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OBJECTIVE To explore the material basis and potential mechanism of Kazakh classic prescription Wuzdekh (WZDK) in the treatment of enteritis. METHODS LC-MS/MS technology was used to analyze the chemical components in WZDK. Through network pharmacology and molecular docking technology, the main chemical components of WZDK were screened and the target was predicted; therapeutic effect and target of WZDK on acute enteritis were verified through in vivo experiments. The acute enteritis model of mice was induced by dextran sulfate sodium salt; the general condition of the mice was observed during administration and the disease activity index (DAI) score was calculated; pathological changes of the intestine and mRNA expression of core target were validated by HE staining and quantitative real-time PCR. RESULTS A total of 316 chemical components were obtained by LC-MS/MS. The core targets of network pharmacological analysis mainly included interleukin 1β(IL- 1β), protein kinase B1 (AKT1), tumor protein p53 (TP53), IL-6, tumor necrosis factor (TNF) and so on. The results of molecular docking showed that chemical components such as mairin, lappadilactone, costunolide and dehydrocostus lactone were stable in binding to the core target. The results of in vivo experiment showed that, compared with model group, high dose (5.00 g/kg) of WZDK could significantly reduce the DAI score (P<0.05), improve inflammatory cell infiltration and mucosal tissue damage of colon tissue, and significantly down-regulated mRNA expressions of IL-6, TNF-α, IL-1β and TP53 in colon tissue(P< 0.05 or P<0.01). CONCLUSIONS Chemical components of WZDK such as mairin, lappadilactone, costunolide and dehydrocostus lactone may play the role of improving the imbalance of local inflammatory factors in the intestine and repairing damage of colonic mucosal tissue by down-regulating mRNA expressions of TNF-α, IL-6, IL-1β and TP53 in colon tissue.
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Objective:To investigate the effects on perceived quality of a management assessment system for orthopedic surgery in elderly patients.Methods:Based on the concept of lean management and process nodes in systems, an information management system was established to evaluate the perceived quality of surgical service for elderly orthopedic patients.Four hundred and two elderly orthopedic patients receiving orthopedic surgery after the implementation of the management system were included in the observation group, while 402 elderly patients undergoing orthopedic surgery before the implementation of management system formed the control group.Results:The incidence of hypothermia in patients admitted to the resuscitation room after surgery was lower in the observation group than in the control group(1.7% vs.4.5%, χ2=5.136、 P=0.023). The incidence of shivering during resuscitation was lower and the satisfaction rate of surgical services was higher in the observation group than in the control group(0.7% vs.3.2%) χ2=5.279、 P=0.022, 96.55(95.58, 97.28)分, vs.87.9(83.2, 89.95)分, Z=-4.16, P=0.00). Conclusions:A perceived quality assessment system composed of lean management and information technology for surgical services can reduce complications and improve satisfaction for patients receiving surgery.
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Objective:To analyze the clinical characteristics and prognosis of coronavirus disease 2019 (COVID-19) patients complicated with pneumothorax.Methods:The clinical data of 7 COVID-19 patients complicated with pneumothorax admitted to Huanggang Central Hospital from January 3 to March 10, 2020 were retrospectively analyzed. The clinical features, diagnosis and treatment were summarized, and experience in the treatment of COVID-19 was shared.Results:① General information: among the 7 patients, 5 were males and 2 were females. Four of them had no underlying disease, and 1 had a history of diabetes and hypertension. One patient had only a history of hypertension. There were 6 cases of right pneumothorax and 1 case of bilateral pneumothorax. The 7 patients had a long hospital stay, all over 4 weeks, mostly complicated with multiple organ dysfunction. ② Imaging examination: 1 case evolved from the early stage to the advanced stage within 1 week and to the severe stage within 2 weeks. Pneumothorax occurred 4 weeks later, and was absorbed within 2 weeks. The remaining 6 patients presented progressive stage on admission, all of them advanced to severe stage within 1 to 2 weeks, and most of them presented diffused consolidation shadows, striation shadows and fibrosis of both lungs, obvious pleural adhesion, and extremely slow lesion absorption. ③ Treatment: 1 severe patient with pneumothorax 4 weeks after onset was given non-invasive mechanical ventilation. The remaining 6 critically ill patients were treated with endotracheal intubation and mechanical ventilation. Five patients were treated with mechanical ventilation within 3 days after the occurrence of pneumothorax, and 1 patient was treated with mechanical ventilation after 11 days. ④ Outcome: 1 patient without endotracheal intubation was continuously given nasal high-flow oxygen therapy, and the condition was stable. Four of the 6 patients complicated with pneumothorax after endotracheal intubation died, and the other 2 patients successfully removed the drainage tube within 2 weeks of closed thoracic drainage, and their condition gradually stabilized.Conclusion:COVID-19 complicated with pneumothorax is a dangerous disease with poor prognosis, and should be paid adequate attention.
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Objective@#To analyze the clinical characteristics of hand, foot and mouth disease (HFMD) caused by coxsackievirus A6 (CV-A6) in different age groups.@*Methods@#From January 2015 to December 2017, throat swabs were collected from children with or suspected of having HFMD then quantitative real-time PCR was performed to detect enterovirus nucleic acid. HFMD cases caused by CV-A6 were divided into different groups according to age for comparison.@*Results@#In total, there were 467 cases of HFMD caused by CV-A6 with the age ranging from 3 months to 16 years. There were 273 cases in the infants and young children group (< 3 years old), 131 cases in the pre-school group (3-6 years old), and 63 cases in the school-age group (> 6 years old). The peak incidence was found between May and November.Fever was the common symptom, and the rate of fever in infant group was the highest (220/273, 80.5%); The proportion of cases with leucocyte elevation in the infant group was the highest (127/273, 46.5%) than that in the school-age group (17/63, 27.0%) with a statistical significance. The skin erythra of the HFMD caused by CV-A6 were diverse in forms. Over two forms of skin erythra accounted for 53.9% (257/476) of all cases, and the cases in the infant group showing more forms of skin erythra (163/273, 59.7%). The oral herpes were mainly distributed in the upper palate and pharyngeal isthmus, but the school age group had the least number of distribution sites (0.89±0.86). The cases in the infant group showed higher incidence of skin rash at the elbow joint (109/273, 39.9%), knee (88/273, 32.6%), thigh (112/273, 41%), buttock (122/273, 44.7%) than the other two groups, However, the school age group showed lower incidence of skin rash in the lower leg (0/63, 0%) and thigh (6/63, 9.5%) than the other two groups. The differences between groups were statistically significant. All cases were cured clinically, no severe cases occurred. Among the 288 cases followed up for 6 months, 33 (33/288, 11.5%) suffered from nail exfoliation.@*Conclusions@#Different age groups of HFMD caused by CV-A6 had different clinical manifestations. In the infant group, more cases had fever and the erythra were more diverse in forms and wider in distribution. In addition, the increased leukocytes in routine blood test was also more common in the infant group.
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In the field of bilingualism research, a key scientific question is how bilinguals process two language systems, particularly the effective switch from one language to another, namely bilingual code switching. With the rapid development of neuroimaging techniques, important progresses have been made in bilingual processing studies, especially in code switching. However, consensus has not been achieved regarding the mechanisms of bilingual code switching. Bilingual switching studies using neuropsychological and neuroimaging techniques have gained insights into the temporal and spatial features of the language switching process and the neurological mechanism, which provide direct evidence for the generation mechanism of bilingual code switching.
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Objective@#To understand the epidemiological and etiological characteristics of enterovirus (EV)-associated diseases among children in Beijing from 2010 to 2016.@*Methods@#This was a repeated cross-sectional study. The throat swabs were collected from children with probable EV-associated diseases at the Children' s Hospital Affiliated to Capital Institute of Pediatrics from 2010 to 2016. The samples were sent for pan-EV, enterovirus 71 (EV-A71) and coxsackievirus A16 (CV-A16) detection by real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR) . The viral types of non-EV-A71 and non-CV-A16 EV-positive samples were identified using modified RT-PCR and sequencing with CV-A6, EV-A/B group and 5 'UTR universal primers. The constituent ratios of the prevalence of different EV types in different age and gender groups were compared.@*Results@#Of the 2 703 throat swabs, 1 992 (73.7%) samples were positive for EV, including EV-A71 (19.1%, 516/2 703), CV-A16 (24.3%, 658/2 703), CV-A6 (22.2%, 600/2 703), CV-A10 (4.5%, 122/2 703) and other types of EV (3.5%, 95/2 703). There was 1 case of EV-A71 and CV-A16 co-infection. The positive detection rate of EV-A group (excluding EV-A71, CV-A16, CV-A6 and CV-A10) increased from 11.3% (7/62) to 95.2% (59/62) after using the modified VP1-specific primers and PCR amplification conditions. During the period between 2010 and 2012, CV-A16 and EV-A71 predominated in EV-positive samples. However, CV-A6 accounted for 60.7% (68/112) in 2013, much higher than CV-A16 (23.2%, 26/112) and EV-A71 (12.5%, 14/112). In 2014, EVs were mainly of CV-A16 and EV-A71, but CV-A6 was the predominant type in 2015 (68.2%, 232/340) and in 2016 (38.6%, 151/391). The epidemic season of EVs was mostly from April to August, but CV-A6 showed a small epidemic peak from October to November. The male-to-female ratio of EV-positive patients was 1.50∶1, and EV-associated diseases mostly occurred in children under 5 years of age. Younger children were more susceptible to CV-A6 than to EV-A71 and CV-A16.@*Conclusions@#From 2010 to 2016, there was a significant change in the spectrum of EVs in children with EV-associated diseases in Beijing. Since 2013, non-EV-A71 and non-CV-A16 increased, and CV-A6 gradually became one of the major pathogens of EV-associated diseases. The modified PCR primers and amplification conditions can effectively improve the reliability of test results.
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Objective@#To investigate the relationship between human bocavirus 2 (HBoV2) infection and acute diarrhea in children younger than 5 years of age in a case-control study.@*Methods@#This was a prospective case-control study. During May 2016 to December 2016, fecal specimens were collected from children ≤5 years of age with acute diarrhea who visited the Affiliated Children's Hospital of Capital Institute of Pediatrics (case group), or from children ≤5 years of age without diarrhea from Longtan Community Medical Service Center, Beijing (control group). The case group (n=240) and the control group (n=240) were divided into 8 age subgroups: ≤1 month old, >1-3 months old, >3-6 months old, >6-12 months old,>1-2 years old,>2-3 years old,>3-4 years old and >4-5 years old, and there were 30 cases in each age subgroup. The specimens were tested for 7 types of diarrhea-associated viruses, especially for HBoV2 by real-time PCR method. The HBoV2 viral load was predicted according to the cycle threshold (Ct). Finally, t-test was used to compare the differences between groups.@*Results@#In the case group (n=240), the positive rate of norovirus was 16.7% (40 cases); rotavirus, 10.8% (26 cases); HBoV2, 7.5% (18 cases); adenovirus, 7.1% (17 cases); astrovirus, 6.3% (15 cases); parachovirus, 3.8% (9 cases); and Aich virus, 0.4% (1 case). The positive rates of HBoV2 in case group (7.5%, 18 cases) and control group (5.0%, 12 cases) showed no significant difference (χ2=1.280, P=0.258), as well as in different age groups (all P>0.05) . However, the mean viral load of the HBoV2 in the case group (1×109copies/L with cycle threshold (Ct) 25.8) was higher than that of control group (1×105copies/L with Ct 33.8), showing a significant difference (t=0.597, P=0.000).@*Conclusions@#Norovirus and rotavirus are still the important viral pathogens in children with acute diarrhea. A higher load of HBoV2 may indicate a higher risk of acute diarrhea in children ≤5 years of age in Beijing.
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Febrile neutropenia (FN) causes a major threat to cancer patients after chemotherapy. Broadspectrum antibiotic treatment is a well-established practice for febrile neutropenia. Piperacillin/Tazobactam (P/T) is the frequently used antibiotic in most of FN cases, whereas the use of cefepime remains unclear regarding its potential risk. However, little systematic analysis has been conducted about comparison between these two drugs. Thus, we undertook this meta-analysis to compare these two monotherapies for febrile neutropenia. Through searching Pubmed, Google scholar, Medline databases, EMBASE, OvidSP, ScienceDirect, Web of science, and China Journal Net (CJN) databases, we used the keywords "(Piperacillin/Tazobactam AND cefepime) AND (febrile neutropenia) AND (cancer or tumor)". Only studies with randomized controlled trials were included in the meta-analysis. We screened out a total number of seven clinical trials. This meta-analysis supported that P/T treatment was superior to cefepime treatment based on the average OR comparison, without statistical significance (OR = 1.27, 95% confidence interval = 0.98 to 1.64, p = 0.07). We further divided the seven studies into two subgroups based on age and treatment time. The young group (age <= 19) showed no significant difference (OR = 1.10, p = 0.65). While the old group (age > 19) showed that P/T treatment was better than cefepime with statistical difference (OR = 1.44, p = 0.05). The short-term group (time <= 3 ds) showed P/T treatment was better than cefepime with statistical difference (OR = 1.40, p = 0.05). While in the long-term group (time > 5 ds), there was no significant difference between P/T and cefepime therapy (OR = 1.06, p = 0.79) Asymmetry in Funnel plots indicated no publication bias (CHI2 = 1.47, I2=0%, and p-value = 0.96) in this meta-analysis. It would be a good clinical trial to use P/T treatment to cure FN in cancer patients compared with cefepime treatment, especially in adult patients or patients with a short-term treatment period. This meta-analysis is practically important during antibiotic treatment in FN management.
A Neutropenia Febril (NF) apresenta-se como uma grande ameaça aos pacientes oncológicos após a quimioterapia. O tratamento antibiótico de amplo espectro é uma prática bem estabelecida para a neutropenia febril. Piperacilina/tazobactam (P/T) é o antibiótico frequentemente na maioria dos casos de NF, enquanto que o uso de cefepima permanece pouco claro em relação ao seu potencial risco. No entanto, pouca análise sistemática foi feita sobre a comparação entre esses dois fármacos. Assim, nós realizamos esta meta-análise para comparar estas duas monoterapias para a neutropenia febril. Através da pesquisa na Pubmed, Google Scholar, nas bases de dados da Medline, EMBASE, OvidSP, ScienceDirect, Web of science e nas bases de dados do China Journal Net (CJN), nós usamos as palavras-chave "(Piperacillin/Tazobactam AND cefepime) AND (febrile neutropenia) AND (cancer or tumor)". Apenas estudos com ensaios clínicos randomizados foram incluídos na meta-análise. Nós selecionamos um número total de sete ensaios clínicos. Esta meta-análise suportou que o tratamento com P/T foi superior ao tratamento com cefepima baseado na média da comparação OU (average OR comparision, em inglês), sem significância estatística (OR = 1.27, 95% confidence interval = 0.98 to 1.64, p = 0.07). Posteriormente, nós dividimos os sete estudos em dois subgrupos baseados na idade e no tempo de tratamento. O grupo jovem (idade <= 19) não mostrou uma diferença significativa (OR = 1.10, p = 0.65). Enquanto que o grupo mais velho (idade > 19) mostrou que o tratamento com P/T foi melhor do que o com cefepima com diferença estatística (OR = 1.44, p = 0.05). O grupo de curto prazo (tempo <= 3 ds*) mostrou que o tratamento com P/T foi melhor do que o com cefepima com diferença estatística (OR = 1.40, p = 0.05). Enquanto isso, no grupo de longo termo (tempo > 5 ds) não houve diferença significativa entre as terapias com P/T e Cefepima (OR = 1.06, p = 0.79). A assimetria nos gráficos de funil (funnel plots, em inglês) não indicaram viés de publicação (CHI2 = 1.47, I2=0%, and pvalue = 0.96) nesta meta-análise. Seria um bom ensaio clínico utilizar o tratamento P/T para curar NF em pacientes oncológicos comparados com o tratamento com cefepima, especialmente em pacientes adultos ou pacientes submetidos a um tratamento de curto prazo. Esta meta-análise é importante na prática durante o tratamento com antibióticos na administração de NF.
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Pipéracilline , Neutropénie fébrile , Tazobactam , TumeursRésumé
Objective To investigate the effects of octreotide on the apoptosis of human hepatic stellate cells (HSCs) and expression of Bcl-2/Bax in HSCs,and to reveal the mechanism underlying octreotide against hepatic fibrosis. Methods HSCs lines (HSC-LX2) were incubated with different concentrations of octreotide for 24 and 48 hours. Cell apoptosis was evaluated by Fitc-tunel fluorescence staining. Bcl-2 and Bax protein exoression in HSC-LX2 was detected by immunocytochemistry. Meanwhile, Bcl-2 protein of HSC-LX2 were detected by Western blot assay. Results Octreotide could promote the apoptosis of HSC-LX2, and the apoptosis rate was significantly increased with the concentration of octreotide(P < 0.05). The HSC-LX2 were incubated with the same concentration of octreotide for 24 and 48 hours, the cell apoptosis rate of 48-hour octreotide treatment was significantly higher than that of 24-hour octreotide treatment (P < 0.05). The immunocytochemistry result indicated that octreotide could significantly decrease Bcl-2 expression and increase Bax expression in HSC-LX2 (P<0.05); Western blot assay showed that octreotide could also significantly inhibit Bcl-2 expression in HSC-LX2 (P<0.05). Conclusions Octreotide could induce the apoptosis of HSCs in a dose-and time-dependent manner, the mechanism of octreotide inducing HSCs apoptosis might be associated with down-regulation of Bcl-2 and upregulation of Bax in HSC.
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Objective To investigate the effect of simulated microgravity on growth , morphology, protein expression and virulence gene expression of Klebsiella pneumoniae (KPN).Methods KPN was divided into simulated microgravity group and control group in the experiment .The former group was in the ambient of simulated microgravity in a clinostat .The bacterial growth curves , morphologyical changes in electron microscopy , and protein expression were detected by SELDI-TOF-MS, and the expression of 4 virulence genes(ureA,wabG,uge and fimH) by real-time fluorescence quantitative PCR (RT-PCR) in both groups.Results Compared with the control group , the growth of KPN under simulated microgravity was accelerated , and the total bacterial count increased in microgravity group .The bacterial morphology in microgravity group was changed under scanning electron microscopy (SEM), and thinner and longer bacteria were increased .The transmission electron microscopy ( TEM) analysis revealed increase in cytoplasmic granular substance in microgravity group .Proteome analysis showed that the expression of 18 proteins was changed , half of which up-regulated and the rest were down-regula-ted.Those 18 proteins were searched in the protein library .And 21 proteins of a similar molecular mass were retrieved ,13 of which,proteins with known functions ,were closely related to bacterial life activities .RT-PCR results showed that four virulence genes of KPN were down-regulated.Conclusion Upon exposure to simulated microgravity , the growth and repro-duction of KPN are accelerated and enhanced .The bacterial morphology is changed .The strain′s protein expression and four virulence genes expressionare also changed .Therefore,microgravity can change the characteristics of KPN .
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Objective To analyze the diagnosis value of Color Doppler Uitrasound for the formation of rectus abdominis muscle laceration hematoma after strenuous exercise in children. Methods 29 cases of children patients with rectus sheath hematoma were selected as the research object from January 2009 to January 2014 in our hospital, all patients received rectus abdominal ultrasonography before treatment,got the two-dimensional ultrasonic images and color Doppler image of patients of abdominal rectus. Results Rectus sheath hematoma in the detection rate of the study group was significantly higher than the control group, the misdiagnosis and missed diagnosis ratio were significantly lower than the control group, the average size of hematoma was significantly higher than the control group, the difference was statistically significant(P<0.05); Ultrasonographic study group in regular shape, boundary definition, sensitivity, specificity, accuracy were significantly higher than that of control group,al the differences were statistically significant(P<0.05). Conclusion Children's blood vessels that supply the prone to rupture after strenuous exercise muscle, the rectus sheath hematoma formation, and color Doppler ultrasound compared with two-dimensional ultrasound in diagno-sis is more suitable to occur after strenuous exercise rectus sheath hematoma.
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Objective To investigate the efficacy of auricular plaster therapy for post-laparoscopic cholecystectomy nausea and vomiting. Methods Two hundred and forty laparoscopic cholecystectomy patients were randomly allocated to treatment and control groups, 120 cases each. Both groups received routine nursing care. In addition, the treatment group received auricular plaster therapy. The incidence of nausea and vomiting was observed in the two groups.Results The incidence of nausea and vomiting was 46.7% in the treatment group and 60.8% in the control group; there was a statistically significant difference between the two groups (P<0.05).Conclusion Auricular plaster therapy can relieve postoperative nausea and vomiting in laparoscopic cholecystectomy patients.
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Objective To evaluate the diagnosic value of miniprobe sonography (MPS) in eminence lesion of upper gastrointes-itinal tract .Methods MPS were performed in 485 patients with eminence lesion of upper gastrointesitinal tract .334 patients were given histologic examination .The pathological examination results were compared with diagnosis of MPS and gastroenterology .Re-sults 80 stromal tumors ,41 leiomyomas ,22 cases of ectopic pancreases ,67 cancers ,28 cysts ,26 lipomas ,25 hemangioma ,42 in-flammatory hyperplasia ,104 polyps ,25 cases of protuberance from outside organs ,1 lymphoma and 8 cases of duodenal Brunner gland hyperplasia were detected .The accurancy rate of MPS was 76 .65% which is higher than gastroscope′s .Conclusion MPS is helpful to scope the extent and location of the eminence lesion of upper gastrointesitinal tract and display security of endoscopic bi-opsy .M PS do benefit to the diagnosis ,differential diagnosis and treatment of eminence lesion of upper gastrointesitinal tract .
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Objective To research expression changes of COX-2 gene in Barrett′s esophageal mucosa before and after argon plasma coagulation (APC)operation and to explore it′s clinical curative effect and possible mechanism .Methods 66 barrett′s e-sophageal diagnosed with gastroscope and pathology were randomly divided into normal esophageal group (the negative control group ,group A) ,the APC with acid suppression therapy group(group B) ,Acid antimicrobial drug treatment group(group C) .Ob-serve symptom relief condition ,barrett esophagus epithelium ablation under gastroscope before and after treatment ,esophageal mu-cosa tissue samples was detected by real-time fluorescent quantitative PCR technique .Expression of COX-2 gene in barrett′s esoph-ageal mucosa were compared among groups .Results Compared with group A ,both Group B and group C could effectively relieve symptoms (P<0 .05) .Symptom remission rate had no no statistically significant difference in Group B and group C .Barrett esopha-gus epithelium was not significantly narrowed under gastroscope follow-up Group A and group C ,while Barrett esophageal mucosal occurred ablation with pink mucous covered in Group B .The expression of Cox-2 mRNA in group B decreased ,which was similar to group A .The expression of Cox-2 mRNA in group C also decrease ,but there was no significant differences before and after treat-ment .Conclusion APC with acid suppression therapy could effectively relieve symptoms ,melting Barrett esophagus epithelium ,and is of a safe and effective treatment on Barrett′s esophagus .
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Objective To analyze the combination characteristics between Tir-IBD( intimin binding domain) and its ligand intimin or mutant intiminN916Y of EHEC O157 ∶H7.Methods The gene of TirIBD (tir-ibd) from EHEC O157 ∶H7 chromosome was cloned into pMD18-T vector.Thereafter,the amplified gene was cloned into prokaryotic expression plasmid pET-21 a (+).The recombinant pasmid pET-21 a( +)-tir-ibd was transformed into E.coli BL21 (DE3).After inducement,the protein Tir-IBD was successfully expressed and analyzed with SDS-PAGE and Western blot.It was purified by affinity chromatography and ionexchange chromatography and was coupled on the Ni-NTA chip of BIACore 3000.Then the ligand intimin and mutant intiminN916Y were flow through the chip and their combination characteristics were detected.Results In the present study,the gene of Tir-IBD(tir-ibd) was successfully cloned into pET-21a(+).The results of SDS-PAGE and Western blot assay showed that the protein was successfully expressed,which accounts for 15% of total expression products,and its molecular weight was about 10×103.The purity was above 95% after purification.After coupled on the Ni-NTA chip of BIACore 3000,their combination characteristics with ligand intimin and mutant intiminN916Y were successfully detected.The equilibrium binding constants Ka was obtained by fitting the data with the BIACore evaluation program ( Version 4.1 ).The result showed that the combination characteristics between Tir-IBD and intimin have some difference compared with that of mutant intiminN916Y and the difference is temperature dependent.Conclusion Tir-IBD of EHEC O157 ∶H7 was successfully constructed and purified.The method to analyze the combination characteristics between Tir-IBD and its ligand intimin or mutant was established.The combination characteristics between Tir-IBD and intimin or mutant intiminN916Y have some temperature dependent difference and the mutated amino acids residue is crucial for their receptor-ligand binding.
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Objective To obtain the specific antigens of the expressed major capsid proteins from Noroviruses with different sub-genotypes in Beijing area. Methods The full-length genes of the major capsid proteins (VP1)were obtained through the amplification of the VP1 encoding gene in the recombinant plasmids pBST-CR2987(G Ⅱ-3)and pBST-CR2932(GⅡ-4),which represented different Norovirus geno-types. The full-length genes were sub-cloned into the expression vector pET-30a(+),resulting in a recombinant plasmid, with which the BL21 competent cells were transformed, and the expression of the gene was induced by adding IPTG to the growth culture. The expression of the major capsid proteins were analyzed with Coomassie blue staining after SDS-PAGE, and assayed by Western blot with serum from human. Results (1)The major capsid protein genes of CR2987 and CR2932 were sub-cloned into expression vector pET-30a(+). The VP1 encoding genes were 1647 bp in length for CR2987 and 1623 bp for CR2932. The open reading frames(ORF)coded for 549 and 541 amino acids for these two proteins, respectively. (2)The expressed VP1s were present primarily as inclusion bodies,and the maximal amount of the expressed proteins occurred at 4-6 h after IPTG induction.(3)These VP1s could be recognized by specific immune serum against VP1 of Norovirus as well as His-tag antibody. Conclusion The VP1s of CR2987 and CR2932 are expressed in BL21 E.coli cells.The expressed VP1s could react with specific immune serum against VP1 of Norovirus, indicating that the expressed VP1s are of antigenicity.
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OBJECTIVE To investigate the nasopharynx carriage status of opportunistic pathogens in non-infectious old people.METHODS The totally of 592 pharyngeal samples from old persons were collected.RESULTS The isolated rates of main opportunistic pathogens were Haemophilus parainfluenzae,Stomatococcus mucilaginosus,Streptococcus pneumoniae and Klebsiella pneumoniaa.The isolated rate of single opportunistic pathogen was 45.8% and that of multiple opportunistic pathogens was 3.9%.The total carriage rate of opportunistic pathogen was 49.7%.The carriage rate of G+ or G-bacteria was 13.2% or 86.8%,respectively.The isolated rates of 2 multiple pathogens,S.pneumoniae and K.pneumoniae were increased with aging.The isolated rate of K.pneumoniae was higher in persons with more than 3 kinds of underlying diseases.CONCLUSIONS Aging,pharyngeal carriaging of S.pneumoniae,H.parainfluenzae,K.pneumoniae,et al and with multiple underlying diseases might be the risk factors for elder people to suffer from infectious respiratory disease.Isolating pharyngeal opportunistic pathogen among non-infection people is necessary and significant for prevention and treatment of infectious respiratory disease.
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OBJECTIVE To investigate the pharyngeal opportunistic pathogens and their drug resistance related to different duty of the hospital staff,in order to improve the control and prevention of hospital infection.METHODS Totally 302 pharyngeal samples were cultured and isolated,and the tests of antibiotic susceptibility were(performed) for isolated strains to 15 kinds of antibiotics by using of the paper diffusion method.RESULTS The(isolated) rate of the opportunistic pathogens among hospital staff was 30.13%,higher in nurses(41.32%) than in technicians(20%,P
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<p><b>OBJECTIVE</b>To define roles of B7-1 co-stimulation factor expressed in human malignant cell lines in mediating anti-tumor T cell immune responses.</p><p><b>METHODS</b>Examining human leucocyte antigen (HLA) and B7 expressions on 8 human blood malignancies cell lines by flow cytometry. Transfecting B7-1 gene to B7-1 negative (B7(-)) Raji and B7(-) Jurkat cell lines by liposome, and comparing the potencies of blood malignant cell lines in the induction of T cell activation by examination of T cell cytokine mRNAs before and after transfection using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>High level of HLA I and II molecules were expressed in most human blood malignant cell lines examined, and the co-stimulatory factor B7-2 was also highly expressed. In contrast, another member of B7 family: B7-1 was either not expressed or very limitedly expressed in most of these hematopoietic malignant cell lines. Most importantly, transfection of B7-1 gene to B7(-). Raji and B7(-). Jurkat cell lines made these cell lines better antigen presenting cells for stimulation of anti-tumor T cell activation, which was demonstrated by up regulation of expression of T cell cytokines IL-2, IL-4 and INF-gamma mRNAs after incubation of these tumor cells with T cells for 24 h.</p><p><b>CONCLUSIONS</b>B7 co-stimulation plays an important role in anti-tumor immunity. Transfection of B7-1 gene to the human hematopoietic malignant cell lines that are deficient in the B7-1 expression empowers their antigen presentation potency for activation of anti-tumor T cells. Our results suggested that repairing the deficiency of B7-1 co-stimulatory pathway in tumor cells might be a novel immunotherapeutic approach for human hematopoietic malignancies.</p>
Sujets)
Humains , Présentation d'antigène , Antigènes CD , Antigène CD80 , Physiologie , Antigène CD86 , Cytokines , Génétique , Cytométrie en flux , Antigène HLA-DR7 , Physiologie , Tumeurs hématologiques , Allergie et immunologie , Cellules K562 , Activation des lymphocytes , Glycoprotéines membranaires , ARN messager , Lymphocytes T , Allergie et immunologie , Transfection , Cellules U937Résumé
<p><b>OBJECTIVE</b>To explore the effects of a novel human chemokine-like factor 1 (CKLF1) on stem cell/progenitor cells.</p><p><b>METHODS</b>Human bone marrow mononuclear cells were separated by Ficoll (1.077 g/ml), and cultured in suspension and semisolid colony culture. The effects of CKLF1 on CFU-GM and CFU-Mix were observed.</p><p><b>RESULTS</b>The number of CFU-GM was significantly increased in 10 groups by addition of CKLF1 plasmid supernatant. The mean value was 234.81 +/- 98.71/1 x 10(5) cells, 2.42 fold of control group (P < 0.05). The mean value of CFU-Mix in groups of negative control, CKLF1, PHA, GM-CSF and G-CSF were 82.00 +/- 20.25, 105.76 +/- 36.70, 146.63 +/- 27.09, 143.33 +/- 60.23 per 1 x 10(5) cells, respectively, no statistical differences were seen between control and CKLF1 groups. The CD(34)(+) cells were detected by FACS. The average percentage in the groups of normal control, CKLF1, PHA and GM-CSF were (0.75 +/- 0.62)%, (1.32 +/- 0.87)%, (0.15 +/- 0.02)%, and (0.29 +/- 0.23)%, respectively. Compared with control, no significant differences were seen between each group (P > 0.05).</p><p><b>CONCLUSION</b>Novel chemokine-like factor 1 can increase the proliferation of CFU-GM, which indicated that CKLF1 could increase the proliferation of human bone marrow hematopoietic progenitor cells and colony formation.</p>