Objective: To study the chemical constituents of Eleutherine americana. Methods: The compounds were isolated by various chromatographic techniques, including silica gel, TLC, sephadex LH-20, and semi-preparative HPLC, and their structures were identified by their physicochemical properties and 1H-NMR and 13C-NMR data. Results: Fifteen compounds were isolated from E. americana, which were germacrenin B (1), phaffiaol (2), 4,8-dihydroxy-3-methoxy-l-methylonthra-9,10-quinone-2-carboxylic acid methyl ester (3), 3-heptadecyl-5-methoxyphenol (4), ethyl linoleate (5), 3,5-dimethoxybiphenyl-4’-ol (6), karwinaphthol A (7), 5-hydroxylkarwinaphthol A (8), 3,4-dimethoxy-8-hydroxy-1-methyl-anthra-9,10-quinone-2-carboxylic acid methyl ester (9), isoeleutherin (10), eleutherin (11), isoeleutherol (12), naphtho-γ-lactone (+)-9-hydroxyeleutherol (13), senkyunone (14), and palmitoleic acid (15). Conclusion: Compounds 2, 4-7, and 13-15 were isolated from the genus for the first time. The compounds 3, 8, and 13 had strong vasodilator effects with diastolic rate of 85.3%, 81.8%, and 89.5%, respectively, which were basically equivalent to the positive drug of tanshinone IIA (86.3%).

Objective: To investigate the effects of antimicrobial peptides merecidin on the biological functions of human lung adenocarcinomaA549 cells and the potential signaling pathways and targets that involved in promoting apoptosis, by studying the changes of phosphorylation levels of proteins in A549 cells after merecidin treatment. Methods: The antibacterial peptide mericidin (9 μmol/L) was applied to treat A549 cells for 6 h, and the total protein was collected and extracted. The peptide was digested by trypsin and labeled with TMT, and then fractionated by HPLC. The phosphorylated peptides were enriched with IMAC-Fe, and finally subjected to mass spectrometry analysis. Library identification and quantification of phosphorylated peptides obtained by mass spectrometry were processed using MaxQuant software, to further analyze the functions and pathways of differentially expressed phosphorylated proteins by combining with bioinformatic analysis. Results: Through IPA analysis of phosphorylated proteins in the normal control group and the antibacterial peptide merecidin treatment group, 753 differentially phosphorylated proteins in mericidin treatment group were screened out under the conditions of |Fold Change|≥2 and P<0.05, including 229 significantly up-regulated genes and 417 down-regulated genes. Among them, the differentially expressed proteins related to apoptosis included RB1, MAPK1, ARAF, PTK2, FOXO, MARCKSandsoon.Theresultsofbiologicalprocessanalysisshowedthatdifferentiallyexpressed phosphorylated proteins were mainly concentrated in cell signal transduction, degradation and transport of nucleic acid, and cellular energy metabolism, protein translation and synthesis, and cytoskeleton formation etc. The enrichment results showed that the differentially expressed phosphorylated proteins were mainly involved in apoptosis-related MAPK, ErbB, PI3K-Akt, and Ras signaling pathways. Protein-protein interaction analysis indicated the associations among apoptosis-related proteins PTK2, PRKCA, MA2PK2, MAPK1, and LMNA. Conclusion: The antibacterial peptide merecidin may induce apoptosis and alteration of other cell functions by affecting a variety of genes and signaling pathways such as RB1, MAPK1,ARAF, PTK2, FOXO and MARCKS etc.

【Objective】 To explore the effects of anti-inflammatory of short-chain fatty acids in human peripheral blood mononuclear cells （THP-1） and chronic obstructive pulmonary （COPD） mice. 【Methods】 We adopted the methods of qRT-PCR， ELISA， Western blotting， HE staining and Sirius staining to detect the changes of inflammatory factors in THP-1 cells and COPD with or without short-chain fatty acids. 【Results】 The ELISA and qRT-PCR experiments showed that the serum inflammatory factors， including interleukin-6 （IL-6）， interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α）， were higher in clinical COPD patients than in healthy people. And the THP-1 cells expressed these inflammatory factors highly after LPS stimulation. Among three short-chain fatty acids treated， butyrate could reduce the levels of inflammatory factors better than acetate and propionate. After COPD mice were treated with butyrate in the drinking water， the mRNA expressions of inflammatory factors IL-6， IL-1β and TNF-α in the lung tissue decreased， and the concentrations of IL-6 and IL-1β in plasma decreased. The protein detection results showed that the phosphorylation expression of NF-κB decreased， and butyrate might inhibit the expressions of inflammatory factors by inhibiting the NF-κB signaling pathway. 【Conclusion】 The SCFA butyrate can inhibit LPS-induced inflammatory response of THP-1 cells and have an inhibitory effect on inflammation in COPD mice.

Objective:To investigate the effect of intervention on oral health of pre-pregnancy women before and after oral health education. Methods:A total of 40 pre-pregnancy women were selected from the Reproductive Medicine Center of General Hospital of Ningxia Medical University according to the inclusion criteria， general conditions， clinical evaluation of plaque and oral health education. Their oral health conditions were evaluated before and after oral health intervention. Results:Based on the oral health status survey， there were significant differences between before and after intervention （all P<0.001） in the following five items： “bleeding from brushing teeth”， “difficulty biting or chewing food”， “sensitivity of teeth or gums to cold， hot， or sweet stimuli”， "restriction of the type and amount of food eaten for dental reasons” and “medication for oral pain or discomfort”. There were significant differences between before and after intervention （all P<0.001） in four items of oral health care behavior including “How often do you brush your teeth？”， “How do you brush your teeth？”， “gargle after meals”， and “floss use or not” but showed no significant difference in toothbrush replacement （P=0.467）. There were significant differences （all P<0.001） in five items of oral health knowledge including “periodontal disease can lead to premature delivery of newborns”， “periodontal disease can lead to low birth weight of newborns”， “need oral examination before pregnancy”， “pregnancy prone to oral diseases”， “mid-pregnancy is the best period for the treatment of oral diseases”. The oral plaque index before intervention was 5.47±1.08 and reduced to 4.37±0.94 after intervention （t=7.93， P=0.001）. Conclusion:Through education intervention， the oral health status of pre-pregnancy women can be improved. The knowledge of oral health can be improved and the level of oral health care can be enhanced. Oral health intervention can effectively reduce the level of plaque in pre-pregnancy women and improve the efficiency of plaque clearance.

[摘 要] 目的：以磷酸化蛋白质组学技术分析抗菌肽merecidin处理人肺腺癌A549细胞后细胞内磷酸化蛋白质表达的差异，探究merecidin对肺腺癌A549细胞蛋白质活性、功能的影响以及涉及的信号通路。方法：采用9 μmol/L merecidin处理肺腺癌A549细胞6 h，收集并提取总蛋白，SDS-PAGE检验全蛋白提取效果，加入胰酶来对蛋白质进行酶解。酶解所获肽段用TMT标记、采用HPLC分级分离、经IMAC磷酸化修饰富集以及液相色谱-质谱联用（HPLC-MS/MS）分离肽段。使用localization probability>0.75的标准对鉴定数据进行过滤，利用GO（Gene Ontology）数据库、KEGG（KyotoEncyclopedia of Genes and Genomes）数据库和STRING数据库对磷酸化蛋白组学数据进行分析。结果：SDS-PAGE 结果显示，经9 μmol/L merecidin处理后的A549细胞全蛋白分离效果清晰、无明显降解，且实验组与对照组条带差异明显；质谱共鉴定出位于3 089个蛋白上的10 320个磷酸化修饰位点，以|Fold change|>或<1.5且P<0.05为阈值从中筛选出差异明显的753个蛋白质及其1 172个磷酸化位点。蛋白质功能富集显示，磷酸化水平显著变化的蛋白质功能主要集中在蛋白质分子结合、代谢活性、分子功能调节、细胞进程、生物功能调节等方面；整合通路生物信息学分析结果显示，差异蛋白与Ras、PI3K/AKT、mTOR、AMPK等多条通路相关联；经过COG数据库筛选，发现差异性磷酸化蛋白主要集中在细胞信号转导、RNA转录、翻译后加工和修饰、核糖体合成蛋白质、细胞骨架蛋白形成及细胞内的物质转运和分泌、囊泡运输等多个方面；蛋白质互相作用层面分析结果显示，merecidin处理后的A549细胞中形成以MAPK1、RPL23A、SRSF3H、NCBP1等为关键蛋白的相互作用网，其中ATG2B、ULK1等蛋白显著上调，MAPK1、AKT1等蛋白显著下调。结论：磷酸化蛋白组学分析结果显示，抗菌肽merecidin可能通过MAPK、RPL23A、SRSF3H和AKT1等关键蛋白质在多方面生物功能和多条信号通路中发挥作用，促进肺腺癌A549细胞凋亡和自噬，从而抑制细胞的增殖。

ObjectiveTo investigate current status of hospital infection management in psychiatric hospitals in the Ningxia Hui Autonomous Region， so as to provide references for improving the level of hospital infection management of psychiatric hospitals. MethodsIn December 2020， on-site supervision was conducted on hospital infection management in all 9 psychiatric hospitals in the Ningxia Hui Autonomous Region， meantime， the self-compiled questionnaire on hospital infection management status was used for investigation. ResultsAmong the selected hospitals， nine （100.00%） psychiatric hospitals had the main hospital leaders in charge of hospital infection management， five （55.56%） hospitals had established a hospital infection management committee， six （66.67%） hospitals had established an independent hospital infection management department， and one （11.11%） hospital had developed all 13 systems mentioned in the questionnaire related to hospital infection management and job responsibilities. In terms of hospital infection management staff， there were 23 staff members in the nine psychiatric hospitals， including 3 in the specialty （13.04%） and 20 in the part-time setting （86.96%）. The score of the implementation of the basic system of hospital infection management in nine hospitals was （3.28±2.22）. ConclusionThe system specification related to hospital infection management in the Ningxia Hui Autonomous Region psychiatric hospitals needs to be improved and further strengthened， the professionalism of hospital infection management personnel needs to be improved.

This study aims to observe the effect and explore the mechanism of Qirong Tablets in the treatment of premature ovarian insufficiency(POI) in mice via the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/hypoxia inducible factor 1(HIF-1) signaling pathway. Sixty SPF female BALB/c mice were randomly divided into normal group, model group, positive control group, Qirong Tablets low-, medium-and high-dose group. The normal group was intraperitoneally injected with the same amount of normal saline, and the other groups were intraperitoneally injected with cyclophosphamide 120 mg·kg~(-1)·d~(-1) once to establish a POI animal model. After the model was successfully established, the low-, medium-and high-dose groups of Qirong Tablets were administered orally with 0.6, 1.2, 2.4 mg·kg~(-1)·d~(-1) respectively. The positive control group was given 0.22 mg·kg~(-1)·d~(-1) Clementine Tablets by intragastric administration, and the normal group and model group were given intragastric administration with the same amount of normal saline, and the treatment was 28 d as a course of treatment. After drug intervention, enzyme-linked immunosorbent assay(ELISA) was employed to measure the levels of estradiol(E_2), follicle-stimulating hormone(FSH), luteinizing hormone(LH), and anti-mullerian hormone(AMH) in peripheral blood, and hematoxylin-eosin(HE) staining to observe the ovarian tissue. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay was used to detect the apoptosis of granulosa cells, and Western blot to determine the expression levels of B-cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), caspase-3, PI3K, Akt, and HIF-1. Compared with the normal group, the modeling of POI caused loose or destroyed ovarian tissue with vacuolar structures, edema and fibrosis in the ovarian interstitium, disordered or loose arrangement of granulosa cells, and reduced normal follicles. Compared with the model group, drug interventions restored the ovarian tissue and follicles at all the development stages and reduced atretic follicles. Compared with the normal group, the modeling of POI lowered the serum level of E_2 and AMH(P<0.01), and elevated the level of FSH and LH(P<0.01). Compared with the model group, high-dose Qirong Tablets elevated the levels of E_2 and AMH(P<0.05), and lowered the levels of FSH and LH(P<0.05). Compared with the normal group, the modeling of POI up-regulated the protein levels of PI3K, Akt, HIF-1, Bax, and caspase-3 and down-regulated the protein level of Bcl-2 in the ovarian tissue(P<0.01). Compared with the model group, low-, medium-, and high-dose Qirong Tablets down-regulated the protein levels of PI3K, Akt, HIF-1, Bax, and caspase-3 proteins and up-regulated the protein level of Bcl-2 in the ovarian tissue(P<0.05). In conclusion, Qirong Tablets can up-regulate the expression Bcl-2, down-regulate the expression of Bax and caspase-3 in POI mice. Qirong Tablets may inhibit the apoptosis of follicular granulosa cells in mice, thereby delaying ovarian aging, improving reproductive axis function, and strengthening ovarian reserve capacity, which may be associated with the inhibition of PI3K/Akt/HIF-1 pathway.
Humans ; Mice ; Female ; Animals ; Proto-Oncogene Proteins c-akt/metabolism* ; bcl-2-Associated X Protein ; Phosphatidylinositol 3-Kinases/metabolism* ; Caspase 3/metabolism* ; Saline Solution/therapeutic use* ; Signal Transduction ; Granulosa Cells ; Primary Ovarian Insufficiency/drug therapy* ; Follicle Stimulating Hormone/therapeutic use* ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Apoptosis

Objective: To investigate the influence of the Rho/ROCK signaling pathway on the anti-cryodamage ability of human sperm and provide some theoretical evidence for the development of high-efficiency semen cryoprotectants. METHODS: We collected semen samples from 25 healthy males, each divided into a fresh, a normal cryopreservation control and an Rho-inhibition group. Before and after freezing, we detected sperm motility, viability, membrane integrity, morphology, DNA fragmentation index (DFI), acrosomal enzyme activity (AEA) and mitochondrial membrane potential (MMP) and determined the expressions of RhoA and ROCK proteins in the sperm by immunofluorescence staining. RESULTS: Compared with the normal cryopreservation control, the frozen-thawed sperm of the Rho-inhibition group showed significantly increased sperm motility （ ［51.20 ± 7.70］% vs ［57.50 ± 6.83］%, P = 0.002), survival rate ( ［52.87 ± 5.07］% vs ［60.24 ± 5.53］%, P = 0.001), membrane integrity (［59.78±5.56］% vs ［67.10 ± 4.43］%, P = 0.001), percentage of morphologically normal sperm (［4.83 ± 1.11］% vs ［7.46 ± 1.28］, P = 0.001) and MMP (56.30 ± 4.28 vs 63.11 ± 2.97, P = 0.001), but decreased DFI (［27.64 ± 6.64］% vs ［18.87 ± 4.07］%, P = 0.001). There was no statistically significant difference in the AEA of the frozen-thawed sperm between the control and Rho-inhibition groups (97.65 ± 9.31 vs 98.30 ± 11.33, P > 0.05). Immunofluorescence staining revealed extensive expressions of RhoA and ROCK proteins in the head and neck of the sperm. CONCLUSIONS The Rho/ROCK signaling pathway plays a role in the cryodamage to human sperm, and inhibiting the activity of Rho/ROCK can significantly improve the ability of sperm to resist cryodamage.

Objective To investigate the expression of mitogen-activated protein kinase kinase 1 (MAP2K1) in gastric cancer and its clinical significance. Methods Immunohistochemistry and Western blotting were used to detect the protein expression of MAP2K1 in gastric cancertissues and cells. The morphology and the expression position of MAP2K1 were observed by immunofluorescence. MAP2K1 mRNA expression in gastric cancer tissues was analyzed by data mining of Starbase database and Oneomine database. The correlation between MAP2K1 mRNA expression and clinicopathological features was analyzed by UALCAN database. Survival analysis was performed using Kaplan Meier-Plotter online analysis tools. GEPIA2 database mining the relationship between MAP2K1 and gastric cancer stem cell related factors and drug resistance related factors. Results Immunohistochemistry, immunofluorescence and Western blotting showed that MAP2K1 protein was highly expressed in gastric cancer tissues and cells, and MAP2K1 was expressed in the cytoplasm of gastric cancer. According to the analysis of various databases, the expression of MAP2K1 mRNA in gastric cancer tissue was higher than that in normal gastric tissue, and the expression of MAP2K1 mRNA was closely related to gastric cancer stage, grade, lymph node metastasis and patient gender, and the overall survival rate of gastric cancer patients in the group with high MAP2K1 mRNA expression was significantly lower than that in the group with low MAP2K1 mRNA expression, which may be related to the characteristics of gastric cancer stem cells and drug resistance. Conclusion MAP2K1 is highly expressed in gastric cancer, and its expression level may affect the poor prognosis of patients by regulating stem cell related factors and drug resistance related factors. MAP2K1 may be a new diagnostic marker to determine the prognosis of gastric cancer patients.

OBJECTIVE: To study the role of gamma-delta T (γδ T) cells and its subsets in the immunopathogenesis of Henoch-Schönlein purpura (HSP) in children, and to provide new ideas for the treatment of HSP in children from the aspect of γδ T cell regulation. METHODS: A total of 33 children with HSP were enrolled as the HSP group, and 21 healthy children were enrolled as the healthy control group. The percentages of γδ T cells and its subsets Vδ1 T and Vδ2 T cells among peripheral blood mononuclear cells (PBMCs) were measured, as well as the apoptosis rate of γδ T cell and plasma level of interleukin-17 (IL-17). RESULTS: Compared with the healthy control group, the HSP group had significantly lower percentages of lymphocytes in PBMCs and Vδ2 T cells in γδ T cells (P<0.05). The HSP group had significantly higher percentage of Vδ1 T cells in γδ T cells and plasma level of IL-17 than the healthy control group. The HSP group had a significantly higher overall apoptosis rate of γδ T cells than the healthy control group (P<0.05), especially early apoptosis. The percentage of Vδ2 T cells was positively correlated with overall apoptosis rate (r=0.615, P<0.05) and was negatively correlated with IL-17 level (r=-0.398, P<0.05). CONCLUSIONS Vδ1/Vδ2 T cell immune imbalance mediated by γδ T cells and over-activation of IL-17 may be involved in the development of HSP, among which the disturbance of immune tolerance induced by Vδ2 T cells plays an important role in the pathophysiology of the disease.
Child ; Humans ; Leukocytes, Mononuclear ; Purpura, Schoenlein-Henoch ; Receptors, Antigen, T-Cell, gamma-delta ; T-Lymphocytes