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Objective:To construct a recombinant bioluminescent bacteriophage (HT7) targeting Escherichia coli, and evaluate its ability to identify Escherichia coli. Methods:Initially, pCRISPR-sg (1-10) and PFN-1000 plasmid strains were constructed by genetic engineering, and the most efficient small guild RNA (sgRNA) were screened by bilayer plate. By the gene editing technique, which comprised homologous recombination and clustered regularly interspaced short palin dromic repeats (CRISPR)-Cas system, the Nanoluc luciferase gene was integrated into the downstream non-coding region of 10A gene of T7 phage, to constructe the bioluminescent phage HT7 successfully. The difference of biological characteristics between HT7 phage and T7 phage was evaluated by plaque assay and liquid amplification assay. In addition, 51 strains of Escherichia coli, 20 strains of Klebsiella pneumoniae, 14 strains of Staphylococcus aureus, 6 strains of Enterococcus faecium, 5 strains of Enterococcus faecalis, 3 strains of Acinetobacter baumannii and 1 strain of Pseudomonas aeruginosa were collected and isolated to evaluate the limit of detection and specificity of HT7 phage. Results:Among the 10 CRISPR-targeted cleavage systems constructed, sgRNA8 exhibited the highest cleavage efficiency, with a cleavage rate of 0.18. After three rounds of recombination screening using the pCas9/pCRISPR/PFN-1000 triple-plasmid system, PCR validation yielded recombinant phage bands at 2 798 bp, indicating the successful construction of the HT7 phage. The recombinant phage showed significant differences in biological characteristics in terms of lysis efficiency ( P<0.001), one-step growth curve ( P=0.001), and infection multiplicity ( P=0.031). Both lysis burst time and log growth node were extended by 10 min, with the optimal infection multiplicity being 0.1. Clinical sample testing identified lysis of 6 strains of Escherichia coli within 4.5 h, while other strains remained unaffected, with detection of pathogenic bacteria below 10 CFU/ml. Conclusions:The developed pCas9/pCRISPR/PFN-1000 triple-plasmid editing system efficiently edits the bacteriophage genome. The constructed HT7 fluorescent bacteriophage enables the detection of Escherichia coli below 10 CFU/ml within 4.5 hours, demonstrating low detection limits and high detection specificity.
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Objective:To explore the clinical value of synovial fluid calprotectin for the diagnosis of periprosthetic joint infection (PJI).Methods:Based on prospective cohort study design, a total of 82 patients suspected of PJI after hip and knee arthroplasty in the First Medical Center of the PLA General Hospital from July 2021 to June 2022 were selected. Patients were divided into infection group (PJI, n=39) and non-infection group (non-PJI, n=43) according to the diagnostic criteria proposed by the Second International Consensus Conference in 2018. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used for double-blind detection of calprotectin and internal reference standard (IRS) in synovial fluid of patients. The peaks of target protein and IRS were recorded for further analysis. Mann-Whitney U test was used to compare the concentrations of S100A8 and S100A9 between the two groups, and receiver operating characteristic curve (ROC) was used to analyze the diagnostic efficacy of S100A8 and S100A9 for PJI. Results:Calprotectin was detected as monomers S100A8 and S100A9. Synovial fluid S100A8 was significantly higher in the PJI group than that in the non-PJI group [1.57 (0.48, 4.17) vs 0.00 (0.00, 0.05), Z=?7.221, P<0.05]. Synovial fluid S100A9 was also significantly higher in the PJI group than that in the non-PJI group [0.74 (0.29, 1.70) vs 0.06 (0.00, 0.10), Z=?6.255, P<0.05]. When using S100A8 and S100A9 to diagnose PJI, the sensitivity were 97.4% and 87.2%, the specificity were 86.0% and 88.4%, and the area under the ROC were 0.964 (95% CI 0.929-0.998) and 0.902 (95% CI 0.924-0.996), respectively. Conclusion:The detection of synovial fluid S100A8 and S100A9 by MALDI-TOF MS can make a satisfactory diagnosis for PJI.
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Objective:A high-throughput assay for the detection of five common clinical Candidaemia pathogens was established by combining polymerase chain reaction (PCR) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).Method:Establishment of methodology. We selected Candida albicans, Candida parapsilosis, Candida glabrata, Candida krusei, Candida tropicalis to be the target pathogens and the internal transcribed spacer (ITS) region as the target gene. Specific single base extension primers were designed to perform single base extension reaction in the same reaction system. MALDI-TOF MS was used to detect the characteristic peaks of each target pathogen. The sensitivity and specificity of the detection system were verified by using spiked blood samples. Totally 108 blood samples from proven or suspected candidaemia patients were collected from October 2021 to September 2022 in a hospital in Beijing. The results of nucleic acid mass spectrometry were compared with those of clinical blood culture. Results:The established nucleic acid mass spectrometry detection system can simultaneously detect five common clinical Candida species. Each strain can produce specific product peaks and there is no mutual interference between the strains. The detection limit of Candida albicans was 100 CFU/ml. The detection limit of Candida parapsilosis, Candida glabrata, Candida krusei and Candida tropicalis was 10 CFU/ml. For the 108 blood samples, the sensitivity, specificity, positive predictive value and negative predictive value of nucleic acid mass spectrometry were 94.74% (36/38), 97.14% (68/70), 92.31% (36/39) and 98.55% (68/69), respectively. The McNemar χ 2 test showed no significant difference between the two methods ( P>0.05), and the Kappa consistency test showed good consistency between the two methods ( Kappa=0.9, P<0.05). Conclusion:A nucleic acid mass spectrometry detection system suitable for clinical candida detection was successfully constructed, and the method validation results were consistent with the clinical blood culture.
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Objective:To investigate the current situations and development requirements of emergency testing among secondary and tertiary hospitals in China.Methods:The data were collected from secondary and tertiary hospitals via online questionnaire across 31 provinces in China from February 1 to March 1, 2021. The questionnaire involves various aspects of emergency testing, including area of emergency laboratory, staffs and equipment configuration, inspection items, Turn-around time (TAT), reagents and consumables management, pre-analysis quality control, laboratory information system, critical values management and biosafety, etc.Results:A total of 2 187 questionnaires were obtained, and 1 503 valid questionnaires from 755 secondary hospitals and 748 tertiary hospitals were finally analyzed. The research data showed that daily average number of patients visiting emergency department exceeding 300 person-time in 29.41% (220/748) tertiary hospitals, but that number was less than 100 person-time in 76.69% (579/755) secondary hospitals; daily average emergency tests exceeding 5 000 was reported in 24.47% (183/748) tertiary hospitals, and less than 2 000 was reported in 93.51% (706/755) secondary hospitals; the area of emergency laboratory was less than 100 m 2 in 68.79% (238/346) tertiary hospitals with independent emergency testing laboratory; there were no fixed staffs of emergency testing in 56.02% (842/1 503) hospitals; the biochemical/immunoassay analyzer in 8.65% (130/1 503) hospitals did not have STAT position; one hundred and twenty-six hospitals (8.38%) did not have stock in and stock out record for reagents and consumes materials; the conventional statistical analysis of unqualified specimen was not carried out in 24.62% (370/1 503) hospitals; priority on emergent specimen was not set in 58.62% (881/1 503) hospitals; whole process monitoring function was not equipped in 48.64% (731/1 503) hospitals; there was no conventional communication working mechanism with clinicians on critical value in 7.32% (110/1 503) hospitals; overall, 50.23% (755/1 503) participants did not consider that biosafety risks exist in their emergency testing laboratory. Conclusions:This survey objectively presents the current situations and future development requirements of emergency testing among secondary and tertiary hospitals in China. The survey also reflects that some important process and concepts need to be improved, and extensive attention should be paid by laboratory and hospital administrator, in the area such as communication with clinician, site construction and staff configuration, administration on the priority of emergency testing, administration on the reagent and consumable materials, laboratory informatization construction, laboratory biosafety, and so on.
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Objective:Based on the high-throughput detection technique of multiplex PCR combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, constructing the characteristic SNP profiles of different strains, and establishing a rapid, accurate and highly sensitive method for the diagnosis of bloodstream infection pathogens.Methods:Seven kinds of pathogens such as common Escherichia coli were selected as target. The multiple PCR reaction conditions was optimized, and the characteristic peaks of each target bacteria were detected by MALDI-TOF MS to establish the joint detect system. Common primer pairs and central homo-sequence primer pairs were designed to analyse the formation of primer dimer. Using simulated bacterial infection blood samples with detection system to determine specificity and sensitivity. One hundred and fifty blood samples from suspected bacteremia patients were collected from June to September 2020 in a hospital in Beijing, and the identification results were compared to traditional identification method of clinical application that are using χ 2 test. Results:The cycle threshold (Ct) value of the central homo-sequence primers that were designed were more than 38, with a delay of 6-10 cycles. The joint mass spectrometry detection system could detect seven kinds of bacteria divided into two groups at the same time. The target bacteria can be detected specific product of the peak, and the clinical strains other than the target strains only had primer peaks. All maps had non-specific miscellaneous peaks. The sensitivity of Escherichia coli could reach 50 CFU/ml, and the detection limit of other bacteria was 100 CFU/ml. The detection results of 150 patients showed that 46 cases were positive by traditional method. The positive rate was 30.67% (46/150), including two cases of mixed infection. Forty-eight cases were positive by mass spectrometry, and the positive rate was 32.0% (48/150), including three cases of mixed infections. The negative coincidence rate was 100% (101/101). The comparison of the two methods showed that the P=0.625>0.01, the Kappa=0.938, the sensitivity and specificity was 97.82%(45/46) and 97.11%(101/104), respectively. There was no significant difference between the two methods, and the results of nucleic acid mass spectrometry could also be used in clinic. Conclusions:The established detection system can not only quickly and accurately detect seven common pathogens causing bloodstream infection, and effectively shorten the time needed for traditional culture and identification, but also can detect multiple bacterial mixed infections at the same time to make up for the possibility of missed detection. Besides, the method can also be used to identify other bacteria.
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Objective:To establish an inductively coupled plasma mass spectrometry (ICP-MS) based immunoassay method for the quantitative detection of human chorionic gonadotropin (HCG), and evaluate the clinical applicability of this method.Methods:Sm was selected as element tags, and the HCG quantitative detection system was established by double antibody sandwich method. The dosage of biotinylated antibody and reaction time were optimized. According to EP documents of Clinical and Laboratory Standards Institute (CLSI) and related standards, the analytical performance was evaluated after the establishment of the assay, including the limit of blank (LOB), linearity, precision, recovery, cross reactivity and interference test. And 88 clinical samples were measured using the new method compared to the electrochemical immunoassay (ECLIA) method.Results:Total process completed within 30 min after optimization, and the optimal biotinylated antibody dosage was 0.5 μl. The LOB was 0.29 mIU/ml. The linearity was good within the range of 1.16-8 365.62 mIU/ml with the linear correlation coefficient greater than 0.995 ( R2=0.998 0), the recovery was 97.53%-102.01%. Both intra-and inter-assay coefficients of variation of the high-value sample and the low-value sample were less than 10%. And there was no significant cross-reaction with Luteinizing hormone (LH), follicular stimulating hormone (FSH), and thyroid-stimulating hormone (TSH). The interference bias caused by different concentrations of interference substances was less than 10%. When compared with the ECLIA method for clinical sample detection, the proposed method showed a significant correlation( R2=0.960 0). Conclusion:The proposed ICP-MS base immunoassay for HCG detection has good accuracy, high sensitivity and specificity, and the results of analytical performance verification meet the clinical requirements, which provides experimental basis for the clinical application of this method.
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Objective:To study the non-target metabolomics analysis and to analyze the metabolomic changesof cerebrospinal fluid (CSF) in patients with tuberculous meningitis.Methods:Case-control study. From July 2018 to July 2019, 20 cerebrospinal fluid specimens of diagnosed patients with tuberculous meningitis were collectedin the department of neurology from the first medical center of the PLA general hospital and the eighth medical center of the PLA general hospital and 20 CSF without tuberculous meningitis as the control. Among them, there were 12 males and 8 femalesin the tuberculous meningitis group, aged (37.9±16.1) years; there were 13 males and 7 femalesin the control group, aged (34.7±14.8) years. Using ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) technology with three different mode, namely reverse phase chromatography positive ion mode, reverse phase chromatography negative ion mode and hydrophilic chromatography positive ion mode,to detectthe metabolic fingerprints of patients′CSF and analyzed by SIMCA software for orthogonal partial least squares discriminant analysis (OPLS-DA). The variable importance projection value of OPLS-DA model (threshold value>1) plus the P value of t-test (P<0.05) was applied to find the differential metabolites in the cerebrospinal fluid of the two groups of patients.Results:Ten differential metabolites were found in CSF, including L-isoleucine, L-phenylalanine, L-kynurenine, L-methionine, L-tyrosine acid, dimethylglycine, L-alanine, L-threonine, L-histidine and L-lysine, and all of them were up-regulated in the tuberculous meningitis group.Conclusion:Changesof the amino acid metabolism found in the cerebrospinal fluid of tuberculous meningitis patients can provide basis for differential diagnosis and basic molecular research of tuberculous meningitis.
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Objective To understand the prevalence of Salmonella and the characteristics of drug resistance genes in General Hospital of People's Liberation Army, and to provide evidence for the prevention and treatment of Salmonella infection. Methods A retrospective study was conducted to collect 78 clinical isolates of Salmonella from 2015 to 2017. The age of the patients was 49 ± 21 years old. The infected patients were mainly young and middle-aged. The clinical samples mainly came from feces and venous blood, accounting for 44.87％(35/78) and 33.33％(26/78), respectively. After serotype identification, drug sensitivity test and whole genome sequencing, multilocus sequence typing and drug resistance genotyping were performed. Cluster of Cefotaxime or Ciprofloxacin resistant Salmonella was analyzed. Results Salmonella group D (53.85％) and Salmonella group C (21.79％) were dominant Salmonella serotype. ST11 was mainly ST type. Drug sensitivity test showed that the multidrug resistance rate of Salmonella was 64.11％ (50/78). The sensitivity to all antimicrobial agents' rate was 25.64 (20/78). The resistance rate of Salmonella to nalidixic acid was 65.38％(51/78). The most common drug resistance gene of Salmonella was extended-spectrum β-lactam drug resistance gene, accounting for 78.21％ (61/78). Conclusions The ST-type and carrying resistance genes of Salmonella in this hospital were diverse. Most pathogens were multi-drug resistant to antimicrobial agents. Molecular typing and drug resistance gene analysis of Salmonella and construction of resistant strains to determine the inheritance of Salmonella relationships have a certain clinical significance.
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Objective To explore the effects of Fufang-Haqing capsule with formoterol in the treatment of elderly pulmonary emphysema combined with chronic wheezing bronchitis. Methods A total of 156 patients with elderly pulmonary emphysema combined with chronic wheezing bronchitis in our hospital were divided into control group (88 cases) and observation group (88 cases). The control group was treated with formoterol, the observation group was treated with Fufang-Haqing capsule combined with formoterol. Both group treatment last three months. In the 2 groups, the levels of lung function indicators (FEV1, FEV1/FVC, PEF and PEFR), inflammatory markers [Matrix metalloproteinase inhibitors-1 (TIMP-1), interleukin-17 (IL-17), matrix metalloproteinase-9 (MMP-9) and interleukin-8 (IL-8)], immune function indicators [T lymphocyte subgroup (CD3+ and CD4+), immunoglobulin (IgA, IgG and IgM)] were detected before and after the treatment. During the treatment, the adverse actions were observed. Results The effect rate in treatment group was 93.2% (82/88), significantly higher than 80.7% (71/88) in control group (Z=3.781, P<0.05). After the treatment, the levels of FEV1 (2.89 ± 0.37 L vs. 2.22 ± 0.33 L, t=3.781), FEV1/FVC (65.10% ± 6.67% vs. 57.56% ± 5.98%, t=3.894), PEF (6.76 ± 0.69 L/S vs. 5.57 ± 0.59 L/S, t=3.351) and PEFR (3.67 ± 0.39 L/S vs. 2.87 ± 0.32 L/S, t=3.561) in the treatment group were significantly higher than those in the control group (P<0.05). The TIMP-1, IL-17, IL-8 and MMP-9 in the treatment group were significantly lower than those in the control group (t were 3.567, 3.692, 3.491, 3.394, all Ps<0.05), while the levels of CD3+, CD4+, IgA, IgG and IgM were in the treatment group were significantly higher than those in the control group (t were 3.791, 3.593, 3.258, 3.682, 3.526, all Ps<0.05). There was no significantly difference between the 2 groups in the adverse actions. Conclusions The Fufang-Haqing capsule with formoterol in the treatment of elderly pulmonary emphysema combined with chronic wheezing bronchitis showd good effect, could promote the levels of lung function, inflammatory and immune function.
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In the context of the rapid development of big data in the healthcare field, deep learning (DL), as a machine learning algorithm that provides a more flexible solution for image and speech recognition as well as natural language processing, has the ability to extract important information from medical data into valuable knowledge and it has received unprecedented attention in many real-world tasks. This paper briefly introduces common network structure of deep learning and its latest research progress in the field of medical laboratory. In addition, this review also exploreed some of the inherent challenges and prospective research directions about deep learning that affecting in the medical laboratory.
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In the context of the rapid development of big data in the healthcare field, deep learning (DL), as a machine learning algorithm that provides a more flexible solution for image and speech recognition as well as natural language processing, has the ability to extract important information from medical data into valuable knowledge and it has received unprecedented attention in many real-world tasks. This paper briefly introduces common network structure of deep learning and its latest research progress in the field of medical laboratory. In addition, this review also exploreed some of the inherent challenges and prospective research directions about deep learning that affecting in the medical laboratory.
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In recent years, mass spectrometry has been playing an increasingly prominent role in clinical testing due to its high specificity,high sensitivity and the ability to detect multiple indicators at one time. This article mainly focuses on different types of mass spectrometry, their superiority technical characteristic and main clinical application areas. We present an overview of challenges and countermeasures of clinical application of mass spectrometry in China and prospect its future developments .
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Objective Toinvestigatestatistically significant peptide peaks as biomarkersto diagnose osteoarticular tuberculosis, matrix-assisted laser desorption/ ionization time of flight mass spectrometry (MALDI-TOF MS) was applied to identify the characteristic fingerprint among the serum of patients with osteoarticular tuberculosis, rheumatoid arthritis and healthy adults.Methods Clinical Study. Serum samples of untreatedpatients with osteoarticular tuberculosis and rheumatoid arthritis were collected from August 2018 to December 2018, and serum samples of healthy adults from physical examination were collected as control. After analysis with MALDI-TOF MS, the serum peptide fingerprint datawas imported into software, and protein polypeptide peaks with obvious differences were screened to establish diagnostic models.Results Established the diagnostic model of osteoarticular tuberculosis and healthy adults with m/z 2943.9, 5929.6, 7615.4 and 9033.8 as differential protein polypeptides, the diagnostic model of osteoarticular tuberculosis and rheumatoid arthritis with m/z 4195.6, 5847.6, 5929.6 and 7748.6 as differential protein polypeptides. To these two models, the sensitivity were 95.00％ and 97.50％, respectively. The specificity were 85.71％ and 88.46％, respectively. The accuracy rates were 89.58％ and 92.39％, respectively. The AUC value of ROC curves were 0.8859 and 0.8709, respectively. Conclusions By mass spectrometry and software analysis, the serum protein polypeptides with statistical difference were found successfully. The related diagnostic modelsarealso established, which has certain reference value for auxiliary diagnosis of osteoarticular tuberculosis.
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The past 70 years witnesses that China's laboratory medicine has realized the development from "medical laboratory" to "laboratory medicine". The construction of the laboratory has undergone tremendous changes from manual workshop to standardization, automation and intelligence, and the test efficiency and test results accuracy has been greatly improved. The professional team has also grown stronger, and the education level has been continuously improved, marking the entry of China's laboratory medicine into the modern era. The article also reviews the important role of the Chinese Medical Association played in promoting the development of China's laboratory medicine in the 40 years since its establishment, and looks forward to the future development of laboratory medicine.
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Objective To investigate the presence of apolipoprotein A 1 (APOA1) and its function in human spermatozoa. Methods The expression assays for APOA 1 were carried out by immunofluorescence and Western blotting in human sperm cells . Set up a blank control group , rabbit polyclonal IgG group , which contain anti-APOA1 antibody 10 μg/ml, 20 μg/ml and 40 μg/ml, to treat normal semen samples and incubated at 37 ℃for 1 h, 2 h and 3 h.The progressive motility of spermatozoa was determined bya computer-assisted motion analyzer ( CASA ) , apoptosis rate by flow cytometry and ultrastructural changes by electron microscopy .Comparisons of sperm progressive motility and apoptosis rate were performed by independent sample t tests.Results The study showed APOA1 protein mainly located in the sperm head , the molecular size was 31000.A significant decline in sperm progressive motility was observed after 1,2 and 3 hours of incubation with APOA1 antibody.There was a statistically significant difference between the blank control group and the APOA 1 antibody concentration of 20 μg/ml and 40 μg/ml groups [ 1 hour after incubation , blank control group ( 68.65％ ±15.70％) with 20 μg/ml group (48.45％±5.20％), 40 μg/ml group(39.25％±7.89％), t=2.442, 3.345 , both P<0.05;2 hours after incubation, blank control group(55.33％±10.12％) with 20 μg/ml group(28.68％±11.70％), 40μg/ml group(18.13％ ±10.52％), t=3.445, 5.097,both P<0.05; 3 hours after incubation, blank control group(35.73％±14.08％) with 20μg/ml group(15.53％±8.42％), 40μg/ml group(9.98％± 7.08％), t=2.462, 3.268,both P<0.05].After incubation 2 hours, the percentage of apoptotic cells increased from 16.02％ ±4.28％ in the blank control group to 21.72％ ±2.67％ ( 20 μg/ml APOA1 antibody)and 28.01％±3.93％(40 μg/ml APOA1 antibody), respectively (t=3.177, 5.834, both P<0.01).Treatment with 40 μg/ml APOA1 antibody for 4 hours resulted in major morphological changes to sperm cells observed by electron microscope , including membrane distension ,vacuole formation and different periods of apoptosis cells .Conclusion APOA1 plays an important role in maintaining sperm motility and survival rate,however the mechanism needs further study .
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Objective To investigate the expressions of leukaemia inhibitory factor (LIF) and regulated upon activation,normal T cell expressed and secreted factor (RANTES) in mice with bloodstream infection by 4 different single pathogen and provide research basis for the early diagnosis of bacteriogenous bloodstream infection.Methods CD-1 (ICR,Institute of Cancer Research) mouse models of bloodstream infection with the standard strains of Staphylococcus aureus (S.aureus),Enterococcus faecalis (E.faecalis),Escherichia coli (E.coli) and Klebsiella pneumonia(K,pneumoniae) were established.The serum samples were collected at the 0.5,1,3,6,12,24 and 48 hours after infection and the concentrations of LIF and RANTES in mouse serum of experimental groups and control were detected by Luminex liquid chip system.Results The median lethal dose (LD50) of S.aureus,E.faecalis,E.coli and K.pneumoniae were 8.1 × 108/mL,9.6 × 108/mL,8.1 × 108/mL and 1.1 × 109/mL,respectively.The concentration of serum LIF was significantly increased in 1 hour after infection.The peak concentrations of LIF in the four groups were (51.6±5.0),(73.2±20.8),(7.3 ±0.9)and (6.1 ± 1.2) pg/mL respectively,and the differences were statistically significant compared with the control group (P ＜ 0.01).The concentrations of RANTES in E.faecalis group,E.coli group and K.pneumoniae group were increased after infection for 1 hour and increased significantly after infection for 3 hours.The increased concentrations of RANTES in E.coli group and K.pneumoniae group were more than those in S.aureus group and E.faecalis group.The peak concentrations of RANTES in S.aureus group,E.faecalis group,E.coli group and K.pneumoniae group were (1 929.0-± 25.2),(1 218.1 ± 227.4),(55.7 ± 10.0) and (179.2 ± 9.2)pg/mL,and the differences were statistically significant compared with the control group (P ＜ 0.01).Conclusion The concentrations of LIF and RANTES increased obviously in 1 h after the bacteria entered bloodstream.After 2 days of infections,the levels of LIF and RANTES in E.coli group and K.pneumoniae group were significantly higher than those in S.aureus group and the E.faecalis group.Combined detections of LIF and RANTES may be of certain values to differentiate the infections caused by the pathogens between gram positive and gram negative bacteria.
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Anesthesia was done for 36 patients undergoing orthotropic heart transplantation in Bei-jing Anzhen Hospital from April 2015 to November 2016. Anesthesia management for orthotropic heart transplantation and related problems were analyzed and investigated. Anesthesia management protocol for patients with end-stage heart disease was aimed at reducing fluctuation of hemodynamics and avoiding malig-nant arrhythmia. Anesthesia was induced by intravenously injecting diazepam 5-10 mg, etomidate 0. 2-0. 3 mg∕kg or ketamine 1 mg∕kg, sufentanil 1. 0-1. 5 μg∕kg or fentanyl 10-15 μg∕kg and rocuronium 0. 6 mg∕kg. Anesthesia was maintained by continuously infusing dexmedetomidine 0. 3-0. 5μg·kg-1 ·h-1 , ci-satracurium 10 mg∕h and sufentanil 0. 5-1. 0 μg·kg-1 ·h-1 . Pulmonary arterial pressure and donor heart function were monitored using the flow-directed pulmonary artery catheter. Dopamine, epinephrine and iso-prenaline were intravenously infused after cardiopulmonary bypass to maintain circulation stable. Nitroglyc-erin and prostacyclin were intravenously infused to decrease pulmonary arterial pressure. Immunosuppressive therapy was performed with methylprednisone, mycophenolate mofetil and cyclosporine∕FK506. Thirty-two patients were discharged from hospital, and 4 cases died. Among the 4 patients died, 1 patient died of pul-monary hypertension ( pulmonary arterial systolic pressure>67 mmHg) and right heart failure and, 1 patient showed difficulty in weaning from cardiopulmonary bypass and 2 patients died of refractory low cardiac outputand multi-organ failure. Anesthetic management for heart transplantation required an appreciation of the pathophysiological mechanism of heart failure. Invasive monitoring, steady anesthesia induction and mainte-nance, stable hemodynamics in the perioperative period and good donor heart protection were the keys to ensuring anesthesia management for orthotropic heart transplantation.
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<p><b>OBJECTIVE</b>To explore the clinical and genetic characteristics of a case with Pallister-Killian syndrome (PKS).</p><p><b>METHODS</b>Chromosomal karyotype of umbilical cord blood sample derived from a 36-year-old pregnant woman was analyzed by G-banding analysis. After birth, the child was further analyzed with single nucleotide polymorphism microarray (SNP array) and fluorescence in situ hybridization (FISH) using 12pter/12qter probes.</p><p><b>RESULTS</b>G-banding analysis showed that the fetus has a karyotype of 46,XY [77]/47,XY,+mar [23]. After birth, Affymetrix CytoScan 750K array analysis showed a segmental tetrasomy of arr [hg19] 12p13.33p11.1(173 786 - 34 835 641)×4 and a 34.6 Mb repeat at 12p13.33p11.1 with in the neonate. FISH analysis confirmed that 39% of cells harbored the 12p tetrasomy.</p><p><b>CONCLUSION</b>Combined clinical examination, G-banded chromosomal karyotyping, FISH and microarray analysis can delineate the origin and fragments of small supernumerary marker chromosomes and diagnose PKS with precision.</p>
Тема - темы
Adult , Female , Humans , Pregnancy , Chromosome Banding , Chromosome Disorders , Diagnosis , Chromosomes, Human, Pair 12 , In Situ Hybridization, Fluorescence , Methods , Karyotyping , Oligonucleotide Array Sequence Analysis , Polymorphism, Single Nucleotide , Prenatal Diagnosis , MethodsРеферат
The emergence of new oral anticoagulant(NOAC)reduced the risk of venous thromboembolism recurrence and major bleeding, with their specificity on inhibiting single coagulation factor,hence avoided frequent dose adjustment of classic anticoagulant therapy.Recently,A new stage was initialed by several new assays with their approval of SFDA in clinical application,either providing a targeted assessment on targeted anticoagulation, or filling the gaps between measurement and evaluating progressive blood coagulation.Clinicians could take stratified way-"global coagulation-effective dose of anticoagulant-thrombin generation-fibrin production"to monitor and treat in the major axis of coagulation,eliminating the confusion of cross-network-like coagulation mechanism previously.In this paper,focusing on characteristics of new oral anticoagulants,we interpret the advantages of anti-activated factor Ⅱand anti-activated factor Ⅹactivity in evaluating anticoagulant efficacy, and then suggest thrombin generation test and thrombin-antithrombin complex these two important parameters to understand the inhibitory action of NOAC, which reflect the degree of thrombin activation directly or indirectly.Moreover,fibrin monomer play a unique role in assessment of thrombin inhibition,as an early marker of fibrinogen formation.Potential value of factor ⅩⅢin bleeding etiology is also discussed for its essentiality in the process of fibrin formation.Different from traditional coagulation tests determining the comprehensive activity of a variety of coagulation proteins, new assays provide information on activity or inhibition of a single target in anticoagulation therapy to analyze the anticoagulant effect or prognosis of thromboembolism accurately,which leads hemostasis and thrombosis tests into a new era.
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Objective To reveal the mechanisms of immunological infertility,the method of coimmunoprecipitation(CO-IP) and liquid chromatogram mass/mass (LC-MS/MS) was used to screen sperm membrane proteins which interacting with antisperm antibodies (ASA).Methods This study was designed as a case-control.The disease group including 56 serum samples from 521 cryptogenic infertile patients were screened ASA positive by ELISA and conformed with mixed antiglobulin reaction(MAR).The controls were 31 serum samples which ASA is negative and already possessed healthy offspring.All subjects were enrolled from September 2015 to December 2015 in China PLA General Hospital.Spermatozoa samples from 48donors with normal sperm parameters were from January 2016 to April 2016 in China PLA General Hospital.The purified human sperm membrane proteins were then mixed with serum from disease group (positive for ASA) and control group (not containing ASA).The binding proteins of antisperm antibodies were enriched using CO-IP assay.The immunoprecipitates were separated on sodium dodecyl sulfate-polyactylamide gel (SDS-PAGE),then the binding proteins were cut from the gel and analyzed by LC-MS/MS after the enzymolysis.These proteins could bc idcntified as definition,biological function (s) and subccllular localization with Uniprot database.Results The serum samples from infertile persons (39 females and 17males) were screened ASA positive by ELISA and conformed with MAR.The healthy controls (17 females and 14 males) were ASA-negative in ELISA.Forty proteins that interact with ASA were obtained from the study and these could be divided into three groups:11 antigens detected by control serum samples only,14antigens recognised by both infertile patients and control sera,and 15 antigens specific for patients with ASA.These 15 proteins are Sperm Cation channcl protein 1,Sperm Cation channel protein 3,Sperm Cation channel protein 4,Sperm associated antigen 9,Apolipoprotein A-I,Dynein heavy chain 14,Cylicin-2,Izumo sperm-egg fusion protein 4,Thioredoxin domain-containing protein 2,IQ domain-containing protein H,IQ domain-containing protein F1,Spermatogenesis-associated protein 5,Spermatogenesis-associated protein 5-like protein 1,Sperm acrosome membrane-associated protein 1,E3 ubiquitin-protein ligase RNF 114.Conclusion Fifteen proteins discovered with CO-IP technology and LC-MS/MS analysis could be referred as male immunoinfertility-related antigens and they may hold the great importance in revealing the secret of immunological infertility.