Objective: To reveal the molecular mechanism underlying the compatibility of Salvia miltiorrhiza Bge (S. miltiorrhiza, Dan Shen) and C. tinctorius L. (C. tinctorius, Hong Hua) as an herb pair through network pharmacology and subsequent experimental validation. Methods: Network pharmacology was applied to construct an active ingredient-efficacy target-disease protein network to reveal the unique regulation pattern of S. miltiorrhiza and C. tinctorius as herb pair. Molecular docking was used to verify the binding of the components of these herbs and their potential targets. An H9c2 glucose hypoxia model was used to evaluate the efficacy of the components and their synergistic effects, which were evaluated using the combination index. Western blot was performed to detect the protein expression of these targets. Results: Network pharmacology analysis revealed 5 pathways and 8 core targets of S. miltiorrhiza and C. tinctorius in myocardial protection. Five of the core targets were enriched in the hypoxia-inducible factor-1 (HIF-1) signaling pathway. S. miltiorrhiza-C. tinctorius achieved vascular tone mainly by regulating the target genes of the HIF-1 pathway. As an upstream gene of the HIF-1 pathway, STAT3 can be activated by the active ingredients cryptotanshinone (Ctan), salvianolic acid B (Sal. B), and myricetin (Myric). Cell experiments revealed that Myric, Sal. B, and Ctan also exhibited synergistic myocardial protective activity. Molecular docking verified the strong binding of Myric, Sal. B, and Ctan to STAT3. Western blot further showed that the active ingredients synergistically upregulated the protein expression of STAT3. Conclusion The pharmacodynamic transmission analysis revealed that the active ingredients of S. miltiorrhiza and C. tinctorius can synergistically resist ischemia through various targets and pathways. This study provides a methodological reference for interpreting traditional Chinese medicine compatibility.

The importance of structural variants(SVs)for human phenotypes and diseases is now recognized.Although a variety of SV detection platforms and strategies that vary in sensitivity and specificity have been developed,few benchmarking procedures are available to confidently assess their performances in biological and clinical research.To facilitate the validation and application of these SV detection approaches,we established an Asian reference material by characterizing the genome of an Epstein-Barr virus(EBV)-immortalized B lymphocyte line along with identified benchmark regions and high-confidence SV calls.We established a high-confidence SV callset with 8938 SVs by integrating four alignment-based SV callers,including 109x Pacific Biosciences(PacBio)continuous long reads(CLRs),22 x PacBio circular consensus sequencing(CCS)reads,104x Oxford Nanopore Technologies(ONT)long reads,and 114×Bionano optical mapping plat-form,and one de novo assembly-based SV caller using CCS reads.A total of 544 randomly selected SVs were validated by PCR amplification and Sanger sequencing,demonstrating the robustness of our SV calls.Combining trio-binning-based haplotype assemblies,we established an SV benchmark for identifying false negatives and false positives by constructing the continuous high-confidence regions(CHCRs),which covered 1.46 gigabase pairs(Gb)and 6882 SVs supported by at least one diploid haplotype assembly.Establishing high-confidence SV calls for a benchmark sample that has been characterized by multiple technologies provides a valuable resource for investigating SVs in human biology,disease,and clinical research.

Objective: To determine the in vitro and in vivo absorption properties of active ingredients of the Chinese medicine, baicalein, to enrich mechanistic understanding of oral drug absorption. Methods: The Biopharmaceutic Classification System (BCS) category was determined using equilibrium solubility, intrinsic dissolution rate, and intestinal permeability to evaluate intestinal absorption mech-anisms of baicalein in rats in vitro. Physiologically based pharmacokinetic (PBPK) model commercial software GastroPlus?was used to predict oral absorption of baicalein in vivo. Results: Based on equilibrium solubility, intrinsic dissolution rate, and permeability values of main absorptive segments in the duodenum, jejunum, and ileum, baicalein was classified as a drug with low solubility and high permeability. Intestinal perfusion with venous sampling (IPVS) revealed that baicalein was extensively metabolized in the body, which corresponded to the low bioavailability predicted by the PBPK model. Further, the PBPK model predicted the key indicators of BCS, leading to reclassification as BCS-II. Predicted values of peak plasma concentration of the drug (Cmax) and area under the curve (AUC) fell within two times of the error of the measured results, highlighting the superior prediction of ab-sorption of baicalein in rats, beagles, and humans. The PBPK model supported in vitro and in vivo evi-dence and provided excellent prediction for this BCS class Ⅱ drug. Conclusion: BCS and PBPK are complementary methods that enable comprehensive research of BCS parameters, intestinal absorption rate, metabolism, prediction of human absorption fraction and bioavailability, simulation of PK, and drug absorption in various intestinal segments across species. This combined approach may facilitate a more comprehensive and accurate analysis of the absorption characteristics of active ingredients of Chinese medicine from in vitro and in vivo perspectives.

Objective:To screen and identify differentially expressed long non-coding RNA (lncRNAs) in peripheral blood of children with sepsis, and to explore the role of lncRNAs in the pathogenesis of sepsis in children.Methods:The peripheral blood samples of 3 children with sepsis admitted to the ICU of Children′s Hospital of Capital Institute of Pediatrics from January to December 2016 and 3 healthy children who underwent physical examination in our hospital during the same period were selected, and the differential expression profiles of lncRNAs and mRNAs were screened by lncRNAs sequencing technology.The target genes of differentially expressed lncRNAs were predicted and the relationship pairs of lncRNA-mRNA related to F 1F O-ATPase activity were constructed according to the results of GO analysis.Further increasing the sample size, we verified the expression of some F 1F O-ATPase activity-related mRNAs and lncRNAs which were differentially expressed in the screening results by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR). Results:Sequencing results showed that there were 252 lncRNAs with significant differential expression in peripheral blood of children with sepsis compared with healthy children, of which 86 were up-regulated and 166 were down-regulated; meanwhile, there were 2 652 mRNAs with significant differential expression, of which 955 were up-regulated and 1 697 were down-regulated.The results of qRT-PCR showed that the expression of lncRNA ENST00000621933.1, ENST00000616950.1 and ENST00000595748.1 in peripheral blood of children with sepsis increased( P<0.05), while the expression of MT-ATP8, ATP5E and ENST00000624705.1, ENST00000615535.1 in peripheral blood of children with sepsis decreased( P<0.05), which was consistent with the sequencing results. Conclusion:lncRNAs are differentially expressed in peripheral blood of children with sepsis compared with healthy children.The expression levels of lncRNA ENST00000621933.1, ENST00000616950.1, ENST00000595748.1, ENST00000624705.1 and ENST00000615535.1 which their target genes are MT-ATP8 and ATP5E may be related to the development of sepsis in children.

Objective: To study the clinical characteristics of children with adenovirus pneumonia and provide evidence for clinical diagnosis and treatment timely. Method: This retrospective study included 89 children who were confirmed to have adenovirus pneumonia in hospital from January 2015 to December 2016. All the immunofluorescence test result of the 89 children showed that the exfoliated nasopharyngeal cells from the 89 children were all adenovirus antigen positive. All the severe type children reached the diagnostic criteria of severe pneumonia by the respiratory group in the society of pediatrics, Chinese Medical Association. The children were divided into 2 groups (severe type group and common type group). Different factors such as epidemiologic feature, clinical manifestation, laboratory examination and imaging data were analyzed. Results: Among the 89 pediatric patients, the male to female ratio was 1.5∶1. The ages ranged from 1 month to 14 years. Children under 5 years of age accounted for 96.6%(86/89). The incidence was 37.1%(33/89)in winter and 30.3%(27/89)in spring. The lengths of hospital stay were 3-48 days and the median length of stay was 8.25±4.75 days. All of these 89 cases had fever and cough. The proportion of severe adenovirus pneumonia was high among male, under 2 years of age, those with dyspnea, hepatosplenomegaly, tachycardia, leukocytosis, elevated C-reactive protein (CRP), PCT, myocardial enzymes, electrocardiogram abnormality and cluster shadow in chest CT. Differences were statistically significant (P<0.05). Conclusions Special attention should be paid to the male children under the 2 years of age with abnormal related indicators to consider severe adenovirus pneumonia. Early detection and treatment are the key to improve treatment and reduce death.

OBJECTIVETo investigate the infection status and virulent genes of Aeromonas in patients with acute diarrhea in Pudong New Area, Shanghai.

METHODSIn 2012, stool samples were collected from diarrhea patients in 12 sentinel hospitals in Pudong for the detections of 13 pathogens causing diarrhea, and the detections of 5 diarrhea related virulent genes were conducted for Aeromonas isolates.

RESULTSA total of 101 patients were infected with Aeromonas in 2533 patients (4.0%). A total of 101 Aeromonas strains were isolated, including 17 Aeromonas hydrophila strains (18.8%), 44 Aeromonas veronii biovar sobria strains (52.5%) and 12 Aeromonas caviae strains (29.7%). And 44 coinfections with other pathogens were detected. Aeromonas infection mainly occurred in summer and in people aged ≥20 years. Among the patients infected with Aeromonas, 71 (70.3%) had watery diarrhea, 20 (19.8%) had vomiting and 11 (10.9%) had fever. Virulent genes detection showed that 95.0% of the Aeromonas. strains carried virulent genes, and the detection rates of hlyA, aerA, act, alt, and ast genes were 5.9%, 6.9%, 67.3%, 42.6% and 13.9%, respectively.

CONCLUSIONSHigh incidence of Aeromonas infection was found in the patients with acute diarrhea in Pudong, and a high proportion of coinfections with other pathogens was detected too. Most Aeromonas strains carried virulent genes, and the distribution varied.

Aeromonas ; genetics ; Aeromonas hydrophila ; genetics ; China ; epidemiology ; Diarrhea ; microbiology ; Gram-Negative Bacterial Infections ; epidemiology ; microbiology ; Humans ; Seasons ; Virulence ; genetics ; Young Adult

Objective To evaluate the performance of VITEK-2 compact,VITEK MS and Bruker MS on the identification of Corynebacterium.Methods This was a methodological evaluation study.The 40 Corynebacterium from bioMerieux were i-dentified with the three methods respectively.16S rDNA gene sequencing was conducted as reference method.Made a de-scriptive analysis of the identification ability,time and cost.Resulets The accuracy of species level of the three methods was 95.0%,88.9% and 97.5%.The mean time was 5~6 h,2~3 min and 2~3 min.The cost of consumable was 50~70 yuan, 15~25 yuan and 10~20 yuan.Conclution Three methods with high accuracy can meet the requirement of clinical diagno-sis,and the identification ability of VITEK MS on Corynebacterium amycolatum need to be further improved.

Objective To construct recobinant pmRNA IRES-hKDR and lay a primary foundation for further study of gene therapy for tumors.Methods pmRNA IRES-hKDR was obtained from pDC520 hkDR and pmRNA IRES-Luc which was reserved by our laboratory through molecular biology technology and was transfected into Hepall-6 via lipsome after identification of PCR,restriction enzyme digesting and DNA ELISA and Western blot.Results The recombiant plasmid had pmRNA IRES-hKD gene which was expressed in Hepall-6//G418 in vitro and the expressed protein was secreted out of the cells could response with specific hKDR antibody,which was identified by Western blot.Conclusion An eukaryotic expression recombiant plasmid pmRNA IRES-hKDR can be constructed successfully which provides the possibility of further researches on its anti-tumor effect.

Objective To investigate the immune protection of dendritic cells(DCs) harboring kinase domain-containing receptor(KDR) fusion gene on mice carrying B16 melanoma.Methods The bone marrow precursor-derived dendritic cells(BMDCs) were induced from bone marrow progenitors of mice by GM-CSF.The KDR fusion gene mRNA was transfected into DCs in vitro.Mice were immunized with the same amount of DCs at 7-day interval and then each mouse was injected with 5?10~5 B16 cells.The mice without tumors were injected with B16 cells again 20 days later.Mice were randomly divided into 4 groups: group A(n=7),group B(n=8),group C(n=8) and group D(n=7).The mice were immunized with DCs one time in group A,2 times in group B,3 times in group C and as controls in group D.Results After one week,all mice in group D had tumors with average survival 15 days.All mice in group A,B and C had no tumors after 20 days later.After the second injection of B16 cells,2 mice in group A and 2 mice in group B had tumors.The mice in group C had no tumor.The average survival periods were calculated from first injection of B16 cells to the study end.The average survival period of group A was 50 days and that of group B was 72 days.Conclusions The dendritic cells vaccine harboring KDR fusion gene has immune protection against melanoma in mice.

Objective To evaluate the clinical value of selective salpingography (SSG) and fallopian tube recanalization (FTR) by home made coaxial catheter. Methods 116 cases of tube obstruction were diagnosed by hysterosalpingogram (HSG). SSG was performed with home made coaxial catheter. If proximal tube complete obstruction was still present, then followed by FTR. Results 116 cases of follow up showed 55 intrauterine pregnancies (IUPS) and 6 ectopic pregnancies. IUP rate was 47.4%. 28 of 116 cases were performed by HSG again, 25 of 28 cases were improved, effective rate was 89.3%.Conclusions SSG and FTR were a safe and effective procedure for the diagnosis and treatment of tube obstruction with home made coaxial catheter.