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@#Abstract: Objective To investigate the differences in epidemiological and clinical characteristics of patients with spotted fever (SF) and severe fever with thrombocytopenia syndrome (SFTS). Methods A total of 86 patients with SF and 113 patients with SFTS who were laboratory-confirmed in the second-level and above hospitals in Lu'an City from January 2017 to January 2022 were selected. The basic data, epidemiological history, clinical data and laboratory test results of the two diseases were retrospectively analyzed for comparison. Results The proportion of male in SF group was 32.56% (28/86), and the proportion of male in SFTS group was 53.98% (61/113), the difference was statistically significant (χ2=9.067, P<0.01). The proportions of abdominal pain and diarrhea in the SF group were (3.49%, 3/83) and (21.24%, 24/113), which were significantly lower than corresponding (6.98%, 6/86) and (46.90%, 53/113) in the SFTS group (χ2=13.121, 37.322, P<0.01). The incidences of rash and eschar in SF group were 95.35% (82/86) and 20.93% (18/86), which were significantly higher than corresponding 1.77% (2/113) and 0.88% (1/113) in SFTS group (χ2=175.311, 22.721, P<0.01). The levels of leukocytes, platelets and C-reactive protein in the SF group were significantly higher than those in the SFTS group, and the levels of transaminase, lactate dehydrogenase and D-dimer were significantly lower than those in the SFTS group, and the differences were statistically significant (all P<0.05). Conclusions The rash and inflammatory reaction are more obvious in SF patients, while the liver function, myocardial function and coagulation function are significantly impaired in SFTS patients.
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Objective To provide anatomical basis for clinical treatment of acromioclavicular joint dislocation by studying the morphology of coracoid process of human scapula. Methods A total of 500 patients with shoulder injury were selected from the Affiliated Hospital of Traditional Chinese Medicine of Southwest Medical University in Sichuan Province, and 300 patients were selected as subjects, including 159 cases of right shoulder and 141 cases of left shoulder. CT scan images and 3D reconstruction results of scapula of the subjects were collected. The basic morphological characteristics of coracoid process CT images of the subjects were observed, and the relevant parameters were measured, including the longest horizontal distance of the coracoid process tip and the thickness of the midpoint (cd, pp’), the distance from the upper part of the coracoid process scapula to the base and the thickness of the midpoint (mn, kk’). The distance from the apex of the coracoid process to the base of the coracoid process (ab), the longest horizontal distance of the recursion part of the coracoid process (ef), the distance of as (point s was the intersection of point a perpendicular to mn), the distance of hj (point h and j were the intersection of the base of the coracoid process and the recursion part respectively), and ik (point i was the intersection of point k perpendicular to mn and the coracoid process retraction). Results According to the morphological characteristics of coracoid process, they were divided into five types, including peanut 29. 7%; Short rod type accounted for 27. 4%; Melon seed type accounted for 12. 6%; Rod type accounted for 17. 0%; Wedge type accounted for 13. 3%. Through data comparison, it was found that the distance ef and distance hj on the left were larger than those on the right, P<0. 05. All types had statistical difference in comparison distance cd, P<0. 05. The melon seed type showed statistical differences with peanut type, wedge type, long stick type and short stick type in thickness pp’, distance ab and as of point p, P<0. 05. In the comparison of point K thickness kk’, there was statistical difference between melon seed type and other four types, P<0. 05. In the distance ab comparison, there was statistical difference between the short bar type and the other four types, P < 0. 05. Conclusion The study on the morphology of coracoid process can provide anatomical basis for clinical reconstruction of coracoid ligament to treat acromioclavicular joint dislocation.
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Liver is one of the most important organs in the human body. Liver diseases are also a major threat to human health and longevity. Hepatic decompensation treatment is quite difficult due to multiple reasons. Extracorporeal liver support devices are unable to solve this problem, and there is a severe shortage of orthotopic liver transplant donors. Study of pluripotent stem cell-derived hepatocytes and organoids can determine not only hepatocyte fate, but also liver development, regeneration mechanisms, and pathophysiology. Furthermore, it can be used for drug screening in order to provide a stable source of functional hepatocytes for future transplantation therapy. Culture of pluripotent stem cell-derived hepatocytes and organoids has a self-organizing process similar to liver development, i.e., starting with changes in several key factors, and eventually forming functionally complex cells/organs. This paper introduces the main methods and progress of pluripotent stem cell-derived hepatocytes and organoids, with hope to provide clues for future research.
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Humans , Cell Differentiation , Hepatocytes , Induced Pluripotent Stem Cells , Liver , Organoids , Pluripotent Stem CellsABSTRACT
OBJECTIVE@#To analyze the acupoint selection rules of post-stroke cognitive impairment (PSCI) treated with acupuncture by ancient and modern medical record cloud platform (V1.5).@*METHODS@#The published randomized controlled trial (RCT) literature of PSCI treated with acupuncture was retrieved from CNKI, Wanfang, VIP, SinoMed, PubMed, EMbase, Cochrane Library and Web of Science, and the data was extracted to establish prescription database. The ancient and modern medical record cloud platform (V1.5) was used to analyze the data.@*RESULTS@#A total of 185 articles was included, involving 91 acupoints, the total using frequency was 1174 times. The top 5 acupoints in frequency of PSCI treated with acupuncture were Baihui (GV 20, 151 times), Shenting (GV 24, 97 times), Sishencong (EX-HN 1, 83 times), Neiguan (PC 6, 69 times) and Sanyinjiao (SP 6, 64 times); the most involved meridian was the governor vessel, and the generally used acupoints were mainly distributed in the head, face and neck. The top 5 acupoint combinations in frequency were Baihui (GV 20)-Shenting (GV 24, 89 times), Baihui (GV 20)-Sishencong (EX-HN 1, 79 times), Baihui (GV 20)-Neiguan (PC 6, 59 times), Baihui (GV 20)-Sanyinjiao (SP 6, 56 times) and Baihui (GV 20)-Zusanli (ST 36, 51 times). The acupoint combination with the strongest association was Shenting (GV 24)→Baihui (GV 20). There were 6 acupoint cluster groups according to the cluster analysis, and the main core prescription was Baihui (GV 20), Shenting (GV 24), Sishencong (EX-HN 1), Zusanli (ST 36), Neiguan (PC 6) and Sanyinjiao (SP 6).@*CONCLUSION@#Acupoints on the governor vessel, and distributed in head, face and neck are the main acupoints for PSCI treated with acupuncture, Baihui (GV 20), Shenting (GV 24), Sishencong (EX-HN 1), Zusanli (ST 36), Neiguan (PC 6) and Sanyinjiao (SP 6) can be used as the main acupoints, but it is still necessary to combine with syndrome differentiation.
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Humans , Acupuncture Points , Acupuncture Therapy , Cloud Computing , Cognitive Dysfunction , Medical RecordsABSTRACT
Objective To evaluate the efficiency of a recombinase-aided amplification (RAA) assay for the detection of Schistosoma japonicum infections in Oncomelania hupensis snails. Methods A group test was employed. Fifty Oncomelania snails were collected as a detection sample. The detection samples without infected snails were designated as negative specimens, while the detection samples that contained different numbers of infected snails were designated as positive specimens. A total of 10 negative specimens, 10 positive specimens containing 1 infected snail, 20 positive specimens containing 2 infected snails and 10 positive specimens containing 3 infected snails were assigned. Following random grouping, 40 specimens were subject to the florescent RAA assay using a blind method. The miradium shedding method served as a gold standard, and the sensitivity, specificity, Youden’s index and coincidence rate of the florescent RAA assay were estimated. In addition, 20 samples consisted of 5 negative specimens and 15 positive specimens with 1, 2 and 3 infected snails respectively were grouped randomly. The same specimens were detected using the crushing method and fluorescent RAA assay with the blind method in a paired-design manner. Then, the test results were compared and analyzed. Results Florescent RAA assay detected 29 positives in the 30 specimens containing different numbers of infected snails, with a sensitivity of 96.67%, and 8 negatives in the 10 detection specimens without infected snails, with a specificity of 80.00%, showing a Youden’s index of 0.77. The coincidence rate was 100% among 10 repeated assays for a detection specimen. In addition, there was no significant difference in the detection of infected snails between the florescent RAA assay and the crushing method (χ2 = 0, P > 0.05), and the actual coincidence rates of the florescent RAA assay and crushing method were 95.00% (19/20) and 90.00% (18/20) with the real results, respectively. Conclusion Fluorescent RAA assay has a favorable efficiency for the detection of S. japonicum infections in Oncomelania snails, which shows a potential in screening of S. japonicum-infected Oncomelania snails.
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Objective:To evaluate the effects of slow breathing training on hypertension. Methods:Articles about slow breathing training for hypertension were retrieved from CNKI, Wangfang Data, PubMed and Web of Science, until March, 2021. The authors, publishing time, subjects, interventions and courses, and outcome indexes and conclusion were extracted. Results:There were 924 articles returned, and 35 included, which published mainly from 2009 to 2020. The subjects were patients with hypertension, and the outcome index was blood pressure. Conclusion:Slow breathing training may work for hypertension, which associates to baroreflex sensitivity, heart rate variability, sympathetic nerve activity and cardiopulmonary diastolic receptors.
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@#AIM:To analyze the effects of corneal surface morphology and tear-film with different incisions guided by corneal topography after phacoemulsification.<p>METHODS: Retrospective study. Totally 56 patients(60 eyes)were selected and divided into clear corneal incision group and angle scleral margin incision group who were treated by phacoemulsification in our hospital from June 2019 to December 2020. Best corrected visual acuity(BCVA), ocular surface disease index(OSDI)questionnaire, tear film break-up time(TBUT), corneal surface regularity index(SRI), corneal surface asymmetry index(SAI)and corneal surface astigmatism(CYL)were measured preoperatively and postoperatively, the results were statistically analyzed.<p>RESULTS: BCVA in both groups increased after operation(<i>P</i><0.01). The OSDI score of the two groups increased significantly after surgery(<i>P</i><0.01). TBUT was shortened to varying degrees after surgery(<i>P</i><0.05). SAI and SRI increased to varying degrees in both groups after surgery, and SAI value of angle scleral margin incision group in 1wk and 1mo postoperatively was significantly lower than that in clear corneal incision group(<i>P</i><0.05). The corneal astigmatism at 1wk postoperatively was significant increased in both groups(<i>P</i><0.05). The astigmatism increase of clear corneal incision group at 3mo postoperatively was still higher than that before surgery.<p>CONCLUSION: Angle scleral margin incision has less effect on stability of tear film and corneal surface morphology, which is reliable on phacoemulsification.
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Objective To establish a nucleic acid assay for detection of Paragonimus skrjabini based on the recombinase-aided isothermal amplification (RAA) technique, and to preliminarily evaluate its detection efficiency. Methods The metacercariae of P. skrjabini, P. westermani and Euparagonimus cenocopiosus were isolated from crabs, and genomic DNA was extracted for molecular characterization. The cytochrome coxidase 1 (cox1) gene sequence of P. skrjabini was selected as the target gene fragment, and the primers and probes were designed, screened and synthesized for RAA assay. The genomic DNA of P. skrjabini metacercariae from Jiyuan City and Yiyang County of Luoyang City, Henan Province were used as templates for verification of the fluorescent RAA assay. The fluorescent RAA assay was performed to detect different concentrations of plasmids containing target gene fragment and P. skrjabini metacercariae genomic DNA to determine the sensitivity. Fluorescent RAA assay was performed with recombinant plasmids containing P. skrjabini cox1 gene sequences at different concentrations and P. skrjabini genomic DNA as templates to evaluate its sensitivity, and the genomic DNA of P. westermani, E. cenocopiosus, Clonorchis sinensis and Schistosoma japonicum was detected with fluorescent RAA assay to evaluate its specificity. Results P. skrjabini, P. westermani and E. cenocopiosus metacercariae were isolated from crabs, respectively. Molecular characterization and phylogenetic analysis confirmed their homology with the genes sequences of standard Paragonimus strains in GenBank. A fluorescent RAA assay was successfully established for nucleic acid detection of P. skrjabini, and the genomic DNA of P. skrjabini metacercariae from Jiyuan City and Yiyang County of Luoyang City, Henan Province was amplified using the fluorescent RAA assay within 5 min, while the negative control was not amplified. If the recombinant plasmid containing P. skrjabini cox1 gene sequences was used as templates, the fluorescent RAA assay showed the lowest detection limit of 10 copies/μL, and positive amplification was observed within 5 min. If genomic DNA was used as templates, the fluorescent RAA assay showed the lowest detection limit of 10 pg/μL, and all positive amplifications were found within 5 to 10 min. In addition, the fluorescent RAA assay was tested negative for P. westermani, E. cenocopiosus, C. sinensis and S. japonicum. Conclusions A rapid, sensitive and specific fluorescent RAA assay is successfully established for nucleic acid detection of P. skrjabini, which has potential values in rapid field detection and species identification in freshwater crabs in areas endemic for P. skrjabini.
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Objective To develop a fluorescent recombinase-aided isothermal amplification (RAA)-based nucleic acid assay for detection of Leshimania. Methods Specific primers and probes were designed targeting Leishmania internal transcribed spacer 1 (ITS1) gene for RAA assay, and a fluorescent RAA assay was developed for detection of Leishmania following screening of primer pairs and optimization of primer and probe concentrations. The sensitivity of RAA assay for detection of Leishmania was evaluated using recombinant plasmid containing Leishmania ITS1 gene sequences at different copies and Leshimania genomic DNA at different concentrations as templates, and the specificity of RAA assay for detection of Leishmania was evaluated using the genomic DNA of transfusion-transmitted parasites, including Babesia microti, Toxoplasma gondii, Plamodium vivax, P. ovale, P. falciparum, P. malariae, L. donovani and L. infantum. Results After the optimal primer pair was screened from 9 pairs of primer combinations, the final primer and probe concentrations were optimized as 0.3 μmol/L and 0.08 μmol/L, respectively. Nucleic acid detection of Leishmania was completed by the fluorescent RAA assay at an isothermal temperature of 39 °C within 20 min. Remarkable florescent signals were seen within 5 min following RAA detection of genomic DNA of L. donovani and L. infantum, and no cross-reactions were observed with B. microti, T. gondii, P. vivax, P. ovale, P. falciparum or P. malariae. The lowest limitation of detection of the fluorescent RAA assay was 10 copies/μL recombinant plasmid containing Leishmania ITS1 gene sequences and 1 fg/μL Leishmania genomic DNA. Conclusions A rapid, simple, sensitive and specific fluorescent RAA assay is successfully developed for detection of L. donovani and L. infantum, which is effective for field screening of leishmaniasis.
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Objective To develop a rapid test for detection of Schistosoma japonicum specific gene fragments based on the recombinase-aided isothermal amplification assay (RAA) and nucleic acid dipstick test. Methods The S. japonicum SjG28 gene fragment was selected as the target gene fragment, and the primers and fluorescent probe were designed and synthesized. Then, a S. japonicum nucleic acid dipstick test was established. The sensitivity of this dipstick test was evaluated by detecting different copies of recombinant plasmids containing the S. japonicum SjG28 gene fragment and different concentrations of genomic DNA from adult worms of S. japonicum, and the specificity of the dipstick test was evaluated by detecting the genomic DNA from Clonorchis sinensis, S. mansoni, Ancylostoma duodenale, S. haematobium, Babesia and Paragonimus westermani. Results The S. japonicum nucleic acid dipstick test based on the S. japonicum SjG28 gene fragment showed the minimum detectable limit of 10 copies/μL of the recombinant plasmid containing the S. japonicum SjG28 gene fragment and the minimum detectable limit of 1 pg/μL of S. japonicum genomic DNA, and the dipstick assay tested negative for the genomic DNA from C. sinensis, S. mansoni, A. duodenale, S. haematobium, Babesia and P. westermani. Conclusion A rapid, simple, and visualized assay is established for detection of S. japonicum specific gene fragments based on RAA and nucleic acid dipstick test.
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Objective:To investigate the epidemiological characteristics of heat stroke cases in Minhang District of Shanghai in 2013-2018 and to explore potential risk factors， so as to provide the evidence for making the preventive and control measures. Methods:Meteorological parameters and heat stroke cases during May-September were included for statistical analysis. Results:A total of 101 heat stroke cases were studied， in which the majority occurred during June-August. Male cases were more than female cases and 51.5% （52/101） of the cases were severe cases. The incidence of severe heat stroke in people aged 40-59， and 60 and over was significantly higher than that in people under 40 years old. The number of people with severe heat stroke was positively correlated with daily maximum temperature and daily minimum temperature， whereas the number of mild heat stroke was only positively correlated with daily maximum temperature. More than half of total cases （86 cases） were documented in 15 heat wave periods， two of which had significantly increased risk. Conclusion:High temperature in summer is dangerous in Minhang District of Shanghai. Men and older people are susceptible to heat stroke， which occurs mainly in the period of high temperature in July and August. The period of daily highest temperature ≥35 ℃， especially during continual high temperature， is the key stage for heat stroke prevention. In addition to focusing on the effect of daily maximum temperature on heat stroke， the effect of daily minimum temperature on severe heat stroke should be considered.
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Objective:To investigate the hygienic condition and maintenance management of air conditioners in observation hotels， and give suggestions on reducing the risk of COVID-19 transmission from the daily use. Methods:This study selected 11 observation hotels chosen by government and 3 observation hotels chosen by large companies in Minhang District. The types and sanitary conditions of the air conditioning system were revealed through the daily supervision. Hotel staffs’ knowledge of air conditioning system and their mastery of how to use and maintain air conditioning system were surveyed through questionnaire. Results:Survey of air conditioning types showed that in 14 hotels， 12 were distributed air conditioning systems and 2 were semi-centralized air conditioning systems （including fresh air systems）. The investigation found that there was dust accumulation in the fresh air ducts in one hotel guest room， dust accumulation in the filter screen of fresh air intake in one hotel， and the sanitary problem of condensate water （without centralized discharge） in two hotels. All of 14 hotels had daily cleaning and disinfection records， but they were not perfect. The hotel health management personnel’ awareness rate of air conditioning was low， although they had a positive attitude towards the cleaning and disinfection of the air conditioning system. They could do the active entrusted testing， cleaning and disinfection of the air conditioning systems. Conclusion:The air conditioning systems of some hotels have hygiene problems， and hotel health management personnel are lack of knowledge of standard operation and maintenance of air conditioning systems. The air conditioning systems of observation hotel should be cleaned and disinfected before use. At the same time， it is necessary to strengthen the training of hotel health management personnel on the use and maintenance of air conditioning systems. So the transmission of the COVID-19 through air conditioning systems can be effectively prevented.
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Particulate matters are primary pollutants and ubiquitous in most urban areas of China and prove to be an acknowledged risk to human health, whose degree of harm is realated to the particle size.Ultrafine particles (UFPs) with its diameter less than 0.1 μm may have greater adverse health effects compared with larger ones as fine particulate matters(PM10)and (PM2.5).However, there have been fewer studies and insuffient evidence in this regard.In this review, we analyze recent population-based studies on UFPs exposure and its health impacts and elucidate associations of UFPs exposure with subclinical endpoints changes so as to promote the awreness of UFPs and provide reference for its further research.
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@#AIM: To investigate the corneal curvature and refractive changes after microscopically extraocular muscle surgery in adults.<p>METHODS: To observe 62 cases of 85 eyes underwent microscopic extraocular muscle surgery in the Second People's Hospital of Yunnan Province from December 2016 to May 2017, age 18 to 43, according to the surgical method, patients were divided into horizontal rectus surgery: Ⅰ group(simple migration group)32 eyes, Ⅱ group(rectus monocular migration combined with its antagonist muscle excision group)34 eyes, the oblique muscle group: Ⅲ group(single oblique muscle tendon broken group)10 eyes, Ⅳ group(oblique muscle transposition group)9 eyes; Subjective optometry was performed preoperatively and postoperatively at 1d, 1wk, 1mo, 3mo and 6mo, respectively, to obtain the refractive state changes and the curvature value of the anterior surface of the cornea(0-3mm, 5mm, 7mm)collected by corneal topography. The dynamic changes of the refractive state and corneal morphology of the patients in each group at different time points were compared. Anova and LSD-<i>t</i> test were used for comparison among time points and within each group.<p>RESULTS: Changes of corneal curvature in horizontal rectus muscle can be induced in a short period after the operation, which is limited to the central 5mm area of the cornea, and basically restored to the preoperative level 1mo after the operation. There was no significant effect on corneal curvature after inferior oblique operation. Astigmatism value increased in the early postoperative period of horizontal rectus muscle; Myopic drift occurred and recovered to preoperative level 1mo after surgery. After the operation of the inferior oblique muscle, there was no significant change in the spheroscopy and astigmatism.<p>CONCLUSION:Corneal curvature and refractive changes may occur after extraocular muscle in adults.
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OBJECTIVE@#To study the effect and mechanism of action of irisin on hypoxic-ischemic brain damage in neonatal rats.@*METHODS@#A total of 248 7-day-old Sprague-Dawley rats were randomly divided into a sham-operation group, a model group, and low- and high-dose irisin intervention groups (n=62 each). The rats in the model and irisin intervention groups were given hypoxic treatment after right common carotid artery ligation to establish a model of hypoxic-ischemic brain damage. Those in the sham-operation group were given the separation of the right common carotid artery without ligation or hypoxic treatment. The rats in the high- and low-dose irisin intervention groups were given intracerebroventricular injection of recombinant irisin polypeptide at a dose of 0.30 µg and 0.15 µg respectively. Those in the model and sham-operation groups were given the injection of an equal volume of PBS. The water maze test was used to compare neurological behaviors between groups. TTC staining, hematoxylin-eosin staining and TUNEL staining were used to observe histopathological changes of the brain. Western blot was used to measure the expression of the apoptosis-related molecules cleaved-caspase-3 (CC3), BCL-2 and BAX.@*RESULTS@#Compared with the sham-operation group, the model group had a significant increase in latency time and a significant reduction in the number of platform crossings (P<0.05). Compared with the model group, the high-dose irisin intervention group had a significant reduction in latency time and a significant increase in the number of platform crossings (P<0.05). Compared with the sham-operation group, the model group had massive infarction in the right hemisphere, with significant increases in karyopyknosis and karyorrhexis. Compared with the model group, the high-dose irisin intervention group had a smaller infarct area of the right hemisphere, with reductions in karyopyknosis and karyorrhexis. The model group had a significantly higher apoptosis rate of cells in the right cerebral cortex and the hippocampus than the sham-operation group. The high-dose irisin intervention group had a significantly lower apoptosis rate than the model group (P<0.05). At 24 and 48 hours after modeling, the sham-operation group had a significantly lower level of CC3 than the model group (P<0.05). Compared with the model group, the high-dose irisin intervention group had a significantly lower level of CC3 and a significantly higher BCL-2/BAX ratio (P<0.05). The low-dose irisin intervention group had similar laboratory markers and histopathological changes of the brain to the model group.@*CONCLUSIONS@#Irisin can alleviate hypoxic-ischemic brain damage in neonatal rats in a dose-dependent manner, possibly by reducing cell apoptosis in the cerebral cortex and the hippocampus.
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Animals , Rats , Animals, Newborn , Apoptosis , Brain , Hypoxia-Ischemia, Brain , Rats, Sprague-DawleyABSTRACT
Objective To establish a recombinase-aided isothermal amplification (RAA) assay for the nucleic acid detection of Angiostrongylus cantonensis. Methods The internal transcribed spacer-1 (ITS1) gene sequence of A. cantonensis was used as the detection target sequence, and the specific primers and probes were designed and synthesized, followed by screening of the primers and probes with the highest specificity, to establish the basic and fluorescent RAA assay for nucleic acid detection of A. cantonensis. The sensitivity of the fluorescent RAA assay was evaluated by using the target gene fragment sequence-contained recombinant plasmids at various copy numbers and the genomic DNA from A. cantonensis as the template DNA samples, and the specificity of the fluorescent RAA assay was evaluated by using the genomic DNA from A. cantonensis, Schistosoma mansoni, Ascaris lumbricoides, Clonorchis sinensis, Echinococcus granulosus and Ancylostoma duodenale, as well as Pomacea canaliculata and Biomphalaria straminea snail tissues as the template DNA samples. Results A fluorescent RAA assay was successfully established for nucleic acid detection of A. cantonensis, which achieved real-time amplification of the specific DNA fragment of A. cantonensis within 20 min at 37 ℃. By using the target gene fragment sequence-contained recombinant plasmids at various copy numbers and the genomic DNA from A. cantonensis as the DNA templates, the lowest detection limits of the fluorescent RAA assay were 10 copies/μL of recombinant plasmids and 100 pg/μL of genomic DNA, respectively. The fluorescent RAA assay was negative for detection of the genomic DNA from A. cantonensis, S. mansoni, A. lumbricoides, C. sinensis, E. granulosus, A. duodenale, and P. canaliculata and B. straminea snail tissues. Conclusions A simple, rapid fluorescent RAA assay has been successfully established, which has a high sensitivity and specificity for the nucleic acid detection of A. cantonensis.
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Objective To establish a novel nucleic acid assay for detection of Giardia lamblia based on the recombinase-aided isothermal amplification (RAA) assay, and evaluate its sensitivity and specificity for detection of G. lamblia. Methods The specific primer sequences and florescent probes were designed and synthesized based on the G. lamblia β-giardin gene as the target gene, and a fluorescent RAA assay was established. The recombinant plasmids at various copies (containing the β-giardin gene target sequence) and the genomic DNA of G. lamblia at various concentrations were used as templates for the fluorescent RAA assay to assess the sensitivity, and the genomic DNA from G. lamblia, Schistosoma japonicum, Clonorchis sinensis, Cryptosporidium parvum, Ascaris lumbricoides, Salmonella and Shigella was used as templates to assess the specificity of the fluorescent RAA assay. Results A novel fluorescent RAA assay was successfully established for detection of G. lamblia, which allowed the rapid and specific amplification of the target gene fragments at 39 ℃ within 20 min. The sensitivities of the fluorescent RAA assay were 102 copies/μL and 1 pg/μL for detection of the recombinant plasmid and G. lamblia genomic DNA, respectively, and the fluorescent RAA assay was negative for detection of the genomic DNA from S. japonicum, C. sinensis, C. parvum, A. lumbricoides, Salmonella and Shigella, which showed a high specificity. Conclusions A fluorescent RAA assay, which is simple, sensitive and specific, is successfully established for nucleic acid detection of G. lamblia.
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Objective To establish a nucleic acid assay for detection of Echinococcus granulosus based on recombinase-aided isothermal amplification (RAA) assay. Methods The 12S rRNA gene of E. granulosus was selected as the target gene, and the specific primers and fluorescent probes for RAA assay were designed, screened and synthesized to establish a fluorescent RAA assay for detection of E. granulosus. The sensitivity of the fluorescent RAA assay was evaluated using different copy numbers of target gene sequence-contained recombinant plasmids and various concentrations of E. granulosus genomic DNA as templates, and the specificity of the fluorescent RAA assay was evaluated using the genomic DNA from E. granulosus, E. multilocularis, Schistosoma japonicum, S. mansoni, Ancylostoma duodenale, Clonorchis sinensis, Taenia saginata, Spirometra mansoni and Taenia solium as templates. Results A fluorescent RAA assay was successfully established for detection of E. granulosus, which achieved specific amplification of E. granulosus genomic DNA within 20 min at 39 ℃. The lowest detection limit of the fluorescent RAA assay was 10 copies/μL of recombinant plasmids and 0.1 ng/μL E. granulosus genomic DNA, which exhibited a high sensitivity, and the fluorescent RAA assay was all negative for the genomic DNA from E. multilocularis, S. japonicum, S. mansoni, A. duodenale, C. sinensis, T. saginata, Spirometra mansoni and T. solium, which exhibited a high specificity. In addition, this fluorescent RAA assay successfully detected genomic DNA from E. granulosus cysts. Conclusions A rapid, sensitive and specific fluorescent RAA assay is successfully established for nucleic acid detection of E. granulosus.
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Objective To establish a recombinase-aided isothermal amplification (RAA) assay for nucleic acid detection of Schistosoma mansoni. Methods The 121 bp highly-repeated sequence of S. mansoni was selected as the target gene fragment to be detected. The primers and fluorescent probes were designed using the Amplfix software, and a fluorescent RAA assay was established and optimized. The fluorescent RAA assay was performed to detect gradient diluent recombinant plasmids containing target gene fragment and different concentrations of S. mansoni genomic DNA to determine the sensitivity, and this assay was applied to detect the genomic DNA of S. japonicum, S. haematobium, Ancylostoma duodenale and Clonorchis sinensis to evaluate the specificity. Results A fluorescent RAA assay was successfully established, which was effective to amplify the specific gene fragments of S. mansoni within 20 min at 39 ℃. The minimum detectable limit of the fluorescent RAA assay was 10 copies/μL using recombinant plasmids as templates and 0.1 fg/μL using S. mansoni genomic DNA samples as templates. The fluorescent RAA assays were all negative for detecting the genomic DNA from S. japonicum, S. haematobium, A. duodenale and C. sinensis. Conclusions A novel fluorescent RAA assay is successfully established, which is simple, rapid, sensitive and specific to detect genomic DNA of S. mansoni.
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OBJECTIVE: To explore the value of water swallow test(WST)and simple two-step swallowing provocation test(SSPT)in the diagnosis of aspiration in patients with acute exacerbation of chronic obstructive pulmonary disease. METHODS:87 hospitalized patients with acute exacerbation of chronic obstructive pulmonary disease were recruited from the First Affiliated Hospital of Guangzhou Medical University during the period between December 2014 to December 2015. RESULTS: The number of patients of grade1,2,3,4 and 5 of water swallow test successively were 44,39,4,0 and 0. Patients with positive aspiration by the first-step(water injection of 0.4 mL)and the second-step(water injection of 2.0 m L)were 16 and 0. Patients with positive aspiration by radionuclide imaging was 35. Comparison of radionuclide imaging, the rate of missed diagnosis applying water swallow test was high 37.3%(31/83). Both the water swallow test and simple two-step swallowing provocation test have poor consistency with radionuclide imaging in the diagnosis of aspiration in patients with acute exacerbation of chronic obstructive pulmonary disease(McNemar consistency test P=0.00).CONCLUSION: There is a high rate of missed diagnosis applying water swallow test and simple two-step swallowing provocation test to diagnosis aspiration in patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD),and combined use of multiple assessment methods can reduce the missed diagnosis rate of aspiration.