Objective To explore the mechanism of Panax notoginsenosides in the treatment of sepsis based on network pharmacology and molecular docking technology.Methods The action targets of total saponins of Panax notoginsenosides were obtained by searching the databases of TCMSP,Swiss Target Prediciton and PharmMapper,and the disease-related targets of sepsis were searched in the databases of Genecard,Drugbank,Disgenet and OMIM.The selected targets of total saponins of Panax notoginsenosides were intersected with the disease-related targets of sepsis,which were used as potential targets for the treatment of sepsis.The protein-protein interaction(PPI)network of potential targets was constructed by STRING database,and the key targets were screened;the potential targets were analyzed by GO function and KEGG pathway enrichment analysis,and the drug-disease-target-pathway network was constructed;the molecular docking of five monomer saponins of Panax notoginsenosides and the key targets of Panax notoginsenosides in the treatment of sepsis was studied by AutoDock Vina software.Results A total of 206 potential targets of Panax notoginsenosides in the treatment of sepsis were obtained,and key targets such as AKT1,TP53,SRC,STAT3,JUN,TNF,IL6,MAPK1,PIK3R1 and IL1B were screened.A total of 2 548 biological process(BP)items,174 molecular function(MF)items and 47 cellular component(CC)items were obtained by GO functional enrichment analysis of potential targets,and 171 signal pathways were obtained by KEGG pathway enrichment analysis.The main active components of Panax notoginsenosides R1,ginsenoside Rg1,ginsenoside Re,ginsenoside Rb1 and ginsenoside Rd have strong binding activity with key targets AKT1,TP53,STAT3,SRC and JUN.Conclusion Panax notoginsenosides may act on the main signal pathways such as PI3K-Akt and AGE-RAGE through the key targets such as AKT1,TP53,SRC,STAT3 and TNF,and then affect the physiological processes such as inflammation,apoptosis and blood coagulation in sepsis,and play a role in the treatment of sepsis.

[摘 要] 目的：探讨抑制素β亚基A反义RNA1（INHBA-AS1）对宫颈癌HeLa细胞EMT和鸟氨酸代谢途径的影响及其机制。方法：体外常规培养HeLa细胞，实验分为10组：对照组、阴性对照（NC）组、sh-INHBA-AS1组、PluriSIn 1[硬脂酰辅酶A去饱和酶（stearyl CoA desaturase，SCD）抑制剂]组、NC+PluriSIn 1组、sh-INHBA-AS1+PluriSIn 1组、10058-F4（c-Myc抑制剂）组、NC+10058-F4组、sh-INHBA-AS1+10058-F4组、sh-INHBA-AS1+OE-c-Myc组。平板克隆实验检测各组细胞的增殖能力，FCM检测各组细胞的凋亡情况，Transwell小室实验检测各组细胞的侵袭、迁移能力，qPCR法检测各组细胞中INHBA-AS1、c-Myc、SCD和EMT相关基因（N-cadherin、TGF-β、ZEB1）mRNA的表达，WB法检测各组细胞中c-Myc、SCD、EMT相关（N-cadherin、TGF-β、ZEB1）、S-腺苷-甲硫氨酸脱羧酶（SAMDC）和亚精胺/精胺N1-乙酰转移酶（SSAT）蛋白的表达，ELISA检测各组细胞上清液中鸟氨酸脱羧酶（ODC）的含量。结果：敲减INHBA-AS1表达使HeLa细胞的增殖、侵袭和迁移能力显著降低（均P<0.05）而细胞凋亡率显著升高（P<0.05），qPCR、WB法检测结果显示，敲减INHBA-AS1均可显著抑制HeLa细胞中c-Myc、SCD、N-cadherin、TGF-β、ZEB1和SAMDC的表达（均P<0.05），而促进SSAT的表达（P<0.05），并降低HeLa细胞上清液中ODC的含量（P<0.05）。与c-Myc抑制剂和SCD抑制剂单独处理相比，其联合敲减INHBA-AS1后上述作用更加显著（均P<0.05）；与sh-INHBA-AS1组相比，进一步过表达c-Myc后HeLa细胞的增殖能力显著升高（P<0.05）、SCD和N-cadherin蛋白表达水平显著升高（P<0.05）、细胞上清液中ODC含量显著升高（P<0.05）。结论：INHBA-AS1可通过c-Myc调控SCD的表达，从而影响HeLa细胞鸟氨酸代谢和EMT进程，进而促进HeLa细胞的增殖、侵袭和迁移能力。

The clinical data of a child with chemical pneumonia caused by kerosene in Hainan Maternal and Children′s Medical Center in June 2019 were retrospectively analyzed.The patient was a 2 years and 1 month old boy with a history of kerosene inhalation and fever.The clinical features included low breath sounds in the left lung and dry and wet rales in both lungs.The white blood cells (WBC) level, C-reactive protein (CRP) level, and erythrocyte se-dimentation rate (ESR) were significantly increased.Chest CT showed inhalation pneumonia.Chest ultrasound suggested medium pleural effusion on the left side.The patient was given antibiotics, nebulization and other treatment.On the 12 th day of the course of the disease, his temperature returned to normal, and breath sounds on the left side were stronger than before.The WBC level, CRP level and ESR were improved according to the re-check results, but pulmonary ventilation was still obstructed mildly to moderately.Fourteen days after hospital discharge, the patient coughed less.Reexamination of chest CT prompted the lesions were further absorbed, but the mild to moderate obstructive lesions were still observed.With the reduction of kerosene use in daily life, kerosene-induced chemical pneumonia is rare, but due to its diverse and complex clinical manifestations and slow absorption of pulmonary inflammation, attention should be paid to its progression into chronic cough.

The E class MADS-box genes SEPALLATA (SEP)-like play critical roles in angiosperm reproductive growth, especially in floral organ differentiation. To analyze the sequence characteristics and spatio-temporal expression patterns of E-function MADS-box SEP-like genes during kale (Brassica oleracea L. var. acephala) flower development, BroaSEP1/2/3 (GenBank No. KC967957, KC967958, KC967960) homologues, three kale SEP MADS-box gene, were isolated from the kale variety 'Fourteen Line' using Rapid amplification of cDNA ends (RACE). Sequence and phylogenetic analysis indicated that these three SEP genes had a high degree of identity with SEP1, SEP2, SEP3 from Brassica oleracea var. oleracea, Brassica rapa, Raphanus sativus and Brassica napus, respectively. Alignment of the predicted amino acid sequences from these genes, along with previously published subfamily members, demonstrated that these genes comprise four regions of the typical MIKC-type MADS-box proteins: the MADS domain, intervening (I) domain and keratin-like (K) domain, and the C-terminal domain SEPⅠ and SEP Ⅱ motif. The longest open reading frame deduced from the cDNA sequences of BroaSEP1, BroaSEP2, and BroaSEP3 appeared to be 801 bp, 759 bp, 753 bp in length, respectively, which encoded proteins of 266, 252, and 250 amino acids respectively. Expression analyses using semi-quantitative RT-PCR and quantitative real-time PCR indicate that BroaSEP1/2/3 are specifically expressed in floral buds of kale during flower development process. The expression levels of the three genes are very different at different developmental stages, also in wild type, mutant flower with increased petals, and mutant flower with decreased petals. These different patterns of gene expression maybe cause the flowers to increase or decrease the petal number.
Brassica/metabolism* ; Flowers/genetics* ; Gene Expression Regulation, Plant ; MADS Domain Proteins/metabolism* ; Phylogeny ; Plant Proteins/metabolism*

Objective: To investigate the regulatory effect of miR-1297 on the malignant biological behaviors of breast cancer cells and its underlying mechanism. Methods: Twenty pairs of breast cancer tissues and para-cancer tissues resected at the Department of Thyroid and Breast Surgery of Leshan People′ s Hospital from May 2016 to May 2018, as well as breast cancer cell lines MCF-7, SW626, HCC1937 and human breast epithelial MCF-10A cells were collected for this study. qPCR was performed to evaluate the expression of miR-1297 in breast cancer tissues and cell lines. The experimental cells were divided into control group, miR-1297 inhibitor group; TET3 over-expression group and simultaneous over-expression of TET3 and miR-1297 group. CCK-8 assay was used to detect the cell proliferation of MCF-7 cells; Transwell assay was carried out to detect the migration and invasion of MCF-7 cells; and WB was used to measure the expressions of TET3 and EMT related proteins (E-cadherin, N-cadherin and vimentin). Dual luciferase reporter gene assay was used to verify the relationship between miR-1297 and TET3. Results: miR-1297 was up-regulated in both breast cancer tissues and cell lines (P<0.01 or P<0.05). Knockdown of miR-1297 dramatically repressed the proliferation, migration, invasion and EMT of MCF-7 cells (P<0.01 or P<0.05). Over-expression of TET3 significantly up-regulated the expression of TET3 in MCF-7 cells (P<0.05). Simultaneous over-expression of TET3 and miR-1297 could reverse the expression level of TET3 in MCF-7 cells and the inhibitory effect of TET3 on the proliferation, migration, invasion and EMT of MCF-7 cells. Dual luciferase reporter gene assay results showed that miR-1297 targetedly bound to the 3' UTR of TET3. Further experiment results demonstrated that miR-1297 targetedly down-regulated TET3 and promoted the malignant biological behaviors of MCF-7 cells. Conclusion: miR-1297 is up-regulated in breast cancer tissues and cells; it promotes the malignant biological behaviors such as proliferation, migration, invasion and EMT through targetedly down-regulating the expression of TET3.

Objective: To detect the single nucleotide polymorphism (SNP) at locus 1 165 of β1-adrenoceptor (β1-AR) and to investigate the association between the SNP and the infection by enterovirus A71(EV-A71). Methods: Polymerase chain reaction (PCR) amplification technique was used to detect the SNP at locus 1 165 of β1-AR between hand, foot and mouth disease (HFMD) and healthy controls by sanger sequencing method . Results: There was a G1165C SNP and three kinds of genotypes (GG, GC, CC) in β1-AR gene in the 77 cases of EV-A71 HFMD patients and 66 cases of healthy controls. For HFMD patients, frequencies of GG, GC and CC genotypes of the G1165C locus were 10%, 47% and 43%, respectively, and alleles frequency of G and C were 34% and 66%, respectively. But in healthy children, GG, GC, CC genotype frequencies were 7%, 41% and 52%, respectively, and G and C allele frequencies were 28% and 72% respectively. Chi-square analysis showed that there were no significant differences in distribution of genotypes (χ²=1.154, df=2, P=0.562) and alleles frequency (χ²=1.091, df=2, P=0.296) between the EV-A71-infected group and the healthy control group. Between mild and severe EV-A71-infected group, there were no significant differences in distribution of genotypes (χ²=3.945, df=2, P=0.139) and alleles frequency (χ²=3.763, df=2, P=0.052). Conclusions The 1 165 SNP in the coding region of β1-AR was not associated with EV-A71 infection and its severity.

Objective To study the clinical value of electronic bronchoscope in diagnosis and treatment of recurrent dyspnea in neonates.Method From October 2014 to October 2017,the clinical data of recurrent dyspnea receiving electronic bronchoscopy examination and treatment in the neonatal intensive care unit of our hospital were retrospectively selected.Their clinical characteristics and treatment effects were summarized and analyzed.Result A total of 171 infants of neonatal recurrent respiratory infections were examined using electronic bronchoscope.The top four causes included endo－tracheo－bronchitis in 78 cases (45.6%), laryngomalacia, and tracheobronchomalacia in 22 cases (12.9%), airway stenosis in 14 cases (8.2%) and esophagotracheal fistula in 12 cases ( 7.0%).The complications of intraoperative and postoperative included decline of percutaneous oxygen saturation and /or heart rate (20.5%, 35/171), mucosal bleeding (12.3%, 21/171 ), and fever after bronchoalveolar lavage.Electronic bronchoscopy examination confirmed all the 171 neonates′diagnosis and some of them recovered after corresponding treatment.78 cases of infants with endo－tracheobronchitis were all cured.22 cases of laryngomalacia and tracheobronchomalacia and nine patients with airway stenosis improved and were discharged after treatment . One patient with subglottic stenosis received bronchoscopic holmium laser ablation therapy and the airway significantly expanded.No re－stenosis was found during follow－up.Conclusion Electronic bronchoscopy is an important method to determine the cause of recurrent dyspnea in newborns , and it′safe,reliable and can play a therapeutic role in some neonates.

Objective Based on the observation of the changes of symptoms, histopathology, visceral sensitivity, mast cell activation, autophagy, and Beclin-1 and Claudin-2 expression in rats, we established and evaluated a new rat model of diarrhea-predominant irritable bowel syndrome (D-IBS) induced by restraint-stress combined with capsaicin (CAP) administration.Methods Forty healthy 5-week old male Sprague Dawley (SD) rats were randomly divided into normal group, model group I, model group II and model group III, with 10 rats in each group.The D-IBS model was established by restraint-stress combined with intragastric administration of CAP (2 mL/100 g body weight, 0.125% in group I, 0.250% in group II, 0.500% in group III), tail clipping and forelimb restriction for 30 minutes every day for 2 weeks.The rats in the control group were treated with saline for 2 weeks.The number of contraction of abdominal wall and arched back were measured by Power Lab instrument.The mast cell activation was detected using aldehyde-magenta-orange G staining.Light and electron microscopic examinations were performed to detect the morphology and autophagy of colonic tissues.The expressions of Beclin-1 and Claudin-2 in the colonic mucosa were detected by streptavidin-biotin complex (SABC) immunohistochemical staining.Results All rats in the model group III died during the experiment.Compared with the control group and model group I, the stool frequency was increased and the visceral sensitivity threshold decreased in the model group II, and there were statistically significant differences between the model group II and the control and model groups I (P < 0.05).The colonic mucosa, mucosal epithelium and glands in each group showed normal morphology and there was no submucosal vasodilatation and diffuse inflammatory cell infiltration.Except for the control group, round purple-reddish staining spots were observed in the rat mucosal stroma or submucosa in the model groups I and II, indicating an increased expression of mast cells.The autophagy, expressions of Beclin-1 and Claudin-2 in the colonic epithelium were significantly increased in the model group II compared with control group and model group I (P< 0.05).Conclusions The model of D-IBS induced by restraint-stress combined with capsaicin is characterized by increased diarrhea, visceral hypersensitivity, increased mast cell expression and autophagy of intestinal epithelial cells, and disruption of the intestinal mucosal barrier.This model is simple to set up and shows similar symptoms of human irritable bowel syndrome.Therefore, it is worthy of popularization and application.

Objective:To detect the neutrophil extracellular traps (NETs) formation in the peripheral blood of the patients with recurrent respiratory tract infection,and to evaluate the effect of sulbactam sodium/cefoperazone sodium on the formation of NETs.Methods:A total of 36 patients with recurrent respiratory tract infection (case group) and 30 healthy volunteers (healthy control group) were selected.The NETs formation of subjects in two groups was detected by confocal microscope and scanning electron microscope (SEM).According to the appearance of neutrophils,the formation of NETs was classified as grade Ⅰ,Ⅱ and Ⅲ,the number of NETs formation cells of subjects in two groups was calculated.The formation of NETs of the patients in case group were detected before and after treated with sulbactam sodium/cefoperazone sodium.Results:The number of NETs formation cells of grade Ⅰ and Ⅱ of the patients in case group was more than that in healthy control group (P<0.05);while the number of NETs formation cells of grade Ⅲ of the patients in case group was less than that in healthy control group (P<0.05).The number of NETs formation cells of grade Ⅰ and Ⅱ of the patients in case group were significantly decreased (P<0.05),while the number of NETs formation cells of grade Ⅲ was significantly increased (P<0.05) after treated with sulbactam sodium/cefoperazone sodium.Conclusion:A lot of NETs with high antibacterial function can be formed in the patients with recurrent lower respiratory tract infection,and sulbactam sodium/cefoperazone sodium can inhibit the formation of NETs.