OBJECTIVETo study the effect of Wuhu Decoction (WHD) on the expression of co-stimulation molecule of peripheral dendritic cells (DC), CD80, CD83 and CD86, in infants with asthma, in order to provide practical basis for further elucidate the action mechanism of WHD in preventing and treating infantile asthma.

METHODSSixty infants with asthma of Fei phlegm-heat accumulation syndrome type were randomized into the treatment group treated with WHD and the control group treated with Western medicine (fluticasone propionate oral taking or inhalation). And 10 healthy infants were set as normal control. With Thomas method adopted, the DC were isolated from peripheral blood of all infants subjected. The expressions of surface co-stimulation molecules of DC, CD80, CD83 and CD86, were detected by flow cytometry. Their changes before and after treatment in different groups were analyzed and compared.

RESULTSExpressions of CD80 and CD86 of peripheral blood DC in asthmatic infants were remarkably higher than those in the normal control (P<0.01). In the treated group, CD80 expression lowered from 18.06 +/- 4.53 before treatment to 13.18 +/- 3.02 after treatment and CD86 expression lowered from 38.61 +/- 10.54 to 29.65 +/- 8.55; while in the control group, the two expressions were lowered from 18.40 +/- 3.86 to 15.34 +/- 3.90, and from 38.29 +/- 11.67 to 35.88 +/- 13.85 respectively, the lowering in both groups were statistically significant (P<0.01 and P<0.05), but it was more significant in the treated group (P<0.05). As for CD83, no significant difference existed between groups and no change was found in either group after treatment (P>0.05).

CONCLUSIONWHD can regulate the co-stimulation molecules of dendritic cells in asthma infants to reduce the expressions of CD80 and CD86.

Antigens, CD ; metabolism ; Asthma ; blood ; drug therapy ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; Child, Preschool ; Dendritic Cells ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulins ; metabolism ; Infant ; Male ; Membrane Glycoproteins ; metabolism ; Phytotherapy

Objective To predict and identify liner B-cell epitopes in the hemagglutinin ( HA) of human-infected avian-origin H7N9 influenza virus and analyze the specificity of H7 subtype.Methods Three serum samples collected at different times from the same patient who was confirmed to be infected with H7N9 influenza virus were provided by Shaoxing People’s Hospital, and one serum sample from healthy person was collected as the control.The extracellular region of HA protein was predicted by TMHMM Sever v.2.0.The potential B-cell epitopes were predicted by DNAStar Lasergene’ s Protean, BcePred and ABCpred tools, and the immunogenicity of the predicted B cell antigen epitopes was assessed by indirect enzyme-linked immunosordent assay ( ELISA ) .H7 subtype specificity was analyzed by comparing HA protein amino acid sequence with H7N9 and H1-H16 subtype influenza virus from Genbank using Clustal X 2.1 software, and Cn3D 4.3.1 software was used to detect the distribution and 3D structure of predicted epitopes on the HA protein of H7N9.Results The potential B-cell epitopes may be located in 172-183, 363-380, 452-472 and 491-506 of extracellular N-terminus of HA protein.ELISA showed that four predicted eptiopes specifically reacted with positive serums from patient.Multi-sequence alignment demonstrated that peptide 172-183 and 363-380 had higher H7 subtype specificity compared with amino acid sequences of other subtypes.Moreover, the predicted linear B-cell epitopes all located on the surface of HA protein according to the 3D structure analysis.Conclusion Four potential B-cell epitopes were identified, in which peptide 172-183 and 363-380 have higher H7 subtype specificity, and may be used in the design of epitope-based vaccines and diagnostics tests.

OBJECTIVETo establish a high performance liquid chromatographic (HPLC) amalgamated to double UV waves method for simultaneous determination of loganin and paeonol in Liuwei Dihuang pills.

METHODA HPLC method was developed. The separation was carried out on a Agilent Zorbax SB C18 column (5 microm, 4.6 mm x 250 mm). The mobile phase consisted of water (A) and acetonitrile (B) with linear linear gradient elution [0-8 min, (B) from 1% to 12%; 8-21 min, B keep 12%; 21-40 min, (B) from 12% to 90%; 40-50 min, B keep 90% for 10 min]. The detection was Photodiode Array with the detection wavelengths were at 236 nm and 274 nm. The column temperature being 30 degrees C and the flow rate was 1.0 mL min(-1). Extracting the chromatergraph from 274 nm and 236 nm, we amalgamated the two chromatographs by matlab programmed.

RESULTThe calibration curves of loganin and paeonol were linear in the ranges of 0.0362-1.09 microg (r =0. 9998) and 0.0450-1.35 microg (r =0.9998), respectively. The average recoveries of loganin and paeonol were 97.3% (RSD 1.4 %) and 103.0% (RSD 1.9%), respectively. Three different batches of Liuwei Dihuang pills were determined with this method.

CONCLUSIONThis is a more convenient, reasonable and credible quality control method for the traditional Chinese medicine.

Acetophenones ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Iridoids ; chemistry ; Reproducibility of Results ; Spectrophotometry, Ultraviolet

OBJECTIVETo investigate the influence of intrauterine infection caused by lipopolysaccharide on Toll-like receptor 4 (TLR4) signaling pathway in fetal and neonatal rat lungs in order to explore immunomodulating activity of innate immunity responding to intrauterine infection and its effect on lung development.

METHODSOn day 17 of pregnancy, 30 pregnant Sprague-Dawley (SD) rats were randomly divided into two groups: LPS group and saline group. For LPS group, LPS (10 microl, 40 microg/ml) was intrauterine injected between every two embryonic sacs of the pregnant rats, while the rats in the control group were injected with the same volume of pyrogen-free saline. Lung tissues of fetal rats and corresponding placental tissues were collected on the embryonic day 18 (E18), E20, and E22. Neonatal lung tissues were also harvested on postnatal day 1 (P1), P3, and P7. Lung sections and placental tissues were stained with hematoxylin and eosin for histological examination. Reverse transcription quantitative polymerase chain reaction (RT-PCR) analysis was performed to test mRNA expression for TLR4, myeloid differentiation 88 (MyD88) and IL-1beta, while immunohistochemistry was used to evaluate TLR4 and MyD88 expression in lung tissues. All data were analyzed with one-way analysis of variance (ANOVA) and q test.

RESULTS(1) Placental hematoxylin-eosin staining showed a great number of neutrophils infiltration, obvious interstitial hyperplasia and narrow capillaries in placental tissues in the LPS group which indicated that intrauterine infection occurred. However, there were no obvious inflammatory cells in the control group. (2) On E18, E20 and E22, the lung of LPS group showed no obvious pathological changes, and there were no apparent neutrophils infiltrated in alveoli, then some structural changes appeared. On P7, we found that the number of alveoli decreased, space of alveoli was larger than ever, septa thickened, but without significant constructive disorder. (3) In the LPS group, the TLR4, MyD88 and IL-1beta mRNA levels increased compared with control group, higher than control group at E20 and E22 (P < 0.05), and peaked at E22. Then the expression levels of these substances decreased slowly. (4) The result of immunohistochemistry showed that in lung tissues of the two groups at E18, there was no remarkable positive staining of TLR4 and MyD88, which mainly expressed in cytoplasm of bronchiole and alveolar epithelial cells, then positive cells increased slowly.

CONCLUSION(1) For perinatal rat lungs, intrauterine LPS infusion can induce an increased expression levels of TLR4 and MyD88 to a certain extent, which then returned to normal level gradually. At the same time, lung tissues showed a mild pathological change and inflammatory reaction. We propose that innate immune system of fetal lungs controls the magnitude of the LPS-induced cytokine response during the perinatal period. (2) The findings confirmed that LPS-activated signaling transduction pathway was the MyD88-dependent pathway.

Animals ; Animals, Newborn ; Female ; Infection ; metabolism ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; administration & dosage ; pharmacology ; Lung ; growth & development ; metabolism ; Myeloid Differentiation Factor 88 ; metabolism ; Pregnancy ; Pregnancy Complications, Infectious ; metabolism ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; metabolism

BACKGROUNDDrug susceptibility assay is very important in tuberculosis therapy. Pyrazinamide is a first line antituberculosis drug and diagnosis of its resistance in Mycobacterium tuberculosis (M. tuberculosis) is difficult and time consuming by conventional methods. In this study, we aimed to evaluate the performance of the microscopic observation drug susceptibility (MODS) assay in the detection of pyrazinamide resistance in M. tuberculosis relative to the conventional Wayne assay and Lowenstein-Jensen (LJ) proportion method.

METHODSM. tuberculosis clinical isolates (n = 132) were tested by the MODS and the Wayne assay: the results were compared with those obtained by the LJ proportion method. Mutations in the gene were identified by direct sequencing of the pncA genes of all isolates in which pyrazinamide resistance was detected by any of the three methods.

RESULTSCompared to the LJ results, the sensitivity and specificity of the MODS assay were 97.8% and 96.5% respectively; the sensitivity and specificity of the Wayne assay were 87.0% and 97.7% respectively. Mutations in the pncA gene were found in 41 of 46 strains that were pyrazinamide resistant (3 tests), in 1 of the 4 strains (LJ only), in 42 of 48 strains (at least 1 test), but no mutations in 1 strain sensitive according to the MODS assay only. The MODS assay, Wayne assay and LJ proportion method provided results in a median time of 6, 7 and 26 days respectively.

CONCLUSIONSMODS assay offers a rapid, simple and reliable method for the detection of pyrazinamide resistance in M. tuberculosis and is an optimal alternative method in resource limited countries.

Antitubercular Agents ; pharmacology ; Microbial Sensitivity Tests ; Microscopy ; methods ; Mycobacterium tuberculosis ; drug effects ; Pyrazinamide ; pharmacology

OBJECTIVETo determine the changes in the levels of endogenous metabolites in rats with chronic immobilization stress (CIS) taking Xiaoyao Powder (XYP) and its modified prescription version, which lacks the volatile oils extracted from Herba Menthae.

METHODSTwenty-four experimental male SD rats were randomly divided into 4 groups of 6 rats each: control, model, XYP-1 (containing volatile oils from Herba Menthae), and XYP-2 (lacking volatile oils). All rats except control group rats were subjected to CIS 3 h per day for 21 consecutive days. Groups XYP-1 and XYP-2 were given the extracted XYS with or without volatile oils (3.854 g/kg; suspended in distilled water) via gavage 1 h before CIS each day for 21 days. Rats were anesthetized using intraperitoneal injection of pentobarbital sodium (40 mg/kg) on the 22nd day. Observations were made using a Varian INOVA 600 MHz NMR spectrometer at 27 °C. Carr-Purcell-Meiboom-Gill (CPMG) and longitudinal eddy-delay (LED) were applied, resulting in spectra showing only the signals from micro- and macro-metabolites.

RESULTSCompared to controls, rats subjected to CIS showed increased levels of plasma metabolites, such as acetic acid, choline, N-glycoprotein (NAC), saturated fatty acid, and blood sugars. Levels of low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), and unsaturated fatty acids were decreased. The biochemical effects of XYS were characterized by elevated levels of VLDL, LDL, threonine, methionine, and glutamic acid in plasma.

CONCLUSIONSome common and characteristic metabolites on the anti-CIS of XYP and its modified prescription were obtained. The metabolomics technology is a valuable tool and may be used to identify the specific metabolites and potential biomarkers of therapeutic effect of Chinese medicinal prescriptions.

Animals ; Blood Proteins ; drug effects ; metabolism ; Chronic Disease ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Male ; Metabolome ; drug effects ; Powders ; Rats ; Rats, Sprague-Dawley ; Restraint, Physical ; Stress, Psychological ; blood ; drug therapy ; metabolism

Objective To establish the PCR amplification and sequencing system of Hsa-miR-17 gene promoter region.Methods To establish the PCR amplification and sequencing system of Hsa-miR-17 gene promoter region and evaluate its performance by analyzing the sequence characteristics,designing PCR and sequencing primers,extracting sample genome DNA from different human cell lines and using optimization strategy of adding DMSO to the final concentration of 5 ％ and making use of 5' tailed PCR primers;Then the established system was further verified in 10 other cell line samples.Results The PCR amplification and sequencing system of hsa-miR-17 gene promoter region,which had good repeatability,good specificity and its detection limit was 10 ng/μl,was established successfully under the condition of using 5' tailed PCR primers and 5 ％ DMSO and that was confirmed by electrophoresis analysis and Sanger sequencing.And fragment of Has-miR-17 gene promoter region in 10 human cell lines were successfully achieved and verified by this system.Conclusion The PCR system of Hsa-miR-17 gene promoter region was established successfully.

Background/Aims: Endoscopic radiofrequency ablation (ERFA) is a treatment option for superficial esophageal squamous cell neoplasia (ESCN), with a relatively low risk of stenosis; however, the long-term outcomes remain unclear. We aimed to compare the long-term outcomes of patients with widespread superficial ESCN who underwent endoscopic submucosal dissection (ESD) or ERFA. Methods: We retrospectively analyzed the clinical data of patients with superficial ESCN who underwent ESD or ERFA between January 2015 and December 2021. The primary outcome measure was recurrence-free survival. Results: Ninety-two and 33 patients with superficial ESCN underwent ESD and ERFA, respectively. The en bloc, R0, and curative resection rates for ESD were 100.0%, 90.2%, and 76.1%, respectively. At 12 months, the complete response rate was comparable between the two groups (94.6% vs 90.9%, p=0.748). During a median follow-up of 66 months, recurrence-free survival was significantly longer in the ESD group than in the ERFA group (p=0.004), while no significant differences in overall survival (p=0.845) and disease-specific survival (p=0.494) were observed.Preoperative diagnosis of intramucosal cancer (adjusted hazard ratio, 5.55; vs high-grade intraepithelial neoplasia) was an independent predictor of recurrence. Significantly fewer patients in the ERFA group experienced stenosis compare to ESD group (15.2% vs 38.0%, p=0.016). Conclusions The risk of recurrence was higher for ERFA than ESD for ESCN but overall survival was not affected. The risk of esophageal stenosis was significantly lower for patients who underwent ERFA.

Background/Aims: Endoscopic radiofrequency ablation (ERFA) is a treatment option for superficial esophageal squamous cell neoplasia (ESCN), with a relatively low risk of stenosis; however, the long-term outcomes remain unclear. We aimed to compare the long-term outcomes of patients with widespread superficial ESCN who underwent endoscopic submucosal dissection (ESD) or ERFA. Methods: We retrospectively analyzed the clinical data of patients with superficial ESCN who underwent ESD or ERFA between January 2015 and December 2021. The primary outcome measure was recurrence-free survival. Results: Ninety-two and 33 patients with superficial ESCN underwent ESD and ERFA, respectively. The en bloc, R0, and curative resection rates for ESD were 100.0%, 90.2%, and 76.1%, respectively. At 12 months, the complete response rate was comparable between the two groups (94.6% vs 90.9%, p=0.748). During a median follow-up of 66 months, recurrence-free survival was significantly longer in the ESD group than in the ERFA group (p=0.004), while no significant differences in overall survival (p=0.845) and disease-specific survival (p=0.494) were observed.Preoperative diagnosis of intramucosal cancer (adjusted hazard ratio, 5.55; vs high-grade intraepithelial neoplasia) was an independent predictor of recurrence. Significantly fewer patients in the ERFA group experienced stenosis compare to ESD group (15.2% vs 38.0%, p=0.016). Conclusions The risk of recurrence was higher for ERFA than ESD for ESCN but overall survival was not affected. The risk of esophageal stenosis was significantly lower for patients who underwent ERFA.

Background/Aims: Endoscopic radiofrequency ablation (ERFA) is a treatment option for superficial esophageal squamous cell neoplasia (ESCN), with a relatively low risk of stenosis; however, the long-term outcomes remain unclear. We aimed to compare the long-term outcomes of patients with widespread superficial ESCN who underwent endoscopic submucosal dissection (ESD) or ERFA. Methods: We retrospectively analyzed the clinical data of patients with superficial ESCN who underwent ESD or ERFA between January 2015 and December 2021. The primary outcome measure was recurrence-free survival. Results: Ninety-two and 33 patients with superficial ESCN underwent ESD and ERFA, respectively. The en bloc, R0, and curative resection rates for ESD were 100.0%, 90.2%, and 76.1%, respectively. At 12 months, the complete response rate was comparable between the two groups (94.6% vs 90.9%, p=0.748). During a median follow-up of 66 months, recurrence-free survival was significantly longer in the ESD group than in the ERFA group (p=0.004), while no significant differences in overall survival (p=0.845) and disease-specific survival (p=0.494) were observed.Preoperative diagnosis of intramucosal cancer (adjusted hazard ratio, 5.55; vs high-grade intraepithelial neoplasia) was an independent predictor of recurrence. Significantly fewer patients in the ERFA group experienced stenosis compare to ESD group (15.2% vs 38.0%, p=0.016). Conclusions The risk of recurrence was higher for ERFA than ESD for ESCN but overall survival was not affected. The risk of esophageal stenosis was significantly lower for patients who underwent ERFA.