[Pharmacodynamic substances and mechanism of action of Xiaoer Chiqiao Qingre Granules in treatment of acute upper respiratory tract infection in children].

This study aimed to forecast the main active components of Xiaoer Chiqiao Qingre Granules(XECQ) in the treatment of children with acute upper respiratory tract infection by UPLC-MS, network pharmacology, molecular docking and cell biology, and explore the mechanism of action, so as to provide certain reference for the research on its pharmacodynamics substances and mechanism of action. The main chemical components of XECQ were comprehensively analyzed by UPLC-Q-TOF-MS combined with UNIFI platform. According to the MS1 and MS2 data of XECQ, comparison and identification were carried out in combination with reference substances and reference articles. On this basis, the chemical components of XECQ were targeted and enriched by network pharmacology, to screen the main pharmacodynamic substances of XECQ in the treatment of acute upper respiratory tract infection in children and discuss the mechanism of action. In addition, the binding degree of core targets and main active components was verified by molecular docking. The results revealed that 202 compounds were identified from XECQ, among which 22 were the main active components, including obovatol, dihydroartemisinin, and longikaurin A. Enrichment analysis of the key target pathways showed that XECQ played its role in the treatment of children with acute upper respiratory tract infection mainly by regulating PI3K/Akt signaling pathway and MAPK signaling pathway. In the experimental verification by Western Blot(WB), it was found that XECQ significantly inhibited the expression of PI3K and Akt, which was consistent with the prediction results of network pharmacology. In conclusion, the potential pharmacodynamic substances of XECQ were obovatol, dihydroartemisinin, longikaurin A and other 19 active components. It treated children with acute upper respiratory tract infection by regulating the PI3K/Akt signaling pathway.

[Digestive properties of flavonoid components in Xiaoer Chiqiao Qingre Granules in simulated infant gastrointestinal environment].

This study aimed to elucidate the digestive characteristics of flavonoid components in Xiaoer Chiqiao Qingre Granules(XECQ) in the gastrointestinal environment of infants. An in vitro model was established to simulate the gastric and intestinal environment of infants. UPLC was used to analyze the content change of flavonoid components in XECQ, and their overall content was integrated through the mass fraction weight coefficient method. UPLC-Q-TOF-MS was employed to determine the digestive products of flavonoid components in gastrointestinal fluids and their metabolic pathways. The results showed that in the process of digestion, 11 digestion products were generated by oxidation, reduction, deglycosylation, methylation and other phase Ⅰ metabolism. From flavonoid content and component changes, it was found that the flavonoid components in XECQ were relatively stable in the gastric fluid, while their content in the intestinal fluid was first increased and then maintained stable. This was mainly because flavonoid components were released from proteins, polysaccharides and other macromolecular substances during gastrointestinal digestion. In addition, phase Ⅰ metabolism occurred, but with relatively low metabolic rate, resulting in their stable content. This study preliminarily explored the digestive characteristics of flavonoid components in XECQ in the infant gastrointestinal environment, which laid a foundation for further studying the absorption, transport and metabolism of pharmacodynamics components in XECQ, and facilitated the study of the biopharmaceutical pro-perties of pediatric Chinese medicine.

[Phenylethanoid glycosides in Xiaoer Chiqiao Qingre Granules and its metabolic transformation in vitro and in vivo].

To clarify the metabolic transformation mechanism of phenylethanoid glycosides in Xiaoer Chiqiao Qingre Granules in vivo, this study extracted and separated the phenylethanoid glycosides in Xiaoer Chiqiao Qingre Granules. Based on UPLC-Q-TOF-MS/MS technology, the retention time and primary and secondary mass spectrometry information were analyzed by UNIFI software, and 11 phenylethanoid glycosides in Xiaoer Chiqiao Qingre Granules were preliminarily identified. Sixty-nine metabolites related to phenylethanoid glycosides were identified from the plasma samples of juvenile rats after administration of Xiaoer Chiqiao Qingre Granules. In addition, this study simulated the transformation system of intestinal flora in children, and discussed the metabolic effects of intestinal flora on the representative components forsythoside A, forsythoside E, and salidroside of phenylethanoid glycosides. The model of gastrointestinal heat retention in children with food accumulation was established to study the differential metabolites of phenylethanoid glycosides. Through the comparative analysis of the representative components absorbed in blood and the intestinal floral transformation products, it was found that the main metabolic pathways of phenylethanoid glycosides were dehydrogenation, oxidation, acetylation, sulfation, and glucuronidation. The findings of this study revealed the transformation law of phenylethanoid glycosides in the gastrointestinal tract. Through the preliminary discussion of the pharmacological mechanism, this study provides references for further clarifying the pharmacodynamic material basis of Xiaoer Chiqiao Qingre Granules and exploring the pediatric Chinese medicine compound.

Long non-coding RNA Hottip modulates high-glucose-induced inflammation and ECM accumulation through miR-455-3p/WNT2B in mouse mesangial cells.

Long non-coding RNAs (lncRNAs) play important roles in the pathogenesis of various diseases, including diabetic nephropathy (DN). However, the detailed mechanism is still largely unknown. High-glucose treated SV40-MES13 cells was used to mimic diabetic nephropathy in vitro. qRT-PCR was introduced to measure Hottip, collagen type I (Col. I), collagen type IV (Col. IV), fibronectin (FN), PAI-1, miR-455-3p and Wnt2B, IL-6, TNF-α mRNA level. Ellisa was used to examine the expression level of IL-6, TNF-α in the cell culture medium. Western blotting was employed to detect the protein level of Col. I, Col. IV, FN, PAI-1, Wnt2B, ß-catenin and cyclin D1. Cell viability was examined by MTT assay, luciferase reporter assay were used to determine the relationship between Hottip, miR-455-3p and Wnt2B. In the results, Hottip and Wnt2B was upregulated in db/db DN mice and high-glucose treated mouse mesangial cells (MMCs) while miR-455-3p was downregulated. High glucose treatment could enhance cell proliferation, and inflammation, increase fibrosis-related protein expression and active Wnt2B/ß-catenin/cyclin D1 pathway, while Hottip silencing reversed all the effects caused by high-glucose treatment. miR-455-3p was a sponge target of Hottip while Wnt2B was a downstream target of miR-445-3p. miR-445-3p inhibitor could suppress the effect of Hottip knockdown in cell proliferation, inflammation and fibrosis-related protein expression. Our data supported lncRNA Hottip/miR-455-3p/Wnt2B axis plays an important role in cell proliferation, inflammation, and extracellular matrix (ECM) accumulation in diabetic nephropathy.

Effectiveness of 10-year vaccination (2001-2010) on Hepatitis A in Tianjin, China.

Vaccination is an effective strategy to prevent and control the transmission of hepatitis A. Hepatitis A immunization program has been taken into effect since 2001 in Tianjin, China. This study evaluated the effectiveness of strategies in the prevention and control of hepatitis A. Data of serological survey, annual hepatitis A incidence, immunization coverage and the positive rate of hepatitis A IgG before and after the immunization program in residents under 15 years old were used to do the analysis. The results indicated that hepatitis A vaccine induced a striking decrease of hepatitis A incidence and a significant increase in the positive rate of anti-HAV IgG among the children younger than 15 years old. Hepatitis A vaccination in children was proved to be effective in the prevention and control of hepatitis A in Tianjin, China.

Interchangeability and tolerability of two inactivated hepatitis A vaccines in Chinese children.

In China, no data are available to evaluate the interchangeability between Chinese domestic inactivated hepatitis A vaccines (Healive) and imported inactivated hepatitis A vaccines (Havrix). A double-blind, randomized controlled study was to compare interchangeability and safety of Healive and Havrix among Chinese children. Vaccine was administered to 303 healthy children at 0 and 6 months in one of four vaccine regimens: Healive-Healive; Healive-Havrix; Havrix-Healive or Havrix-Havrix. We collected sera samples at 0 (before vaccination), 6 (before second dose) and 7 months (after second dose), and compared groups in terms of proportion of sero-conversions which is defined as ≥ 20 mIU/ml, and geometric mean concentrations (GMCs) of anti-hepatitis A virus (HAV) antibody. Seroconversion rates were 133/133 (100%) for those received one dose of Healive and 105/131 (80.2%) for those received one dose of Havrix at 6 months, respectively (P<0.001), GMCs for Healive and Havrix were 126.1 and 40.9 mIU/ml (P<0.001), respectively. At 7 months, the seroconversion rate was 100% among all groups. The GMC after two doses of Healive was 8905.5 mIU/ml compared with 1900.9 mIU/ml after two doses of Havrix (P<0.001). The GMC in the Healive-Havrix group was 3275.8 mIU/ml compared with 4165.8 mIU/ml in the Havrix-Healive group (P=0.058). There is not different of reported adverse reactions across the groups. The present study indicated that both vaccines can be recommended for interchangeable using of immunization among Chinese healthy children.

[Cross immunity study of two different influenza A3 virus vaccine strain].

OBJECTIVE: To explore the cross immunity response between two similar strains of influenza A3 virus vaccine from 2007 to 2008. METHODS: Healthy adults aged 18-60 years old without history of flu vaccination were inoculated Anflu ™( 52 cases) or VAXIGRIP ® (137 cases) influenza split vaccine. A micro-hemagglutination inhibition (HI) assay was used to test the serum specimens collected from the subjects before and after vaccination. The seroconversion rate, geometric mean titer (GMT) and antibody protective rate were used to evaluate the effect. RESULTS: The seroconversion rates of Anflu ™ and VAXIGRIP ® tested by A/Hiroshima/52/2005 virus antigen were 82.7% (95%CI: 69.2% - 91.8%) and 80.3% (95%CI: 72.4% - 86.5%) respectively and there was no significant difference (χ(2) = 0.141, P > 0.05). The seroconversion rates of Anflu™ and VAXIGRIP ® tested by A/Wisconsin/67/2005 (H3N2)-like virus antigen were 71.2% (95%CI: 56.7% - 82.8%) and 73.7% (95%CI: 65.4% - 80.8%) respectively and there was no significant difference observed (χ(2) = 0.126, P > 0.05). GMT of Anflu™ and VAXIGRIP ® tested by A/Hiroshima/52/2005 virus antigen after vaccination increased 11.5 (95%CI: 7.5 - 17.5) times and 13.0 (95%CI: 10.0 - 16.9) times without significant difference (F = 0.497, P > 0.05). GMT of Anflu ™ and VAXIGRIP ® tested by A/Wisconsin/67/2005 (H3N2)-like virus antigen after vaccination increased 9.5 (95%CI: 6.3 - 14.3) and 10.9 (95%CI: 8.5 - 13.7) times, and there was no significant difference either (F = 0.554, P > 0.05). The antibody protective rate of two vaccines before and after immunity tested by A/Hiroshima/52/2005 virus antigen were 48.1% and 54.7% before vaccination and 98.1% and 95.6%after vaccination respectively without significant difference (χ(2) = 0.135 - 0.673, P > 0.05). The antibody protective rates of two vaccines tested by A/Wisconsin/67/2005 (H3N2)-like virus antigen were 11.5% and 13.9%before vaccination and 80.8% and 86.1%after vaccination respectively, and there was no significant difference (χ(2) = 0.178 - 0.834, P > 0.05). But the results tested by A/Hiroshima/52/2005 virus antigen were higher than those of A/Wisconsin/67/2005 (H3N2)-like virus antigen (χ(2) = 7.111 - 52.155, P < 0.01). CONCLUSION: The two similar seasonal influenza vaccine strains recommended by WHO had a good cross immunity response, but the systematic error of test existed in two similar stains and the same strains should be used.

[Safety and immunogenicity on three lots of influenza split vaccines among adults].

OBJECTIVE: To evaluate the immunogenicity, safety and stability of the manufacture process regarding three consecutive lots of influenza split vaccines (Anflu). METHODS: A double-blind, randomized and controlled clinical trial was conducted in healthy volunteers. A total of 566 subjects aged 18 to 60 years were recruited and stratified into four age groups before randomly assigned into four groups. Each group would receive one dose of influenza vaccine from either one of the three lots of Anflu or one lot of the licensed control vaccine. Each dose of the vaccines contained 15 microg of each of the H1N1, H3N2 and B type antigen. Safety was assessed through 30-minute observation for immediate allergic reaction and three-day observation after vaccination. HI antibody titers were determined before vaccination and on day 21, after vaccination. RESULTS: Mild adverse reaction was reported and the overall incidence rates on fever of the four groups were from 1.4% to 2.8% but no significant difference was observed between groups. Seroconversion rates of the three viral strains in four groups were 80.3% and above with fold increase as > or = 11.1 and protection rate was > or = 93.4%. For the three lots of investigated vaccines, all of the indexes of the three viral strains in four groups exceeded the standards on EMEA and FDA for influenza vaccine. CONCLUSION: The three consecutive lots of Anflu appeared to be good, with both consistent immunogenicity and safety, indicating the stability of manufacture process.

[Evaluation on the role of hepatitis A vaccine in the prevention and control of hepatitis A in Tianjin city].

OBJECTIVE: To evaluate the effects of prevention and control strategies on hepatitis A. METHODS: Surveillance data on hepatitis A from 1990 to 2006 in Tianjin was analyzed, and the coverage rate of hepatitis A vaccine among targeted population was estimated, to compare the anti-HAV IgG level of children younger than 15 years old in 1999 and in 2005. RESULTS: Results showed that a) the morbidity of hepatitis A decreased from 25.26/10(5) in 1990 to 0.82/10(5) in 2006; b) the ratio of hepatitis A in viral hepatitis decreased from 30.43% in 1990 to 1.05% in 2006; c) the estimated coverage rate was 72.7%; d) the positive rate of anti-HAV among children younger than 15 years old in 2005 was distinctly higher than that in 1999. CONCLUSION: Positive results showed that it was successful to use hepatitis A vaccine as the strategy to prevent and control hepatitis A in the past five years in Tianjin.

[Epidemiological characteristics of severe acute respiratory syndrome in Tianjin and the assessment of effectiveness on measures of control].

OBJECTIVE: To analyze the epidemiologic characteristics of severe acute respiratory syndrome (SARS) and to evaluate the effectiveness on its major control measures in Tianjn. METHODS: Adopting two case reports 1 and 2, designed by the Tianjin Centers for Disease Control and Prevention to develop a unified case-tracing table including the map of the distribution of close contacts to SARS patients. With those methods, investigation on patients and their close contacts at hospital wards, families, communities and institutions of the patients were carried out. RESULTS: From April 13 through May 8, 2003, there were 175 SARS cases including imported ones, were identified with an incidence rate of 1.9 cases per 100,000. Among them, 14 died with a fatality of 8.0%. The whole process of epidemic in Tianjin was less than one month with the following features: (1) 93.7% of the total SARS cases in Tianjin were directly or indirectly transmitted by a super-spreader. (2) 68.6% of the total SARS patients were concentrated in 3 hospitals A, B and C which was menifastated in 'clustering'. Through study on the rest of the SARS patients, results showed that 16.8% of them were transmitted through family close contact and 2.3% due to contact to colleagues. However, 12.6% of the patients were not able to show evidence that they had any contact to a diagnosed SARS patient. At the early stage of the epidemic, a number of medical practitioners were infected, taking up 38.2% of the total SARS cases. Among the total number of 1 975 medical workers who participated in the SARS medical cares and treatments, 3.4% of them got infected. During the outbreak, all index cases and chains of transmission seemed to be clear, with only 3 patients not able to be traced for the source of infection, taking up 2% of the total SARS patients in Tianjin. Among the 10 index cases, only the super spreader and another one index case transmitted the virus to their contacts but the rest of index cases did not cause any secondary infection. CONCLUSION: Though SARS is clinically severe and can be spreaded quickly, the epidemic can be under control within a short period of time if chains of SARS transmission are broken down and effective measures as isolation and quarantine against patients as well as underscoring awareness among the publics in a scientific way, being carried out.