Objective To develop an ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） method to simultaneously determine 10 main components, including berberine, phellodendrine, specnuezhenide, mangiferin, loganin, paeoniflorin, geniposide, baicalin, and acteoside in Compound Dihuang oral solution. Methods An UPLC-MS/MS method was established with an ACQUITY UPLC BEH-C18 （2.1 mm×100 mm, 1.7 μm）column and mobile phase of 0.1% formic water（A）-methanol solution（B） in a gradient elution manner. The flow rate of mobile phase was 0.2 ml/min.The temperature of column was 30℃. The injection volume was 2 μl. The MS detection was in MRM mode. Results 10 components in Compound Dihuang oral solution had a good linear relationship within their concentration range,and the precision, repeatability, stability and recovery met the requirements. The contents of berberine, phellodendrine, specnuezhenide, mangiferin, loganin, paeoniflorin, geniposide, baicalin, and acteoside in 7 batches of samples were （89.7-95.6） μg/ml, （164.0-177.7） μg/ml, （540.0-610.0） μg/ml, （408.7-429.0） μg/ml, （726.0-825.0） μg/ml, （503.7-572.0） μg/ml, （1380.0-1540.0） μg/ml, （2596.7-2896.7) μg/ml, （4866.7-5520.0) μg/ml, （61.8-67.2) μg/ml, respectively. Conclusion The established method was easy to be repeated and operated. The contents of 10 components in Compound Dihuang oral solution could be determined quickly and accurately, which provided a reference for the quality control of hospital preparation.

OBJECTIVES: To evaluate the association of GGN repeat polymorphism of androgen receptor (AR) with ovarian reserve and ovarian response in controlled ovarian stimulation (COS). METHODS: This genetic association study was conducted among a total of 361 women aged ≤40 years with basal FSH≤12 U/L undergoing the GnRH-agonist long protocol for COS in a university-affiliated IVF center. GGN repeat in the AR gene was analyzed with Sanger sequencing. The primary endpoint was the number of antral follicle counts (AFCs), and the secondary endpoints were stimulation days, total dose of gonadotropin (Gn) used, total number of retrieved oocytes, ovarian sensitivity index, and follicular output rate. RESULTS: The GGN repeat in exon 1 of the AR gene ranged from 13 to 24, and the median repeat length was 22. Based on the genotypes (S for GGN repeats <22, L for GGN repeats ≥22), the patients were divided into 3 groups: SS, SL, and LL. Generalized regression analysis indicated that the number of AFCs in group SS was significantly lower than those in group SL (adjusted β=1.8, 95% CI: 0.2-3.4, P=0.024) and group LL (adjusted β=1.5, 95% CI: 0.2-2.7, P=0.021). No significant difference was observed in the number of AFCs between group SL and group LL (P>0.05). Generalized regression analysis indicated no significant differences in ovarian stimulation parameters among the 3 groups, either before or after adjusting for confounding factors (P>0.05). CONCLUSIONS GGN repeat length on the AR gene is associated with AFC but not with ovarian response in Chinese women, indicating that AR gene polymorphisms may affect ovarian reserve.
Adult ; Female ; Humans ; Genotype ; Ovarian Follicle/cytology* ; Ovarian Reserve/genetics* ; Ovulation Induction/methods* ; Polymorphism, Genetic ; Receptors, Androgen/genetics* ; East Asian People/genetics*

Objective To investigate the expression of HOXB4 gene in patients with myelodysplastic syn-dromes(MDS)and its clinical significance in disease progression.Methods mRNA expression of HOXB4 gene in bone marrow mononuclear cells was detected by real-time fluorescence quantitative PCR(RT-qPCR),and the difference in HOXB4 expression was compared between 49 patients with MDS(MDS group)and 35 patients without MDS(group C).The relationship of mRNA expression of HOXB4 with disease characteristics and clinical prognosis was explored in MDS patients.Results mRNA expression level of HOXB4 gene was higher in MDS group than that in group C(P＜0.05).The patients were divided into a high-and a low-expression group according to the median expression level of HOXB4.Leukocyte count was lower in the high-expression group in the low-expression group at the time of initial diagnosis.The proportion of patients with subtypes of primitive cellular hyperplasia,poor prognostic staging and leukemic transformation was higher in the high-expression group than in the low-expression group.Conclusions mRNA expression level of HOXB4 gene has certain relation with AML transformation in MDS patients.

Background::The goal of the assisted reproductive treatment is to transfer one euploid blastocyst and to help infertile women giving birth one healthy neonate. Some algorithms have been used to assess the ploidy status of embryos derived from couples with normal chromosome, who subjected to preimplantation genetic testing for aneuploidy (PGT-A) treatment. However, it is currently unknown whether artificial intelligence model can be used to assess the euploidy status of blastocyst derived from populations with chromosomal rearrangement.Methods::From February 2020 to May 2021, we collected the whole raw time-lapse videos at multiple focal planes from in vitro cultured embryos, the clinical information of couples, and the comprehensive chromosome screening results of those blastocysts that had received PGT treatment. Initially, we developed a novel deep learning model called the Attentive Multi-Focus Selection Network (AMSNet) to analyze time-lapse videos in real time and predict blastocyst formation. Building upon AMSNet, we integrated additional clinically predictive variables and created a second deep learning model, the Attentive Multi-Focus Video and Clinical Information Fusion Network (AMCFNet), to assess the euploidy status of embryos. The efficacy of the AMCFNet was further tested in embryos with parental chromosomal rearrangements. The receiver operating characteristic curve (ROC) was used to evaluate the superiority of the model. Results::A total of 4112 embryos with complete time-lapse videos were enrolled for the blastocyst formation prediction task, and 1422 qualified blastocysts received PGT-A ( n = 589) or PGT for chromosomal structural rearrangement (PGT-SR, n = 833) were enrolled for the euploidy assessment task in this study. The AMSNet model using seven focal raw time-lapse videos has the best real-time accuracy. The real-time accuracy for AMSNet to predict blastocyst formation reached above 70% on the day 2 of embryo culture, and then increased to 80% on the day 4 of embryo culture. Combing with 4 clinical features of couples, the AUC of AMCFNet with 7 focal points increased to 0.729 in blastocysts derived from couples with chromosomal rearrangement. Conclusion::Integrating seven focal raw time-lapse images of embryos and parental clinical information, AMCFNet model have the capability of assessing euploidy status in blastocysts derived from couples with chromosomal rearrangement.

Objective:To investigate the protective effects of an anti-inflammatory mixture on acute lung injury (ALI) induced by sepsis in rats, as well as its possible mechanisms.Methods:A total of 40 Sprague-Dawley (SD) rats were randomly divided into the sham group, septic ALI model group (model group), 3-methyladenine (3-MA) control group, and anti-inflammatory mixture pretreatment group, with 10 rats in each group. Cecal ligation and perforation (CLP) was performed to reproduce a septic ALI model. The rats in the sham group only underwent opening and closing the abdomen without perforation and ligation. Both groups were given saline gavage and intraperitoneal injection for 3 consecutive days before surgery. The 3-MA control group was given intraperitoneal injection of saline and autophagy inhibitor 3-MA 15 mg/kg for 3 consecutive days before modeling. The anti-inflammatory mixture pretreatment group was given 8.8 mL/kg of anti-inflammatory mixture by gavage [the composition of anti-inflammatory mixture: rhubarb 15 g (after the next), coptis chinensis 15 g, baical skullcap root 12 g, magnoliae cortex 12 g, dahurian patrinia herb 30 g] and saline intraperitoneal injection for 3 consecutive days before modeling. The rats in each group were anesthetized 24 hours after surgery and died due to abdominal aortic blood collection. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum inflammatory cytokines interleukins (IL-1β and IL-6). Lung tissue was taken and then the bronchoalveolar lavage fluid (BALF) was collected, and the levels of IL-1β and IL-6 were detected by ELISA. Lung wet/dry weight (W/D) ratio was measured. After hematoxylin-eosin (HE) staining, the histopathological changes of the lungs were observed under light microscopy. Western blotting was used to detect the expression of autophagy markers microtubule-associated protein 1 light chain 3-Ⅱ/Ⅰ (LC3-Ⅱ/Ⅰ) and Beclin-1 protein in lung tissue. Autophagosomes in lung tissue were observed with transmission electron microscopy.Results:Compared with the sham group, the rats in the model group exhibited severe destruction of lung tissue structure, with significant infiltration of inflammatory cells, the lung W/D ratio and the levels of IL-1β and IL-6 in serum and BALF were significantly increased, the expressions of LC3-Ⅱ/Ⅰ and Beclin-1 protein were down-regulated, the autophagosomes were more. The rats in the 3-MA control group exhibited more severe lung tissue injury as compared with the model group, the lung W/D ratio and the levels of inflammatory cytokines in serum and BALF were further increased, the expressions of LC3-Ⅱ/Ⅰ and Beclin-1 protein still showed a decrease tendency as compared with the sham group, and the autophagosomes were less than that in the model group. Compared with the model group, the anti-inflammatory mixture pretreatment group showed milder lung tissue injury with a minimal amount of inflammatory cell infiltration, the lung W/D ratio was significantly reduced (7.07±1.02 vs. 11.33±1.85, P < 0.05), the levels of IL-1β and IL-6 in both serum and BALF were significantly decreased [IL-1β (ng/L): 26.04±3.86 vs. 40.83±5.46 in serum, 17.75±2.02 vs. 26.86±4.32 in BALF; IL-6 (ng/L): 91.28±10.15 vs. 129.44±13.05 in serum, 76.06±7.51 vs. 120.91±7.47 in BALF, all P < 0.05], and the ratio of LC3-Ⅱ/Ⅰ and Beclin-1 protein expression were significantly increased [LC3-Ⅱ/Ⅰ ratio: 1.23±0.02 vs. 0.60±0.02, Beclin-1 protein (Beclin-1/GAPDH): 2.37±0.33 vs. 0.62±0.05, both P < 0.05]. Furthermore, an increase in the number of autophagosomes was observed. Conclusion:The anti-inflammatory mixture improves lung injury in rats with sepsis induced by CLP and reduce inflammation levels, potentially through upregulation of Beclin-1-mediated autophagy.

Objective:To investigate the value of large-scale brain network research based on independent component analysis (ICA) in discovering the changes of intrinsic functional connections within and between resting-state networks (RSNs) in first-episode untreated adult patients with major depressive disorder (MDD).Methods:From January 2019 to June 2021, twenty-three patients with MDD (MDD group) and 30 healthy volunteers (HC group) matched with gender, age and years of education were selected. All participants underwent resting-state brain function imaging (rs-fMRI), and the MDD group completed the 17-item Hamilton depression scale(HAMD-17). The independent component analysis (ICA) method was used to analyze rs-fMRI data, and meaningful RSNs were obtained. SPM12 and Gift softwares were used to compare the strength of intrinsic functional connection within and between the RSNs of the MDD group and HC group, and the Pearson correlation analysis was conducted by IBM SPSS statistics 25.0 to evaluate the correlation between the functional connection strength and HAMD-17 scores in MDD group.Results:Compared with the HC group, intrinsic functional connection strength of medial prefrontal cortex (mPFC) (MNI: x, y, z=-6, 54, 25)in MDD group was significantly enhanced, while the intrinsic functional connection strength of the left angular gyrus (AG) (MNI: x, y, z=-48, -66, 21), the left precuneus (PCu) (MNI: x, y, z=-6, -63, 33), the left dorsolateral prefrontal cortex (dlPFC) (MNI: x, y, z=-36, 12, 51)and the right anterior insula (AI) (MNI: x, y, z=36, 21, 0)were significantly weakened. Compared with the HC group, functional connection strength between posterior default mode net work(pDMN) and anterior default mode network(aDMN) in MDD group was significantly weakened ( t=-2.206, P=0.032), and function connection strength between pDMN and left frontal parietal network(lFPN) was significantly strengthened ( t=2.318, P=0.025). In MDD group, intrinsic functional connection strength of mPFC and the functional connection strength of pDMN-lFPN were positively correlated with the HAMD-17 score ( r=0.524, P=0.010; r=0.441, P=0.035). Conclusion:Large-scale brain network study based on the ICA can find abnormal functional connections within and between RSNs in first-episode untreated adult patients with MDD, and provide objective imaging markers for the clinical diagnosis and treatment of MDD.

The dihydrofolate reductase (dhfr) and dihydropteroate synthetase (dhps) genes of Plasmodium vivax, as antifolate resistance-associated genes were used for drug resistance surveillance. A total of 375 P. vivax isolates collected from different geographical locations in China in 2009-2019 were used to sequence Pvdhfr and Pvdhps. The majority of the isolates harbored a mutant type allele for Pvdhfr (94.5%) and Pvdhps (68.2%). The most predominant point mutations were S117T/N (77.7%) in Pvdhfr and A383G (66.8%) in Pvdhps. Amino acid changes were identified at nine residues in Pvdhfr. A quadruple-mutant haplotype at 57, 58, 61, and 117 was the most frequent (57.4%) among 16 distinct Pvdhfr haplotypes. Mutations in Pvdhps were detected at six codons, and the double-mutant A383G/A553G was the most prevalent (39.3%). Pvdhfr exhibited a higher mutation prevalence and greater diversity than Pvdhps in China. Most isolates from Yunnan carried multiple mutant haplotypes, while the majority of samples from temperate regions and Hainan Island harbored the wild type or single mutant type. This study indicated that the antifolate resistance levels of P. vivax parasites were different across China and molecular markers could be used to rapidly monitor drug resistance. Results provided evidence for updating national drug policy and treatment guidelines.
Antimalarials/pharmacology* ; China/epidemiology* ; Drug Combinations ; Drug Resistance/genetics* ; Folic Acid Antagonists/pharmacology* ; Humans ; Mutation ; Plasmodium vivax/genetics* ; Prevalence

Streptomyces are major sources of bioactive natural products. Genome sequencing reveals that Streptomyces have great biosynthetic potential, with an average of 20-40 biosynthetic gene clusters each strain. However, most natural products from Streptomyces are produced in low yields under regular laboratory cultivation conditions, which hamper their further study and drug development. The production of natural products in Streptomyces is controlled by the intricate regulation mechanisms. Manipulation of the regulatory systems that govern secondary metabolite production will strongly facilitate the discovery and development of natural products of Streptomyces origin. In this review, we summarize progresses in pathway-specific regulators from Streptomyces in the last five years and highlight their role in improving the yields of corresponding natural products.
Biological Products ; Multigene Family ; Secondary Metabolism ; Streptomyces/genetics*

Angucyclines/angucyclinones are a large group of polycyclic aromatic polyketides and their producers are widely distributed in nature. This family of natural products attracts great attention because of their diverse biological activities and unique chemical structures. With the development of synthetic biology and the exploitation of the actinomycetes from previously unexplored environments, angucyclines/angucyclinones-like natural products with new skeletons were continuously discovered, thus enriching the structural diversity of this family. In this review we summarize the new angucyclines/angucyclinones analogues discovered in the last decade (2010-2020) by using different strategies, such as changing cultivation conditions, genetic modification, genome mining, bioactivity-guided compound isolation, and fermentation of actinomycetes from underexplored environments. We also discuss the role of synthetic biology in the discovery and development of new compounds of the angucycline/angucyclinone family.
Anthraquinones ; Biological Products ; Polyketides ; Streptomyces

Objective To explore how to optimize management process of pressure injury for surgery patients by applying information system.Methods We built an integrated surgery patients pressure injury management information system based on chain-type process along with controlling key links and realizing monitoring in a whole process and many links for surgery patients from be hospitalized to leave hospital.This research took the surgery patients pressure injury chain-type process as a breakthrough and selected 1240 patients as subjects in experimental operating room.A total of 578 patients were in control group in May 2016 before implementing information system and 662 patients were in observation group in May 2017 after implementing information system.We compared the qualified rate of pressure injury assessment,accuracy rate of screen for patients with high-risk pressure injury and being with pressure injury,accordance rate of description in skin condition during transportation and handover,the incidence of pressure injury in hospital and surgery patients in the first stage,the second stage and above.Results The qualified rate of pressure injury assessment,accuracy rate of screen for patients with high-risk pressure injury,accordance rate of description in skin condition during transportation and handover of observation group was 99.24％,100％and 100.00％significantly higher than those of control group(P< 0.05).The incidence of pressure injury in hospital and in the first stage of surgery patients was 2.95％and 2.65％in observation group with significant differences compared with those in control group(6.51％,5.82％)(P< 0.05).Conclusions The integrated surgery patients pressure injury management information system based on chain-type process realizes a full,continuous,accurate,dynamic evaluation and monitoring for patients' skin.It can effectively improve the nursing quality of pressure injury and is convenientfor managers to carry out a continuous improvement.