Analysis on Material Basis of Anti-COPD Effect of Euphorbia helioscopia Based on Serum Pharmacochemistry and Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo analyze the migrating components absorbed into blood of the aqueous extract of Euphorbia helioscopia， and to explore the pharmacodynamic material basis of the aqueous extract of E. helioscopia against chronic obstructive pulmonary disease（COPD）. MethodUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detecte the migrating components absorbed into blood of rats after intragastric administration of aqueous extract of E. helioscopia. An Agilent RRHD SB-C18 column（3 mm×100 mm， 1.8 μm） was used with 0.1% formic acid aqueous solution（A）-acetonitrile（B） as the mobile phase for gradient elution（0-15 min， 5%-30%B； 15-20 min， 30%-50%B； 20-30 min， 50%-95%B； 30-35 min， 95%-5%B）， and the detection wavelength of 190-800 nm， column temperature of 40 ℃， flow rate of 0.3 mL∙min-1 and injection volume of 4 μL. The electrospray ionization（ESI） was used in positive and negative ion modes， and the detection range was m/z 50-1 250. Network pharmacology was used to screen out the key components and the key targets of COPD through the interaction analysis. Metascape database was used to predict the molecular function， biological process， cellular composition and signal pathways mainly involved in the anti-COPD effect of E. helioscopia. Molecular docking technique was used to determine the affinity of key targets with key components. ResultA total of 29 migrating components absorbed into blood of rats were identified after intragastric administration of aqueous extract of E. helioscopia， 9 of which were prototype components and 20 were metabolites. Network pharmacological analysis showed that luteolin， quercetin， apigenin， naringenin and helioscopinolide C were the key components of E. helioscopia against COPD， and vascular endothelial growth factor A（VEGFA）， albumin（ALB）， protein kinase B1（Akt1）， tumor necrosis factor（TNF） and interleukin-6（IL-6） were the key targets. Molecular docking results showed that one diterpene lactone（helioscopinolide C） and three flavonoids（naringenin， luteolin， apigenin） in the migrating components absorbed into blood all had strong binding activity to the key targets of E. helioscopia against COPD. ConclusionNaringenin， helioscopinolide C， luteolin and apigenin may be the main anti-COPD active substances of E. helioscopia.

Effect of Euphorbia helioscopia Aqueous Extract on Chronic Obstructive Pulmonary Disease and Precancerous Lesion-associated Proteins of Lung Tissues in Mice Induced by Cigarette Smoke / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect and mechanism of Euphorbia helioscopia aqueous extract （EHE） on mice with chronic obstructive pulmonary disease （COPD） and its influence on precancerous lesion-associated proteins in lung tissues induced by cigarette smoke （CS）. MethodThe COPD model was induced by CS in 60 mice and the model mice were randomly divided into control group， model group， positive drug group （dexamethasone， 2 mg·kg-1）， and low-， medium-， and high-dose EHE groups （1.875， 3.75， 7.5 g·kg-1）. The high-performance liquid chromatography （HPLC） method was used to determine the related components in EHE. The changes in end-expiratory pause （EEP）， airway resistance （Penh）， expiratory flow at 50% vital capacity （EF50）， and other pulmonary function indexes were detected by the spirometer. The levels of inflammatory factors， such as interleukin （IL）-2， IL-5， IL-18， IL-17A， and IL-27 in bronchoalveolar lavage fluid （BALF） of mice were detected by high-throughput liquid protein chip technology. Hematoxylin-eosin （HE） staining was used to detect the pathological changes in lung tissues in mice. The content of malondialdehyde （MDA）， myeloperoxidase （MPO）， and glutathione peroxidase （GSH-Px） in lung tissues was determined by the colorimetric method. The mRNA relative expression of tumor necrosis factor-α （TNF-α）， transforming growth factor-β （TGF-β）， matrix metalloproteinase-2 （MMP-2）， matrix metalloproteinase-9 （MMP-9）， and matrix metalloproteinase-12 （MMP-12） was detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Immunohistochemistry （IHC） was used to detect the expression of tumor protein （P53） and cell proliferation-associated antigen （Ki67） in lung tissues， and Western blot was used to detect the relative expression of tumor suppressor protein （P16）， DNA （cytosine-5）-methyltransferase 1 （DNMT1）， and fragile histidine triad （FHIT） in lung tissues. ResultThe results showed that the main compounds in EHE included phenols （gallic acid and protocatechuic acid） and flavonoids （such as hyperoside， rutin， myricetin， naringenin， quercetin， luteolin， kaempferol， and licorice chalcone A）， among which gallic acid and rutin were the highest in content. Compared with normal group， model group showed increased levels of EEP， EF50， and Penh （P<0.05）， and showed increased MDA and MPO levels （P<0.01） and decreased GSH-Px （P<0.01）， and the model group displayed increased levels of IL-2， IL-5， IL-18， IL-17A， IL-27， TNF-α， TGF-β， MMP-2， MMP-9， and MMP-12 （P<0.05）. And the model group exhibited up-regulated expression of P53， Ki67， and FHIT in lung tissues （P<0.01） and down-regulated expression of DNMT1 and P16 （P<0.01）. Compared with model group， the EHE groups showed decreased EEP and EF50 levels （P<0.05）. The pathological injury of lung tissues in mice of the model group was observed under HE staining， and the pathological injury of basal cell hyperplasia of lung tissues was gradually improved after treatment with EHE. The EHE groups showed reduced levels of MDA and MPO （P<0.01） and increased GSH-Px （P<0.01）. The EHE groups displayed decreased levels of IL-2， IL-5， IL-18， IL-17A， IL-27， TNF-α， TGF-β， MMP-2， MMP-9， and MMP-12 （P<0.05）. And the EHE groups showed down-regulated Ki67 and FHIT in lung tissues （P<0.05） and up-regulated expression of P53 and DNMT1 （P<0.05）. ConclusionEHE can protect mice from COPD and inhibit precancerous lesions， and the mechanism may be related to the inhibition of inflammation and oxidative stress response， regulation of protease and antiprotease imbalance， and regulation of epithelial cell growth.

Clinical analysis of 50 cases of tracheal bronchus in children / 中华全科医师杂志

The clinical data of 50 children with tracheal bronchus, 35 males and 15 females with a mean age of 1.3 months (9 days to 10 years), diagnosed by bronchoscopy in the First Affiliated Hospital of Xiamen University from July 2017 to September 2021 were collected, including gender, age, symptoms, signs, bronchoscopy manifestations, imaging manifestations, associated diseases, and outcomes. There were 26 cases (52%) of severe pneumonia, 14 cases (28%) of recurrent wheezing, 8 cases (16%) of recurrent cough, and 2 cases (4%) of foreign bodies. The clinical symptoms were cough in 45 cases (90%), phlegm in 37 cases (74%), asthma in 28 cases (56%), and fever in 25 cases (50%). The physical signs were wet rale in 33 cases (66%), dry rale in 24 cases (48%), shortness of breath in 23 cases (46%), and triple concave sign in 21 cases (42%). The site of occurrence of tracheal bronchi was the right wall of the lower segment of the trachea. All 50 patients underwent complete pulmonary imaging examinations, but the presence of tracheal bronchus was reported in only 3 cases (6%). There were 24 cases (48%) with other types of tracheal malformations, including 9 cases (18%) of single malacia, 6 cases (12%) of single tracheal stenosis, 1 case (2%) of external tracheal compression, and 8 cases (16%) of multiple tracheal malformations. There were other underlying diseases in 27 cases (54%), congenital heart disease was the most common (17 cases, 34%), followed by premature infants (9 cases, 18%). In addition, there were 3 cases (6%) of chromosomal diseases, esophageal atresia, tracheoesophageal fistula, and polydactyly. All children were discharged after anti-infection and bronchoscopy alveolar lavage. It is suggested that tracheobronchial malformation can cause infection, as well as other tracheal deformities such as tracheal softening and stenosis.

Identification of Dalbergiae Odoriferae Lignum and Its Counterfeits by 1H-NMR Combined with Multivariate Statistical Analysis / 中国实验方剂学杂志

ObjectiveTo establish the identification method of Dalbergiae Odoriferae Lignum（DOL） and its counterfeits by nuclear magnetic resonance hydrogen spectrum（1H-NMR） combined with multivariate statistical analysis. Method1H-NMR spectra of DOL and its counterfeits were obtained by NMR， and the full composition information was established and transformed into a data matrix， and the detection conditions were as follows：taking dimethyl sulfoxide-d6（DMSO-d6） containing 0.03% tetramethylsilane（TMS） as the solvent， the constant temperature at 298 K（1 K=-272.15 ℃）， pulse interval of 1.00 s， spectrum width of 12 019.23 Hz， the scanning number of 16 times， and the sampling time of 1.08 s. Similarity examination and hierarchical cluster analysis（HCA） were performed on the data matrix of DOL and its counterfeits， and orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to analyze the data matrix and identify the differential components between them. In the established OPLS-DA category variable value model， the category variable value of DOL was set as 1， and the category variable value of the counterfeits was set as 0， and the threshold was set as ±0.3， in order to identify the commercially available DOL. The OPLS-DA score plot was used to determine the types of counterfeits in commercially available DOL， and it was verified by thin layer chromatography（TLC）. ResultThe results of similarity analysis and HCA showed that there was a significant difference between DOL and its counterfeits. OPLS-DA found that the differential component between DOL and its counterfeits was trans-nerolidol. The established category variable value model could successfully identify the authenticity of the commercially available DOL. The results of the OPLS-DA score plot showed that there were heartwood of Dalbergia pinnata and D. cochinchinensis in the commercially available DOL， and were consistent with the TLC verification results. ConclusionThere is a phenomenon that heartwood of D. pinnata and D. cochinchinensis are sold as DOL in the market. 1H-NMR combined with multivariate statistical analysis can effectively distinguish DOL and its counterfeits， which can provide a reference for the identification of them.

Effects of bisphenols on proliferation and oxidative stress of BRL 3A rat liver cells and their mutagenicities / 预防医学

Objective@#To examine the effects of bisphenol A (BPA), bisphenol S ( BPS ), bisphenol F ( BPF ) and bisphenol AF ( BPAF ) on the proliferation and oxidative stress of BRL 3A rat liver cells, and to preliminarily evaluate their mutagenicities.@*Methods@#In vitro cultured BRL 3A rat liver cells were treated with BPA, BPS, BPF and BPAF at concentrations of 0, 5, 10, 25, 50, 100, 150 and 200 μmol/L for 48 h, respectively. Then, the cell viability was determined using the CCK-8 assay, and the half maximal inhibitory concentration ( IC50 ) was calculated. The minimum inhibitory concentration for BRL 3A cell proliferation was screened, and the intracellular reactive oxygen species ( ROS ) was measured in BRL 3A cells using the 2',7'-dichlorodihydrofluorescein diacetate ( DCFH-DA ) assay. In addition, the effects of BPA, BPS, BPF and BPAF at concentrations of 1 000, 200, 40, 8 and 1.6 μg/plate on the mutant colonies of histidine-deficient Salmonella typhimurium ( TA1535, TA97a, TA98, TA100 and TA102 ) were tested using the Ames test.@*Results@#Treatment with BPA and BPF at concentrations of 100 to 200 μmol/L and with BPAF at concentrations of 25 to 200 μmol/L inhibited BRL 3A cell survival at a concentration-dependent manner, while exposure to BPS at concentrations of 5 to 200 μmol/L resulted in no changes in BRL 3A cell survival. The IC50 values of BPA, BPS, BPF and BPAF were 131.7, >200, 187.5 and 21.6 μmol/L against BRL 3A cells, respectively. Treatment with BPS at 100 μmol/L or BPAF at 25 μmol/L caused no significant changes in the ROS level; however, exposure to BPA at 100 μmol/L and BPF at 100 μmol/L significantly increased the ROS level. Ames test showed that BPA, BPS, BPF and BPAF did not induce mutagenicity in TA1535, TA97a, TA98, TA100 or TA102 strains.@*Conclusions@#BPAF shows the highest cytotoxicity to BRL 3A cells, and low-concentration exposure to BPS has few effects on BRL 3A cells. The cytotoxicity of bisphenols against BRL 3A cells may be associated with the induction of oxidative stress. None of the four bisphenols show mutagenic effects under the present experimental conditions.

Analysis and follow-up of home mechanical ventilation in 11 children with chronic respiratory failure / 中华全科医师杂志

Objective:To explore the feasibility and safety of long-term home mechanical ventilation(HMV) in children with chronic respiratory failure.Methods:Clinical data of 11 children with chronic respiratory failure, who underwent HMV with the care of the First Affiliated Hospital of Xiamen University from January 2013 to December 2019, were retrospectively reviewed. The clinical manifestation, growth and development, quality of life, adverse events and prognosis of HMV children were analyzed.Results:There were 8 boys and 3 girls with the onset age of 26 days to 13 years old; and the age at starting HMV was 3 months to 13 years old. Eight children were diagnosed as neuromuscular diseases, and 3 children were diagnosed as respiratory diseases. The duration of institutional mechanical ventilation was 2 weeks to 8 months. Six patients underwent invasive HMV via a tracheostomy, and 5 received non-invasive ventilation via nasal and face masks. Bi-level positive airway pressure ventilation mode was applied in all the patients. The duration of HMV was 3 months to 27 months. During follow-up, no HMV related adverse events were observed. Both the quality of life and nutritional status were improved in all cases. One patient lost follow-up 9 months later and 1 patient died of severe adenovirus pneumonia during hospitalization for examination, the remaining 9 cases survived. Liberation from HMV was obtained in 4 patients. The frequency of readmission was 1 to 2 times.Conclusion:It is suggested that long-term HMV is safe and feasible for children with chronic respiratory failure.

Comparison of clinical features of Mycoplasma pneumoniae pneumonia and adenoviral pneumonia in children / 中华全科医师杂志

Objective:To compare the clinical characteristics of Mycoplasma pneumoniae pneumonia and adenoviral pneumonia in children. Methods:Clinical data of 151 children with pneumonia admitted to our hospital from January 2019 to June 2019 were retrospectively analyzed, including 106 cases of Mycoplasma pneumoniae pneumonia (MP group) and 45 cases of adenoviral pneumonia (ADV group). The general conditions, clinical manifestations, laboratory results, pulmonary imaging, proportion of severe cases, respiratory support style and length of hospital stay were compared between two groups. Results:The MP group had shorter fever time compared to the ADV group ( t=15.910, P<0.01); and the maximum temperature in the MP group was lower than the ADV group ( Z=3.561, P<0.01). In the comparison of shortness of breath, wet rales in the lungs, tri-concave sign, hypoxemia, the differences were all significant between two groups (χ 2=11.203, 6.807, 36.746, 21.177, all P<0.01). The WBC in the MP group was lower than that in the ADV group ( t=33.960, P<0.01); the PCT, IL-6, LDH and 25-(OH) D 3 levels in the MP group were lower than those in the ADV group ( Z=5.986, 3.146, 4.203, 2.094, all P<0.05); while there was no significant difference in CRP levels between two groups ( Z=1.360, P>0.05). Pulmonary imaging in the MP group mainly involved unilateral lung, and the ADV group mainly involved bilateral lungs (χ 2=27.055, P<0.01). There was no significant difference in pulmonary patchy exudation between two groups(χ 2=0.298, P>0.05). There were 30 patients (28.3%) with severe pneumonia in MP group and 33 patients (73.3%) in the ADV group ( t=26.345, P<0.01). Twelve patients (11.3%) in the MP group were given non-invasive respiratory support, 1 patient (0.9%) was given invasive respiratory support; while 31 patients (68.9%) were given non-invasive respiratory support in the ADV group, and 5 patients (11.1%) were given invasive respiratory support (χ 2=66.439, P<0.01). The MP group had shorter hospital stays than ADV group ( t=31.014, P<0.01). Conclusion:The conditions of disease are more severe, and length of hospital stay is longer in children with adenoviral pneumonia than those with Mycoplasma pneumoniae pneumonia.

Multicenter research on the compliance of clinical pathway of bronchopneumonia in pediatrics of tertiary class A hospitals / 中华实用儿科临床杂志

Objective:To evaluate the enrollment rate, mutation rate and causes of variability the clinical pathway of bronchopneumonia.Methods:The enrollment rate, completion rate, variation and reasons of the clinical pathway in Beijing Children′s Hospital, Capital Medical University from January 2012 to December 2016 were retrospectively collected.Data of patients after the clinical pathway of bronchopneumonia in other tertiary class A hospitals were gathered by questionnaires, and the enrollment rate, completion rate, variation rate and reasons were analyzed.Results:(1)At the end of 2016, 11 of the 13 hospitals included in this study had implemented the clinical pathway for 5 years, 1 hospital for 3 years, and 1 hospital for 2 years.(2) Eleven hospitals provided their enrollment rates.The enrollement rate of 2 hospitals was<50%, and that of 9 hospitals was>80%.The annual completion rate of Beijing Children′s Hospital was ≥75%, and the completion rates offered by 8 hospitals were basically >70%.(3) Since the implementation of the clinical pathway for 5 years in Beijing Children′s Hospital, a total of 427 cases were enrolled of which 93 cases were mutated (variability 21.78%). The variability of 5 hospitals was maintained at <15%.The variability of 3 hospitals decreased with the implementation years, and became qualified.The variability of 1 hospital first rebounded and then controlled; 1 hospital increased by 27.65%; 1 hospital was first controlled and rebounded; 1 hospital was always >15%.The main cause of the mutation was coexisting diseases, complications, progression of the disease, or correction of the first diagnosis, etc.Conclusions:The completion rate of tertiary class A hospitals meets the requirements of national policy.However, the enrollment rate needs to be improved, and the variation rate among different hospitals differs a lot.Further implementation of the clinical pathway should be strengthened and monitored.

Analysis of genetic characterization on hemagglutinin gene of influenza B virus isolated in Fujian province, 2010-2015 / 中华实验和临床病毒学杂志

Objective@#To understand the epidemiological and virological features of influenza B viruses and the difference between the vaccine strains and epidemic strains, the antigenic and genetic characteristics on hemagglutinin (HA) gene of influenza B viruses circulating in Fujian during 2010-2015.@*Methods@#The representative strains were selected randomly according to the lineage of influenza B viruses isolated from network laboratory in Fujian, 2010-2015. Viral RNA was extracted and gene fragments were amplified by reverse transcription polymerase chain reaction (RT-PCR ) and the PCR products were sequenced. The complete HA gene sequence was obtained and analyzed via bioinformatics.@*Results@#Compared to the vaccine strains recommended by WHO, there were significant changes in genetic and antigenic characteristics on HA gene of B Yamagata lineage viruses from 2010 to 2015, especially in 2010, 2014 and 2015. There were major five amino acid residues substitutions (116, 150, 165, 196 and 202) involved in antigenic determinants, and the variable sites gradually increased as time on over. However, the variability of B Victoria lineage viruses on HA gene was less and there was no obvious trend over time. The results showed that the B Yamagata vaccine strains of 2010 and 2015 recommended by WHO had poor protective effect on influenza virus infection, while the B Victoria vaccine strain still play a satisfactory protective effect on humans in Fujian.@*Conclusions@#With time on, influenza B Yamagata lineage viruses had gradually mutated, causing a poorly match with vaccine strains in part of year, and emerging antigenic drift phenomenon. Strengthening further surveillance of mutations of B influenza virus remains essential to allow for early warning of influenza epidemic.

Effect of EDTA combined with ciprofloxacin on Pseudomonas aeruginose biofilm infection in guineapig / 中华微生物学和免疫学杂志

Objective To investigate the effect of EDTA combined with ciprofloxacin on Pseudomonas aeruginose biofilm in vivo.Methods Pseudomonas aeruginosa was inhaled into the lung of guinea pigs and colonized,formed biofilm.After 7 days,the model was treated with ciprofloxacin,EDTA alone,or a combination of both for 7 days.The number of colony in the lungs is measured by agar plate.The pathological change of the lung is observed by hematoxylin and eosin (HE) staining and scanning electron microscope.Results EDTA combined with ciprofloxacin make the number of bacteria in the lungs reduced from l05 CFU/g to 10 CFU/g(t =24.67,P＜0.05),the lung lesion was less-sever histophathologically.Conclusion The combination of EDTA with ciprofloxacin has significant activity to remove mucoid PA biofilm in vivo.

Saturation mutagenesis of three amino acid positions consisting of the active site of an endoglucanase from termite Coptotermes formosanus / 生物工程学报

Functional improvement to one component of the cellulase, endo-beta-1, 4-glucanase, has been a focus of the recent research in this area. We report here the saturation mutagenesis of the active site of an endoglucanase (CfEG) from termite Coptotermes formosanus. First, three dimensional structure of CfEG was built via homology modeling by using a close-related (79% homology in sequence) endo-beta-1,4-glucanase (NtEG PDB id = 1ks8) from higher termite Nasutitermes takasagoensis as a template. Second, we identified three corresponding amino acid positions at the active site of CfEG by structural superposition onto NtEG. These three putative amino acids for the active site of CfEG, i.e., Asp53, Asp56 and Glu411, were subjected to saturation mutagenesis using degenerate primers. Among the mutants, Asp53Glu and Asp56Cys showed somewhow higher activities than the wildtype, with the latter having more than 3-fold decrease in Km. Double mutation Asp53Leu/Asp56IIe showed nearly 2-fold increase in specific activity and in the same time 2-fold decrease in Km. Saturation mutagenesis to the position Glu411 produced no active mutant, even changing Glu411 explicitly into its similar amino acids such as Glu411Asp and Glu411Gln could not result in any active mutant. These imply that position Glu411 could be extremely important and therefore indispensable for CfEG functionality.

A STUDY OF THE SOMATIC CHROMOSOME COMPLEMENT AMONG THE CHINESE / 解剖学报

Leukocytes from peripheral blood of 70 normal Chinese,40 males and 30 females, were cultured in vitro according to a modification of Moorhead's method.Chromosome studies were carried out.Table I lists the results obtained including age group,metaphase plates counted and analysed and the chromosome number of each cell.92.4 per cent of 8031 counted cells had the diploid number(2n=46).Chromosome measurements were made from camera lucida drawings.Measurements were estimated to the nearest 0.1 mm on a fine metric scale.The relative length,arm ratio and centromere index of each chromosome were calculated from measurements of 18 haploid chromosome sets of 11 metaphase plates in 6 males and 4 females.The averages,range of variation and 99% confidence limits are listed in Table Ⅲ.Systematic studies of individual chromosomes under the light microscope revealed the following morphological characteristics.The short arms of all 10 acrocentric chromosomes from groups 13—15 and 21—22 could be shown to possess satellites,but in general only 2—8 of these chromosomes at a time carried satellites.In many cells the satellite-carrying chromosomes tended to associate with each other through their satellites.The number of chromosomes involved in each association varied between 2 to 5,occasionally 6.It was not rare to find secondary constrictions to occur regularly in certain regions of chromosomes 1,4,9,13,and 16.Attachments of satellites to such regions of chromosomes 1,4,9,and 13 were also observed.Karyotypic analysis of 357 metaphase plates led us to conclude that in most cases the unequivocal identification of chromosome pairs 4—5,6—12+X,13—15,19—20,and 21—22 was im- possible.Accurate characterisation of these chromosome pairs will have to await further technical advances. The length of the Y chromosomes was systematically measured in order to deter- mine its variability and heritability.Among our sample of 40 male subjects 29 had a Y chromosome slightly larger than chromosome 21,8 slightly smaller or larger than chromo- some 18(long Y chromosome),and 3 smaller than chromosome 21(short Y chromo- some).The heritability of the size of the Y chromosome was established(Fig.Ⅳ). “Spontaneous”aberrations were discovered in 34 of 70 cases.Aberration rates varied from 0.88 to 13 per cent.The relation between aberrations and radiation injuries was briefly discussed.