Rapid identification of components in Wuzhuyu Decoction using UHPLC Q-Exactive Orbitrap MS~n and molecular network technology / 中国中药杂志

Wuzhuyu Decoction, the classical formula recorded in the Treatise on Febrile Diseases(Shang Han Lun), has been included in the Catalogue of Ancient Classic Prescriptions(the First Batch). Consisting of Euodiae Fructus, Ginseng Radix et Rhizoma, Zingiberis Rhizoma Recens, and Jujubae Fructus, it is effective in warming the middle, tonifying deficiency, dispelling cold, and descending adverse Qi, and is widely applied clinically with remarkable efficacies. For a classical formula, the chemical composition is the material basis and an important premise for quantity value transfer. This study aimed to establish a rapid identification method of chemical components in Wuzhuyu Decoction by high-resolution mass spectrometry(HR-MS) and molecular network. AQUITY UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was used for sample separation, and acetonitrile-0.1% formic acid in water was used as mobile phases for gradient elution. Q-Exactive Orbitrap MS data were collected in positive and negative ion modes, and GNPS molecular network was plotted according to the similarity of MS/MS fragmentation modes. Cytoscape 3.6.1 was used to screen molecular clusters with similar structures. Finally, the chemical components of Wuzhuyu Decoction were rapidly identified according to the controls, as well as the information of retention time, accurate relative molecular weight of HR-MS, and MS/MS multistage fragments. A total of 105 chemical components were identified in Wuzhuyu Decoction. This study can provide data for the follow-up quality control, standard substance research, and pharmacodynamic material research on Wuzhuyu Decoction, as well as references for the rapid qualitative analysis of the chemical components of Chinese medicine.

Chloroplast DNA reveals genetic population structure in Sinomenium acutum in subtropical China / 中草药·英文版

OBJECTIVE@#The population density and diversity of Sinomenium acutum (Menispermaceae) have been greatly reduced recently by overharvesting for medicinal purposes in China. Therefore, it is urgent that the remaining populations are investigated, and that strategies for the utilization and conservation of this species are developed. This study aimed to find the possible glacial refugia and define the genetic diversity of S. acutum for its proper utilization and conservation.@*METHODS@#A total of 77 S. acutum samples were collected from four locations, Qinling Mountains, Daba Mountains, Dalou Mountains, and Xuefeng Mountains, in subtropical China. Genetic diversity among and between these populations were phylogenetically analyzed using four chloroplast DNA molecular markers (atpI-atpH, trnQ-5'rps16, trnH-psbA and trnL-trnF).@*RESULTS@#A total of 14 haplotypes (C1 to C14) were found in collected samples. Haplotypes C1 and C3 were shared among all populations, with C3 as the ancestral haplotype. Haplotypes C11 and C12 diverged the most from C3 and other haplotypes. No obvious phylogeographic structure was found in four locations using the GST/NST test. There is no evidence of rapid demographic expansion in S. acutum based on the mismatch distribution, and the results of Tajima's D test, and Fu's FS test. Our analyses of molecular variance revealed a high level of genetic variation within populations. In contrast, the genetic differentiation among S. acutum populations was low, indicating frequent gene flow.@*CONCLUSION@#Xuefeng, Dalou, and Daba Mountains were possible glacial refugia for the populations of S. acutum. C1, C3, C11 and C12 haplotypes of S. acutum should be carefully preserved and managed for their genetic value.

Interleukin-1 alpha induces osteoclast activation and bone loss / 中国组织工程研究

BACKGROUND: Interleukin-1 is an important pro-inflammatory cytokine that has been documented in the regulation of bone inflammation and bone remodeling. A previous study has demonstrated that interleukin-1α can induce apoptosis while inhibiting osteoblast differentiation in MC3T3-E1 cells. OBJECTIVE: To investigate the role and mechanism of interleukin-1α on osteoclast activation and bone loss in mice. METHODS: (1) Cell test: RAW264.7 cells were either treated with interleukin-1α alone or with receptor activator of nuclear factor-κB ligand (RANKL) for 1 and 4 days. Cell viability was tested by cell counting kit-8 assay. The number of multinuclear osteoclasts was detected by tartrate resistant acid phosphatase assay. The mRNA and protein levels of osteoclast-specific genes and genes related to nuclear factor-κB pathway and Wnt/β-catenin pathway were tested by real-time fluorescence quantitative PCR, immunofluorescence staining or western blot. Bone marrow-derived macrophages were either treated with interleukin-1α alone or with RANKL and macrophage colony-stimulating factor for 7 days. The number of multinuclear osteoclasts was detected by tartrate resistant acid phosphatase assay. The protein levels of osteoclast-specific genes were tested by western blot. (2) Animal test: Twenty-four male C57BL/6J mice (6-8 weeks old) were assigned into two groups at random: control group and test group. Mice were subsequently treated with interleukin-1α solution or PBS by intraperitoneal injection twice a week for 5 weeks. Bone tissues from the femurs were performed with micro-computed tomography analysis and hematoxylin-eosin staining, tartrate resistant acid phosphatase, and immunofluorescence analysis. RESULTS AND CONCLUSION: Cell test: Interleukin-1α alone significantly increased RAW264.7 cell proliferation, but stimulated cell differentiation into osteoclasts in combination with RANKL (P < 0.05). Interleukin-1α significantly increased the expression of osteoclast-related markers and the number of tartrate resistant acid phosphatase-positive multinuclear cells in RAW264.7 cells and bone marrow-derived macrophages in the existence of RANKL or RANKL+macrophage colony-stimulating factor (both P < 0.05). Interleukin-1α was found to significantly enhance the nuclear factor-κB and Wnt/beta-catenin signaling in RAW264.7 cells (P < 0.05). Blocking of nuclear factor-κB or Wnt3 signaling not only reversed the activation of nuclear factor-κB and Wnt3 signaling but also weakened the enhanced expression of osteoclast-specific genes induced by interleukin-1α in RAW264.7 cells (P < 0.05). Animal test: interleukin-1α induced bone loss in mice while also upregulating the expression of osteoclast-specific markers, RANK, TRAF6 and p65, and Wnt3 in vivo (P < 0.05). The findings indicate that interleukin-1α can induce osteoclast activation and bone loss by promoting the nuclear factor-κB and Wnt signaling pathways.

Pathological Change of Chronic Hepatitis B Patients with Different Tongue Coatings by Circular Multi-Omics Integrated Analysis / 中国结合医学杂志

OBJECTIVE@#To compare the circular pathological changes of chronic hepatitis B (CHB) patients according to the tongue diagnosis.@*METHODS@#Totally 41 CHB patients with typical white tongue coating (WTC) or yellow tongue coating (YTC) were enrolled and 14 healthy volunteers with normal tongue manifestation served as controls. The mRNA expression of peripheral leukocytes was detected by GeneChips, and 9 genes were randomly selected for expression validation. Circular metabolites were detected by gas chromatographymass spectrometry. Biological information was analyzed based on ingenuity pathways analysis or metabolomics database and the integrated networks were constructed by ClueGO.@*RESULTS@#A total of 945 and 716 differentially expressed genes were found in patients with WTC and YTC relative to healthy volunteers respectively. The biological information analysis indicated that CHB patients had obviously increased functions in cell death, apoptosis and necrosis (Z-score ⩾2, P<0.05) and decreased activation in T lymphocytes (Z-score ⩽-2, P<0.05), regardless of the tongue manifestation. Compared to patients with WTC, the YTC patients were predicted to be more active in functions related to virus replication (Z-score ⩾2, P<0.05), and the content of circular fatty acids, such as oleic acid (P=0.098) and lauric acid (P=0.035), and citric acid cycle-related metabolites were higher in the YTC patients (P<0.1). The integrated analysis based on differential genes and metabolites indicated that the most difference in the biological function network between the WTC and YTC patients was tumor necrosis factor receptor associated factor 6 mediated-nuclear factor kappa-B activation process.@*CONCLUSIONS@#CHB patients with YTC had more severe inflammation and fatty acids metabolism aberrant than patients with WTC. The results facilitate the modern pathological annotation of Chinese medicine tongue diagnosis theory and provide a reference for the interpretation of pharmacological mechanisms of Chinese medicine treatment.

Anti-tumor and Phenotypic Regulation Effect of Matrine on Dendritic Cells through Regulating TLRs Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate the effects of matrine on antigen presentation of dendritic cells (DCs), and to explore the pharmacological mechanism of matrine on anti-tumor effect.@*METHODS@#Different concentrations (0, 1, 2, 4, 8 and 16 µ g/mL) of matrine were co-cultured with DCs, the harvested DCs were co-cultured with antigens of Lewis lung cancer (LLC) cells, and then DCs and T cells were co-cultured to produce DCs-activated killer (DAK) cells, which have significant tumor-killing activity. The expression of cytokines, mRNA and protein of toll-like receptors (TLRs) in DCs were detected by enzyme linked immunosobent assay, polymerase chain reaction and Western blot, respectively. And the killing effect of DAK were measured by MTT assay.@*RESULTS@#Matrine significantly increased the mRNA expression of TLR7, TLR8, myeloid differentiation factor 88 (MyD88), tumor necrosis factor receptor-associated factor 6 (TRAF-6) and I κ B kinase (IKK), as well as the protein expression of TLR7 and TLR8, and up-regulated the levels of interleukin-12 (IL-12), IL-6 and tumor necrosis factor-α (TNF-α), meanwhile, it also increased the expressions of MHC-II, CD54, CD80 and CD86 in DCs. DCs-activated effector T cells had significant tumor-killing activity. When the concentration of matrine was more than 4 µg/mL, all indices had significant difference (P<0.01 or P<0.05).@*CONCLUSION@#Matrine plays an anti-tumor role by regulating TLRs signal transduction pathway, promoting the secretion of inflammatory cytokines and enhancing immune function.

Spectrum-effect relationship of hemostatic effects of Notoginseng Radix et Rhizoma with different commodity specifications / 中国中药杂志

This article aims to establish the fingerprints, determine the hemostatic pharmacodynamic indicators, and explore the spectrum-effect relationship of Notoginseng Radix et Rhizoma in 12 different specifications. Firstly, HPLC and liquid chromatography-mass spectrometry(LC-MS) were employed to establish the fingerprints of Notoginseng Radix et Rhizoma. The rat plasma recalcification experiment and the rat gastric bleeding experiment were conducted to determine the pharmacodynamic indicators, including plasma recalcification time(PRT), thrombin time(TT), prothrombin time(PT), and activated partial thromboplastin time(APTT). Afterwards, the partial least squares method was employed to explore the spectrum-effect relationship of Notoginseng Radix et Rhizoma in different specifications. Twenty-six common peaks were detected in the HPLC fingerprints of different specifications of Notoginseng Radix et Rhizoma, and 11 out of the 26 common peaks represented saponins. The content of dencichine was determined by LC-MS. The rat experiments showed that the pharmacodynamic indicators were significantly different among different specifications of Notoginseng Radix et Rhizoma. The spectrum-effect relationship was explored between 27 common components and pharmacodynamic indicators. Among them, 16 components had positive effects on the pharmacodynamic indicators of Notoginseng Radix et Rhizoma, and 11 exerted negative effects. This study provides a basis for the precision medication and quality control of Notoginseng Radix et Rhizoma.

Effect of Valeriana jatamansi extract on fecal UPLC-MS/MS metabolomics in rats with diarrheal irritable bowel syndrome / 中国中药杂志

The purpose of this study was to understand the pharmacodynamic effect of Valeriana jatamansi extract in diarrhea predominant irritable bowel syndrome(IBS-D) rat model induced by maternal separation combined with three kinds of stress, and observe the changes of endogenous metabolites in feces after intervention to find potential biomarkers and related metabolic pathways. The animal model of IBS-D was established by maternal separation combined with restraint, ice swimming and tail clamping. The therapeutic effect of each dose group of V. jatamansi extract was evaluated in terms of abdominal withdrawal reflex pressure threshold, fecal water content and immobility time of forced swimming test. In addition, rat feces were collected for detection of metabolic profiles of small molecular metabolites with UPLC-LTQ-Orbitrap MS platform, so as to find the biomarkers of differential metabolism with multivariate statistical analysis methods such as principal component analysis(PCA) and orthogon partial least squares discrimination analysis(OPLS-DA). The results showed that as compared with the normal group, the threshold of abdominal withdrawal reflex pressure was decreased, the fecal water content was increased, and the immobility time of forced swimming test was prolonged in the model group. The results of fecal metabonomics showed that the levels of 39 metabolites were down-regulated and those of 37 metabolites were up-re-gulated. Further analysis showed that these metabolites were related to bile acid metabolism, unsaturated fatty acid metabolism, amino acid metabolism, ceramide metabolism and other metabolic pathways. This study proved that the extract of V. jatamansi had definite pharmacodynamic effect on IBS-D model rats, and the mechanism was discussed from the perspective of fecal metabonomics.

Efficacy and Material Basis of Shuangjin Sanjie Granules in Treatment on Hyperplasia of Mammary Gland / 中国实验方剂学杂志

Objective:The SD rat model of hyperplasia of mammary gland（HMG） and the ultrahigh-performance liquid chromatography and mass spectrometry (UHPLC-LTQ-Orbitrap MS) technology were used to explore the pharmacological material basis of Shuangjin Sanjie granules (SJSJG) for the treatment on HMG.Method:SD rat models of HMG were administered in groups, and the nipple height and the diameter were measured; the levels of estradiol (E2), progesterone (P) and prolactin (PRL) in serum were detected, pathological examination was conducted for the hyperplasia of breast tissue. Histochemical methods were used to detect the expressions of estrogen receptor α (ERα), androgen receptor (AR), progesterone receptor (PR), tumor necrosis factor-α (TNF-α) proteins. Finally, UHPLC-LTQ-Orbitrap MS technology was used to detect the main chemical constituents of SJSJG, and the pharmacodynamic substance basis was analyzed based on the pharmacological effect.Result:The results of animal experiments showed that compared with the normal group, nipple height and diameter of the model group increased remarkably (P<0.01), serum E2 significantly increased (P<0.01). Pathological examination showed abnormal hyperplasia of breast tissue， expressions of ERα, AR, PR and TNF-α increased， compared with the model group, the nipple height and diameter of the SJSJG group decreased remarkably (P<0.01), serum E2 was decreased significantly (P<0.01)， pathological examination showed weakened abnormal hyperplasia of breast tissue, ERα, AR, PR and TNF-α protein expressions were significant decreased (P<0.01). The results of basic material study showed that 85 chemical components were identified from SJSJG, including 16 alkaloids, 7 flavonoids, 15 terpenes, 9 phenolic acid compounds, 3 coumarin compounds, 10 esters and lactone compounds, 7 fatty acids compounds, 4 amino acids compounds, and 14 other types of ingredients， among them, alkaloids and terpenoids chemical drug substances were closely related.Conclusion:SJSJG can effectively improve the condition of breast hyperplasia, and its medicinal substance basis may include saikosaponin A, Saikosaponin D, verticinone, peimine.

Ovarian Stimulation and In Vitro Maturation for Fertility Preservation in Female Cancer Patients / 中山大学学报(医学科学版)

@#【Objective】To determine the effect of ovarian stimulation or in vitro maturation for fertility preservation in female cancer patients. 【Methods】 A retrospective study was conducted in 27 females who underwent fertility preservation procedures in our center.【Results】Female patients were included in this study with an average age of 27.1. Patients spent on average for 6.8 d to retrieve oocytes since their attendance day. Total amount of Gn was on average 910 U per patient and for patients with breast cancer，the average estrogen level on trigger day reached 360 pg/mL. The maturation rate of oocytes from ovarian stimulation cycles was 82.6% ，which of that in emergency in- vitro maturation cycles was 38.1%.【Conclusion】The development capability of oocytes from cancer patients are comparable with those of other infertility patients. Peak estradiol levels were controlled by the administration of letrozole. In vitro maturation of oocytes performed at random time of the menstrual cycle may result in a lower maturation rate ，which is associated with the time limit of the follow- up cancer treatment. In conclusion，clinicians should consider a more holistic approach for female cancer patients，which focuses not only on the characteristic of the primary cancer but also on the phase of the menstrual cycle at their attendance day.

Chinese Herbal Medicine Combined with Entecavir for HBeAg Positive Chronic Hepatitis B: Study Protocol for a Multi-Center, Double-Blind Randomized-Controlled Trial / 中国结合医学杂志

<p><b>BACKGROUND</b>The domestic prevalence of chronic hepatitis B (CHB) in China is 7.18% in 2006, imposing great societal healthcare burdens. Nucleot(s)ide analogues (NUCs) anti-hepatitis B virus (HBV) therapies are widely applied despite the relatively low rate of seroconversion and high risk of drug-resistant mutation. More effective treatments for CHB deserve further explorations. Combined therapy of NUCs plus Chinese herbal medicine (CHM) is widely accepted in China, which is recognized as a prospective alternative approach. The study was primarily designed to confirm the hypothesis that Tiaogan-Yipi Granule (, TGYP) or Tiaogan-Jianpi-Jiedu Granule (, TGJPJD) plus entecavir tablet (ETV) was superior over ETV monotherapy in enhancing HBeAg loss rate.</p><p><b>METHODS</b>The study was a nationwide, large-scale, multi-center, double-blind, randomized, placebo-controlled trial with a designed duration of 108 weeks. A total of 16 hospitals and 596 eligible Chinese HBeAg positive CHB patients were enrolled from November 2012 to September 2013 and randomly allocated into 2 groups in 1:1 ratio via central randomization system: experimental group (EG) and control group (CG). Subjects in EG received CM formulae (TGYP or TGJPJD, 50 g per dose, twice daily) plus ETV tablet (or ETV placebo) 0.5 mg per day in the first 24 weeks (stage 1), and CHM granule plus ETV tablet (0.5 mg per day) from week 25 to 108 (stage 2). Subjects in CG received CHM Granule placebo plus ETV tablet (0.5 mg per day) for 108 weeks throughout the trial. The assessments of primary outcomes (HBV serum markers and HBV-DNA) were conducted by a third-party College of American Pathologists (CAP) qualified laboratory. Adverse effects were observed in the hospitals of recruitment.</p><p><b>DISCUSSION</b>The study was designed to compare the curative effect of CM plus ETV and ETV monotherapy in respect of HBeAg loss, which is recognized by the European Association for the Study of the Liver as "a valuable endpoint". We believe this trial could provide a reliable status for patients' "journey" towards durable responses after treatment discontinuation. The trial was registered before recruitment on Chinese Clinical trial registry (No. ChiCTR-TRC-12002784, Version 1.0, 2015/12/23).</p>

Behavioral Characteristics and Medicolegal Identification of Infanticide / 法医学杂志

Due to the concealment and incidence higher than reported statistics, infanticide should attract the attention of legal medical experts.The infanticide process has common behavioral characteristics.The perpetrators are most frequently biological mothers who are unmarried, with inferior education, and lack of routine pregnancy tests.Medicolegal identification of infanticide cases includes identification of live births and causes of death.Besides the classical lung floating test, there are other methods to identify live births such as pulmonary interstitial emphysema, CT imaging examination, immunohistochemical examination of umbilical cord vitality markers.Neonatal line measurement is the only way to identify live births in corpse with severe decomposition.The main causes of death in infanticide are mechanic asphyxia and fatal head trauma.Shaken baby syndrome, as a common abusive head trauma, is pathologically characterized by the triad of encephalopathy, subdural haemorrhage (SDH) and retinal haemorrhage (RH).During the autopsy of suspected shaking baby syndrome, in addition to carefully examining cervical cord, the possibility of pathological temporal lobe hemorrhage should also be ruled out.

A preliminary study on the disappearance time of influenza virus antigen / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the antigen clearance time, time to symptom disappearance, and the association between them using immunofluorescence assay for dynamic monitoring of influenza virus antigen in children with influenza.</p><p><b>METHODS</b>A total of 1 063 children suspected of influenza who visited the Hunan People's Hospital from March to April, 2016 were enrolled. The influenza A/B virus antigen detection kit (immunofluorescence assay) was used for influenza virus antigen detection. The children with positive results were given oseltamivir as the antiviral therapy and were asked to re-examine influenza virus antigen at 5, 5-7, and 7 days after onset.</p><p><b>RESULTS</b>Of all children suspected of influenza, 560 (52.68%) had an influenza virus infection. A total of 215 children with influenza virus infection were followed up. The clearance rate of influenza virus antigen was 9.8% (21 cases) within 5 days after onset. The cumulative clearance rate of influenza virus antigen was 32.1% (69 cases) within 5-7 days, and 98.1% (211 cases) within 7-10 days after onset. Among these children, 6 children (2.8%) achieved the improvement in clinical symptoms within 3 days after onset. The cumulative rate of symptom improvement was 84.7% (182 cases) within 3-5 days after onset, and 100% achieved the improvement after 5 days of onset.</p><p><b>CONCLUSIONS</b>The time to improvement in symptoms after treatment is earlier than antigen clearance time. Almost all of the children achieve influenza virus antigen clearance 7-10 days after onset. Therefore, it is relatively safe for children to go back to school within 7-10 days after onset when symptoms disappear.</p>

Content Determination of Four Triterpenes in Caragana sinica by HPLC and α-glucosidase Activity Assay in Vitro / 中国药学杂志

OBJECTIVE: To establish HPLC methods for the determination of betulinic acid, oleanolic acid, ursolic acid, and lupeol in Caragana sinica roots, and investigate the α-glucosidase activity of the four compounds in vitro. METHODS: An Agilent Eclipse XDB-C18 column (4.6 mm × 250 mm, 5 μm) was applied with methanol-water-phosphoric acid (85:15:0.1) as the mobile phase at a flow rate of 0.6 mL·min-1 to separate betulinic acid, ursolic acid, and oleanolic acid. An Inert Sustain C18 column (4.6 mm × 150 mm, 5 μm) was applied with acetonitrile-water (70:30) as the mobile phase at a flow rate of 0.8 mL·min-1 to separate lupeol. RESULTS: Good linearities were achieved for betulinic acid, oleanolic acid, ursolic acid, and lupeol within the range of 0.56-5.58, 0.42-4.20, 0.19-1.92, and 6.10-61.00 μg, respectively. The average recoveries were 99.79%, 98.51%, 98.05%, and 99.47%, and the RSDs were 1.53%, 1.74%, 1.78%, and 1.94%, respectively. CONCLUSION: The developed methods are accurate and can be used for the quality control of Caragana sinica. In addition to betulinic acid, the other three compounds show good α-glucosidase inhibitory activity and are expected to be developed as new hypoglycemic drugs.

Expressions of interleukin-1 beta and matrix metalloproteinase-13 in articular cartilage and synovial fluid of osteoarthritis model in rabbits / 西安交通大学学报(医学版)

Objective To investigate the feasibility of osteoarthritis (OA) model in rabbits by injuring medial meniscus so as to understand the role of interleukin-1 beta (IL-1β) and matrix metalloproteinase-13 (MMP-13) in pathological mechanism of articular cartilage and synovial fluid of OA model in rabbits.Methods We randomly divided 40 New Zealand white rabbits into experimental group (n =30) and control group (n =10).Pathological changes in articular cartilage of the femoral condyle were scored at weeks 2,6 and 12 after surgery.We detected the expressions of IL-1β and MMP-13 by immunohistochemistry.The cell fractions of IL-1β and MMP-13 were recorded by ELISA.Results The articular cartilage score and HE staining significantly differed at various time points of gross and pathological observation between control group and experimental group (P＜ 0.05).Immunohistochemistry showed that IL-1β was expressed in both groups and that the cell fraction differed significantly at weeks 6 and 12 (P＜0.05),but not at week 2 between the two groups.MMP-13 protein expression was not detected in articular condrocytes in control group,but was detected in experiment group with a significant difference (P＜0.05).IL-1β expression was consistent in articular cartilage and synovial fluid.Conclusion The reasonable rabbit animal OA model could be established by knee meniscus injury caused by surgical method.Expressions of IL-1β and MMP-13 change obviously in the pathomechanism of OA.Further clinical studies are needed to determine whether they can be used as markers in early diagnosis of OA.

Analysis of influence factors for spontaneous closure of atrial septal defect and ventricular septal defect in the northern of Xinjiang / 新乡医学院学报

Objective To analyse the risk factors for spontaneous closure of atrial septal defect(ASD) and ventricular septal defect(VSD) in children in the northern of Xinjiang.Methods Ninety-two ASD and sixty-five VSD children in Department of Pediatrics,the First Affiliated Hospital of Medical College,Shehezi University from January 2010 to May 2014 were selected as research subject.According to the spontaneous closure condition of children,the children with ASD were divided into ASD spontaneous closure group and ASD non spontaneous closure group;the children with VSD were divided into VSD spontaneous closure group and VSD non spontaneous closure group.The risk factors for spontaneous closure of ASD and VSD were analysed by single factor and multiple factor logistic regression analysis.Results In 92 ASD children,12 cases (13.04％) were spontaneous closure.In 65 VSD children,9 cases (13.85％) were spontaneous closure.Single factor analysis result showed that there was statistic difference in defect diameter,defect type and the age between spontaneous closure group and non spontaneous closure group in ASD and VSD children (P ＜ 0.05);but there was no statistic difference in defect number and complications between spontaneous closure group and non spontaneous closure group in ASD and VSD children (P ＞ 0.05).Multivariate logistic regression analysis showed that the defect diameter,defect type and the age were the independent risk factors for the spontaneous closure of ASD and VSD (P ＜ 0.05).Conclusion The age ＜ 2 years old,central ASD,perimembranous VSD and defect diameter ＜ 5 mm are important factors for promoting spontaneous closure of ASD and VSD.

Correlation between Constitution of Yin Deficiency Syndrome and Polymorphism of HLA-DQA1/Treatment Response of Peg-IFNalpha Therapy in HBeAg Positive Chronic Hepatitis B Patients / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the correlation between constitution of yin deficiency syndrome (YDS) and polymorphism of HLA-DQA1/treatment response of Peg-lFNalpha therapy in HBeAg positive chronic hepatitis B (CHB) patients, and to explore constitution of Chinese medicine (CM) in response of interferon therapy.</p><p><b>METHODS</b>Totally 120 HBeAg positive CHB patients who were treated with Peg-IFNalpha were enrolled, and assigned to YDS group (59 cases) and non-YDS group (61 cases) according to classification of CM constitutions. All patients were subcutaneously injected with Peg-IFNalpha-2b (1.0 microg/kg body weight) or Peg-IFNalpha-2a (180 microg), once per week. Effective efficacy was primarily judged when complete response (CR) or partial response (PR) was obtained at month 6. Those with CR or PR completed 1 year therapeutic course. HLA-DQA1 gene types were detected by polymerase chain reaction sequence specific primers (PCR-SSP). The distribution difference of CM constitutions in patients with CR or PR and their inter-group HLA-DQA1 allele frequency were compared.</p><p><b>RESULTS</b>Different treatment responses of Peg-IFNalpha were observed in CHB patients of two different CM constitutions. The ratio of CR + PR was 61.0% (36/59) in YDS group, obviously lower than that in NYDS group [78.7% (48/61), P < 0. 05]. Patients with CR had a lower allele frequency of HLA-DQA1 * 0501 than those with no-response [14.8% (8/54) vs. 30.6% (22/72)] with statistical difference (P < 0.05). Patients with CR had a higher allele frequency of HLA-DQA1 * 0601 than those with no-response [18.5% (10/54) vs. 5.6% (4/72)] with statistical difference (P < 0.05). The allele frequency of HLA-DQA1 * 0301 was lower in YDS group than in non-YDS group [2. 5% (3/118) vs. 9.8% (12/122)] with statistical difference (P < 0.05). The allele frequency of HLA-DQA1 * 0501 was higher in YDS group than in non-YDS group [33.9% (40/118) vs. 18.9% (23/122)] with statistical difference (P < 0.05). Yet statistical significance was lost after adjustment (Pc > 0.05 for both).</p><p><b>CONCLUSIONS</b>Both constitutions of CM and HLA-DQA1 gene polymorphism af- fect HBeAg positive CHB patients' response to Peg-INFalpha. Constitutions of YDS and HLA-DQA1 * 0501 was not favorable to response, their association needed to be further studied.</p>

Clinical value of fluorescence lateral flow immunoassay in diagnosis of influenza A in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the clinical value of a new type of fluorescence lateral flow immunoassay in rapid detection of influenza A virus.</p><p><b>METHODS</b>A total of 378 samples of nasopharyngeal secretions were collected from 378 children with influenza-like symptoms to detect the influenza A virus by fluorescence lateral flow immunoassay, colloidal gold immunoassay, and RT-PCR between July 2015 and August 2015.</p><p><b>RESULTS</b>Of the 378 samples, 81 (21.4%) were positive for influenza A virus by RT-PCR. Compared with RT-PCR, the sensitivities of fluorescence lateral flow immunoassay and colloidal gold immunoassay were 90.1% (73/81) and 75.3% (61/81), respectively, and the specificities were 99.3% (295/297) and 98.3% (292/297), respectively. The average threshold cycle (Ct) value for the positive samples detected by the fluorescence lateral flow immunoassay (30.6) was higher than that for the positive samples detected by the colloidal gold immunoassay (28.7).</p><p><b>CONCLUSIONS</b>Compared with colloidal gold immunoassay, fluorescence lateral flow immunoassay has higher sensitivity, specificity, and concordance rate with RT-PCR, suggesting that it can be used for early screening and diagnosis of influenza A.</p>

Quality Consistency Evaluation of Omeprazole Enteric-coated Capsules / 中国药师

Objective:To establish a method for the determination of omeprazole delayed release capsules and investigate the be-havior of reference preparations to provide experimental basis for generic drugs quality consistency evaluation. Methods:According to the first dissolution method (basket method) stated in 0. 931 of Chinese Pharmacopeia (2015 edition), the type of release media, solu-bility and stability of omeprazole in different media, effect of different treatment methods on the drug adsorption in the solution, differ-ent rotation speed and the methodology of ultraviolet spectrophotometry were investigated. Results:The rotation speed was 75 r·min-1 with the dissolution medium volume of 900 ml. The dissolution profiles of omeprazole delayed release capsules in three different media ( pH 6. 0 phosphate buffer, pH 6. 8 phosphate buffer and water containing 3% Tween-80) were determined with online filteration. The solubility of omeprazole in the different media was 0. 123, 0. 078 and 0. 275 mg·ml-1 , respectively. The results showed that ome-prazole was degraded 44%, 8% and 14% in 2 h in the above three release media, and degraded up to 43% in 6 h in water containing 3% Tween-80. The linear of omeprazole was 0. 209 4-20. 94, 0. 204 8-20. 48 and 0. 2016-20. 16 μg·ml-1 with the average recovery of 99.3% (RSD=0.7%,n=12), 99.7% (RSD=0.9%,n=12) and 99.5% (RSD=0.6%,n=12) respectively in the three media. Conclusion:The method is accurate and reliable, which can be used to study the quality consistency of omeprazole delayed re-lease capsules.

Effect of the same mechanical loading on osteogenesis and osteoclastogenesis in vitro / 中华创伤杂志(英文版)

<p><b>PURPOSE</b>To investigate the influence of the same mechanical loading on osteogenesis and osteoclastogenesis in vitro.</p><p><b>METHODS</b>Primary osteoblasts, bone marrow-derived mesenchymal stem cells (BMSCs, cultured in osteoinductive medium) and RAW264.7 cells cultured in osteoclast inductive medium were all subjected to a 1000 μstrain (μs) at 1 Hz cyclic mechanical stretch for 30 min (twice a day).</p><p><b>RESULTS</b>After mechanical stimulation, the alkaline phosphatase (ALP) activity, osteocalcin protein level of the osteoblasts and BMSCs were all enhanced, and the mRNA levels of ALP and collagen type I increased. Additionally, extracellular-deposited calcium of both osteoblasts and BMSCs increased. At the same time, the activity of secreted tartrate-resistant acid phosphatase, the number of tartrate-resistant acid phosphatase-positive multinucleated cells, matrix metalloproteinase-9 protein levels of RAW264.7 cells and the extracellular calcium solvency all decreased.</p><p><b>CONCLUSION</b>The results demonstrated that 1000 μs cyclic mechanical loading enhanced osteoblasts activity, promoted osteoblastic differentiation of BMSCs and restrained osteoclastogenesis of RAW264.7 cells in vitro.</p>

Determination of oleanic acid and paeoniflorin in Paeonia lactiflora by ultrasound-assisted ionic liquid-reversed phase liquid chromatography / 中国中药杂志

Four kinds of ionic liquids [BMIM] Br, [BMIM] BF4, [BMIM] PF6, [HMIM] PF6 were used to analyze the content of oleanic acid and paeoniflorin in Paeonia lactiflora with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 μm), was used. Acetonitrile and water (90:10) as mobile phase was used to determine the content of oleanic acid with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 210 nm, chromatographic column temperature at room temperature. Paeoniflorin content was determined using acetonitrile and water (18:82) as mobile phase with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 250 nm, the chromatographic column temperature at room temperature. The result show that oleanic acid has the highest extraction yield when the conditions are solid-liquid ratio of 1:80 (g · mL(-1)), and the [BMIM] Br methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, the content of oleanic acid from 0.24 to 3.76 μg showed a good linearity (r = 0.9999), the average recovery was 97.20%. Paeoniflorin has the highest extraction yield when the conditions are solid-liquid ratio of 1:130 (g · mL(-1)), and the [C4 MIM] PF6 methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, paeoniflorin content from 0.42 to 4.20 μg showed a good lin- earity (r = 1.000), the average recovery was 98.84%. This method is simple and reliable, its repeatability is also very good. It has important significance in the study P. lactiflora of ionic liquid microextraction.