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Objective:To investigate the biological characteristics of Kerstersia gyiorum and to support the rapid and accurate identification of Kerstersia gyiorum on mass spectrometry by using Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) for self-built libraries. Methods:From November 2020 to February 2022, thirty-eight strains of Kerstersia gyiorum isolated from clinical patients of the General Hospital of Southern Theatre Command were collected and identified by the fully automated microbial analysis system (Vitek-2 Compact), the automatic microbial mass spectrometry detection system (Vitek-MS) and the 16S ribosomal RNA sequencing. Thirteen strains were randomly selected and mass spectra were obtained by using Vitek-MS. The SARAMIS software was used to construct a Kerstersia gyiorum library, and the remaining 25 strains were used to validate the constructed library. Results:The Vitek-2 Compact and Vitek MS were unable to identify Kerstersia gyiorum; 13 strains were successfully built into a self-built library of Kerstersia gyiorum by SARAMIS software, and 25 validated strains were identified as Kerstersia gyiorum with a confidence level of more than 99.0% and 100% (25/25) accuracy. Conclusion:Kerstersia gyiorum has unique mass spectrometry profile, which can be identified as species quickly and accurately by the establishment of the self-constructed library of profiles.
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Objective:To investigate the application of a video laryngoscope combined with a fiberoptic bronchoscope in emergency endotracheal intubation and its effects on pulmonary infection in patients with craniocerebral trauma.Methods:A total of 105 patients with craniocerebral trauma who received treatment in Yiwu Central Hospital from January 2020 to December 2020 were included in this study. They were randomly allocated to undergo endotracheal intubation with a video laryngoscope (control group, n = 50) or a video laryngoscope combined with a fiberoptic bronchoscope (observation group, n = 55). Glottic exposure, intubation, vital signs, lung infection rate during hospitalization, and incidence of complications were monitored/determined in each group. Results:Glottic exposure in the observation group was superior to that in the control group ( Z = 4.29, P < 0.001). First-pass success rate was significantly higher in the observation group than in the control group [96.36% (53/55) vs. 82.00% (41/50), χ2 = 5.76, P < 0.05]. The number of intubation attempts and the time to successful intubation were (1.07 ± 0.53) times and (85.12 ± 15.36) seconds, respectively in the observation group, which were significantly less or shorter than those in the control group [(1.92 ± 0.74) times, (106.13 ± 16.34) seconds, t = 6.81, 6.79, both P < 0.001). The changes in mean arterial pressure and heart rate during intubation were less in the observation group than in the control group (both P < 0.05). The amplitude of increase in blood oxygen saturation after intubation was greater in the observation group than in the control group ( P < 0.05). Lung infection rate was significantly lower in the observation group than in the control group [10.91% (6/55) vs. 30.00% (15/50), χ2 = 5.96, P < 0.05]. The incidence of complications was significantly lower in the observation group than in the control group [5.45% (3/55) vs. 18.00% (9/50), χ2 = 4.07, P < 0.05]. Conclusion:Application of a video laryngoscope combined with a fiberoptic bronchoscope in emergency endotracheal intubation can increase the first-pass success rate, reduce repeated intubation attempts, shorten time to successful intubation, help to maintain stable vital signs, prevent lung infection and complication. Therefore, the combined method is of clinical application value.
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Objective@#To establish finite element models of skull, fronto-orbital advancement and fronto-orbital distraction osteogenesis of craniosynostosis, to analyze the mechanical characteristics of skull base and fronto-orbital operation area, so as to guide the later application of tractors.@*Methods@#One 6-year-old male patient with unilateral coronal synostosis was enrolled in October 2015. Three-dimensional (3D) computed tomography (CT) scan of skull was performed. DICOM data was imported into Mimics 17.0 for contour extraction and cranial 3D reconstruction. The skull model was processed by Mimics, Geomagic Studio 12.0, Hypermesh 12.0 and other software to establish a three-dimensional finite element model. The unilateral and bilateral fronto-orbital anterior osteotomy models were simulated respectively. The mechanical analysis was performed at point A in forehead area and point B in temporal area. Three different groups of traction forces were loaded: (1) 50 Newton for point A, 50 Newton for point B; (2) 80 Newton for point A and 50 Newton for point B; (3) 100 Newton for point A and 50 Newton for point B, to obtain the optimized traction force.@*Results@#Stress analysis was performed on established cranial finite element model, as well as unilateral and bilateral fronto-orbital advancement procedures. The stress distribution of the anterior and middle cranial fossae was found to be concentrated. After unilateral fronto-orbital advancement, the stress of anterior cranial fossa, especially the affected side, was decreased. The stress on both side in anterior cranial fossa was decreased after bilateral fronto-orbital advancement. After force was applied to point A and point B, the optimum deviation result at supraorbital notch point, midpoint of supraorbital margin, frontal temporal point and frontal zygomatic suture point in 3D (Deviation result of X value: -29.4%, -20.5%, -8.6%, -9.3%; Deviation result of Y value: 20.9%, 31.5%, 73.0%, 539.4%; Deviation result of Z value: 4.4%, 1.9%, 0.1%, 11.8%) demonstrated the application of traction force can inwardly, downwardly and forwardly move the bone flap. The optimized traction was 80 Newton at point A and 50 Newton at point B by preliminary assessment.@*Conclusions@#The finite element analysis of the fronto-orbital advancement can be used for more accurate preoperative simulation, to clarify the influence of fronto-orbital advancement on craniofacial morphology and development, as well as skull base. It also facilitates surgical decision and predicts the postoperative distraction vectors.
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Objective@#The purpose of this study was to explore the surgery-first approach in sequential combined orthodontic-orthognathic treatment to shorten total treatment duration and improve the clinical outcome.@*Methods@#This study included 185 patients with Angle classⅢ malocclusion. The patients were divided into 3 different types according to cephalometry analyses and facial features. ①Type Ⅰ: mandibular prognathism or asymmetry mandibular prognathism; ②Type Ⅱ: mandibular prognathism and maxillary retrusion; ③ Type Ⅲ: mild Angle′s Class Ⅲ malocclusion, cross bite in anterior teeth, or normal overlap and overbite relation with midfacial hypoplasia. All of patients received surgery first approach therapy. The surgical procedures were chosen according to different malformation types. Type Ⅰ was treated with the sagittal split ramus osteotomy (SSRO). Type Ⅱ was treated by Le Fort Ⅰmaxillary osteotomy combined with SSRO. Type Ⅲ underwent anterior subapical osteotomy combined pyriform aperture augmentation with biomaterials as well as maxillary anterior orthodontics. All patients received postoperative rapid orthodontic treatment for 6-12 month after 2 weeks of operation. Using the straight arch wire techniques and the class Ⅲ intermaxillary traction, we removed the overcrowding upper and lower teeth, the compensatory axial tilt of teeth, and the deviation of the dental arch and maintained the neutral relationship of the molar. The mandible Hawley retaining devices were used during the maintaining stage.@*Results@#The cases in study acquired satisfactory clinical outcome, which included the shortened overall treatment duration, the significantly improved facial features, the corrected occlusion relationship, and the restored function of mastication and temporomandibular joint. There were some complications as follows: intraoperative fracture (6 cases, 3.24%), the inferior alveolar nerve bundle injury (2 cases, 1.1%), and temporary open-bite that diminished by inter-maxillary elastic distraction one month after operation (19 cases, 10%). All cases in this study accepted postoperative orthodontic treatment. Follow-up time ranged from 6 months to 5 years. The cephalometric analysis results of 126 cases who had complete image data and over 6 months of follow-up showed that hard and soft tissue indexes were restored to normal range after combined orthognathic-orthodontic treatment. The stability of the maxillary and occlusive relationship of SFA(surgery-first approach) was similar to that of the COS(conventional orthodontics-first system) [relapse ratio=(T2-T1)/(T1-T0)×100%]. Over six months of follow-up , SNB and ANB showed that the average relapse ratio were 22% and 19.8%, whereas the relapse angle are less than 2°.@*Conclusions@#The Surgery-first approach could be used to treat most patients with Angle skeletal Class Ⅲ malocclusion, but the indications and the surgical procedures should be noticed and chosen.
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<p><b>OBJECTIVE</b>To investigate the differences in the expression profiles of circular RNA (circRNA) between luminal breast cancer cells and normal breast cells.</p><p><b>METHODS</b>Total RNA extracted from luminal breast cancer cells MCF7 and normal breast cells MCF10A was digested with Rnase R to remove linear RNAs and enrich circRNAs. The enriched circRNAs were amplified and transcribed into fluorescent cRNAs using a random priming method, and were hybridized onto the circRNA hybridization array. The circRNA expression profiles of MCF7 and MCF10A cells were analyzed using Agilent Feature Extraction software. Quantile normalization and subsequent data processing were performed, and volcano plot filtering and hierarchical clustering were utilized to analyze the circRNA expression patterns. The expressions of 3 circRNAs with significant log fold changes were validated using qPCR.</p><p><b>RESULTS</b>The hybridization array data revealed significant differences in the circRNA expression profiles between MCF7 and MCF10A cells. Compared with those of MCF10A cells, the 12910 circRNAs expressed in MCF7 cells showed 5964 up-regulated, 81 consistently regulated, and 6865 down-regulated circRNAs; 343 circRNAs showed a log fold change by more than 2 folds, among which 213 circRNAs were up-regulated and 130 were down-regulated. Nine circRNAs showed differential expressions by more than 2 folds, including 8 up-regulated ones, namely hsa_circRNA_061260 (6.02 folds), hsa_circRNA_103933 (5.96 folds), hsa_circRNA_005239 (5.84 folds), hsa_circRNA_100689 (5.69 folds), hsa_circRNA_004087 (5.60 folds), hsa_circRNA_104420 (5.25 folds), hsa_circRNA_104421 (5.13 folds) and hsa_circRNA_101222 (5.03 folds); only one circRNA was down-regulated, namely hsa_circRNA_104864 (5.09 folds). The expressions of hsa_circRNA_100689, hsa_circRNA_005239 and hsa_circRNA_104864 were further validated by qPCR, which yielded consistent results with the microarray data.</p><p><b>CONCLUSIONS</b>The circRNA expression profiles differ significantly between luminal breast cancer cells and normal breast cells. These differentially expressed circRNAs may serve as potential novel targets for the diagnosis of luminal breast cancer.</p>
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Objective To investigate the deformity of cranial base in patients with craniosynostosis and to evaluate the effect of modified fronto-orbital advancement (FOA) for craniosynostosis using three-dimensional digital technology.Methods From January 2010 to December 2016,10 cases of craniosynostosis were included in the study.All patients underwent modified FOA,we quantitatively analysed the pre-operative deformity and the surgical outcome of FOA performed in ten patients with craniosynostosis.The measurements of skull base were performed based on 3D computed tomography (CT) scan.We assessed the symmetry of skull base and the deflection of anterior cranial fossa.Results The mean patients' age at FOAs was 3.2 years.A significant asymmetry of skull base was found in all patients.The growth of anterior and middle fossae was restricted.On the follow-up CT images 23.6 months after surgery,there was prominent change (P<0.05) between the two hemibases (CSO(-)ratio) and between the lengths and angles of anterior and middle cranial hemi-fossae (CX,CSX(-),XSM(-) ratio).Anterior cranial vault asymmetry index obviously decreased from 12.4% to 1.05% before surgery (P<0.05).In addition,anterior cranial vault deflection angle proved to also be less after surgical correction (20.33°±9.19° versus 13.13°±5.28°,P<0.05).Conclusions The asymmetry of skull base can be presented by intracranial view at over 1 year of age.FOA can correct the skeletal malformation of frontotemporal region and the asymmetry of skull base is improved after surgical treatment.
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Objective CRISPR/Cas9 genome-editing technique provides an novel method for whole genome editing in eukaryotic cells.Recently,we found that gene subtype library with smaller size and focused pur-pose is more economical and practical. In this study,we aimed to target kinases,a group of pivotal cell signal transducers,to construct a kinase knock-out library using CRISPR/Cas9 technique.The construction strategy wll al-so be discussed. Methods 10 sgRNA was designed for each kinase target.After oligo pool synthesis by semicon-ductor chip,the oligos were eluted from the chip. The oligo templates were amplified and cloned into Cas9 vector and transformed into Stble3 competent cells.Monoclonal colonies were selected for DNA sequencing. Results(1) GO analysis of 507 cell kinases showed that the cell kinases took part in a wide range of cell signaling.(2)The sgRNA pool with about 140 bp in length was successfully amplified by using oligo pool as the template and univer-sal PCR primers.(3)In 40 identified library clones,34 clones were sequenced successfully. Among them,the DNA sequencing results of 25 samples were completely consistent with the designed target sequences.But there are some mutations in the primers of 9 samples.Failure in bacteria shaking,DNA sequencing and other factors were ex-isted in the other clones. Conclusion The CRISPR/Cas9 kinase knock-out library can be widely used for screen-ing the important kinases which may mediate cell proliferation,metastasis,drug resistance and autophagy.This li-brary will play an important role in clarifying the development of disease associated with kinases.
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Objective To investigate the effect of biofilm cleaning agents and multienzyme cleaning agents in the cleaning of tubular instruments. Methods About 200 pieces of intramedullary nail used in tibial fracture and intramedullary nailing were selected and divided into multienzyme cleaning group and biomembrane cleaning group according to the order of recovery. According to the random number table method, each group had 100 pieces. The biofilm cleaning group was cleaned with biological membrane cleaning agent, and the multienzyme cleaning group was cleaned with multienzyme cleaning agent. The cleaning effect of luminal instruments was observed by means of microscopy, dipstick test and ATP bioluminescence assay. Result The cleaning quality and biofilm removal effect of the biofilm cleaning group were better than those of the multienzyme cleaning group (P<0.05). Conclusion Cleaning the luminal instruments with biomembrane cleaning agent can improve the cleaning quality and prevent the formation of bacterial biofilm.
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Objective To observe the effect of magnetic attachments application on oral denture restoration .Methods A to-tal of 50 patients with overdenture restoration treatment in this department from April 2012 to April 2014 were enrolled .Abutment tooth were given magnetic attachments restoration after preparations .The masticatory efficiency ,patient satisfaction and post-resto-ration complication were observed and recorded .Results The immediate masticatory efficiency after magnetic attachments restora-tion (0 .65 ± 0 .09) was significant higher than that before restoration(0 .22 ± 0 .04) ,t= 20 .812 ,P 0 .05) .There were 49 cases of immediate satisfaction and 1 case of satisfactory after restoration ,the patient satisfaction rate was 100% ,it was significant higher than that of before restoration(46% ) ,χ2 = 31 .815 ,P< 0 .05 .There were 45 ca-ses of satisfaction and 5 case of satisfactory after restoration ,the patient satisfaction rate was 100% ,it was significant higher than that of before restoration (χ2 = 31 .815 ,P< 0 .05) .Conclusion The masticatory efficiency could be improved significantly by mag-netic attachments to restore denture depletion or dentition defect ,compare with traditional denture ,it can significant shorten adapt time for patients ,and it can enhance patient satisfaction .It has wide prospect for future clinical application .
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Objective To study the influence factors of infertility by analysis of semen sample and reference for clinical treat-ment.Methods 3 527 cases of semen sample were collected from Jan 2012 to Jun 2014.All samples were analysed by SQA-V analyzer and compared with 80 cases of normal semen.Results There were 358 normal samples (10.2%)and 3 169 ab-normal samples (89.8%).Among the abnormal samples low sperm motility had the highest ratio (2.7%)while abnormal pH had the lowest ratio (61.5%).All the indexes had significance difference to normal sample expect pH value (t=0.065, P =0.969).Among them,the comparison of rate had statistical significance(χ2 =3.214~24.712,P <0.05).The compari-son of mean also had statistical significance(t=2.523~15.324,P <0.05).Conclusion Infertility male almost has abnormal index of semen volume,liquefaction time,sperm motility,sperm density,sperm morphology,sperm viability.Accurately sperm analysis can provide objective basis to clinical diagnosis and treatment.
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<p><b>OBJECTIVE</b>To prepare a monoclonal antibody (mAb) against the fusion protein preM/EIII of West Nile virus (WNV) for clinical detection of WNV.</p><p><b>METHODS</b>Sp2/0 cells were fused with the spleen cells of BALB/c mice immunized with the recombinant fusion protein preM/EIII expressed in E. coil to obtain the hybridoma cell line that secreted preM/EIII mAb. The hybridoma cells were injected into the peritoneal cavity of BALB/c mice and the ascites was collected and purified. The specificity and titer of the obtained mAb were determined using ELISA and Western blotting.</p><p><b>RESULTS</b>One hybridoma cell line secreting preM/EIII mAb, named ME1, was obtained. The titer of the purified mAb was 10(-6). Identified as a mAb of the Ig subclass IgG1, ME1 was capable of specific reactions with preM/EIII protein and WNV without cross-reactions with other viruses such as JEV, SLEV, YFV and DENV. The accuracy of clinical testing of MNV with ME1 was 97.78%.</p><p><b>CONCLUSION</b>The mAb against preM/EIII obtained have a high specificity and accuracy in clinical testing of MNV and can be used in clinical diagnosis of MNV infection.</p>
Subject(s)
Animals , Mice , Antibodies, Monoclonal , Allergy and Immunology , Blotting, Western , Cross Reactions , Hybridomas , Allergy and Immunology , Mice, Inbred BALB C , Viral Fusion Proteins , Allergy and Immunology , West Nile virus , Allergy and ImmunologyABSTRACT
Objective To detect of clostridium perfringens by qPCR in mouse models and a clinical case in order to offer early diagnosis.Methods 40 Kunming mice were randomly grouped and intramuscular injected clostridium perfringens type A in leg 0.1 ml(3.5 × 109cfu/ml or 3.5 × 108cfu/ml or 3.5× 107cfu/ml,diluted with saline),while control group was injected with 9% sodium chloride 0.1ml.The mouse models and a clinical case were detected by qPCR.Results The death rate of 3.5 × 109,3.5 × 108,3.5 × 107cfu/ml and the blank group were 90%,70%,10% and 0% after intramuscular injection for 72 h spectively.The mean Ct values among these groups were 21.21 ±2.69,28.45 ±2.74,32.49 ±2.87 and 0.00 ± 0.00(P < 0.05).The Ct values of the patient were 30.67 and 30.44.Conclusions Cclostridium perfringens could be successful identified with qPCR in mouse models when the mice still did not show any symptoms.