Clinical evaluation of laboratory diagnosis of Mycoplasma pneumoniae / 中华实用儿科临床杂志

Objective:To compare the laboratory diagnostic methods of Mycoplasma pneumonia(MP) and evaluate its clinical value.Methods:A prospective study.Throat swabs and double sera of children with MP infection were collected from December 2016 to January 2017 in Shengjing Hospital Affiliated to China Medical University; throat swab samples of healthy children aged 3 to 5 in Chaoyang District, Beijing were collected from March to May 2017.Passive agglutination (PA) was used to detect the double serum.Taking the 4-fold increase or decrease of the specific antibody titer of the double serum as the gold standard, the receiver operating characteristic curve (ROC) was drawn, and the laboratory methods for detecting MP infection were compared and evaluated.Results:(1)A total of 93 children with MP infection were clinically diagnosed, including 42 males (45.2%) and 51 females (54.8%), with an average age of 5.5 years.Sixty cases (64.5%) of MP infection were diagnosed.There were 349 healthy children, 198 males and 151 females, with an average age of 4.3 years.The positive rate of throat swab culture was 0.6% (2 cases), and the positive rate of fluorescent quantitative PCR(qPCR) was 18.9% (66 cases). (2) The culture specificity was the highest (100.0%) and the sensitivity was the lowest (65.0%). PA and enzyme linked immunosorbent assay (ELISA) were used to detect a single serum in the acute phase, the sensitivity was 71.7% and 86.5% respectively.ROC curve suggested that the current clinical diagnostic threshold MP specific antibody IgM ≥ 1∶160 was not the best diagnostic threshold.Molecular biological diagnostic methods were the most sensitive, RNA simultaneous and testing (SAT) was 85.0% and qPCR was 93.0%; while the specificity was low, 75.7% (SAT) and 63.6% (qPCR), respectively.(3) At the same time, MP nucleic acid (SAT, PCR) of throat swabs and a single serum (ELISA, PA) of children in acute phase were detected, the sensitivity was increased to 95.0%-100.0%, and the specificity was 63.6%-75.7%.Conclusions:Molecular biology is highly sensitive in diagnosing MP infection.It has asymptomatic infection or is carried after infection.Whether it needs treatment needs to be combined with clinical practice, when MP detection is positive.The detection of a single serum in the acute phase with a course of about 1 week has high sensitivity and is of reference value for the diagnosis of MP infection, but the diagnosis needs to be combined with clinical practice.The sensitivity and accuracy of detecting MP infection by single serological test combined with SAT in acute phase are higher than that by single application.

Common bacteria in oropharynx of children with Mycoplasma pneumoniae pneumonia and its clinical significance / 中华实用儿科临床杂志

Objective:To investigate the common bacteria in the oropharynx of children with Mycoplasma pneumoniae pneumonia (MPP) and its clinical significance.Methods:A total of 134 children with MPP who were hospitalized in the Department of Pediatric Respiratory, Shengjing Hospital of China Medical University from December 2016 to June 2017 were selected as the research subjects, and 42 healthy children in the same hospital were selected retrospectively as the healthy control group during the same period.Fluorescent quantitative polymerase chain reaction Taqman probe was used to detect common oropharyngeal bacteria[ Streptococcus pneumoniae(SP), Moraxella catarrhalis(CTA), Haemophilus influenza(HI)] for the enrolled children.Firstly, the bacterial detection rate of MPP children and healthy children was compared.Then, according to age(<1 years old, 1-<3 years old, 3-<6 years old and 6-14 years old), bacterial detection[Mycoplasma pneumoniae(MP), MP+ bacteria]and bacterial species(MP+ SP, MP+ CTA, MP+ HI), 134 children with MPP were divided into groups to compare.Moreover, the relevant clinical datas were retrospectively analyzed by rank sum test and chi- square test. Results:Among 134 children with MPP, 79 (58.96%) children were detected bacteria, and 17 (40.48%) children were detected bacteria among 42 healthy children, with statistically significant differences( χ2=4.404, P<0.05). Compared with the MP group, the level of white blood cell (WBC)[8.5(6.7, 12.0)×10 9/L vs.7.8(5.8, 9.3)×10 9/L, Z=-2.232], C reactive protein(CRP)[19.2(7.2, 35.0) mg/L vs.8.4(3.4, 24.6) mg/L, Z=-2.810], lactate dehydrogenase(LDH)[286(244, 365) U/L vs.250(210, 302) U/L, Z=-2.474] and the incidence of lobar pneumonia[40.51%(32/79 cases) vs.18.18%(10/55 cases), χ2=7.510], pleural effusion[13.92%(11/79 cases) vs.3.64%(2/55 cases), χ2=3.917], refractory Mycoplasma pneumoniae pneumonia (RMPP)[34.18%(27/79 cases) vs.18.18%(10/55 cases), χ2=4.151] in MP+ bacteria group were higher; the course of fever[10(7, 12) d vs.8(6, 10) d, Z=-2.706] and duration of antibiotic use[16(13, 19) d vs.12(9, 16) d, Z=-3.747] in MP+ bacteria group were longer (all P<0.05). The level of WBC in MP+ SP group[12.20(7.80, 17.30)×10 9/L] was higher than that in MP+ HI group [6.75(5.37, 9.44)×10 9/L], and the differences were statistically significant( Z=11.574, P<0.05), and the incidence of lobar pneumonia in MP+ SP group [56.67%(17/30 cases)]was higher than that in MP+ CTA group [0(0/3 cases)]and MP+ HI group[18.75%(3/16 cases)], and the differences were statistically significant( χ2=9.770, P<0.05). Conclusions:Bacterial colonization or infection is more likely to occur in the oropharynx of children with MPP.When WBC, CRP, and LDH are significantly increased and the image shows a large consolidation or pleural effusion, it may indicate mixed bacterial infection, longer course of fever and higher incidence of RMPP, and the common mixed bacteria is SP.

Clinical analysis of early damage in multiple extra-pulmonary organs in COVID-19 / 南方医科大学学报

OBJECTIVE@#To analyze the clinical manifestations of heart, liver and kidney damages in the early stage of COVID-19 to identify the indicators for these damages.@*METHODS@#We analyzed the clinical features, underlying diseases, and indicators of infection in 12 patients with COVID-19 on the second day after their admission to our hospital between January 20 and February 20, 2020.The data including CK-MB, aTnI, BNP, heart rate, changes in ECG, LVEF (%), left ventricular general longitudinal strain (GLS, measured by color Doppler ultrasound) were collected.The changes of liver function biochemical indicators were dynamically reviewed.BUN, UCR, eGFR, Ccr, and UACR and the levels of MA, A1M, IGU, and TRU were recorded.@*RESULTS@#The 12 patients included 2 severe cases, 8 common type cases, and 2 mild cases.Four of the patients presented with sinus tachycardia, ECG changes and abnormal GLS in spite of normal aTNI and LVEF; 1 patient had abnormal CKMB and BNP.On the first and third days following admission, the patients had normal ALT, AST and GGT levels.On day 7, hepatic function damage occurred in the severe cases, manifested by elevated ALT and AST levels.Abnormalities of eGFR, Ccr and UACR occurred in 8, 5 and 5 of the patients, respectively.Abnormal elevations of MA, A1M, IGU and TRU in urine protein were observed in 4, 4, 5, and 2 of the patients, respectively.@*CONCLUSIONS@#In patients with COVID-19, heart damage can be identified early by observing the GLS and new abnormalities on ECG in spite of normal aTNI and LVEF.Early liver injury is not obvious in these patients, but dynamic monitoring of the indicators of should be emplemented, especially in severe cases. In cases with normal CR and BUN, kidney damage can be detected early by calculating eGFR, Ccr and UACR and urine protein tests.

The antibacterial effect of Fusidic acid on Mycoplasma pneumoniae and antibiotic resistant Mycoplasma pneumoniae in vitro / 中华实用儿科临床杂志

Objective To investigate the antibacterial effect of Fusidic acid on Mycoplasma pneumoniae and antibiotic resistant Mycoplasma pneumoniae in vitro.Methods Twenty-eight clinical strains of Mycoplasma pneumoniae isolated from patients with respiratory tract infection at Beijing Friendship Hospital Affiliated to the Capital University of Medical Sciences from January to December 2016 and 2 Mycoplasma pneumoniae reference strains were enrolled.The minimum inhibitory concentration (MIC) of Fusidic acid and Azithromycin were determined by using micro-dilution ration method.The chessboard method was used to check the antibacterial effect of combination between Fusidic acid and Azithromycin.The antibacterial activity of the Fusidic acid was evaluated by measuring the antibacterial rate of different concentrations.Results One isolate showed no mutation in 23SrRNA,26 isolates had one point mutation in loci 2063 and 1 isolate had one point mutation in loci 2064 among the 28 clinical isolates.The findings by micro-dilution method results showed that the MIC values of all the clinical isolates with mutations associated with macrolide resistance to Azithromycin were ＞ 1.000 0 mg/L,and the MIC values of all the clinical isolates with no mutations to azithromycin were ＜ 0.500 0 mg/L.The findings by micro-dilution method results showed that the MIC value of Fusidic acid for Mycoplasma pneumonia and drug resistance Mycoplasma pneumoniae was 1.000 0 mg/L.The Fractional Inhibitory Concentration index of Fusidic acid and Azithromycin combination was ≤0.500 0 mg/L.When the concentration of the Fusidic acid was lower than or equal to 32 MIC,the antibacterial effect of Fusidic acid against Mycoplasma pneumoniae increased with its higher concentration.When the concentration of the Fusidic acid was lower than or equal to 8 MIC,the longer the strain was exposed to the drug,the stronger antibacterial effect was against Mycoplasma pneumoniae.Conclusion If the treatment of Mycoplasma pneumoniae infection is not effective or the infection of patient is combined with bacteria,the application or combination of Fusidic acid may inhibit pathogenic bacteria effectively.Of course,how to use Fusidic acid in clinical treatment needs further study and discussion.

Effect of left ventricular hypertrophy and deformation on cardiac function in patients with uremia cardiomyopathy by using quantitative speckle tracking technology / 中南大学学报(医学版)

Objective:To evaluate the effect of left ventricular hypertrophy and deformation on cardiac function in patients with uremic cardiomyopathy (UCM) by using the technology of two dimensional speckle tracking imaging (2D-STI).Methods:A total of 67 UCM patients were randomly divided into the normal cardiac function group (subgroup A,32 cases) and the abnormal cardiac function group (subgroup B,35 cases)according to the New York Heart Association points (NYHA-P).A total of 30 healthy subjetcs served as the control group.Parameters including left ventricular ejection fraction (LVEF),left ventricular mass index (LVMI),left ventricular spherical index (LVSI),left ventricular myocardial mean radial strain (MRS),mean radial strain rate (MRSR),mean longitudinal strain (MLS),local systolic twist angle (STA),and mitral annulus maximum displacement (TMAD) were detected.Results:MLS,MRS,MRSR,LVSI,STA and TMAD in the Group A and Group B were lower than that in the control group (P＜0.05),and LVMI in the Group A and Group B was increased than those in the control group (P＜0.05);LVEE MLS,MRS,MRSR,LVSI and STAin the Group B was decreased than that in the Group A (P＜0.05).MLS in the Group A and B were positively correlated with LVEF and LVSI,but negatively correlated with LVMI.Using the point of 14.10％ for MLS to evaluate UCM patients with NYHA-P＞4 points,the sensitivity,the specificity and Yuedden index were 90.5％,71％ and 0.585,respectively.STA in UCM patients were lower than that in the control (P＜0.05).Conclusion:2D-STI possesses a unique advantage in detecting left ventricular strain and strain rate on left ventricular regional function in UCM with left ventricular hypertrophy and ventricular deformation.There is no direct correlation between the left ventricular hypertrophy and ventricular deformation,but the ventricular hypertrophy and deformation are correlated with regional cardiac function and clinical cardiac function.Left ventricular regional dysfunction may occur before cardiac hypertrophy and deformation.

Application of two dimension speckle tracking imaging in evaluating on cardiac dysfunction in patients with malignant lymphoma after anthracycline treatment / 吉林大学学报(医学版)

Objective:To assess the changes of myocardial biomechanical parametres of left ventricle of the patients with non-Hodgkin lymphoma (NHL)after anthracycline (ANTH)treatment by using two dimension speckle tracking imaging (2D-STI),and to study the value of 2D-STI in evaluating and monitoring the early cardiac dysfunction of the NHL patients induced by ANTH treatment.Methods:A total of 37 hospital patients who were firstly diagnosed as NHL (ANTH chemotherapy group)and 20 healthy volunteers (normal control group)were selected.The global longitudinal strain (GLS),global radial strain (GRS),global circumferential strain (GCS) and left ventricle twist (LVtw)of the subjects in two groups were detected before chemotherapy and 1,2,3 weeks after chemotherapy with 2D-STI;the GLS×LVtw was calculated.Results:There were no significant differences in the general clinical parameters of the subjects between normal control group and ANTH chemotherapy group before chemotherapy (P >0.05);the values of GLS,GRS and GCS of the patients in ANTH chemotherapy group after ANTH chemotherapy were decreased than those in normal control group and before ANTH chemotherapy (P <0.05),and GLS had the most obvious change.The GLS, GRS, GCS and GLS × LVtw of the patients after 3 cycles of chemotherapy in ANTH chemotherapy group were decreased (P > 0.05 ). Four patients meet the diagnostic criteria of cardiac toxicity in the couse of chemotherapy.The sensitivity was 94%,the specificity was 66.7%,and the maximal Youden index was 0.667 when the △ GLS × LVtw =-64.53% ×°was used as the cut-off point in predicting the occurrence of myocardial toxicity.Conclusion:2D-STI can detect the early changes of biomechanical parameters of left ventricular myocardium with highly sensitive in predicting early myocardial toxicity and early cardiac dysfunction caused by ANTH chemotherapy.It may be an effective way to predict the early myocardial toxicity of ANTH chemotherapy in the future.

Development of real-time fluorescence quantitative PCR for the detection of Moraxella catarrhalis / 国际儿科学杂志

Objective To developed A laboratory diagnosis of Moraxella catarrhalis by an laboratories diagnostic method real-time fluorescence quantitative PCR assay. Methods The specific primers and probes were designed based on the sequence of outer membrane protein CopB(copB)gene in Moraxella catarrhalis,and the Taqman probe RT-PCR method was developed to detect the Moraxella catarrhalis.The standard plasmids ex-tracted from the Moraxella catarrhalis standard strains were used to constitute the standard samples,and compared with these standard samples,the sensitivity of the fluorescence quantitative PCR assay was tested by the estab-lished standard curves.The specificity of the fluorescence quantitative PCR assay was tested by the DNA samples of other bacterias in the laboratory.Meanwhile,321 throat swab samples from inpatient and outpatient child pa-tients,with asthma infection were collected as clinical samples to validate the fluorescence quantitative PCR as-say.Results The standard curve was drawn in the real-time PCR by the Taqman fluorescence reporter.During the sensitivity tests,the newly-developed real-time fluorescence PCR could detect at least 10 copies of Moraxella catarrhalis,and could successfully distinguish several DNAs of the pathogens.On the basis of the validation result of the 321 throat swab samples,there are 25 Moraxella catarrhalis with 7.79 % positive rate.Conclusion The fluorescence quantitative PCR assay is of great sensitivity and specificity,and it can be widely used for the detec-tion of Moraxella catarrhalis.

Development and clinical application of real-time quantitative PCR for the detection of mycoplasma pneumonia / 国际儿科学杂志

Objective To develope a new Real-time quantitative PCR assay using SYBR green as fluorescence reporter, which is rapid, specific, sensitive, cheap and accurate for the detection of mycoplasma pneumoniae(MP), and evaluated its clinical application value.Methods The sequence of the 23S rRNA gene in MP type strain FH was selected as amplified regions, and specific primers were designed.Then the related plasmids were extracted as standards,and the absolute quantitative standard curve was established.The sensitivity ,specificity of the fluorescence quantitative PCR assay was compared with the nest-PCR and kit;To calculate correlation coefficient, coincidence rate and kappa coefficient, clinical samples were detected using above-mentioned methods and cultivation,respectively.Results The detection sensitivity of the new real-time PCR and nest-PCR was 10 copies of FH DNA,while the kit 100 copies.In the specificity tests,the MP sample was positive,while mycoplasma hominis and other four bacteria were all negative.We applied this real-time PCR assay ,nest-PCR, kit and cultivation to 182 clinical specimens, and the detection rates were 55.49％, 52.75％, 47.25％ and 39.01％ ,respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and nest-PCR were 89.6％ ,0.790, respectively;while those of the new method and cultivation were 83.5 ％ ,0.678, respectively.The total consistency rate and Kappa coefficient of the new real-time PCR method and the kit were 89.6％ ,0.792,respectively;and the correlation coefficient of these two methods was 0.923,P ＜ 0.001.Conclusion Compared with other methods, the new real-time PCR assay could be used to detect mycoplasma pneumoniae quickly and economically, with high sensitivity and specificity ,revealing great utility value on varied instrumentation platforms.

Determinationofleftventricularbiomechanicalparametersbyusing velocity vector imaging technology and its application in assessing coronary artery stenosis in coronary artery disease / 吉林大学学报(医学版)

Objective To determine the biomechanical parameters of left ventricular by using velocity vector imaging (VVI),and to indirectly assess the coronary artery stenosis with VVI in the patients with coronary artery disease (CAD).Methods 52 patients who had one coronary artery lesions at least diagnosed by coronary angiography (CAG)were divided into coronary artery mild stenosis group and severe stenosis group;2 1 patients in mild stenosis group had one coronary stenosis <75%;31 patients in severe stenosis group had one coronary artery stenosis ≥75% at least.At the same time,20 cases of normal people without coronary artery stenosis showed by CAG were selected as normal control group. VVI was used to detect the left ventricular wall segments of the overall longitudinal strain (GLS ), the overall circumferential strain (GCS ) and the overall radial strain (GRS ). Results The absolute values of GLS,GRS,GCS of the patients in mild and severe coronary artery stenosis groups were significantly decreased than those in normal control group(P<0.05),and the strain parameters in severe stenosis group were decreased more significantly, there were significant differences compared mild stenosis group (P<0.05 ). The distribution of the segments with decreased longitudinal strain matched the LV myocardial segment with the coronary stenosis rate ≥ 75%,and GLS had the most sensitivity.The GLS in normal control group,mild stenosis group and severe stenosis group were negatively correlated to left ventricular ejection fraction (LVEF)(r=-0.58,P<0.05;r=-0.51,P<0.05;r=-0.43,P<0.05).GLS-16.14 % was used to assess the severe coronary artery stenosis with requiring the implementation of PTCA treatment as the diagnostic cut-off point with sensitivity 96.8%, specificity of 70%, the highest Yuedden index 0.668. Conclusion The decreasing of left ventricular strain could be detected by VVI, which suggests that severe coronary artery stenosis exists in coronary artery;the distribution of the segments with significantly decreased strains can be used to assess the coronary lesions and stenosis degrees.

The study on the change of left ventricular geometry shape and left ventricular regional function in mitral insufficiency (MI) / 中国医师杂志

Objective To evaluate the left ventricle geometry shape and left heart regional function in mitral insufficiency (MI) by quantitative tissue velocity imaging (QTVI) and the left ventricle systolic and diastolic geometry spbericity index. Methods Thirty normal subjects and 62 MI patients underwent QTVI and color Doppler imaging (CDFI) were enrolled in this study in order to measure the left ven-tricular geometry shape and left ventrieular regional function along LV apical long-axis view. Off-line LV regional velocity images along LV apical long-axis view were synchronously obtained. Peak tissue velocities of LV regional muscular tissue during systole(Vs),systolic acceler-ation(a), early diastole(Ve), LA contraction(Va)were synchronously measured as the index of left ventricular regional systolic and dias-tolic function. The left ventricle geometry shape index were reflected from the systolic and diastolic geometry sphericity index (SIs and Sid) and the left ventrieular ejection fraction (LVEF) and the peak D and peak a (PVd/PVa) of pulmonary veins flowing spectrum reflected the global left ventricular function index. The Vs, Ve, Va, a, PVd/PVd ratio, LVEF, SIs, SId were recorded and their correlation between normal subjects and patients with MI were compared. Result Vs, Ve,Va,a,PVd/ PVa ratio,SIs,SId in patients with MI were significantly re-duced. There was positive relation between Sis and a(r=0.602)and Ve/Va and SId(r=0.635). Conclusion There was negative rela-tion between regional cardiac function and LV spberieity, the higher cardiac function was accompanied with the lower SI.

The clinical analysis of hypoxic spells on patients with tetralogy of fallot / 中国基层医药

Objective To explore the prophylaxis and treatment of hypoxic spells on patients with TOF.Methods Retrospectively analysed and compared the case history of the inpatients with TOP in our hospital,to analyse the epilepticus cause of hypoxic spells and study the hypoxia mechanism and summarize the prevention and control measures.Results 4(17%)cases of TOF died after hypoxic spells,the epilepticus cause of hypoxic spells included suckling,early getting up,crying,standing abruptly after stool,scorching climate,infection fever and anemia,the hypoxia mechanism was likely to the increase of infundibulum cardiac muscle contraction,the systemic circulation resistance drop suddenly,hypezventilation,high-viscosity syndrome.Conclusion To control the epilepticus cause and perform operative treatment early can reduce the hypoxic spells.The prevention and control measures were strengthening the live nurse,disconnected oxygen inhalation,oral beta-blocker therapy and venons injection 5% sodium bicarbonate.Early operative treatment was the best redical treatment.

Evaluation of Right Ventricular Volume and Systolic Function by Real-time Three-dimensional Echocardiography / 华中科技大学学报（医学）（英德文版）

The optimal plane for measurement of the right ventricular (RV) volumes by real-time three-dimensional echocardiography (RT3DE) was determined and the feasibility and accuracy of RT3DE in studying RV systolic function was assessed. RV "Full volume" images were acquired by RT3DE in 22 healthy subjects. RV end-diastolic volumes (RVEDV) and end-systolic volumes (RVESV) were outlined using apical biplane, 4-plane, 8-plane, 16-plane offline separately. RVSV and RVEF were calculated. Meanwhile tricuspid annual systolic excursion (TASE) was measured by M-mode echo. LVSV was outlined by 2-D echo according to the biplane Simpsons rule. The results showed: (1) There was a good correlation between RVSV measured from series planes and LVSV from 2-D echo (r=0.73; r=0.69; r=0.63; r=0.66, P＜0.25-0. 0025); (2) There were significant differences between RVEDV in biplane and those in 4-, 8-, 16-plane (P＜0. 001). There was also difference between RV volume in 4-plane and that in 8-plane (P＜0.05), but there was no significant difference between RV volume in 8-plane and that in 16-plane (P＞0.05); (3) Inter-observers and intro-observers variability analysis showed that there were close agreements and relations for RV volumes (r=0. 986, P＜0. 001; r=0.93, P＜0. 001); (4) There was a significantly positive correlation of TASE to RVSV and RVEF from RT3DE (r=0.83; r=0.90). So RV volume measures with RT3DE are rapid, accurate and reproducible. In view of RVs complex shape,apical 8-plane method is better in clinical use. It may allow early detection of RV systolic function.

Evaluation of right ventricular volume and systolic function by realtime three-dimensional echocardiography / 华中科技大学学报（医学）（英德文版）

The optimal plane for measurement of the right ventricular (RV) volumes by real-time three-dimensional echocardiography (RT3DE) was determined and the feasibility and accuracy of RT3DE in studying RV systolic function was assessed. RV "Full volume" images were acquired by RT3DE in 22 healthy subjects. RV end-diastolic volumes (RVEDV) and end-systolic volumes (RVESV) were outlined using apical biplane, 4-plane, 8-plane, 16-plane offline separately. RVSV and RVEF were calculated. Meanwhile tricuspid annual systolic excursion (TASE) was measured by M-mode echo. LVSV was outlined by 2-D echo according to the biplane Simpson's rule. The results showed: (1) There was a good correlation between RVSV measured from series planes and LVSV from 2-D echo (r = 0.73; r = 0.69; r = 0.63; r = 0.66, P 0.05); (3) Inter-observers and intro-observers variability analysis showed that there were close agreements and relations for RV volumes (r = 0.986, P < 0.001; r = 0.93, P < 0.001); (4) There was a significantly positive correlation of TASE to RVSV and RVEF from RT3DE (r = 0.83; r = 0.90). So RV volume measures with RT3DE are rapid, accurate and reproducible. In view of RV's complex shape, apical 8-plane method is better in clinical use. It may allow early detection of RV systolic function.