Imprinted genes play a key role in regulating mammalian placental and embryonic development. Here, we generated glutaminyl-peptide cyclotransferase-knockout (Qpct^-/-) mice utilizing the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) platform and identified Qpct as a novel anti-angiogenic factor in regulating mouse placentation. Compared with Qpct^+/+ mice, placentae and embryos (Qpct^-/+ and Qpct^-/-) showed significant overgrowth at embryonic Day 12.5 (E12.5), E15.5, and E18.5. Using single-cell transcriptome analysis of 32 309 cells from Qpct^+/+ and Qpct^-/- mouse placentae, we identified 13 cell clusters via single-nucleus RNA sequencing (snRNA-seq) (8880 Qpct^+/+ and 13 577 Qpct^-/- cells) and 20 cell clusters via single-cell RNA sequencing (scRNA-seq) (6567 Qpct^+/+ and 3285 Qpct^-/- cells). Furthermore, we observed a global up-regulation of pro-angiogenic genes in the Qpct^-/- background. Immunohistochemistry assays revealed a notable increase in the number of blood vessels in the decidual and labyrinthine layers of E15.5 Qpct^-/+ and Qpct^-/- mice. Moreover, the elevation of multiple pairs of ligand-receptor interactions was observed in decidual cells, endothelial cells, and macrophages, promoting angiogenesis and inflammatory response. Our findings indicate that loss of maternal Qpct leads to altered phenotypic characteristics of placentae and embryos and promotes angiogenesis in murine placentae.
Animals ; Female ; Pregnancy ; Mice ; Placentation/genetics* ; Single-Cell Analysis ; Gene Expression Profiling ; Mice, Knockout ; Transcriptome ; Placenta/blood supply* ; Neovascularization, Pathologic/genetics* ; Genomic Imprinting ; Single-Cell Gene Expression Analysis ; Angiogenesis

ObjectiveTo explore the suitability of four qualitative research methods in the development of TCM scale. MethodsTaking the development of "Seven Emotions Scale" as an example， we conducted semi-structured interviews with 31 patients of emotional disorders and 10 healthy people by objective sampling， and collected psychological feelings and emotional cognition data related to seven emotions according to the interview outline. Two qualitative methods， descriptive qualitative research and descriptive phenomenology， were used to analyze the data and construct the item library of the scale. The conceptual framework of the scale was constructed by using commonly used grounded theory and frame analysis. ResultsDuring data analysis， it is found that the themes extracted from descriptive phenomenology were not easily understood by the interviewees， and it is difficult for the researchers to truly achieve the "suspension" required by phenomenology. Considering the feasibility and convenience of the researchers' actual operation， as well as whether the initial purpose of the scale research can be intuitively included in the interviewees' views and feelings， descriptive phenomenology is not suitable for the formation of scale items. Using descriptive qualitative research method to analyze the interview data of healthy people and patients with emotional disorders， 306 and 476 scale items were obtained respectively. Through grounded theory， five selective codes were obtained： physical symptoms， external manifestations， psychological feelings， behaviors and emotional control. Using frame analysis， four themes including physical symptoms， psychological feelings， behavior and emotional cognition were constructed. Both methods can be used to construct the conceptual frame of scale， but the framework analysis is more convenient and can better ensure the transparency of the research. ConclusionDescriptive qualitative research methods can be used to form the item library of TCM scales. Framework analysis is more suitable for the construction of the conceptual framework of the scale than grounded theory， while descriptive phenomenology is not suitable for the development of TCM scales.

Objective To analyze the causal relationship of gluteal tendinitis and primary coxarthrosis with the occurrence of iliotibial band syndrome using Mendelian randomization.Methods The GWAS data of gluteal tendinitis,primary coxarthrosis and iliotibial band syndrome were screened for high correlation single-nucleotide polymorphisms(SNPs).Mendelian randomization analysis was performed using random-effects inverse variance weighting(IVW),MR-Egger regression,and weighted median method to determine whether gluteal tendinitis and primary coxarthrosis were causally related with iliotibial band syndrome.Heterogeneity test,multiple validity test and sensitivity analysis,and clinical data analysis were used to verify the reliability of the results.Results Both gluteal tendinitis[IVW:OR(95%CI)=1.32(1.03-1.68),P=0.026]and primary coxarthrosis[IVW:OR(95%CI)=1.40(1.06-1.84),P=0.017]was positively correlated with iliotibial band syndrome.Conclusion Gluteal tendinitis and primary coxarthrosis may increase the risk of iliotibial band syndrome.

Objective To analyze the causal relationship of gluteal tendinitis and primary coxarthrosis with the occurrence of iliotibial band syndrome using Mendelian randomization.Methods The GWAS data of gluteal tendinitis,primary coxarthrosis and iliotibial band syndrome were screened for high correlation single-nucleotide polymorphisms(SNPs).Mendelian randomization analysis was performed using random-effects inverse variance weighting(IVW),MR-Egger regression,and weighted median method to determine whether gluteal tendinitis and primary coxarthrosis were causally related with iliotibial band syndrome.Heterogeneity test,multiple validity test and sensitivity analysis,and clinical data analysis were used to verify the reliability of the results.Results Both gluteal tendinitis[IVW:OR(95%CI)=1.32(1.03-1.68),P=0.026]and primary coxarthrosis[IVW:OR(95%CI)=1.40(1.06-1.84),P=0.017]was positively correlated with iliotibial band syndrome.Conclusion Gluteal tendinitis and primary coxarthrosis may increase the risk of iliotibial band syndrome.

Objective:To classify the etiology of secondary hemophagocytic syndrome (sHLH) and explore its clinical, laboratory and therapeutic characteristics in order to deepen the understanding of the disease.Method:A retrospective observational study was conducted on sHLH patients who were treated at Peking University People's Hospital from January 2016 to December 2021. Patients under the age of 18 and those with missing clinical data were excluded. The distribution of departments visited and etiologies of sHLH were analyzed. Baseline data, clinical characteristics, complications, laboratory data, treatment, and in-hospital outcomes of sHLH were collected. The sHLH patients were then divided into 3 groups including malignancy group, macrophage activation syndrome (MAS) group and other etiologies (mainly infection) group. Intergroup comparisons were performed using chi-square tests, analysis of variance, Mann-Whitney tests, and other statistical methods.Results:A total 169 patients were enrolled, among these patients, 27.8% were malignancy-related HLH, 47.9% were MAS, and 24.3% were other etiologies related HLH. Statistical analysis revealed that the clinical characteristics of other etiological group was highly consistent with the malignancy group, including more and severer peripheral blood cell reduction, higher sCD25 levels, more Epstein-Barr virus infection, and the prognosis was similar, both were with more than 50% in-hospital mortality. And the incidence of hemophagocytosis was highest in other etiological groups (65.9%). In contrast, MAS group was with an obviously lower mortality of 17.3% ( P<0.05). Meanwhile, treatments including methylprednisolone pulse, cyclosporine A and interleukin-2 were used frequently in MAS group. Conclusion:Malignancy related HLH and other etiologies related HLH exhibit more similar clinical characteristics and prognosis, while the MAS group, has a milder overall condition and better prognosis.

Alcoholic liver disease (ALD) is a growing global health concern, and its early pathogenesis includes steatosis and steatohepatitis. Inhibiting lipid accumulation and inflammation is a crucial step in relieving ALD. Evidence shows that puerarin (Pue), an isoflavone isolated from Pueraria lobata, exerts cardio-protective, neuroprotective, anti-inflammatory, antioxidant activities. However, the therapeutic potential of Pue on ALD remains unknown. In the study, both the NIAAA model and ethanol (EtOH)-induced AML-12 cell were used to explore the protective effect of Pue on alcoholic liver injury in vivo and in vitro and related mechanism. The results showed that Pue (100 mg·kg^-1) attenuated EtOH-induced liver injury and inhibited the levels of SREBP-1c, TNF-α, IL-6 and IL-1β, compared with silymarin (Sil, 100 mg·kg^-1). In vitro results were consistent within vivo results. Mechanistically, Pue might suppress liver lipid accumulation and inflammation by regulating MMP8. In conclusion, Pue might be a promising clinical candidate for ALD treatment.

【Objective】   To explore the prescriptions related to the treatment of spleen and stomach diseases in Prescriptions of Peaceful Benevolent Dispensary (Tai Ping Hui Min He Ji Ju Fang,《太平惠民和剂局方》, TPHMHJJF), and investigate the medication and prescription rules. 【Methods】   The prescriptions of TPHMHJJF for treating spleen and stomach diseases were screened, and the data set was established by entering the prescriptions and standardized drug names using WPS Excel 2019. Herb frequency statistics, efficacy categorization, property, flavor, meridian tropism, association rules, cluster analysis, factor analysis, and complex network analysis were performed using Python 3.6.8 programming language and Gephi 0.9.2 visualization software. 【Results】   A total of 239 prescriptions were included after the screening, and the dosage forms were mainly pills and powders. The study involved 200 herbal medicines, among which 40 herbs had a frequency of ≥ 12. The herb with the highest frequency was Ganjiang (Zingiber Rhizoma). The herb properties were mainly warm in nature, with the most pungent herbs. Most herbs were attributed to the spleen and stomach meridians, and tonifying medicine were the main species. The association rule analysis identified 26 second-order association rules and 16 third-order association rules, with "Renshen (Ginseng Radix et Rhizoma) → Fuling (Poria)" as the leading rule in the former and "Fuling (Poria) + Baizhu (Atractylodis Macrocephalae Rhizome) → Renshen (Ginseng Radix et Rhizoma)" as the leading rule in the latter. Also, cluster analysis divided the top 30 herbs into six herb combinations that can warm the middle, move Qi, and dry dampness effectively. Factor analysis extracted 13 common factors, with Renshen (Ginseng Radix et Rhizoma), Fuling (Poria), and Baizhu (Atractylodis Macrocephalae Rhizome) as the highest contributing factors. Lastly, complex network analysis yielded the core prescription of 14 herbs, with Ganjiang (Zingiber Rhizoma), Renshen (Ginseng Radix et Rhizoma), Gancao (Glycyrrhizae Radix et Rhizome), and Chenpi (Citri Reticulatae Pericarpium) among the herbs with a higher weighting degree. 【Conclusion】  TPHMHJJF is mainly based on the use of warming herbs to treat spleen and stomach diseases, and its medication rule can be summarized into three aspects: (i) primarily using warming and tonifying to warm the middle and dissipate cold, (ii) using the method of moving Qi with aromatic nature to delight the spleen and appetize the stomach, and (iii) emphasizing the regulation of Qi and blood to calm the five zang-organs.

Objective:To compare the application of three different glomerular filtration rate(eGFR)-estimating equations in the elderly aged 60 years and over.Methods:The patients aged ≥ 60 years in our hospital from January 2012 to October 2017 were included as research subjects who underwent three GFR detection(as measured GFR i. e.mGFR)of serum creatinine(Scr), serum cystatin C(sCysC)and GFR by 99mTc-DTPA renal dynamic imaging.The advantages and disadvantages of each GFR-estimating(eGFR)equation in the elderly patients were compared.Results:A total of 122 patients were enrolled, including 90 males(73.8%), with a median age of 77 years.The median Scr and sCysC were 109 μmol/L and 1.39 mg/L, respectively, with their average of mGFR being(45.70±18.91)ml·min -1·1.73m -2.Overall, each eGFR-Scr equation over-estimated the GFR in varying degrees.In eGFR-Scr, full age spectrum equation(FAS)-Scr had the smallest bias(2.34)and the best accuracy(P30 75.4%), followed by Berlin Initiative Study(BIS)-Scr(P30 71.3%). In eGFR-CysC, the P30 of FAS-CysC and chronic kidney disease(CKD)-Epidemiology Collaboration(EPI)-CysC were 75.4% and 71.3%, respectively, and the accuracy of eGFR-CysC was comparable to that of BIS-Scr and FAS-Scr.In eGFR-combi, the bias of FAS-combi was the smallest(1.10), and the accuracy of FAS-combi, BIS-combi and CKD-EPI-combi was comparable, the P30 was 81.1%, 79.5% and 74.6%, respectively, and the P30 of FAS-combi and BIS-combi was higher than that of eGFR-Scr and eGFR-CysC.In the 60-80 age group, the accuracy of FAS-combi and BIS-combi was the best, P30 was 77.8% and 76.4% respectively, while, the accuracy of other equations were poor.In patients ≥ 80 years old, except for the modification of diet in renal disease(MDRD)equations and CKD-EPI-scr equation, the accuracy of other equations was acceptable, among which the accuracy of eGFR-combi equation was the best, and the P30 of FAS-combi, BIS-combi and CKD-EPI-combi were 86.0%, 84.0% and 80.0%, respectively. Conclusions:In elderly patients aged 60 years and over, the application of eGFR-combi equation is better than that of the respective eGFR-Scr and eGFR-CysC equations.The FAS equation has the best bias and accuracy, followed by the BIS equation.

Objective:To compare the advantages and disadvantages of several formulas for estimated glomerular filtration rate (eGFR) based on serum creatinine in elderly patients with chronic kidney disease (CKD) over 60 years old.Methods:CKD patients aged≥60 years old in Beijing Hospital from January 2012 to October 2017 were selected as subjects. Measured glomerular filtration rate (mGFR) was detected by 99mTc-DTPA renal dynamic imaging and used as a reference standard. According to the mGFR value, the patients were divided into 4 groups: mGFR<30 ml·min -1·(1.73 m 2) -1 group, 30≤mGFR<45 ml·min -1·(1.73 m 2) -1 group, 45≤mGFR<60 ml·min -1·(1.73 m 2) -1 group and mGFR≥60 ml·min -1·(1.73 m 2) -1 group. The deviation of each formula was compared by Bland-Altman scatter chart, and the accuracy of each formula was evaluated by the proportion of eGFR within mGFR (1±30%) ( P30) and root mean square error ( RMSE). Wilcoxon paired rank sum test was used to compare the deviation of each formula, and McNemar test was used to compare the difference of P30 among these formulas. Results:A total of 628 patients with CKD were enrolled in this study. The median age was 76.0(71.0, 81.0) years old. The median serum creatinine and mGFR were 110.0(86.0, 152.0) μmol/L and 42.90(29.88, 55.68) ml·min -1·(1.73 m 2) -1, respectively. Each eGFR formula based on serum creatinine overestimated glomerular filtration rate in varying degrees. Among them, the accuracy of Berlin Initiative Study (BIS) formula and full age spectrum (FAS) formula was the best ( P30 were 68.3% and 68.0% respectively), followed by the Chinese race coefficient of Chronic Kidney Disease Epidemiology Collaboration (C-CKD-EPI) formula ( P30 was 65.4%). The accuracy of the other formulas was poor. In terms of deviation, C-CKD-EPI formula was the best (0.27). In the group of mGFR<30 ml·min -1·(1.73 m 2) -1, the accuracy of all formulas was poor, and the accuracy of FAS formula was slightly better than that of other formulas ( P30 was 51.0%). In the group of 30≤mGFR<45 ml·min -1·(1.73 m 2) -1, the deviation of C-CKD-EPI formula was the smallest (3.11). In terms of accuracy, BIS and FAS formulas were better than others, and the P30 were 64.6% and 63.0% respectively. In the group of 45≤mGFR<60 ml·min -1·(1.73 m 2) -1, the deviation of C-CKD-EPI formula was also the smallest (0.72), and the accuracy of BIS formula was the best ( P30 was 82.5%), followed by FAS formula ( P30 was 79.7%). In the group of mGFR≥60 ml·min -1·(1.73 m 2) -1, the deviation and accuracy of Xiangya formula were the best (the deviation and P30 were -0.53 and 96.5% respectively), and the P30 of BIS and C-CKD-EPI formulas were 87.6% and 87.6%, respectively. Conclusions:In the elderly patients with CKD over 60 years old, the accuracy of eGFR based on serum creatinine increases with the increase of mGFR. BIS and FAS formulas are recommended first. The accuracy of each formula is poor in patients with severe renal insufficiency.