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Bifunctional thiourea-catalyzed asymmetric [3 + 2] annulation reactions of 2-isothiocyanato-1-indanones with barbiturate-based olefins have been developed to afford chiral dispiro[indene-pyrrolidine-pyrimidine]s. Through this strategy, the target products could be obtained in good to excellent yields with excellent stereoselectivities. In addition, the synthetic utility was verified through a gram-scale synthesis, one-pot three-component reactions and further transformation experiments of the products.

Chemie - Ingenieur - Technik 95(6), 874-876 (2023). doi:10.1002/cite.202300016 special issue: "Special Issue:ACHEMA‐Nachberichte", Published by Wiley-VCH Verl., Weinheim

In the era of precision medicine, many biomarkers have been discovered to be associated with drug efficacy and safety responses, which can be used for patient stratification and drug response prediction. Due to small sample size and limited power of randomized clinical studies, meta-analysis is usually conducted to aggregate all available studies to maximize the power for identifying prognostic and predictive biomarkers. Since all available data are already aggregated, it is often challenging to find an independent study to replicate the discoveries from the meta-analysis (e.g., in meta-analysis of pharmacogenomics genome-wide association studies (PGx GWAS)), which seriously limits the potential impacts of the discovered biomarkers. To overcome this challenge, we develop a novel statistical framework, MAJAR (Meta-Analysis of Joint effect Associations for biomarker Replicability assessment), to jointly test prognostic and predictive effects and assess the replicability of identified biomarkers by implementing an enhanced Expectation–Maximization algorithm and calculating their posterior-probability-of-replicabilities (PPR) and Bayesian false discovery rates (Fdr). Extensive simulation studies were conducted to compare the performance of MAJAR and existing methods in terms of Fdr, power, and computational efficiency. The simulation results showed improved statistical power with well-controlled Fdr of MAJAR over existing methods and robustness to outliers under different data generation processes while considering both prognostic and predictive effects in the model. We further demonstrated the advantages of MAJAR over existing methods by applying MAJAR to the PGx GWAS summary statistics data from a large cardiovascular randomized clinical trial (IMPROVE-IT). Compared to testing main effects only, MAJAR identified 12 novel variants associated with the treatment-related LDL cholesterol (LDL-C) reduction from baseline.

Introduction Severe hand, foot, and mouth disease (HFMD) may lead to serious complications, which cause child mortality during outbreaks. The aim of this study was to determine whether neutrophil-to-lymphocyte ratio (NLR) can predict death risk in severe HFMD. Methods Medical records for 664 severe HFMD patients were retrospectively examined, and NLR was calculated from blood counts. Youden's index was calculated to determine the optimal NLR cutoff. Uni- and multivariate logistic regression were used to determine death risk factors associated with severe HFMD. Results An NLR cutoff value of 2.01 and 2.50 respectively predicted mortality among all 664 severe HFMD and 137 critical HFMD. Among all 664 patients, the multivariate model identified the following as independently associated with death risk: high fever (OR 3.342, 95% CI 1.736-6.432), EV71 infection (OR 3.200, 95% CI 1.529-6.698), fasting glucose (OR 37.343, 95% CI 18.616-74.909), and NLR (>2.01) (OR 2.142, 95% CI 1.125-4.079). Among 137 critical HFMD, EV71 infection (OR 3.441, 95% CI 1.132-10.462), fasting glucose (OR 14.173, 95% CI 4.920-40.827), and NLR (>2.50) (OR 4.166, 95% CI 1.570-11.051) were associated with death risk. Conclusion In conclusion, NLR (>2.01) in severe HFMD and NLR (>2.50) in critical HFMD patients may be associated with increased death risk.

Little is known about the treatment of patients with hepatitis B surface antigen (HBsAg) recurrence after being clinically cured by peginterferon alpha(peg-IFN-α)-based regimens. This study aimed to investigate the efficacy and safety of peg-IFNα-2b in re-treating patients with HBsAg recurrence after stopping peg-IFN α-based regimens. In this two-center, prospective observational study, 33 patients with HBsAg recurrence after stopping peg-IFN α-based regimens were enrolled and re-treated with an individualized course of peg-IFN α-2b. The hepatitis B virus (HBV) vaccine could be injected immediately after HBsAg clearance, according to patients’ willingness. All patients were monitored and followed-up for 48 weeks after peg-IFN α-2b re-treatment stop. The primary endpoint was HBsAg clearance at the end of follow-up. At baseline, all patients had HBsAg levels of

SUMMARYMenthol is widely used in tobacco products. This study compared the effects of menthol on human bronchial epithelium using submerged cultures, a VITROCELL® cloud chamber that provides air liquid interface (ALI) exposure without solvents or heating, and a Cultex ALI system that delivers aerosol equivalent to that inhaled during vaping. In submerged culture, menthol significantly increased calcium influx and mitochondrial reactive oxygen species (ROS) via the TRPM8 receptor, responses that were inhibited by a TRPM8 antagonist. VITROCELL® cloud chamber exposure of BEAS-2B monolayers increased mitochondrial protein oxidation, expression of the antioxidant enzyme SOD2, activation of NF-κB, and secretion of inflammatory cytokines (IL-6 and IL-8). Proteomics data collected following ALI exposure of 3D EpiAirway tissue in the Cultex showed upregulation of NRF-2-mediated oxidative stress, oxidative phosphorylation, and IL-8 signaling. Across the three platforms, menthol adversely effected human bronchial epithelium in a manner that could lead to respiratory disease. Graphical Abstract. Mechanism of action of menthol on human bronchial epithelium.Three in vitro platforms were used to study the effect of menthol on bronchial epithelium. In submerged culture (using BEAS-2B cells), menthol produced rapid calcium influx followed by an increase in oxidative stress and inflammatory cytokines. ALI exposure of BEAS-2B cells to unheated menthol in a cloud chamber caused activation of an inflammatory transcription factor (NF-κB) and oxidative stress. Proteomics analysis of human EpiAirway tissues exposed at the ALI to heated menthol EC aerosols identified changes in the expression of proteins involved in oxidative stress and in an inflammatory response.

Various medical treatments for COVID-19 are attempted. After patients are discharged, SARS-CoV-2 recurring cases are reported and the recurrence could profoundly impact patient healthcare and social economics. To date, no data on the effects of medical treatments on recurrence has been published. We analyzed the treatment data of combinations of ten different drugs for the recurring cases in a single medical center, Shenzhen, China. A total of 417 patients were considered and 414 of them were included in this study (3 deaths) with mild-to-critical COVID-19. Patients were treated by 10 different drug combinations and followed up for recurrence for 28 days quarantine after being discharged from the medical center between February and May, 2020. We applied the Synthetic Minority Oversampling Technique (SMOTE) to overcome the rare recurring events in certain age groups and performed Virtual Twins (VT) analysis facilitated by random forest regression for medical treatment-recurrence classification. Among those drug combinations, Methylprednisolone/Interferon/Lopinavir/Ritonavir/Arbidol led to the lowest recurring rate (0.133) as compared to the average recurring rate (0.203). For the younger group (age 20-27) or the older group (age 60-70), the optimal drug combinations are different, but the above combination is still the second best. For obese patients, the combination of Ribavirin/Interferon/Lopinavir/Ritonavir/Arbidol led to the lowest recurring rate for age group of 20-50, whereas the combination of Interferon/Lopinavir/Ritonavir/Arbidol led to lowest recurring rate for age group of 50-70. The insights into combinatorial therapy we provided here shed lights on the use of a combination of (biological and chemical) anti-virus therapy and/or anti-cytokine storm as a potentially effective therapeutic treatment for COVID-19.

Hepatocellular carcinoma (HCC), accounting for approximately 90% of liver cancer, is the most lethal malignant tumors in the world. Large amount of evidence indicate that microRNAs (miRNAs) contribute to the tumorigenesis and progression of HCC. Among them, miR-376c-3p was recently identified as a tumor-related miRNA and is up-regulated in HBV-related HCC. But, the clinical significance of miR-376c-3p and its biological function in HCC progression are still unclear. Here, we confirmed that miR-376c-3p expression level in HCC was markedly higher than that in noncancerous tissues. Up-regulation of miR-376c-3p was detected in four different HCC cell lines. High miR-376c-3p expression correlated with poor prognostic features, such as large tumor size and venous infiltration. Follow-up data indicated that high miR-376c-3p level evidently correlated with poor clinical outcomes of HCC patients. Moreover, knockdown of miR-376c-3p repressed HCC cell growth, migration and invasion in vitro. miR-376c-3p overexpression facilitated these malignant behaviors of Bel-7402 cells. Mechanistically, miR-376c-3p posttranscriptionally repressed ARID2 expression by directly interacting with its 3'-UTR. Furthermore, an obvious negative correlation between miR-376c-3p and ARID2 mRNA expression in HCC tissues was confirmed. Notably, miR-376c-3p knockdown suppressed HCC growth and metastasis in nude mice. Gain-of-function experiments showed that ARID2 inhibited cell growth and mobility of Hep3B cells. Subsequently, ARID2 knockdown rescued miR-376c-3p silencing attenuated Hep3B cell proliferation and mobility. Our results suggest that miR-376c-3p exerts an oncogenic role in HCC progression.

Caffeic acid phenethyl ester (CAPE) has been reported to possess the hepatoprotective effect. This study was to investigate the mechanism underlying CAPE against liver fibrosis in a liver fibrosis model induced by toxic carbon tetrachloride (CCl4) in male Sprague-Dawley rats and in vitro in CAPE (5 μM, 10 μM, 15 μM) treated hepatic stellate cells (HSC-T6). We found that CAPE treatment remarkably attenuated CCl4-induced liver fibrosis by blocking the activation of HSCs as determined by the expression alternation of transforming growth factor (TGF)-β1, phosphorylated Smad3 (p-Smad3), collage I, α-smooth muscle actin (α-SMA), matrix metalloproteinases (MMPs) 2, tissue inhibitor of matrix metalloproteinases (TIMPs) 1. The hepatoprotective effects of CAPE were also associated with upregulation of autophasomes in HSCs as determined by transmission electron microscopy (TEM) detection. The in vitro study further confrimed that CAPE attenuated liver fibrogenesis via inducing authophagic markers including LC3, ATG5, Beclin 1 expressions, while inhibiting AKT/mTOR signaling in HSC-T6 cells. Thus, the protective effects of CAPE against liver fibrosis might due to the inhibition of TGF-β1/Smad3 signaling and induction of authophagy in HSCs.

Coxsackievirus A16 (CA16) remains the most common causative agent of hand, foot, and mouth disease (HFMD), and is related to high incidence and critical complications. Vitamin D receptor (VDR) activity might affect the outcome of CA16 infection. Our case-control research aims to evaluate the relationship between VDR polymorphisms in the gene encoding and susceptibility to and severity of HFMD due to CA16. Three single-nucleotide polymorphisms (SNPs) of VDR gene were selected according to functional prediction and linkage disequilibrium, and were examined utilizing the SNPscan method to identify possible associations with HFMD caused by CA16. A significant relationship was found in the HFMD cases of polymorphism rs11574129 (GA vs GG: odds ratio (OR) = 0.068, 95% confidence interval (CI) = 0.007-0.693, P = .023; GA + AA vs GG: OR = 0.322, 95%CI = 0.106-0.984, P = .047), and vitamin D levels in genotype AA were significantly higher than those in genotype GG (P

Hepatitis C virus (HCV) is closely associated with insulin resistance (IS), acting primarily by interfering with insulin signaling pathways, increasing cytokine-mediated (tumor necrosis factor α, interleukin 6) inflammatory responses and enhancing oxidative stress. In the insulin signaling pathways, the insulin receptor substrate (IRS) is one of the key regulatory factors. The present study constructed gene regulatory sub-networks specific for IRS1 and IRS2 in Huh7 cells and HCV-infected Huh7 (HCV-Huh7) cells using linear programming and a decomposition algorithm, and investigated the possible mechanisms underlying the function of IRS1/2 in HCV-induced IS in Huh7 cells. All data were obtained from {"type":"entrez-geo","attrs":{"text":"GSE20948","term_id":"20948"}}GSE20948 of the Gene Expression Omnibus database from the National Center for Biotechnology Information. Genes which were significantly differentially expressed between Huh7 and HCV-Huh7 cells were analyzed using the significance analysis of microarray algorithm. The top 50 genes, including IRS1/2, were used as target genes to determine the gene regulatory networks and next the sub-networks of IRS1 and IRS2 in HCV-Huh7 and Huh7 cells using Gene Regulatory Network Inference Tool, an algorithm based on linear programming and the decomposition process. The IRS1/2 sub-networks were divided into upstream/downstream groups and activation/suppression clusters, and were further analyzed using Molecule Annotation System 3.0 and Database for Annotation, Visualization, and Integrated Discovery software, two online platforms for enrichment and clustering analysis and visualization. The results indicated that in Huh7 cells, the downstream network of IRS2 is more complex than that of IRS1, indicating that the insulin metabolism in Huh7 cells may be primarily mediated by IRS2. In HCV-Huh7 cells, the downstream pathway of IRS2 is blocked, suggesting that this may be the underlying mechanism in HCV infection that leads to insulin resistance. The present findings add a further dimension to the understanding of the pathological mechanisms of HCV infection-associated insulin resistance, and provide novel concepts for insulin resistance and glucose metabolism research.

This study aimed to explore the mechanism of p47phox-induced increase of reactive oxygen species (ROS) in peripheral blood mononuclear cells (PBMCs) from premature infants after oxygen therapy, and determine a new target for oxidative stress injury alleviation in clinical setting.First, ROS levels as well as p47phox translocation and expression in PBMC samples were evaluated after treatment of premature infants with different concentrations of oxygen. Then, changes of all various parameters were detected after in vitro treatment of PBMCs with diphenyleneiodonium (DPI), apocynin, and high oxygen levels.In premature infants, ROS levels increased significantly after treatment with oxygen, in a concentration-dependent manner (p 0.05); meanwhile, p47phox translocation and expression were significantly enhanced (p 0.05) as well. In agreement, PBMCs cultured in vitro showed increased ROS levels after treatment with high oxygen concentrations; p47phox translocation, and expression increased as well (p 0.05). However, treatment with DPI or apocynin resulted in opposite effects.Treatment with oxygen increases p47phox translocationand expression, which in turn induce ROS production. DPI and apocynin have the opposite effects.

Porous poly(L-lactide), PLLA, and poly(D-lactide), PDLA, films were fabricated by the electric breath figure (EBF) method. The pore sizes of both the PLLA and PDLA films were reduced with the increase in electric voltage; however, the former followed a non-linear fit, whereas the latter followed a linear fit. Under 0 V condition, the average diameter, da, was 15.0 ± 0.50 μm for the PLLA film and 6.66 ± 0.54 μm for the PDLA film, whereas under 3 kV condition, da was 1.65 ± 0.35 μm for the PLLA film and 0.73 ± 0.07 μm for the PDLA film. FTIR spectra analysis showed that the EBF process did not change the structure of PLLA or PDLA.

This work demonstrated that the superhydrohpobic polystyrene, PS, film can be one step fabricated by electric-assisted phase separation, EAPS, method. Experimentally, a linear PS was dissolved in a THF/ethanol mixing solvent and casted on a glass substrate under an electric field condition to start electric-assisted phase separation to form PS film. FESEM images showed that the PS film surface presented numrous micropheres while the size was reduced with the applied voltage increase. Water contact angles on these PS surfaces was increased with the voltages increase, e.g. at 126.5° for 0 V sample and at 151.3° for 3000 V sample.

To apply SiO2 nanoparticles to assist the formation of polystyrene, PS, film via the electric breath figure, EBF, method in relation to various voltages, the obtained PS/SiO2 films showed highly ordered pore structures and the pore diameter was gradually reduced with the applied voltage increase. On these PS/SiO2 film surfaces, water contact angle was found at 110.3° for 0 V sample and 131.4° for 3000 V sample, however, on chlorotrimethylsilane deposited film surface was greatly enhanced to at 129.2° for 0 V sample and 150.1° for 3000 V sample. This indicated that this physical and chemical combined method can form highly ordered porous structure with superhydrophobic behavior.

Small for gestational age (SGA) infants have an increased risk of necrotizing enterocolitis (NEC), but SGA has been found to not be a risk factor for the deterioration of NEC in previous literature. Few studies have focused on correlative factors of the progression of NEC in SGA newborns. The present retrospective observational study was performed in 64 SGA infants with Bell’s stage II NEC. The dependent variable was Bell’s stage II NEC that progressed to stage III after diagnosis. A stepwise forward multivariate logistic regression model was used to select potential correlative factors for the progression of NEC in SGA newborns. The results showed that elevation of CRP after NEC diagnosis (aOR 39.21, 95% CI 6.62–249.2) has an increased risk for deteriorating Bell’s stage II NEC. In contrast, NEC in infants with congenital heart disease had a decreased risk of deterioration (aOR 0.11, 95% CI 0.01–0.92). Our findings indicated that serial CRP measurements post NEC diagnosis may be useful in predicting the deterioration of NEC.

Objective To investigate whether the polymorphisms of TLR7/MyD88 signaling pathway is associated with the susceptibility to and severity of hand, foot and mouth disease (HFMD) caused by enterovirus 71 (EV71) in children. Methods We collected 180 EV71 HFMD cases and 201 healthy controls from both the Second Affiliated Hospital of Xi'an Jiaotong University and Xi'an Children's Hospital. The genotypes including rs3853839, rs179010 of TLR7, and rs7744 of MyD88 were detected in the 381 samples by SNPscan kit. Results The susceptibility risk (OR=2.343, 95%CI:1.516-3.621) and severity risk (OR=1.939, 95%CI: 1.064-3.521) of TLR7 rs3853839 allele C significantly increased in the male children with EV71 HFMD. Also, the susceptibility risk (OR=1.701, 95%CI: 1.142-2.535) and severity risk (OR=1.852, 95%CI: 1.038-3.305) of TLR7 rs179010 allele T significantly increased in the male children with EV71 HFMD. But there was no significant difference in the distribution of TLR7 rs179010 and rs3853839 genes between female children with EV71 HFMD and female controls. There was no correlation between the genetic polymorphisms of MyD88 rs7744 and the susceptibility to and severity of EV71 HFMD in the children. Conclusion Polymorphisms of TLR7 rs3853839 and rs179010 are correlated to the susceptibility to and severity of EV71 HFMD in male children.

Two new guaiane-type sesquiterpenoids, named 4α,5α-epoxy-8β-hydroxy-1α-hydro-α-guaiene (1) and 4α,5α-epoxy-1-hydroxy-α-guaiene (2), were isolated from the whole plants of Valeriana hardwickii. Their structures were elucidated on the basis of spectroscopic analysis. Compounds 1 and 2 showed weak cytotoxicity against the lung adenocarcinoma (A549) and hepatoma (Bel7402) cell lines with IC50 values of 9.2 and 8.5 μM, respectively.

Background and aims Enterovirus 71 (EV71) has caused great morbidity, mortality, and use of health service in children younger than five years in China. Vaccines against EV71 have been proved effective and safe by recent phase 3 trials and are now available in China. The purpose of this study was to evaluate the health impact and cost-effectiveness of a national EV71 vaccination program in China. Methods Using Microsoft Excel, a decision model was built to calculate the net clinical and economic outcomes of EV71 vaccination compared with no EV71 vaccination in a birth cohort of 1,000,000 Chinese children followed for five years. Model parameters came from published epidemiology, clinical and cost data. Results In the base-case, vaccination would annually avert 37,872 cases of hand, foot and mouth disease (HFMD), 2,629 herpangina cases, 72,900 outpatient visits, 6,363 admissions to hospital, 29 deaths, and 945 disability adjusted life years. The break-even price of the vaccine was $5.2/dose. When the price was less than $8.3 or $14.6/dose, the vaccination program would be highly cost-effective or cost-effective, respectively (incremental cost-effectiveness ratio less than or between one to three times China GDP per capita, respectively). In one-way sensitivity analyses, the HFMD incidence was the only influential parameter at the price of $5/dose. Conclusions Within the price range of current routine vaccines paid by the government, a national EV71 vaccination program would be cost-saving or highly cost-effective to prevent EV71 related morbidity, mortality, and use of health service among children younger than five years in China. Policy makers should consider including EV71 vaccination as part of China’s routine childhood immunization schedule., Author summary Enterovirus 71 (EV71) has caused great morbidity, mortality, and use of health service in children younger than five years in China. Recently, effective and safe vaccines against EV71 have been approved. Whether EV71 vaccination should be included as part of China’s routine childhood immunization schedule is unknown. In this study, we built a decision model to evaluate the health impact and cost-effectiveness of a national EV71 vaccination program in China. We find that vaccination would annually avert 567,500 cases of hand, foot and mouth disease (HFMD), 40,000 herpangina cases, 1,093,500 outpatient visits, 95,500 admissions to hospital, 435 deaths, and 14,000 disability adjusted life years based on the current Chinese birth cohort size. The break-even price of the vaccine was $5.2/dose. Within the price range of current routine vaccines paid by the government, a national EV71 vaccination program would be cost-saving or highly cost-effective. Policy makers should consider including EV71 vaccination as part of China’s routine childhood immunization schedule.

A novel electric breath figure (EBF) method was developed by applying an electrostatic generator to assist the common breath figure (BF) method, and it was applied to fabricate polystyrene (PS) film. FESEM images showed that these PS films have a porous structure with controlled pore size because the average pore diameter, da, of these PS films was about 2.30 μm at 0 V, and it gradually reduced to about 0.35 μm at 1000 V, i.e. it followed the relationship: da = a − b ln(ρ + c), where ρ is the voltage and a, b and c are three constants. Wetting results showed that the water contact angle, θw, on these PS films was also controlled because the 0 V PS film showed a θw value at 95.07°, and it increased to 147.09° when ρ increased to 1000 V. The reason that this EBF method can control formation of porous film was found to be a reduction in water surface tension with increasing voltage.

AIM To investigate the antioxidant effect of caffeic acid phenethyl ester (CAPE) in hepatic stellate cell-T6 (HSC-T6) cells cultured in vitro and the potential mechanisms. METHODS HSC-T6 cells were cultured in vitro and treated with various concentrations of CAPE for 24, 48 and 72 h, respectively. Cell proliferation was investigated using the MTT assay, and cell ultrastructural alterations were observed by transmission electron microscopy. Flow cytometry was employed to investigate the effects of CAPE on apoptosis and the levels of reactive oxygen species in HSC-T6 cells cultured in vitro. An enzyme immunoassay instrument was used to evaluate antioxidant enzyme expression. The effect on α-smooth muscle actin was shown using immunofluorescence. Gene and protein levels of Nrf2, related factors, and mitogen activated protein kinases (MAPKs), in HSC-T6 cells were detected using RT-PCR and Western blot, respectively. RESULTS CAPE inhibited the proliferation and activation of HSC-T6 cells cultured in vitro. CAPE increased the antioxidant levels and the translocation of Nrf2 from the cytoplasm to the nucleus in HSC-T6 cells. Moreover, the phosphorylation of MAPKs in cells decreased in response to CAPE. Interestingly, CAPE-induced oxidative stress in the cells was significantly attenuated by pretreatment with MAPKs inhibitors. CONCLUSION CAPE inhibits cell proliferation and up-regulates the antioxidant levels in HSC-T6 cells partly through the Nrf2-MAPKs signaling pathway.

Human enterovirus 71 (EV71) is one of the major causes of hand, foot and mouth disease (HFMD), which leads to significant mortality in infected children. A prophylactic vaccine is urgently needed. However, little is known about the protective T-cell immunity in individuals infected with the EV71 virus. In this study, we performed a comprehensive ex vivo interferon-γ ELISPOT analysis in 31 children infected with EV71 as well as in 40 healthy adult controls of the CD4(+) and CD8(+) T-cell responses to overlapping peptides spanning the VP1 structural protein and RNA-dependent RNA polymerase (RdRp) non-structural protein. EV71-specific CD4 T-cell responses were detected in most of the acute patients and were mostly CD4-dependent RdRp-specific responses. CD8-dependent VP1 and RdRp-specific responses were also detected in a small proportion of recently infected children. There was no significant association between the strength of the T-cell responses and disease severity observed during the acute EV71 infection phase. Interestingly, an RdRp-specific, but no VP1-specific, CD4-dependent T-cell response was detected in 30% of the adult controls, and no T-cell responses were detected in healthy children. In addition, 24 individual peptides containing potential T-cell epitope regions were identified. The data suggest that CD4-dependent RdRp-specific T-cell responses may play an important role in protective immunity, and the epitopes identified in this study should provide valuable information for future therapeutic and prophylactic vaccine design as well as basic research.

To study the compatible mechanisms and compatible proportion of Shaoyao Gancao decoction, the intestinal absorption of main ingredients in Shaoyao Gancao decoction SG11 (Baishao-Zhigancao 1: 1) , SG31 (Baishao-Zhigancao 3: 1), Baishao water decoction S and Zhigancao (G) were investigated and compared using in vitro everted intestinal sac model and in situ single pass intestinal perfusion (SPIP) model. The concentration of paeoniflorin (PF), liquiritin (LQ) and mono-ammonium glycyrrhizinate (GL) in test samples and samples of intestinal sac and intestinal perfusion was determined by HPLC. The intestinal absorptive amount and absorption parameters were calculated. Results showed that in the everted intestinal sac model, three ingredients could be absorbed by duodenum, jejunum and ileum, and the absorption in the jejunum was best for all 3 ingredients. The absorption rate of three ingredients in SG11 was significantly higher than that in single decoction (P0.05), but had no significant difference compared with SG31. In SPIP model, the absorption rate constant K(a), the apparent absorption coefficient P(app) and the absorption rate of three ingredients in SG11 were significantly higher than those in single decoction. Parameters of PF and GL in SG11 were significantly higher than those in SG31, but had no differences of LQ. It proved that the compatibility of Baishao and Zhigancao could improve the intestinal absorption of PF, LQ and GL. The absorption of each ingredient in SG11 was better than that in SG31.

Chemical investigation of the whole plants of Valeriana hardwickii has led to the isolation of 11 flavones and 2 monoterpe- noids by using various chromatographic techniques including column chromatography on silica gel and Sephadex LH-20, preparative TLC, and preparative HPLC. Their structures were identified by spectroscopic data analysis as syzalterin (1), 6-methylapigenin (2), 5-hydroxy-7,4'-dimethoxyflavone (3), genkwanin (4), acacetin (5), apigenin (6), quercetin (7), tricin (8), (-)-farrerol (9), sosakuranetin (10), 5,3',4'-trihydroxy-7-methoxyflavanone (11), (-)-bornyl ferulate ( 12) , and (-)-bornyl caffeate ( 13). All compounds were isolated from this plant for the first time, while compounds 1, 9-13 were obtained from this genus for the first time.

Activating transcription factor 3 (ATF3) is an adaptive‑response gene of the ATF family. ATF3 activity may be induced in response to a number of different stress-associated signals and ATF3 is involved in a variety of cellular processes. However, the functions of ATF3 and its molecular networks in human hepatoma cells lines and hepatitis C virus-infected Huh7 (HCV-Huh7) cells are not well understood. In the present study, ATF3 regulatory networks in Huh7 and HCV-Huh7 cell lines were established using the linear programming-based GRNinfer software and molecule annotation system 3.0 software. The gene expression omnibus dataset, GSE20948, was analyzed. The resulting network consisted of clusters located upstream and downstream of ATF3 in Huh7 and HCV-Huh7 cell lines. Using the annotation, visualization and integrated discovery (DAVID) software, 10 activation and 2 inhibition enriched functional annotation clusters were identified downstream of ATF3 in HCV-Huh7 cells. However, there were no enriched functional annotation clusters identified upstream of ATF3 in HCV-Huh7 cells. Furthermore, no clusters were identified downstream nor upstream of ATF3 in Huh7 cells. Gene ontology term and Kyoto encyclopedia of genes and genomes pathway analyses demonstrated that ATF3 may be involved in a number of biological processes, in particular, in metabolism regulation in HCV-Huh7 cells. It is hypothesized that the ATF3 pathway may be activated in Huh7 cells following HCV infection and that it is a potential 'hub' in the network of HCV-Huh7 cells.

City roads are playing vital roles in the economic development. And both highway roadbeds take place geological disasters frequently. This article evaluates geological disasters with the method of AHP. The indexes include lithology, geological structure, slope, high slope, slope structure types, groundwater, slope size, rainfall, river erosion, slope back excavation, earthquake, etc. Through the analysis of the formation mechanism and impact factors in landslide, the model of landslide hazard assessment is established. Then landslide risk factors are partitioned into different levels, and seven part of geological disasters area are calculated. At last determine the risk level of landslide areas comparing with the risk classification table.

OBJECTIVE: To explore the effect of resveratrol on the levels of sirtuin 1 (SIRT1) and reactive oxygen species (ROS) in peripheral blood mononuclear cells (PBMCs) of premature infants exposed to hyperoxia. METHODS: Peripheral blood and isolated PBMCs from premature infants (gestational age < 32 weeks) without oxygen supplement were collected and were randomly assigned into four groups: control, air+resveratrol, hyperoxia, and hyperoxia + resveratrol. The PBMCs were cultured in vitro for 48 hours, then the ROS content in PBMCs was measured by laser scanning confocal microscopy. Malondialdehyde (MDA) content in the medium was measured by the whole spectrum spectrophotometer. SIRT1 positioning was assessed by immunofluorescence. SIRT1 expression levels in PBMCs were measured by Western bolt. RESULTS: Compared with the control group, the level of SIRT1 in the air+resveratrol group increased significantly (P < 0.05). The levels of ROS and MDA and the SIRT1 transposition rate in the hyperoxia group increased significantly, while the expression level of SIRT1 decreased significantly compared with the control group (P < 0.05). The levels of ROS and MDA and the SIRT1 transposition rate decreased significantly (P < 0.05), and the expression level of SIRT1 increased significantly in the hyperoxia+resveratrol group (P < 0.05). CONCLUSIONS: Resveratrol can increase SIRT1 expression in PBMCs and inhibit SIRT1 shuttle from nucleus to cytoplasm in order to increase the ability of antioxidative stress in premature infants exposed to hyperoxia, thereby reducing the oxidative stress injury in premature infants.

To determine the degree of HIV-1-specific cytotoxic-T-lymphocyte (CTL) cross-responses to the clade B and C consensus sequences at the single peptide level. We assessed CTL responses in 46 HIV-1 clade B chronically infected individuals using an interferon-gamma Elispot assay with a total of 826 overlapping peptides spanning HIV-1 clade B and C consensus sequences. In general, 583 peptides were recognized by HIV-1-specific T cells in the study subjects (292 clade B, 291 clade C respectively), of which 204 peptides in both clades were recognized simultaneously. The HIV-1-specific CTL responses to both clade peptides contributed 54.23% (954/1759) to the total responses. No significant difference was observed between the overall magnitude or frequency of CTL responses to clade B proteins and those to clade C proteins. According to the profiles of CTL magnitude and CTL frequency, the top 44 and 35 synthetic peptides were identified as immunodominant regions in the clade B and C consensus sequences respectively and 27 corresponding peptides in two immunodominant regions were cross-reactive. These peptides with cross-reactivity had a significantly higher ability to elicit CTL responses (P0.01) and preferentially had a trend of lower entropy and higher inter-clade homology. A wide degree of cross-clade reactivity of HIV-1-specific T cells exist in clade B and clade C variants. Most of immunodominant peptides with cross-reactivity are vigorous to elicit CTL responses and preferentially be conservative. This result may make future HIV-1 vaccines including multiple copies of CTL epitopes in these immunodominant peptides effective for this population.

Chinese patent medicine with double identity was a special phenomenon, and many preparations not only were prescription drugs but also over the counter ( OTC) drugs, which brought a lot of trouble. Based on statistics of list of OTC medicines of CFDA, related varieties, route of administration and functions of these drugs were searched. The causes of insufficient were analyzed and the potential risk was investigated. To ensure the safety of drug usage for the patient, risk management system should be set up by improving the technical requirements for registration, improving the drug labels and manuals, playing the role of pharmacists in pharmacy services and raising awareness of doctor and patient for these drugs.

To explore the effect of silence of Pin1 expression on hyperoxia-induced apoptosis in alveolar epithelial cells A549.A549 cells were divided into four groups: control, hyperoxia, negative lentivirus and Pin1-shRNA hyperoxia. The hyperoxia group was exposed to a mixture of 95%O2 and 5%CO2 for 10 minutes. Then cells were cultured in a closed environment. After 24 hours, the changes of morphology were observed under an inverted microscope. Cell apoptosis was detected by flow cytometry (FCM). The expression of X-linked inhibitor of apoptosis protein (XIAP) and Caspase-9 were detected by immunohistochemistry. The production of reactive oxygen species (ROS) and cellular mitochondria membrane potential (△Ψm) were determined by fluorescence microscopy.Under the inverted microscope, the A549 cells grew slowly and the changes in morphology of the cells were most obvious in the hyperoxia and negative lentivirus groups. The changes in morphology of A549 cells were obviously improved in the Pin1-shRNA hyperoxia group. The FCM results showed that the apoptosis rate of A549 cells increased, Caspase-9 expression increased, XIAP expression decreased, mitochondrial ROS production increased and mitochondrial membrane potential decreased in the hyperoxia and negative lentivirus groups compared with the control group (P0.05). Compared with the hyperoxia and negative lentivirus groups, the apoptosis rate of A549 cells decreased, Caspase-9 expression decreased, XIAP expression increased, mitochondrial ROS production decreased and mitochondrial membrane potential increased in the Pin1-shRNA hyperoxia group (P0.05), although the levels of the indexes did not reach to those of the control group.Silencing of Pin1 could suppress hyperoxia-induced apoptosis of A549 cells.

AIM: To investigate the hepatoprotective effects and antioxidant activity of caffeic acid phenethyl ester (CAPE) in rats with liver fibrosis. METHODS: A total of 75 male Sprague-Dawley rats were randomly assigned to seven experimental groups: a normal group (n = 10), a vehicle group (n = 10), a model group (n = 15), a vitamin E group (n = 10), and three CAPE groups (CAPE 3, 6 and 12 mg/kg, n = 10, respectively). Liver fibrosis was induced in rats by injecting CCl4 subcutaneously, feeding with high fat forage, and administering 30% alcohol orally for 10 wk. Concurrently, CAPE (3, 6 and 12 mg/kg) was intraperitoneally administered daily for 10 wk. After that, serum total bilirubin (TBil), aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured to assess hepatotoxicity. To investigate antioxidant activity of CAPE, malondialdehyde (MDA), glutathione (GSH) levels, catalase (CAT) and superoxide dismutase (SOD) activities in liver tissue were determined. Moreover, the effect of CAPE on α-smooth muscle actin (α-SMA), a characteristic hallmark of activated hepatic stellate cells (HSCs), and NF-E2-related factor 2 (Nrf2), a key transcription factor for antioxidant systems, was investigated by immunohistochemistry. RESULTS: Compared to the model group, intraperitoneal administration of CAPE decreased TBil, ALT, and AST levels in liver fibrosis rats (P < 0.05), while serum TBil was decreased by CAPE in a dose-dependent manner. In addition, the liver hydroxyproline contents in both the 6 and 12 mg/kg CAPE groups were markedly lower than that in the model group (P < 0.05 and P < 0.001, respectively). CAPE markedly decreased MDA levels and, in turn, increased GSH levels, as well as CAT and SOD activities in liver fibrosis rats compared to the model group (P < 0.05). Moreover, CAPE effectively inhibited α-SMA expression while increasing Nrf2 expression compared to the model group (P < 0.01). CONCLUSION: The protective effects of CAPE against liver fibrosis may be due to its ability to suppress the activation of HSCs by inhibiting oxidative stress.

A new method (fuzzy factor method based on the fuzzy sets theory) for the dynamic response analysis of fuzzy truss system under non-stationary stochastic excitation is presented. Considering the fuzziness of the structural physical parameters and geometric dimensions simultaneously, the fuzzy correlation function matrix of the structural displacement response in time domain is derived using the fuzzy factor method and the optimisation method; then from the structural non-stationary stochastic response in the frequency domain, the fuzzy mean square values of the displacement and stress response are developed by the fuzzy factor method. The influences of the fuzziness of the physical parameters and geometric dimensions on the fuzziness of the mean square values of the structural displacement and stress response are illustrated via two engineering examples and some important conclusions are obtained.

CD4+ T cell counts and CD4+:CD8+ T cell ratios represent key determinants of HIV disease progression and infectivity. However, the relationship between the HIV-1-specific cytotoxic T lymphocyte (CTL) response and these determinants has not been elucidated for all HIV-1B and HIV-1C proteins. In the present study, virusspecific T cell responses to HIV-1B and HIV-1C proteins were analyzed with interferon gamma (IFN-gamma) enzyme- linked immunospot (ELISpot) assays using synthetic overlapping peptides corresponding to naturally occurring HIV-1B and HIV-1C consensus sequences. For Gag/Gag p24/Gag p17, a correlation between T cell responses and CD4+ T cell count in HIV-1 clade B and clade C was seen: elevated T cell response resulted in higher CD4+ T cell production. A statistically significant correlation between the Pol-specific T cell response and CD4+ T cell counts was also found in HIV-1 subtype C. For all HIV-1B and HIV-1C proteins, a correlation between the HIV-1-specific T cell response and CD4+:CD8+ T cell ratios was found for Tat and Pol proteins. CD4+ T cell counts in patients with Tat and/or Rev T cell response were higher than in patients without Tat and/or Rev T cell response. We suggest that this correlation within HIV-1B and HIV-1C Gag p24/Gag p17 responses makes the Gag p24/Gag p17 region a potential vaccine candidate and that HIV-1-specific CTL epitopes toward Pol are important in controlling HIV-1 infection; we emphasize that future vaccination strategies should include these early antigens, Tat and Rev.

Ethnic medicine industry is facing many problems such as narrow market, exhaustion of resource, decline of ethnic medicine and no qualified successors. Sustainable development theory was utilized to analyse the elements and problems of ethnic medicine industry, and the counter measures were put forward to get rid of the predicament and to realize the sustainable development which depends on the ethnic medicine resources, national medicine, industrial policy, personnel training and modern technology. The development issues of ethnic medicine industry can be solved by the coordination of enterprise, government and public. Finally the ethnic medicine can provide better services for society.

Transforming growth factor (TGF)-β1 expression is induced upon liver injury, and plays a critical role in hepatic fibrosis. Antibodies against TGF-β1 represent a novel approach in the treatment of hepatic fibrosis. However, TGF-β1 is not a suitable antigen due to immunological tolerance. In the current study, we synthesized a multiple antigenic peptide (MAP) vaccine against the dominant B-cell epitope of TGF-β1. The immunogenicity and potential therapeutic effects of this vaccine were examined using a rat model of hepatic fibrosis. Dominant B-cell epitopes of TGF-β1 were identified using bioinformatic program. An MAP vaccine corresponding to the 90–98 amino acid domain of TGF-β1 and containing four dendritic arms was synthesized using a 9-fluorenylmethoxycarbonyl solid phase method. Hepatic fibrosis which was induced in male Sprague-Dawley rats received a high-fat diet and ethanol (1.8 g/kg). Starting from the third week, rats were exposed to 40 % carbon tetrachloride (CCl4; 150 μl/100 g body weight twice weekly, initially 200 μl/100 g) treatment for a duration of 8 weeks. Rats received the MAP vaccine (100 μg) or Freund’s adjuvant at weeks 1, 3, 5. A group of rats receiving the fibrosis-inducing regimen alone and a group of healthy rats (receiving an olive oil vehicle alone) were included as controls. At the conclusion of the experiment, serum titre of TGF-β1 antibody was measured using ELISA and a standard liver functional test panel was examined. The extent of hepatic fibrosis was determined by measuring hydroxyproline content in the liver as well as hematoxylin–eosin (HE) and van Gieson (VG) staining. The expression of TGF-β1 and alpha-smooth muscle actin (α-SMA) was examined using immunohistochemistry, and presented as positive staining cells. The MAP purity was >90 % upon reverse phase high-performance liquid chromatography, with apparent molecular weight at 4.77 kDa. Serum TGF-β1 antibody titre was 1:256. The fibrosis-inducing treatment produced significant liver damage, as reflected by increases in liver functional test, HE and VG staining. The MAP vaccine attenuated such damage, as reflected by decreased alanine aminotransferase, aspartate aminotransferase, total bilirubin, and hepatic hydroxyproline (116.78 ± 23.76 vs. 282.71 ± 136.94 IU/L; 319.78 ± 82.48 vs. 495.29 ± 137.13 IU/L; 2.02 ± 0.27 vs. 4.01 ± 0.52 μmol/L; 263.67 ± 41.18 vs. 439.14 ± 43.29 μg/g vs. in model rats, respectively; p