayed via real-time fluorescence quantitative PCR, and the results showed that all of them were highly expressed in leaves and also has tissue differences. The expression level of AlWRKY65 gene was down-regulated within 48 h of methyl jasmonate (MeJA) induction; subcellular localization and transcriptional activation assay suggested that AlWRKY65 was located in the nucleus and had no transcriptional activation activity. This study provides a reference for further elucidating the biological function of AlWRKY65 in Atractylodes lancea.

Objective:To explore the correlation of early urinary continence(UC)after laparoscopic radical prostatectomy(LRP)with relevant preoperative MRI parameters of the urinary tract structure and provide some theoretical evidence for screening the high-risk population with postoperative urinary incontinence.Methods:This study included 49 PCa patients aged 50-78 years trea-ted by LRP in Beijing Tongren Hospital from January 2015 to February 2021,and all followed up for 12 months.We collected the com-plete baseline data on the patients,the clinical data possibly related to early postoperative UC,the MRI anatomical parameters associat-ed with UC,and the data on the recovery of early postoperative UC.We recorded the number of urinary pads used,submitted the data obtained to SPSS 23.0 statistical analysis,and identified the possible relevant factors by univariate correlation analysis,followed by R40.3 or SPSS 23.0 multivariate logistic regression analysis of the included factors and the results of UC.Results:MRI images mani-fested that the prostate anteroposterior diameter averaged(4.0±1.11)cm,the transverse diameter(4.6±0.83)cm,the cephalo-caudal diameter 2.4-6.4 cm,the membranous urethral length(MUL)(13.16±3.52)mm,and the thickness of the urethral rhab-dosphincter(URS)1.08-4.37 mm.Multivariate analysis showed that age was significantly correlated with the recovery of UC at 1 month after LRP(P=0.035,OR=0.16),and so was the URS thickness at 3 months(P=0.011,OR=0.02),9 months(P=0.014,OR=0.039)and 12 months(P=0.014,OR=0.039).Urinary incontinence with the URS thickness ≤1.6 mm at 12 months after operation was found of a high severity(P=0.010,OR[95％CI]=0.858-6.240).The MUL was positively correla-ted with the recovery of UC at 9 months(P=0.024,OR=0.508)and 12 months(P=0.024,OR=0.508)postoperatively.Correlation analysis revealed that the prostate volume,prostate diameter and other factors included in this study were not significantly correlated to postoperative UC.Conclusion:The thickness of the URS is positively correlated with the recovery of early UC,the thinner the URS,the severer the UC,and so is MUL.Age is an independent risk factor for the recovery of UC at 1 month after LRP.These findings need to be further verified by more prospective studies with long-term follow-ups.

Objective:To identify the key genes involved in the development and progression of prostate cancer(PCa)and those associated with the prognosis of the malignancy.Methods:We obtained the single-cell sequencing data on 4 cases of PCa from the GSE156632 database.Using R language and the Seurat package,we performed cell clustering and annotation,selected the subpop-ulations of epithelial cells for differential analysis after quality control and cell type identification,and conducted enrichment analysis of the identified differential genes using the Hiplot website.Then we downloaded the single nucleotide polymorphism(SNP)loci corre-sponding to the expression quantitative trait loci(eQTL)of these genes from the UK Biobank(UKB)database,and the clinical data and corresponding gene expression data on PCa patients from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO),followed by univariate COX regression analysis of the impact of the genes on the prognosis of the patients after Mendelian ran-domization.Results:A total of 1 566 genes were identified and subjected to enrichment analysis,which indicated that the differenti-al genes might be enriched in the Ras,apoptosis and oxidative phosphorylation signaling pathways.Subsequent Mendelian randomiza-tion revealed 74 potential causal genes among the 1 566 genes,and univariate COX regression analysis of the 74 genes identified 4 pos-sibly related genes FAM3B,JUNB,TMEM59,and KRT5.Comparison of the results of Mendelian randomization and univariate COX regression showed that KRT5 might be the most important gene influencing PCa.Conclusion:FAM3B,JUNB,TMEM59 and KRT5 may play a role in the progression of PCa,and KRT5 may potentially serve as a prognostic predictor and therapeutic target for the malig-nancy.

Cerebrospinal fluid morphology examination is an important method of diagnosing central nervous system diseases, but manual microscopy has shortcomings such as low efficiency, long staff training period, and poor homogeneity of test results. In recent years, the application of artificial intelligence in the medical field has developed rapidly, providing new technical means for cerebrospinal fluid morphology examination. In the future, AI-assisted morphological examination of cerebrospinal fluid will not only realize digitalization and networking, but also improve the level and efficiency of intelligent diagnosis of cerebrospinal fluid morphology, which has a broad application prospect in the intelligent assisted diagnosis of cerebrospinal fluid.

Objective: To investigate the relationship between serum lysophosphatidylcholine (LPC) level and the health index of the elderly. Methods: A total of 251 subjects were selected from the 2016 baseline survey of the Yongfu Longevity Cohort in Guangxi Province among whom 66, 63 and 122 were in the young and middle-aged group (≤59 years old), the young group (60-89 years old) and the longevity group (≥90 years old), respectively. Demographic data were collected and related indicators of height, weight, blood pressure and lipid metabolism were measured. The cognitive and physical functions of the elderly were assessed by the results of the simple mental state scale and the daily living activity scale to construct the health index of the elderly. The serum levels of LPC16∶0, LPC18∶0, LPC18∶1 and LPC18∶2 were determined by liquid chromatography-tandem mass spectrometry, and the differences among different ages and health status groups were compared. The logistic regression model was used to analyze the relationship between the serum LPC level and the health index of the elderly. Results: With the increase in age, the proportion of female subjects increased, and the rate of smoking and drinking decreased. BMI, TC, TG, LDL-C, diastolic blood pressure, and the four LPCs levels decreased with the increase of age, and systolic blood pressure levels increased with the increase of age (all P values<0.05). There was no significant difference in HDL-C levels among age groups (P>0.05). With the decline of health status in the elderly, serum levels of LPC16∶0, LPC18∶0, LPC18∶1 and LPC18∶2 showed a downward trend (all P values<0.001). After adjusting for age and gender, only LPC18∶0 was associated with the health status in old age [OR (95%CI): 0.48 (0.25-0.92)]. For every 1 standard deviation (16.87 nmol/L) increase in serum LPC18∶0 concentration, the risk of poor health status in old age decreased by 52%. Conclusion: Serum LPC18∶0 was associated with the health status in old age independent of age and sex.
Aged ; Middle Aged ; Humans ; Female ; Aged, 80 and over ; Lysophosphatidylcholines ; Risk Factors ; China ; Longevity ; Surveys and Questionnaires ; Triglycerides

International research on healthy life expectancy (HALE) focuses on inequality of socioeconomic status and individual natural attributes. With the acceleration of population ageing and the increase in average life expectancy, the extension of unhealthy life expectancy and the increase of social and economic burden caused by diseases have gradually attracted the attention of countries around the world. Therefore, the evaluation of disease factors affecting HALE is a meaningful direction in the future. This study introduces the development process and commonly used measurement methods of HALE. According to the definition of health from the Global Burden of Disease Study and World Health Organization, physical and mental diseases such as cardiovascular and cerebrovascular diseases, chronic respiratory diseases, diabetes, malignant tumors and depression were selected to summarize the impact of these diseases and pre-disease states on HALE. It is expected to provide a theoretical basis for the formulation of relevant public health policies and the improvement of quality of life in China.
Humans ; Healthy Life Expectancy ; Quality of Life ; Life Expectancy ; Causality ; Social Class