Objective To observe the effect of continuous incision infusion different concentra-tion of ropivacaine for postoperative analgesia after radical mastectomy.Methods One hundred pa-tients under radical mastectomy,aged 40-70 years,ASA Ⅰ or Ⅱ,were randomly divided into four groups (n =25 each):0.2% (group R1),0.3% (group R2),0.4% (group R3)ropivacaine incision continued infiltration group and patient-controlled intravenous analgesia (group PCIA)as control group.VAS pain scores,sedation Ramsay score and side effects were recorded at each time point in rest and turning over 90°,2 h (T1 ),4 h (T2 ),8 h (T3 ),12 h (T4 ),24 h (T5 ),48 h (T6 )after the operation.Results VAS scores in group R1 at T1-T6 in rest and turn over 90°were significantly high-er than that of group PCIA (P <0.05).There were no significant differences among the group PCIA, group R2 and group R3.Sedation score in PCIA group was significantly higher than that in the other three groups (P <0.05),and the adverse reactions,such as nausea and vomiting,in group PCIA (2 cases)were more serious than that in the other groups (0 cases ).There were no significant differences among the other groups.Conclusion Ropivacaine plays an effective role in infiltration an-algesia when its concentration reaches 0.3% subcutaneous after radical mastectomy.

Objective To investigate the predictive value of left ventricular diastolic function on mechanical ventilation weaning in patients with left ventricular ejection fraction (LVEF) > 0.50.Methods A retrospective case control study was conducted. Sixty-five patients with LVEF > 0.50 undergoing mechanical ventilation for more than 48 hours admitted to surgery intensive care unit (ICU) of China-Japan Friendship Hospital from June 2014 to December 2016 were enrolled. The clinical data and parameters of echocardiography before spontaneous breathing trial (SBT) were collected. The possible relationship between left ventricular diastolic function and the results of weaning was analyzed according to analysis of blood flow filling parameters of mitral valve orifice. According to the grading standard of left ventricular diastolic function, the patients were divided into normal, mild (level 1) and moderate to severe (level 2-3) groups, and the outcomes of weaning were compared among the groups. Then patients were also divided into two groups of weaning successfully and weaning failure, and the clinical data and left ventricular diastolic function parameters of patients were compared between the two groups. The predictive value of left ventricular diastolic function on results of weaning was evaluated with receiver operating characteristic curve (ROC).Results Sixty-five patients were enrolled and 28 patients (43.1%) failed weaning, 22 patients failed the first SBT and 6 required reintubation within 48 hours, 31 of the patients presented normal left ventricular diastolic function, 9 of patients presented mild diastolic dysfunction, and 25 of them presented moderate to severe diastolic dysfunction. So with the gradual increase of the severity of diastolic dysfunction, the rate of weaning failure was gradually increased, which was 16.1%, 44.6% and 76.0% respectively (χ2 = 20.240,P = 0.001). Patients who failed weaning presented evidence of increased left ventricular filling pressures at pre-SBT, by demonstrating decreased deceleration time of E (DTE, s: 180.4±5.1vs. 196.8±4.0,t = 2.567,P = 0.013), increased left ventricular mitral valve diastolic early and late filling velocity ratio (E/A: 1.47±0.08 vs. 1.14±0.05, t = 3.827,P = 0.000), increased lateral, septal and averaged left ventricular mitral valve diastolic early velocity and maximal velocity of mitral annulus in early diastolic velocity ratio (E/Em: 10.26±0.52 vs. 7.28±0.41,t = 4.535, P = 0.000; 10.08±0.58 vs. 8.16±0.40,t = 2.797,P = 0.007; 10.17±0.48比7.72±0.35,t = 4.231,P = 0.000), and the rapid shallow breathing index (RSBI) was also increased significantly (61.7±3.6 vs. 50.6±2.7,t = 2.507, P = 0.015). It was shown by ROC curve analysis that the basic left ventricular diastolic function at pre-SBT had the diagnostic performance in predicting the outcome of weaning from mechanical ventilation, especially E/A and lateral E/Em. Pre-SBT values of E/A greater than 1.2 and lateral E/Em greater than 7.9 predicted weaning failure with an area under the ROC curve (AUC), sensitivity, and specificity of 0.81±0.06 and 0.85±0.06, 82.6% and 91.3%, 81.4% and 80.7%, respectively, and the AUC was higher than RSBI (0.70±0.07). The AUC of combination of E/A > 1.2 and lateral E/Em > 7.9predicting weaning failure was 0.86±0.05 with a sensitivity of 78.3% and a specificity of 93.6%.Conclusions The results suggest that left ventricular diastolic dysfunction is significantly associated with weaning outcome in critical patients with LVEF > 0.50. The combination of E/A ratio greater than 1.2 and E/Em ratio greater than 7.9 may identify patients at high risk of weaning failure.

Objective To understand the dietary nutrients among rural stranded children.Methods 2551 children aged 2 to 7, including 1278 stranded children in the rural areas and another 1273 children served as controls were selected, using multistage stratified cluster random sampling. Dietary survey was performed with three-day weighing dietary method and questionnaire on food intake. Data on diet were analyzed and evaluated by the Dietary Reference Intakes (DRIs) recommend by Chinese Nutrition Society, to evaluate the levels on energy and nutrient intake among stranded children in the rural areas. Results The dietary pattern among rural stranded children mainly consisted of grains and vegetables, but the intakes of animal products, fruits, and snacks were significantly less than in the control group. The intakes of three major energy-producing nutrients were below the recommend nutrient intake. Minerals as calcium, zinc, selenium, kalium and vitamins as vitamin A, B1, B2were insufficient.Most of the rural stranded children took nutrients insufficiently, with 50% lack of adequate energy and 80% of protein, 90% of minerals (calcium, zinc etc.) and vitamins (vitamin B1 and vitamin B2 etc.).Sources from high quality protein was insufficient, only consisting 35% of the total protein, but overabundant (over 64 % ) from the plants. The intake of plant-sourced iron was overabundant, accounted for 87%. Conclusion The dietary pattern was unsatisfactory with insufficient intake of energy-sourced proteins and some nutrients. The sources of energy, protein, and iron were mostly obtained from underbalaneed foods. It is necessary to improve the dietary nutrients status among rural stranded children aged 2-7 years.

In this study, we explored the mechanism of Huganning tablet (HGNP) in the treatment of nonalcoholic fatty liver disease (NAFLD) based on network pharmacology and computer-aided drug design. Firstly, the potential ingredients and targets of HGNP were identified from TCMSP database, Swiss Target Prediction database, Chinese pharmacopoeia (2015) and literatures, and then the targets of HGNP intersected with NAFLD disease targets that obtained in GeneCards database to acquired potential targets. The bioconductor bioinformatics package of R software was used for gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. The network of “potential ingredient-key target-pathway” was formed in Cytoscape software to study the interactions between potential ingredients of HGNP, key targets, pathways and NAFLD. Based on the results of network pharmacology, the molecular docking analysis of the key targets and potential active ingredients in HGNP tablets with top degree in the network was conducted using Discovery Studio 2020 software, followed by molecular dynamics simulations, binding free energy calculation, drug-likeness properties analysis and ADMET (absorption, distribution, metabolism, excretion and toxicity) properties prediction. In vitro, HepG2 cells were used to establish steatosis model, and the effects of five key compounds on hepatocyte steatosis were analyzed by oil red O staining and triglyceride (TG) content determination. The results showed that 141 ingredients and 151 potential targets were obtained. A total of 2 526 items and 151 pathways were identified by GO and KEGG enrichment analysis. The molecular docking suggested that five components, isorhamnetin, salvianolic acid B, emodin, resveratrol and rhein, exhibited strong binding ability with key targets [retinoic acid receptor RXR-alpha (RXRA), tumor necrosis factor (TNF), glycogen synthase kinase-3 beta (GSK3B), serine/threonine-protein kinase 1 (AKT1)]. It was further verified that isorhamnetin and salvianolic acid B bind to key targets with good structural stability and binding affinity based on molecular dynamics simulations and binding free energy calculations. The drug-likeness properties, pharmacokinetic properties and toxicity of five key compounds were more comprehensively analyzed through drug-likeness properties analysis and ADMET properties prediction. In vitro, all five compounds, isorhamnetin, salvianolic acid B, emodin, resveratrol, and rhein, improved hepatocyte steatosis of HepG2 cells, confirming the reliability of the present study. In conclusion, based on network pharmacology, computer-aided drug design and in vitro validation, this study investigated the mechanism of HGNP for the treatment of NAFLD at multiple levels and provided a basis for its clinical application.

Aging can cause degenerative changes in the function of multiple tissues and organs in the body. Gastrointestinal diseases and intestinal dysfunction are very common in the elderly people. The purpose of this study is to explore the effect of the total extract of Astragalus membranaceus (Fisch.) Bge. on intestinal function and gut microbiota homeostasis in natural aging mice, which will provide clues for further mechanism study. The natural aging mice model is established and animal experiments follow the regulations of the Animal Ethics Committee of Nanjing University of Traditional Chinese Medicine. The overall health of the mice was evaluated by the "frailty index" scoring method. The intestinal absorption and transport function were measured by detecting intestinal glucose absorption capacity, transport time, lipase and amylase activities of aging mice. Intestinal inflammation was assessed by detecting inflammatory cytokines by enzyme-linked immunosorbent assay (ELISA). The pathological changes in the intestines of aging mice were tested by hematoxylin-eosin (H&E) staining and alizarin blue (AB) staining. The qRT-PCR method was used to explore the gene transcription level related with the proliferation and differentiation of intestinal stem cells. Microbiota analysis based on 16S rDNA were used to evaluate the composition of gut microbiota. The results showed that Astragalus had a tendency to reduce the "frailty index" of aging mice, but did not show a significant difference. In some indicators of aging phenotype, Astragalus has the most significant effect on hair loss and physical fitness. In terms of intestinal function, Astragalus could increase intestinal glucose absorption capacity, shorten intestinal transportation time and promote lipase secretion in aging mice. The levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-‍α) in the aging intestinal tissue were reduced after Astragalus administration. Astragalus also ameliorated the pathological degeneration of the intestinal tissue of aging mice by increasing the length of small intestinal villi, the thickness of colonic mucosa and goblet cell number. In addition, Astragalus elevated the expression of genes associated with the proliferation and differentiation in jejunum and modulated gut microbiota, especially restoring the abundance of Lachnospiraceae. Taken together, the above research results demonstrate the total extract of Astragalus as a key factor improving the intestinal function and gut microbiota homeostasis of aging mice.

In this study, the ameliorative effects of Flos Abelmoschus manihot on mice with chronic inflammatory bowel disease (IBD) were investigated and its effects on the structure of the intestinal flora as well as the lipid profile in feces of IBD mice were analyzed. All animal welfare and experimental procedures followed the regulations of the Animal Ethics Committee of Nanjing University of Chinese medicine. A mouse model with chronic IBD induced by dextran sulfate sodium (DSS) was used to evaluate changes in body weight, disease activity index (DAI), colonic histopathological damage as well as gene expression levels of inflammatory factors in the colon. Fecal samples from mice in each group were collected and subjected to Illumina high-throughput sequencing to detect the abundance of intestinal flora; samples were analyzed by UHPLC-Q-Exactive^® HF Quadrupole-Orbitrap^® of untargeted lipidomics, which detects lipid content in feces. Administration of Flos Abelmoschus manihot could significantly restore the body weight and ameliorate colonic histopathological damage in IBD mice. Sequencing of the gut microbiota revealed that the species diversity and richness of the gut microbiota in IBD mice were decreased, with a significant increase in the abundance of Verrucomicrobia and a significant decrease in the abundance of Bacteroidetes; Flos Abelmoschus manihot significantly increased the richness and diversity of intestinal microbiota in IBD mice, increased the number of taxa species at each level, and restored the abundance of bacteria in the phylum Bacteroidetes. Analysis of fecal lipid profiles identified the most significant changes in sphingolipid and glycerophospholipid metabolic pathways in IBD mice, with Flos Abelmoschus manihot inhibiting ceramide and sphingomyelin synthesis in sphingolipid metabolism. In summary, Flos Abelmoschus manihot can effectively improve the disease condition of mice with chronic IBD, and it has the effect of regulating intestinal flora homeostasis and lipid metabolism, but the related mechanism between the two still needs to be deeply explored.

OBJECTIVETo study the effect of Bushen Tiaojing Recipe (BTR) and Xiaoyao Pill (XP) on cathepsin-L (Cat-L) mRNA in mice.

METHODSImmature mice were randomly divided into the normal group, the control group, the BTR group and the XP group, three in each group. Cat-L mRNA expression in mice was detected using reverse transcription polymerase chain reaction (RT-PCR) at 0, 4, 8 and 12 h after injecting 5 IU (human chorionic gonadotropin, HCG).

RESULTSCat-L mRNA expression increased gradually after HCG injection, the relative levels in the control group at 0, 4, 8 and 12 h were 0.066 +/- 0.005, 0.383 +/- 0.045, 0.737 +/- 0.024 and 1.036 +/- 0.073 respectively, comparisons between different time-points showed significant difference (P < 0.01). Compared with the control group, the Cat L mRNA expression was higher at 4 h in both BTR and XP groups (P < 0.01), at 8 h in the XP group (P < 0.05), and at 12 h in BTR group after injecting HCG (P < 0.05). Compared with the control group, cat L mRNA expression showed no statistic difference at 8 h in BTR group and at 12 h in XC group.

CONCLUSIONSBTR promoted the ovulation by enhancing the expression of CatL gene, and that of XP by advancing the peak of CatL gene expression.

Animals ; Cathepsin L ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gonadotropins ; administration & dosage ; Humans ; Mice ; Mice, Inbred Strains ; Ovulation ; drug effects ; RNA, Messenger ; genetics

Objective To investigate the role of mitochondrial permeability transition pore (mPTP) of hippocampal neurons in process of hydrogen-rich saline attenuating global cerebral ischemia-reperfusion (I/R) injury in rats.Methods Seventy-two male Sprague Dawley rats,weighing 250-300 g,were randomly divided into six groups ( n =12 each):sham operation group (group S),cerebral ischemia-reperfusion group (group IR),normal saline group (group NS),hydrogen-rich saline group (group H),atractyloside group (group A) and hydrogen-rich saline + atractyloside group (group HA).Global cerebral I/R injury was produced by four-vessel occlusion method.Bilateral vertebral arteries were cauterized.Then bilateral common carotid arteries were occluded for 15min and followed by reperfusion.In groups H and HA,hydrogen-rich saline 5 ml/kg was injected intraperitoneally immediately after reperfusion,while equal volume of normal saline was injected in the other four groups.The rats in groups A and HA received intracerebroventricular injection of atractyloside 15 μl 10 min before reperfusion,while groups NS and H received intracerebroventricular injection of equal volume of normal saline.After the neurological behavior was evaluated at 24 h of reperfusion,8 rats in each group were sacrificed and the hippocampi were immediately isolated and homogenized followed by density gradient centrifugation.The opening degree of mPTP was assayed with spectrophotometry and the mitochondrial membrane potential (MMP) was detected with Rhodamine 123 method.Four rats in each group were killed at 72 h of reperfusion and the brains were removed for microscopic examination of the area CA1 of the hippocampus and determination of the number of normal pyramidal neurons.Results Compared with group S,the neurological behavior was compromised,MMP was decreased and mPTP opening degree was enhanced in the other five groups ( P ＜ 0.05).The neurological behavior was better,MMP was increased and mPTP opening degree was decreased in groups H and HA as compared with group IR ( P ＜ 0.05).Compared with group H,the neurological behavior was compromised,MMP was decreased and mPTP opening degree was enhanced in group HA ( P ＜ 0.05).Compared with group IR,the number of normal pyramidal neurons at 72 h of reperfusion in the CA1 region of the hippocampus was higher in group HA ( P ＜0.05).The injury of the CA1 region of the hippocampus at 72 h of reperfusion was attenuated in group H as compared with groups IR,NS,A and HA.Conclusion Hydrogen-rich saline can attenuate global cerebral I/R injury throngh inhibiting the mPTP opening and reducing the dissipation of MMP,thus maintaining the mitochondrial function.

Objective To investigate the inhibitory effect of RNA interference (RNAi) on Gli1,Bcl-2, Bax and cycin D1 gene expressions in U251 cell line and the proliferation of U251 cells.Methods Small interfering RNA (siRNA, at locus of 58, 59, 60 and 61) targeted for Gli1 gene was designed and transfected into U251 cells. RT-PCR was emplyed to detect the mRNA expression of Gli1 gene to select the siRNA interference fragment (siRNA-Gli1) that could most efficiently inhibit the mRNA expression of Gli1 gene. The mRNA and protein expressions of Gli1 gene at different times after siRNA-Gli1 transfection were detected to determine the time law of this interference. U251 cells at logarithmic phase were divided into 3 groups: siRNA-Gli1 group (transfection of selected siRNA-Gli1 fragments), siRNA-NC (transfection of siRNA fragments) and siRNA-N group (blank controls). The mRNA and protein expressions of Bcl-2, Bax and cycin D1 gene were assessed by RT-PCR and Western blotting. Proliferation of cells was measured by MTT assay, and cell apoptosis and cell cycles were detected by flow cytometry (FCM). Results Transfection efficiency of interference fragments (at locus of 58, 59, 60 and 61, and NC) reached 69.2％; RT-PCR indicated that no obvious Gli1 mRNA expression was noted at U251-60 cells 48 h after the transfection therefore, locus 60 was the best interference fragment and 48 h was the best time. The mRNA and protein expressions of Bcl-2 and cycin D1 genes were obviously suppressed by siRNA, and the mRNA and protein expressions of Bax gene were significantly up-regulated in the siRNA-Gli1 group as compared with those in the siRNA-N and siRNA-NC groups 48 h after transfection (P＜0.05). Silencing Gli1 by RNAi significantly inhibited the proliferation and induced the apoptosis of U251 cells as compared with siRNA-N and siRNA-NC groups 24, 48 and 72 h after transfection (P＜0.05). Cells at G0 and G1 phases were obviously increased and those at S phase were significantly decreased in the siRNA-Glil group as compared with those in the siRNA-N and siRNA-NC groups (P＜0.05). Conclusion Expression of Gli1 gene can be effectively inhibited by specific siRNA targeting Gli1 gene in U251 cells and the proliferation of U251 cells can be significantly inhibited, which may possibly be related to that siRNA-Gli1 decreases the expressions of Bcl-2 and cycin D1 and alters the ratio of Bcl-2/Bax.

Aged ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; immunology ; Prognosis ; T-Lymphocyte Subsets ; immunology