Chinese herbal medicine combined with Western medicine for Mycoplasma pneumoniae pneumonia in children:An overview of systematic reviews

Objective:To summarize the characteristics and evaluate the quality of the methodology and evidence within systematic reviews(SRs)of Chinese herbal medicine(CHM)for Mycoplasma pneumoniae pneumonia(MPP)inchildren.Methods:SRs of randomized controlled trials were searched using PubMed,the Cochrane Library,Embase,the Chinese National Knowledge Infrastructure Databases(CNKI),the Chinese Scientific Journals Database(VIP),Wanfang,and the SinoMed Database.SRs on the use of CHM alone or in combination with Western medications for MPP in children were included.The study compared the effects of Western medicine alone with those of CHM.The evidence quality using the A Measurement Tool to Assess Systematic Reviews(AMSTAR)2,the Preferred Reporting Items for Systematic Reviews and Meta-Analyses(PRISMA)2020,and the Grading of Recommendations,Assessment,Development,and Evaluation(GRADE)criteria.The primary indicators were the total effective rate,fever subsidence time,and cough disappearance time.The secondary outcomes were pulmonary rale disappearance time,average hospitalization time,lung X-ray infiltrate disappearance time,immunological indices,and inflammatory cytokine levels.Results:Twelve relevant SRs were included;75%(9/12)were assessed as very low quality,and 25%(3/12)Were rated as low quality using the AMSTAR 2 criteria.According to the PRISMA 2020 checklist,the average SR score was 20.3 out of a 27 point maximum.In all SRs,CHM demonstrated improvement in symptoms and signs among children with MPP.The evidence quality using the GRADE criteria ranged from"very low"(>50%)to"moderate"(<5%).The most common downgrading factor was imprecision,followed by publication bias and inconsistency.Conclusion:This overview highlights the limited quality of the methodology and evidence of the included SRs.Although the included studies showed the beneficial effects of CHM on MPP in children,it was difficult to draw firm conclusions owing to methodological flaws.

Evaluating the role of interleukin-2 and interleukin-12 in pediatric patients with concurrent Mycoplasma pneumoniae and Epstein-Barr virus infections

BACKGROUND Mycoplasma pneumoniae(MP)frequently causes respiratory infections in children,whereas Epstein-Barr virus(EBV)typically presents subclinical manifestations in immunocompetent pediatric populations.The incidence of MP and EBV coinfections is often overlooked clinically,with the contributory role of EBV in pulmonary infections alongside MP remaining unclear.AIM To evaluate the serum concentrations of interleukin-2(IL-2)and interleukin-12(IL-12)in pediatric patients with MP pneumonia co-infected with EBV and assess their prognostic implications.METHODS We retrospectively analyzed clinical data from patients diagnosed with MP and EBV co-infection,isolated MP infection,and a control group of healthy children,spanning from January 1,2018 to December 31,2021.Serum IL-2 and IL-12 levels were quantified using enzyme-linked immunosorbent assay.Logistic regression was employed to identify factors influencing poor prognosis,while receiver operating characteristic(ROC)curves evaluated the prognostic utility of serum IL-2 and IL-12 levels in co-infected patients.RESULTS The co-infection group exhibited elevated serum IL-2 and C-reactive protein(CRP)levels compared to both the MP-only and control groups,with a reverse trend observed for IL-12(P<0.05).In the poor prognosis cohort,elevated CRP and IL-2 levels,alongside prolonged fever duration,contrasted with reduced IL-12 levels(P<0.05).Logistic regression identified elevated IL-2 as an independent risk factor and high IL-12 as a protective factor for adverse outcomes(P<0.05).ROC analysis indicated that the area under the curves for IL-2,IL-12,and their combination in predicting poor prognosis were 0.815,0.895,and 0.915,respectively.CONCLUSION Elevated serum IL-2 and diminished IL-12 levels in pediatric patients with MP and EBV co-infection correlate with poorer prognosis,with combined IL-2 and IL-12 levels offering enhanced predictive accuracy.

Serum inflammatory markers in children with Mycoplasma pneumoniae pneumonia and their predictive value for mycoplasma severity

BACKGROUND Mycoplasma pneumoniae pneumonia(MPP)significantly impacts pediatric health,necessitating markers for early severe disease identification.AIM To investigate the correlation between serum inflammatory marker and the severity of MPP in children.METHODS A prospective study was carried out from January 2023 to November 2023.A total of 160 children with MPP who underwent treatment were selected:80 had severe MPP and 80 had mild MPP.Clinical and laboratory data were collected at the time of hospital admission and during hospitalization.Receiver operating characteristic curves were utilized to assess the diagnostic and prognostic for severe MPP.RESULTS Fever duration and length of hospitalization in pediatric patients with severe MPP exceeded those with mild MPP.The incidence of pleural effusion,lung consolidation,and bronchopneumonia on imaging was markedly elevated in the severe MPP cohort compared to the mild MPP cohort.In contrast to the mild cohort,there was a notable increase in C-reactive protein(CRP),procalcitonin(PCT),erythrocyte sedimentation rate,lactic dehydrogenase,D-dimer,and inflammatory cytokines[interleukin(IL)-6,IL-8,IL-10 and tumor necrosis factor(TNF)-α]in the severe MPP group were significantly higher.CONCLUSION Serum inflammatory markers(CRP,PCT,IL-6,D-dimer,IL-10 and TNF-α)were considered as predictors in children with severe MPP.

Preparation of Ephedra houttuynia Granule and Its Therapeutic Mechanism of Anti-Mycoplasma gallisepticum Infection Based on Network Pharmacology

The use of Chinese herbal medicines can replace antibiotics that cause drug-resistance problems,which are currently necessary for disease control.In this paper,a traditional Chinese medicine compound named Ephedra houttuynia granule for the treatment of Mycoplasma galliscepticum(MG)infection was prepared.Furthermore,its action mechanism was explored through network pharmacology.The optimal extraction and granulation processes of the compound were determined by high performance liquid chromatography(HPLC)method and L9 orthogonal test,and in the treatment experiment,Ephedra houttuynia granule showed a significant therapeutic effect on MG infection.In the study of network pharmacology,the results showed that the core targets of Ephedra houttuynia granule against MG infection were vascular endothelial growth factor(VEGFA),fos proto-oncogene(FOS),prepro-coagulation factor II(F2),etc.,the gene ontology/kyoto encyclopedia of genes and genomes(GO/KEGG)analysis results indicated that the signaling pathways of neuroactive ligand receptor interaction,cAMP,IL-17,T cell receptor,and tumor necrosis factor(TNF)might involve in anti-MG infection.In conclusion,this study would provide a new idea for elucidating the action mechanism of other diseases in veterinary clinic,which had a certain guiding significance.

Research Progress on Combined Chinese and Western Medicine Treatment of Mycoplasma pneumoniae Pneumonia in Children

With the continuous development of medical technology,combined treatment of Chinese and Western medicine has gradually become a research hotspot.As a common disease in pediatrics,the treatment of Mycoplasma pneumoniae pneumonia(MPP)in children is also being explored and improved.This article summarizes the research progress of combined Chinese and Western medicine treatment of MPP in children in recent years,aiming to provide a useful reference for the combined treatment of MPP in children.The article firstly introduces the etiology and pathogenesis of MPP in children,thereafter briefly introduces the Western anti-infective treatment and traditional Chinese medicine(TCM)diagnosis and treatment of MPP in children,and lastly introduces the methods of combined treatment of TCM and Western medicine in detail.The article points out that the combination of Chinese and Western medicine can give full play to the overall regulation of Chinese medicine and the precise treatment advantages of Western medicine,improve the therapeutic effect,reduce the use of antibiotics,and lower the recurrence rate of the disease,which is worthy of further research and promotion.

Clinical characteristics of community-acquired pneumonia in children caused by mycoplasma pneumoniae with or without myocardial damage:A single-center retrospective study

BACKGROUND Mycoplasma pneumoniae(MP)is a prevalent pathogen that causes respiratory infections in children and adolescents.AIM To assess the differences in the clinical features of MP-associated communityacquired pneumonia(CAP)in children who presented with mild or severe mycoplasma pneumoniae pneumonia(MPP);to identify the incidence of myocardial damage between the two groups.METHODS This work is a retrospective study.We identified children between 2 mo and 16 years of age with clinical and radiological findings consistent with CAP.We admitted patients to the inpatient department of the Second Hospital of Jilin University,Changchun,China,from January 2019 to December 2019.RESULTS A total of 409 hospitalized patients were diagnosed with MPP.Among them were 214(52.3%)males and 195(47.7%)females.The duration of fever and cough was the longest in severe MPP cases.Similarly,plasma levels of highly sensitive Creactive protein(t=-2.834,P<0.05),alanine transaminase(t=-2.511,P<0.05),aspartate aminotransferase(t=-2.939,P<0.05),and lactate dehydrogenase(LDH)(t=-2.939,P<0.05)were all elevated in severe MPP cases compared with mild MPP cases,and these elevations were statistically significant(P<0.05).Conversely,the neutrophil percentage was significantly lower in severe MPP cases than in mild MPP cases.The incidence of myocardial damage was significantly higher in severe MPP cases than in mild MPP cases(χ^(2)=157.078,P<0.05).CONCLUSION Mycoplasma pneumoniae is the main cause of CAP.The incidence of myocardial damage was higher and statistically significant in severe MPP cases than in mild MPP cases.

Isolation and Progeny Transmission of Non-Temperature-Sensitive MS-H Vaccine Strains of Mycoplasma synoviae from Temperature-Sensitive MS-H-Vaccinated Laying Breeder Hens

This pilot study reports the vertical transmission and reverse thermosensitivity of the MS-H vaccine strain of Mycoplasma synoviae (MS) by RAPD in commercial breeders and their progeny. At two weeks of age, breeders were vaccinated with the ts+ MS-H strain. At 9 weeks of age, an outbreak of infectious synovitis (IS) was detected in the progeny. Tracheal swab samples were collected from breeders at 24, 39, 48, and 70 weeks of age. At 9 weeks, pullets swab from the elbow joints were collected. RAPD was performed on the isolates at 39.5°C, and the same ts- MS-H strains were identified in the breeder hens and their progeny. Tracheal swabs from breeder hens were negative to MS isolation at 37°C and 39.5°C at 24- and 39-weeks. MS isolation was recovered from tracheal swabs from 9/10 and 10/10 breeders at 48- and 70- week. At 9 weeks of age in the progeny, MS was isolated from tracheal swabs of 10/10 from non-IS pullets. MS was isolated from 9/10 joints samples. The isolates from breeder hens and their progeny showed non-significant differences in five antimycoplasmic MIC100 values;otherwise, enrofloxacin presented a significant difference in MIC100 value (p < 0.05). This investigation demonstrated the reversal of the thermosensitivity, pathogenicity, and vertical transmission of the MS-H strain. Consequently, it is crucial to contemplate the danger of reversing pathogenicity and transmission to progeny when applying the MS-H vaccine strain.

Antibiotics Resistance of Urogenital Mycoplasma in Sexually Active Women Attending Gynecologic Consultation in Douala (Cameroon)

Introduction: Mycoplasmas are bacteria commonly found in the commensal flora of humans and can occasionally be pathogenic. The population and the geographical area determine their susceptibility to various antibiotics. Methodology: We carried out a cross-sectional study from January to June 2022 at the Douala General Hospital. All women who were sexually active and willing to participate were included. Endo-cervical swabbing was used to collect the samples. Culture and antibiotic susceptibility testing were performed in a liquid medium using Mycoplasma-SystemPlus Gallery (Liofilchem). All samples that degraded urea (threshold > 103 for Ureaplasma urealyticum) and arginine (threshold > 104 for Mycoplasma hominis) were deemed positive. Results: A total of 107 women with a median age of 33 ± 8.3 years and a predominance of 30-40 years (41.1%) were included. The frequency of mycoplasma infection was 73.8% with 34.1%, 11.4% and 54.4% for Ureaplasma urealyticum, Mycoplasma hominis and co-infection respectively. Resistance rates to Minocycline were low, 1.3% for Ureaplasma urealyticum and 3.7% for co-infection;for Pefloxacin the resistance rates were 3.7% (Ureaplasma urealyticum, Mycoplasma hominis) and 22.8% (co-infection). Ureaplasma urealyticum had a resistance rate of 3.7% for Erythromycin, 1.3% for Clarithromycin and 5% for Azithromycin. There was no significant correlation between risk factors and infection. Vaginal cleansing and the development of resistance in mycoplasma infection were found to be significantly correlated (OR = 6.915 [1.52 - 31.55];(p = 0.013)). Conclusion: Minocycline was the most active antibiotic and Ureaplasma urealyticum was the species with the lowest rate of resistance. Antibiotic resistance was more common in co-infected people than mono-infected. Antibiotic resistance was independently correlated with vaginal douching.

The Effect of Azithromycin in Treating Mycoplasma Pneumonia in Children

Objective:To discuss and analyze the effect of azithromycin in the treatment of mycoplasma pneumonia in children.Methods:A total of 120 children with mycoplasma pneumonia who were admitted to the Department of Pediatrics of our hospital from January 2022 to December 2022 were selected as the research subjects.They were divided into an azithromycin group and a reference group according to the random number drawing method,with 60 cases in each group.The azithromycin group was treated with azithromycin,and the reference group was treated with conventional treatment.The efficacy of treatment,laboratory indicators,platelet count and D-dimer,and adverse reactions of both groups were compared.Results:The efficacy of the azithromycin group was significantly higher than that of the reference group(P<0.05).Before treatment,there were no significant differences in the laboratory indicators like ferritin,procalcitonin(PCT),and erythrocyte sedimentation rate(ESR)between the two groups(P>0.05);after treatment,the laboratory indicators of the azithromycin group were significantly better than those of the reference group(P<0.05).Before treatment,there was no statistically significant difference in platelet count and D-dimer between the groups(P>0.05);after medication,the platelet count,and D-dimer in the azithromycin group were significantly better than those in the reference group(P<0.05).The total incidence of adverse reactions in the azithromycin group was significantly lower than that in the reference group(P<0.05).Conclusion:Azithromycin is more effective in treating mycoplasma pneumonia in children,and has certain clinical value.

基于16S rRNA基因的鼠Mycoplasma haemomuris的分子鉴定和种系发育分析

为了从分子水平揭示野生鼠类血原体的种类特征和系统发生关系,无菌采集32份野生白腹巨鼠血样,抽提全血基因组DNA,采用真细菌的通用引物进行16SrRNA基因的扩增,对扩增产物进行克隆和测序。结果从其中21只血样中成功地扩增出目的片段大小的核苷酸序列。对阳性产物进行克隆、测序,获得了两条代表性序列(GenBank登录号HQ183731和HQ183732)。序列分析显示,获得的两条序列与GenBank中收录的鼠Mycoplasma haemomuris 16SrRNA基因(AB758435)相似性最高,分别为95%和97%。两个样本间16SrRNA基因序列相似性达98.3%。种系发育分析表明,获得白腹巨鼠血原体的两条序列形成了独立的进化分支,并与来自野鼠的Haemobartonella muris(HMU82963)和来自家鼠M.haemomuris(AB758435)所形成的进化分枝为姊妹枝。上述研究证明,白腹巨鼠具有较普遍的M.haemomuris感染,并与已报道的啮齿动物M.haemomuris有一定的遗传差异,是一种新基因型的血原体。对进一步研究人和动物的亲血性支原体的流行病学、种群生物学等研究具有一定价值。

Preventive Effects of Five Drugs on Mycoplasma Pneumonia of Swine (MPS)

The paper was to explore the preventive effects of five drugs on mycoplasma pneumonia of swine （MPS） and to provide reference for clinical medication of pig farms in Hainan Province. A total of 444 health piglets were randomly divided into 6 groups, including five medication groups （72 piglets in group A, 74 pig- lets in group B, 72 piglets in group C, 76 piglets in group D, 76 piglets in group E） and one control group （74 piglets）. The piglets in experimental groups were treated drugs once a day for successive 5 days at 30, 60, 90, 120 and 150 of age. The piglets in control group were free of medication. At 70 and 140 days of age, 15 piglets of each group were randomly selected to collect their blood sermn. The Mycoplasma hyopneumoniae （M-Hyo） antibodies in serum were measured by en- zyme-linked immunosorbent assay （ELISA）. During the experiment, the incidence rates of respiratory disease, lung lesion, feed conversion rate, average daily gain （ADG）, and mortality rate of pigs were also observed and recorded. The results showed that the five drugs had significant difference in preventative effects. Group C （Zhiyuanjing group） received the best preventive effect and the highest economic benefits. Compared with control group, the ADG and feed conversion rate in group C were increased by 7.53% and 9.09%, respectively; the incidence rate of respiratory disease was reduced by 13.44% and lung lesion was alleviated by 81.43% ; and the earnings of each pig could rise by 132.70 yuan. The preventative effect and economic benefit of the drugs was sequenced by Chansu Kechuanling and Bingchan Kechuanwang. Wante Feilin and amoxicillin had weaker preventive effects against MPS but greatly influenced growth performance of pigs, so they should be used alternatively with other drugs.

Standardization of a Real-time PCR System for Quantitative Detection of Mycoplasma hyopneumoniae

This study was conducted to develop a method for accurate quantification of Mycoplasma hyopneumoniae during vaccine production or experimental research. Primer and probe concentration that gave the highest ΔRn and the lowest Ct were selected to establish the real-time PCR system for the detection of M. hyopneumoniae. Template DNA of M. hyopneumoniae was extracted by boiling under different conditions and detected by real-time PCR to determine the optimal conditions for DNA extraction. Thereafter, intra-and inter-batch reproducibility tests were carried out using a standard plasmid to evaluate the stability of the PCR system. Subsequently, the effect of medium composition on the quantitative detection was evaluated. Finally, the correlation between real-time PCR and CCU method was explored. The optimal primer and probe concentration for real-time PCR were 0.4 and 0.2 μmol/L, respectively. The intra-and inter-batch coefficients of variation(CV) in Ct value of 10~4-10~9 copies/μl standard plasmid were <5%, indicating good reproducibility of the real-time PCR system. Following incubation in a boiling water bath for 10 min, M. hyopneumoniae samples can be used directly as a template in subsequent real-time PCR assays,and good intra-batch and inter-batch reproducibility was observed. The working concentration of KM2 medium should be less than the 1/10 of the concentration of the stock solution to minimize its influence on the quantitative detection. Spearman's correlation analysis revealed that the log of CCU and the log of DNA copy number had a significant positive relationship(r=0.797,P=0.000). Thus, the two methods can be used in combination in the quantitative detection of M. hyopneumoniae. In summary, a rapid, stable and accurate quantitative PCR system for detecting M. hyopneumoniae culture was established in this study, which provides a technical means for accurate quantification of M. hyopneumoniae in vaccine production and laboratory tests.

Detection of Antibodies against Mycoplasma mycoides subsp. capri in Goats with the Complement Fixation Test

Mycoplasma mycoides subsp. capri is the causative agent of severe and acute respiratory problems in goats, which spreads rapidly and represents high mortality. The serological profile of the goat population, from nine regions in seven states of Mexico, was screened by the Complement Fixation test (CF) in sera from asymptomatic goats and animals with mild respiratory symptoms. Sera and nasal swabs of 827 goats were collected for the isolation of the organism. An antiserum was prepared against a previously isolated field strain of Mycoplasma mycoides subsp. capri. CF Antibody titers were associated with the results of the isolates to determine the cutoff point. The CF was considered as positive if its result was ≥1/16. The CF registered 251 positive goats (30.35%) and 576 (69.65%) negative;the test showed high sensitivity (93.33%) and specificity (72.27%). In the specific case of diagnosis for mycoplasmosis associated with respiratory problems in goats, the CF proved to be a good diagnosis test, this study determined that 30% of the goat population showed antibody titers against Mycoplasma mycoides subsp. capri and revealed those animals who have had contact with this microorganism during their lives regardless of the presence or absence of respiratory symptoms.

Adherence of a Clinical Strain of Mycoplasma fermentans to Human Cultured Epithelial Cells

This study compares the kinetics of adherence of both the prototype strain PG-18 and the recently clinically isolated strain P-140 of Mycoplasma fermentans to HEp-2 cells. M. fermentans may induce respiratory and genitourinary tract infections. We investigated the presence of putative adhesins from strain P-140. A surface protein of 47 kDa, labeled SP47, isolated from M. fermentans P-140, seems to be involved in acterial adherence on HEp-2 cells;this study could allow helping understand the interaction between mycoplasmas and their hosts. Anti-SP47 antibodies inhibited the formation of bacterial clusters and adherence to cultured cells, quantified by ELISA. M. fermentans P-140 was more efficient at adhering to cultured HEp-2 cells than PG-18.

氟罗沙星对禽败血霉形体(Mycoplasma gallisepticum)病的药效研究

体外抑菌试验测得氟罗沙星（Ｆｌｅｒｏｘａｃｉｎ）对禽败血霉形体（Ｍｙｃｏｐｌａｓｍａｇａｌｉｓｅｐｔｉｃｕｍ）的最小抑菌浓度（ＭＩＣ）为０．０１５ｍｇ／Ｌ。体内药效试验表明，氟罗沙星以２５、５０、１００ｍｇ／Ｌ饮水给药和以５、１０、１５ｍｇ／ｋｇ肌注给药（１次／ｄ），连续用药５ｄ，对人工气囊接种禽败血霉形体培养液（０．２ｍＬ，约含１０８ｃｆｕ）的雏鸡，保护率均为１００％（３０／３０），而感染不给药组雏鸡存活率为９３．３％（２８／３０）；相对增重率分别为９０．８％、９１．５％、９２．３％和９１．５％、９１．８％、９２．９％，显著高于感染不给药组（７３．２％）；气囊损伤分分别为２．９３、２．６２、０．９３和１．８、１．１、１．０，而感染不给药组为６．５７，差异显著（Ｐ＜０．０５）；血清玻板凝集试验阳性率分别为２０％（２／１０）、１０％（１／１０）、０（０／１０）和３０％（３／１０）、１０％（１／１０）、１０％（１／１０），均极显著低于感染不给药组１００％（１０／１０）（Ｐ＜０．０１）。氟罗沙星不同给药途径及不同剂量间药效差异不显著。

Mycoplasma infections and different human carcinomas

AIM To explore relationships between human carcinomas and mycoplasma infection.METHODS Monoclonal antibody PD4, which specifically recognizes a distinct protein from mycoplasma hyorhinis, was used to detect mycoplasma infection in different paraffinembedded carcinoma tissues with immunohistochemistry. PCR was applied to amplify the mycoplasma DNA from the positive samples for confirming immunohistochemistry.RESULTS Fifty of 90 cases (56%) of gastric carcinoma were positive for mycoplasma hyorhinis. In other gastric diseases, the mycoplasma infection ratio was 28% (18/49) in chronic superficial gastritis, 30% (14/ 46) in gastric ulcer and 37% (18/ 49) in intestinal metaplasia. The difference is significant with gastric cancer (X2=12.06, P＜0.05). In colon carcinoma, the mycoplasma infection ratio was 55.1% (32/58), but it was 20.9% (10/49) in adenomarous polyp (X2=13.46, P＜0.005).Gastric and colon cancers with high differentiation had a higher mycoplasma infection ratio than those with low differentiation (P＜ 0.05). Mycoplasma infection in esophageal cancer, lung cancer, breast cancer and glioma was 50.9% (27/53), 52.6% (31/ 59), 39.7%(25/63) and 41% (38/91), respectively. The mycoplasma DNA was successfully amplified with the DNA extracted from the cancer tissues that were positive for mycoplasma infection (detected with antibody PD4).CONCLUSION There was high correlation between mycoplasma infection and different cancers, which suggests the possibility of an association between the two. The mechanism involved in oncogenesis by mycoplasma remains unknown.

Immune Responses to the Attenuated Mycoplasma hyopneumoniae 168 Strain Vaccine by Intrapulmonic Immunization in Piglets

To investigate the immune responses to the attenuated Mycoplasma hyopneumoniae 168 strain vaccine, 8-15 d old piglets were immunized with M. hyopneurnoniae 168 strain vaccine by intrapulmonic route. And the specific IgG antibody in serum, lymphoproliferation, IFNT, and specific secretory IgA （SIgA） antibody in bronchoalveolar lavage fluid were detected on 30 and 60 d post-immunization （DPI）, respectively. On 60 DPI, all the pigs except for those in health control group were challenged with a field M. hyopneumoniae strain JS. Necropsy was performed on 30 d post-challenge （DPC）. The results showed that IFN7 and specific SIgA were stimulated on surface of respiratory tract after immunization. And peripheral blood mononuclear cells could also be proliferated about 1.81 and 2.12 fold on 30 and 60 DPI when stimulated by M. hyopneumoniae protein in vitro. However, no serum IgG antibody against M. hyopneumoniae was detected during the whole immune phage. After challenge, vaccinated pigs were observed with only very slight histological lesion in individual lobes. None of vaccinated pigs showed any clinical signs. While the unvaccinated pigs from challenge control group showed varying degrees of clinical sign and severe macroscopical lesion of mycoplasmal pneumonia of swine （MPS）. The result suggested that the attenuated M. hyopneumoniae 168 strain vaccine inoculated by intrapulmonic route could activate the systemic cellular immunity, the local mucosal immunity and IFNγ secretion in respiratory tract to against M. hyopneumoniae infection in piglets.

Evaluation of a New Real-time PCR Assay for Detection of Mycoplasma Pneumoniae in Clinical Specimens

Objective To establish and evaluate a real-time PCR assay to detect Mycoplasma pneumoniae （M.pneumoniae） in clinical specimens. Methods By analysing the whole pl gene sequence of 60 M.pneurnoniae clinical isolates in Beijing of China, an optimized real-time PCR assay （MpP1） using pl gene conserved region was designed. The specificity and sensitivity of this assay were evaluated and compared with other two reported assays （RepMpl and Mp181） using 40 positive and 100 negative clinical specimens. Results The detection limit of the new assay was 8.1 fg （about 1-3CFU） M.pneumoniae DNA. The sensitivity of MpP1, RepMpl, and Mp181 assays appeared to be 100%, 100%, and 85%, respectively. Conclusion MpP1 assay is suitable for the detection of M.pneumoniae in Chinese clinical specimens.

Correlation Between the Clinical Severity, Bacterial Load, and Inflammatory Reaction in Children with Mycoplasma Pneumoniae Pneumonia

Given the lack of defining features in the clinical manifestations and radiographic findings for children with mycoplasma pneumoniae pneumonia(MPP),quantitative polymerase chain reaction(qPCR)has become a useful diagnostic method.This study was performed to explore the relationship between the qPCR findings,clinical symptoms,and inflammatory markers in children with MPP.Four hundred children with MPP have been enrolled in this retrospective analysis.All clinical and analytical information,including mycoplasma pneumoniae(MP)PCR results,has been collected.Based on the PCR results,the patients were divided into groups with load values(copy number)<105(54 cases),2105 and<106(71 cases),2106 and<107(112 cases),>107 and<108(114 cases),and>108(49 cases).The clinical features(including symptoms and signs)and inflammatory indicators were compared among the groups.The incidence of high fever(above 39℃),thermal peak during the entire hospitalization period,fever duration,days of hospitalization,and plasma lactate dehydrogenase(LDH)levels were statistically correlated with the MP PCR load value in children with MPP.The analysis of relevance degree showed the correlative order as a thermal peak of hospitalization>duration of fever>period of hospitalization>LDH value>C-reactive protein value.The host immune response was significantly greater in the complication group than in the non-complication group.

Mycoplasma pneumoniae Macrolide Resistance and MLVA Typing in Children in Beijing,China,in 2016:Is It Relevant?

Objective The aim of this study is to investigate the macrolide resistance rate and molecular type with multiple-locus variable-number tandem-repeat analysis(MLVA)of Mycoplasma pneumoniae of Beijing in 2016 in pediatric patients.Methods Real-time quantitative polymerase chain reaction(PCR)was used to identify M.pneumoniae,and MLVA was performed.The domain V of the 23 S rRNA was sequenced to detect macrolide-resistant point mutations.We also investigated the activities of antibiotics against M.pneumoniae isolates in vitro.Results The PCR detection rate of M.pneumoniae in children in Beijing was 40%,and the macrolide resistance rate was 66%.The A2063 G mutation in the 23 S rRNA V region is the dominant mutation(137/146,93.84%),whereas the A2064 G mutation is rare(9/146,6.16%).Seventy-three samples were typed successfully by MLVA typing,including 86.3%(63/73)were MLVA type 4-5-7-2,and 13.7%(10/73)were MLVA type 3-5-6-2.No other types were found.No strains were resistant to levofloxacin or tetracycline.Conclusion In 2016,a specific decrease in the macrolide resistance rate occurred in Beijing.The detection rate and macrolide resistance rate of outpatients are lower than those of inpatients.The A2063 G mutants M.pneumoniae have high levels of resistance to erythromycin and azithromycin.The primary MLVA type is 4-5-7-2,followed by 3-5-6-2.No other MLVA types were detected.No strains resistant to tetracycline or levofloxacin were found in vitro.