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Objective@#To analyze the molecular characteristics of Listeria monocytogenes strains from ready-to eat food in China.@*Methods@#A total of 239 Listeria monocytogenes strains isolated from ready-to-eat food in 2017, all strains underwent whole-genome sequencing (WGS) , and comparisons uncovered population structure derived from lineages, clonal complex, serogroups, antimicrobial susceptibility and virulence, which were inferred in silico from the WGS data. Core genome multilocus sequence typing was used to subtype isolates.@*Results@#All strains were categorized into three different lineages, lineage Ⅱ was the predominant types in food, and IIa was the main serogroups. CC8, CC101 and CC87 were the first three prevalent CCs among 23 detected CCs, accounting for 49.4%. Only 4.6% (11 isolates) of tested strains harbored antibiotic resistance genes, which were mostly trimethoprim genes (7 isolates, 2.9%). All strains were positive for LIPI-1, and only a part of strains harbored LIPI-3 and LIPI-4, accounting for 13.8% (33 isolates) and 14.2% (34 isolates), respectively. ST619 carried both LIPI-3 and LIPI-4. 51.5% (123 isolates) of strains carried SSI-1, and all CC121 strains harbored SSI-2. Different lineages, serogroups and CCs can be separated obviously through cgMLST analysis, and 24 sublineages were highly concordant with CCs.@*Conclusion@#Ⅱa was the main serogroups in ready-to-eat food isolates in China; CC8, CC101 and CC87 were the prevalent CCs, and CC87 isolates was hypervirulent isolates, cgMLST method can be adopted for prospective foodborne disease surveillance and outbreaks detection.
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Objective@#To investigate the antimicrobial resistance status and pulsed field gel electrophoresis (PFGE) patterns of Salmonella enteritidis (S.enteritidis) in Shanxi Province in order to provide references for the treatment of Salmonella infection and for tracing the source of outbreaks of foodborne diseases.@*Methods@#Sixty-four S. enteritidis strains were collected by monitoring sites for foodborne diseases from April 2015 to March 2018. Biochemical identification system and serotyping analysis were used for bacterial identification. Drug susceptibility patterns were analyzed with micro-broth dilution method. PFGE was used for molecular typing.@*Results@#The antimicrobial resistance rate of 64 S. enteritidis strains to nalidixic acid (95.31%) was the highest, followed by that to ampicillin (90.63%) and to ampicillin/sulbactam (81.25%). They had lower resistance rates to cefazolin, cefotaxime, tetracycline, ceftazidime, trimethoprim/sulfamethoxzole and ciprofloxacin (3.13%-23.44%) and were all sensitive to chloramphenicol, cefotaxime, azithromycin, imipenem and gentamicin. No statistically significant difference in drug resistance rates was found between the sporadic strains and the outbreak strains (P>0.05). All 64 S. enteritidis strains digested with XbaⅠwere divided into 33 molecular patterns by PFGE. The numbers of bacteria contained in each pattern ranged from 1 to 10 strains. The similarity among patterns was between 54.6% and 100%.@*Conclusion@#More attention should be paid to the drug resistance status of S. enteritidis in Shanxi Province. It is necessary to strengthen the standardized administration of antibiotics. The PFGE patterns of S. enteritidis reveal the presence of significant genetic polymorphism. PFGE is of great significance in analyzing the genetic relationship among S. enteritidis strains and in identifying and tracing the source of outbreaks of foodborne diseases.
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Anatomy and surgery teaching has always been a difficulty in clinical medical education. In particular, the anatomy and surgery teaching under the endoscope is more difficult to learn and under-stand because of the visual differences with the direct vision. If the students misunderstand, it may lead to the severe surgical complications. We applied the image-guidance technology in the teaching of rhinology endoscope surgery, which can help teachers to teach the sinonasal anatomical structure and give an accurate explanation of surgical procedures. According to the questionnaire survey, 96.23% (51/53) of the students thought the method was effective and clear, and 98.11% (52/53) of them thought the method was better than the traditional teaching method of nasal endoscope. By this technology, the students can acquire a deeper understanding of the anatomy and surgery procedure. It especially fits the surgery teaching of the sinonasal occupying lesion.
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OBJECTIVE To evaluate the clinical outcome of submucosal inferior turbinectomy and outfracture surgery of inferior turbinates. METHODS All patients receiving two different operations were measured by acoustic rhinometry and questionnaire of QOL at preoperative 1 week and postoperative 12 months, seperately. RESULTS Forty-seven patients with inferior turbinate hypertrophy were enrolled in this study. Evaluation of SNOT-20 discovered both surgeries could improve patients' QOL with similar outcome. Preoperative '5 important items' in patients with inferior turbinate hypertrophy were 'lack of a good night's sleep', 'need to blow nose', 'thick nasal discharge', 'fatigue' and 'dizziness'. Postperative '5 important items' were 'postnasal discharge', 'runny nose', 'sneezing', 'reduced concentration' and 'reduced productivity'. Both surgeries could make acoustic rhinometry parameters change obviously, such as minimal cross-sectional area, 0-5 cm nasal volume(NV) and 2-5 cm NV. Furthermore, submucosal inferior turbinectomy produced more volume in nasal cavity than outfacture surgery, (7.28±2.01)cm3 vs (6.01±1.22)cm3, (5.99±1.87)cm3 vs (4.23±1.08)cm3(P<0.05), seperately. There was no correlation between the data of SNOT-20 and acoustic rhinometry. CONCLUSION We recommend outfracture surgery of inferior turbinate as the preferred surgical choice for patients with mild inferior turbinate hypertrophy.
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Objective To observe the clinical effect of Guizhi Fuling pill in the treatment of interstitial tubal pregnancy.Methods80 cases with interstitial tubal pregnancy in Hangzhou obstetrics and gynecology hospital from August 2015 to December.2016 were randomly divided into the observation group and the control group, 40 cases in each group.The control group were given methotrexate and mifepristone, at this basis, the observation group were given Guizhi Fuling pill.The effect was analyzed in the two groups.Results①β-HCG negative time in the observation group were significantly shorter than that in the control group(P<0.05).②Adverse reactions rate in the observation group was significantly lower than that in the control group(P<0.05).ConclusionIt can shorten the treatment time, reduce the adverse reaction which Guizhi Fuling pill was used in the adjuvant treatment of interstitial tubal pregnancy.
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Objective To investigate the effect of ovulation induction of letrozole and clomiphene on the treatment of polycystic ovary syndrome.Methods From February 2014 to February 2014 80 cases with polycystic ovary syndrome in Hangzhou maternity hospitalas the research object were divided into two groups: the control group and the observation group.The control group were treated with clomiphene;the observation group were treated with letrozole.The clinical effect in the two groups were compared.Results The total effective rate was 95.0% in the observation group, which was higher than 67.5% in the control group, the difference was statistically significant(P<0.05).The thickness of endometrium in the observation group was(9.10±1.32)mm and(5.38±0.61)mm in the control group at the HCG injection day, the difference between the two groups was statistically significant (P<0.05).Conclusion Letrozole can be used in the treatment of polycystic ovary syndrome, can get high quality treatment effect, worthy of promotion.
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Objective:To establish the methods for the qualitative identification and content determination of aurantio-obtusin and chrysophanol in Zeju Jiangzhi tablets. Methods:A TLC method was adopted for the qualitative identification, and an HPLC method was used for the content determination. The determination was performed on a Wondasil C18 (250 mm × 4. 6 mm, 5 μm ) column with the mobile phase of acetonitrile -0. 1% phosphonic acid with gradient elution at the flow rate of 1. 0 ml?min-1 , the detection wave-length was 286 nm, the column temperature was 30℃ and the injection volume was 10μl. Results:The TLC spots of aurantio-obtusin and chrysophanol were clear and well-separated without any negative interference. The HPLC experiment results showed the good line-arity within the range of 1. 03-25. 72μg?ml-1(r=0. 999 9) for aurantio-obtusin, and 0. 48-11. 92μg?ml-1(r=0. 999 9) for chry-sophanol. The average recovery was 99. 21% and 98. 85%, and RSD was 0. 70% and 0. 73%, respectively (n=9). Conclusion:The method is simple, accurate and repeatable, which can be used for the qualitative identification and content determination of auran-tio-obtusin and chrysophanol in Zeju Jiangzhi tablets.
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Anthrax is a highly lethal infectious disease caused by the spore-forming bacterium Bacillus anthracis. The major virulence factor of B. anthracis consists of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA binds with LF to form lethal toxin (LT), and PA binds with EF to form edema toxin (ET). Antibiotics is hard to work in advanced anthrax infections, because injuries and deaths of the infected are mainly caused by lethal toxin (LT). Thus, the therapeutic neutralizing antibody is the most effective treatment of anthrax. Currently most of the anthrax toxin antibodies are monoclonal antibodies (MAbs) for PA and US FDA has approved ABTHRAX humanized PA monoclonal antibody for the treatment of inhalational anthrax. Once B. anthracis was artificially reconstructed or PA had mutations within recognized neutralization epitopes, anti-PA MAbs would no longer be effective. Therefore, anti-LF MAbs is an important supplement for anthrax treatment. Most of the anti-LF antibodies are murine or chimeric antibodies. By contrast, fully human MAbs can avoid the high immunogenicity of murine antibodies. First, we used LF to immunize the transgenic mice and used fluorescent cell sorting to get antigen-specific memory B cells from transgenic mice spleen lymphocytes. By single cell PCR method, we quickly found two strains of anti-LF MAbs with binding activity, 1D7 and 2B9. Transiently transfected Expi 293F cells to obtain MAbs protein after purification. Both 1D7 and 2B9 efficiently neutralized LT in vitro, and had good synergistic effect when mixed with anti-PA MAbs. In summary, combining the advantages of transgenic mice, fluorescent cell sorting and single-cell PCR methods, this study shows new ideas and methods for the rapid screening of fully human monoclonal antibodies.
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Objective:To establish a method for the determination of related substances in enoxacin gluconate injection by HPLC with gradient elution. Methods:HPLC was performed on a DiamonsilTM C18 column (250 mm × 4. 6 mm, 5 μm), the mobile phase A was 0. 025 mol·L-1 phosphonic acid solution (adjusting pH to 3. 0 with triethylamine)-methanol- acetonitrile (80∶10∶10) and the phase B was 0. 025 mol·L-1 phosphonic acid solution (adjusting pH to 3. 0 with triethylamine)-methanol-acetonitrile (350∶325∶325) with gradient elution at a flow rate of 1. 1 ml·min-1 , the detection wavelength was 269 nm and 284 nm, the injection volume was 20μl, and the column temperature was 40℃. Results:Under the HPLC conditions, the samples had good stability and separation. The calibration curve was linear within the concentration range of 19. 80 ng·ml-1-19. 80 μg·ml-1(r=0. 999 9) for 5-hydroxymethylfur-fural with the detection limit of 0. 18ng, and the average recovery was 99. 68% with RSD of 0. 12% (n=9). Conclusion:The method is accurate, sensitive, specific and reproducible, and can be used in the determination of related substances in enoxacin gluconate in-jection.
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Objective To explore the sensitivity and the molecular mechanism of cisplatinresistance ovarian cancer cell line C13 to proteasome inhibitors and the combination with cisplatin. Methods After different treatments, methyl thiazolyl tetrazolium (MTT) assay was applied to examine the cell viability, annexin-V/propidium iodide(PI) apoptosis detection kit was used to determine the apoptosis rate of different groups, western blot assay was introduced to evaluate the expression levels of Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein (cFLIPs), and the activity of caspase-8 was examined. Results MTT assay shown that the cell viability ratios of combination group at serial time points from 12, 24, 36, 48, 60, 72 hours were ( 56.0 ± 8.4 ) %, (44.7 ± 7.3 ) %, ( 33.7 ±11.2) %, (27.6 ± 8.0) %, (27. 6 ± 7.6) % and (28.1 ± 2.4) %, which were much lower than those of cisplatin group (P <0.05). After treated for 24 hours, apoptosis rates of cisplatin group, bortezomib group and combination group were ( 16.7 ± 1.7) %, (23.4 ± 2.1 ) % and (26.9 ± 1.6) %, respectively. The rate of combination group was much higher than that of non-treated group and that of cisplatin group or bortezomib group ( P < 0.05 ). Western blot assay showed the changes of expression levels of cFLIPs, which were downregulated seriously after cisplatin, bortezomib or combination treatment [ (43.2 ± 2.3 )% vs( 75.7 ± 3.0)%vs (67.9 ± 2.1 ) %, P < 0.05 ]. The caspase-8 activity of combination group was (5.6 ± 1.6) folds than that of non-treated group, which was higher than those of other two groups [ ( 2.3 ± 1.0) and (4.2 ± 0.9 ) folds,P < 0.05 ]. Conclusions The tumor cell lethal effect of cisplatin could be increase significantly by the combination application of proteasome inhibitors, bortezomib. And the cFLIPs/caspase-8 signaling pathway may be play an important role in the molecular mechanism of the combination treatment.
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To investigate the relationships between constitutional types of traditional Chinese medicine (TCM) and hypertension so as to provide epidemiological evidence for the theory of correlation between constitution and disease.
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Objective To explore the semitivity of ovarian cancer cell line SKOV3 to paclitaxel,oroteasome inhibitors,bortezomib,and their combination.Methods The methyl thiazolyl tetrazolitim (MTT)assay was applied to examine the cell viability after treatment.The annexin V-propidium iodide apoptosis detection kit was used to determine the apoptosis rate of different groups.Western blot assay was used to evaluate the expression levels of phosphorylated protein kinase B(AKT)and glycogen synthase kinase-3 beta(GSK-3β).Results In MTT assay,the cell viability ratios of the combination group at serial time points from 12,24,36,48 and 72 hours Were(65.2±5.8)%,(58.3±14.4)%,(35.3±5.0)%,(19.2±1.5)%,and(11.4 ±2.5)%,which were significantly lower than those of the paclitaxel group (P<0.05).After arug treatments,apoptosis rates of paclitaxel group,bortezomib group and the combination group were (14.7±0.5)%,(15.1±0.8)%and(20.5±0.7)%respectively.The rate of the combination group was significantly higher than that of non-treated group and paclitaxel group(P<0.05).Western blot assay showed the changes in expression levels of phosphorylated AKT and GSK-3β,which were decreased significantly after paclitaxd and bortezomib combination treatment [(3.2±0.8)%,(19.3±0.4)%;P<0.05].Conclusions The lethal effect of paclitaxel on tumor cells could be increased significantly by its combination with proteasome inhibitors,bertezomib.The AKT/GSK-3β signaling pathway plays an important role in the molecular mechanism of the combination treatment.
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[Objective] To optimize the extraction process of Weiyanping granules. [Method]The orthogonal test method was adopted to explore the influences of 4 factors(quantity of water,soaking time,extraction time and times) on the yielding rate of berberine and dry paste rate. [Results]Analysis by synthesis,the optimum condition for extraction was A2B3C1D3.The medicinal substances should be soaked in 10 times water for 2 hours and distilled for 3 times,with 1 hour each time.[Conclusion]The experiment result showed that the optimum extraction process condition was stable,reasonable and economical,which could be used to guide industrialized production.
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OBJECTIVE To explore the valuable information for studying human nasal mucociliary transport system by measuring the nasal ciliary beat frequency(CBF)in vitro.METHODS CBF of cultured human nasal epithelial cells was measured by highspeed digital microscopy in HBSS.RESULTS CBF of nasal epithelial cells from inferior turbinate and uncinate process of same patient has no significant difference in vitro(t=-0.087,P=0.931).CBF of normal mucosa and uncinate process from patients of sinusitis has no significant difference in vitro(t=0.419,P=0.678).CONCLUSION CBF of nasal mucosa of sinusitis can replace the CBF of normal nasal mucosa in vitro for for nasal ciliary movement studying.Because the mucosa of uncinate process is easier to obtain,it can replace the mucosa of inferior turbinate for cell culture in vitro.
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OBJECTIVE To find the safer anesthesia induction methods in children with laryngeal papilloma combined by dyspnea.METHODS 50 laryngeal papilloma children with II degree laryngeal obstruction were randomly divided into 2 groups:inhalational group and intravenous group,no muscle relaxant was used in either of the groups.MAP,and SpO2 were observed at one minute before and after induction.The intubation condition was also assessed and compared between the 2 groups.RESULTS MAP and HR in intravenous group were significantly higher than those of inhaled group after intubation [(68.7?6.4)mmHg vs.(64.0?8.0)mmHg;(142.6? 13.8)bpm vs.(124.6?12.5)bpm;P0.05].For the intubation condition,the satisfactory rate in inhaled group was higher than that of intravenous group,P
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OBJECTIVE To establish a cell culture of human nasal epithelial cells on glass cover slides coated with rat tail collagen and explore a method to research human mucociliary system. METHODS The rat tail collagen and collagen coated cover slides were prepared for explant culture of human nasal epithelial cells. RESULTS The collagen on cover slides must be thin and ? at. It was about 1mm thick. The cultured epithelial cells were monolayer and polygonal cells. There was tight junction between the cells. Ciliary beat frequency of different cilia was equal on the same cell,and it was also equal of on cells from uncinate process and inferior turbinate. CONCLUSION This successful cell culture model of human nasal epithelail cell could be helpful to research the physiology and function of human nasal mucociliary system.
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OBJECTIVE To develop an explant outgrowth culture system for airway ciliated cells and explore the usefulness of the system.METHODS Mucosal specimens from rabbit tracheal epithelium were seeded into collagen-coated cover-slips. After 7 days,the specimens were characterized by Photo- contrast microscopy,transmission and Scanning electron microscope. They were also examined with hematoxylin-eosin staining and mucin-5AC immunohistochemistry. Ciliary beat frequency was measured with high-speed digital microscopy. RESULTS Many ciliated cells with good viability and goblet cells were found in the culture system. Immunohistochemistry analysis revealed that mucin- 5AC antibody labeled mainly in the goblet cells.The ciliated cells were differentiated well detected by electron microscope. At (30?1)℃,the basal ciliary beat frequency was (13.2?0.9) Hz. CONCLUSION The method may be useful in establishing a culture system for ciliated cells and the system will be suitable for the long-term studies of mucociliary transport system.
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Aim To study gyrA and parC mutations of clinical Pseudomonas aeruginosa strains. Methods MIC values of 55 clinical P.aeruginosa isolates were determined by agar dilution test and 1 sensitive strain and 8 resistant strains were selected with standard sensitive strain ATCC27853 as control, the quinolone determining region (QRDR) of the gyrA and parC genes were amplified by PCR, the lengths of PCR products were 351 bp and 397 bp. The gyrA PCR products(351 bp) were digested with enzyme sacⅡ. The gyrA and parC gene were sequenced. Results In this study, gyrA genes of all resistant strains had an ACC to ATC mutation in codon 83, leading to the amino acid substitution of an isoleucine for a threonine, and three high level resistant strains also showed a GAC to GGC mutation in codon 87, leading to the substitution of a glycine for an aspartic acid. In addition, four resistant strains also had an TCG to TTG mutation in codon 87 of parC gene, leading to the amino acid substitution of a serine for a leucine. The strains with both gyrA and parC mutations were two to sixteen times more resistant than the strains which had only gyrA mutations. At the same time, a silent mutation (CAC to CAT) in codon 132 of gyrA gene and a silent mutation(GCT to GCG) in codon 115 of parC gene occured, which did not lead to amino acid change. Conclusion The mutations of 83 and 87 codons of gyrA and the mutatations of 87 codon of parC gene were related to fluroquinolone resistance, and the mutations of the 83 codon of gyrA gene were more important.
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OBJECTIVE To study the correlation between mutations of gyrA and parC genes and ciprofloxacin-(resistant) clinical isolates of Acinetobacter baumannii from Chengdu.METHODS The genes of gyrA and parC DNA in 28 ciprofloxacin-resistant and 2 ciprofloxacin-susceptible isolates of A.baumannii were amplified by PCR and then analyzed by restriction fragment length polymorphism(PCR-RFLP) and DNA sequencing.(RESULTS) Hinf Ⅰ digestion of the gyrA gene products of susceptible isolates(generated) two fragments,but resistant isolates(generated) one fragment.The parC gene products generated 2 fragments.DNA sequencing of 5 resistant isolates revealed mutations in gyrA gene that resulted in amino acid substitutions: Ser83→Leu and Ala88→Thr,especially,Ser83 point mutation accounting for the disappearance of sequence of Hinf Ⅰ.There was not any mutation in(gyrA) of 1 susceptible isolate;the substitution of Ser108→Ile in parC(gene) of 1 susceptible and 1 resistant isolates were identified,the remaining four isolates had more nosense mutations.CONCLUSIONS Compared with parC gene mutations,(gyrA) gene mutations of A.baumannii appears to be the main molecular mechanism responsible for ciprofloxacin resistance.The high-level ciprofloxacin resistance in A.baumannii probably needs a variety of mutations.
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AIM: To use qualitative and quantitative RP HPLC for the sepatation and determination of flavonoids found in the leaves of Acer truncatum bunge in spring, summer and autumn. METHODS: The determination of three kinds of flavonoid aglycones was carried out with Diamonsil TM C 18 column (4.6 mm?250 mm,5 ?m), using methanol water(1% acetic acid) as mobile phase at a flow rate of 1.0 mL?min -1 and detected at the wavelength of 360 nm. RESULTS: The average recovery of quercetin was 100.4%, RSD =1.61%( n =5). The average recovery of kaempferol was 102.5%, RSD =1.77%( n=5 ). The average recovery of isorhamnetin was 102.3% , RSD =1.63%( n=5 ) CONCLUSION: Isorhamnetin and kaempferol in the leaves are reported here for the first time. The methods were simple, rapid and reliable and could be used to control the quality of the herbal medicines prepared from the leaves of Acer Truncatum Bunge.