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.

Study on the Correlation between Human Papillomavirus and Mycoplasma genitalium Combined with TCT Detection

Objective: This study aims to explore the correlation between human papillomavirus (HPV) and Mycoplasma genitalium (CT) combined with TCT detection in cervical cancer screening. Method: A cross-sectional study design was adopted, and a total of 609 women who came to seek medical treatment were recruited as the study subjects. Combination testing was evaluated on cervical cancer screening by testing the women for HPV, CT with TCT detection and analyzing the relationship of cervical lesions with HPV and CT infection. Results: The study results showed that 21.57% of the subjects were infected with both HPV and CT, and 48.42% of the cases had abnormal TCT results at the same time. Further data analysis showed that HPV infection was significantly associated with abnormal TCT outcomes (p < 0.05), suggesting a possible synergistic effect of the two infections in cervical lesions. The combined sensitivity and specificity of HPV, CT and TCT detection were 21.57% and 48.42%, respectively, which were significantly higher than that of single detection. Conclusion: In summary, the results of this study support the importance of combined HPV, CT, and TCT testing in cervical cancer screening, and propose the hypothesis that combined testing may improve screening effectiveness. However, further large sample studies are needed to confirm this conclusion and explore the prospects of combined testing in clinical practice.

The Predictive Value of NLR, IL-6, CRP, and PCT in Mycoplasmal Pneumonia with Complicated Myocardial Injury

Objective:To evaluate the dynamic changes in neutrophil-to-lymphocyte ratio(NLR),interleukin-6(IL-6),C-reactive protein(CRP),and procalcitonin(PCT)levels in children with Mycoplasma pneumoniae pneumonia(MPP)complicated by myocardial injury and to determine their predictive value both individually and in combination.Methods:150 children diagnosed with MPP at Jiujiang Maternal and Child Health Hospital between June 2023 and June 2024 were selected.Patients were divided into the myocardial damage group(MD group,n=65)and the non-myocardial damage group(non-MD group,n=85),based on the presence of myocardial injury.Ninety hospitalized children without MPP served as the control group(Con group).Myocardial enzyme profile indicators,including lactate dehydrogenase(LDH),α-hydroxybutyrate dehydrogenase(α-HBDH),aspartate aminotransferase(AST),high-sensitivity cardiac troponin I(hs-cTnI),creatine kinase(CK),and creatine kinase-MB(CK-MB),were measured using a chemiluminescent immunoassay analyzer.Serum NLR,IL-6,CRP,and PCT levels were determined using appropriate analyzers.The correlation between these markers and myocardial enzyme indicators was analyzed using Spearman correlation analysis.Multivariate logistic regression was applied to identify risk factors for myocardial injury in MPP patients.Results:Serum levels of NLR,IL-6,CRP,and PCT in the MD and non-MD groups were significantly higher than in the Con group(P<0.05),with the MD group showing higher levels than the non-MD group(P<0.05).These markers were positively correlated with myocardial enzyme indicators.Logistic regression identified elevated NLR,IL-6,CRP,PCT,LDH,α-HBDH,AST,hs-cTnI,CK,and CK-MB as risk factors for myocardial injury in MPP patients(P<0.05).Conclusion:Elevated levels of NLR,IL-6,CRP,PCT,and myocardial enzymes are significant risk factors for myocardial injury in children with MPP,offering valuable insights for prevention and prognosis.

Analysis of the Role of D-Dimer,Interleukin-6,and Interleukin-18 in Differential Diagnosis of Pediatric Refractory Mycoplasma pneumoniae Pneumonia

Objective:To analyze the value of D-dimer(D-D),interleukin-6(IL-6),and IL-18 in the differential diagnosis of children with refractory Mycoplasma pneumoniae pneumonia(RMPP).Methods:The medical records of 92 children with Mycoplasma pneumoniae pneumonia(MPP)treated in the hospital were selected for retrospective analysis from January 2023 to January 2024.After comprehensive examinations such as computed tomography examination of the chest,48 children with general Mycoplasma pneumoniae pneumonia(GMPP)were put in the GMPP group and 44 children with RMPP were grouped in the RMPP group.The IL-6,IL-18,and D-D levels were compared between the two groups,and the receiver operating characteristic(ROC)curves were plotted to analyze their value for differential diagnosis of RMPP.Results:The levels of IL-6,IL-18,and D-D in the RMPP group were higher than those in the GMPP group(P<0.05);the ROC curves showed that the specificity of the differential diagnosis of IL-6,IL-18,and D-D was higher,and their diagnostic value was significant.Conclusion:Determination of IL-6,IL-18,and D-D levels in children with MPP can further diagnose the children’s condition,which can help physicians formulate targeted treatment plans,and is of great significance to the improvement of the children’s condition,which is worthy of attention.

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.

Identification and antibiotic susceptibility evaluation of Mycoplasma synoviae isolated from chickens in central China

Mycoplasma synoviae(M.synoviae)infections have become an increasingly serious concern in China because they cause huge economic losses to the poultry industry.Antibiotic treatment is one of control strategies that can be used to contain clinical outbreaks in M.synoviae-free flocks,especially because the bacteria can be transmitted through eggs.To understand M.synoviae infection status in farms of central China and the antibiotic susceptibility of the circulating strains in vivo and in vitro,485 samples were collected from five provinces from 2019 to 2021.Fifty-two strains were isolated and identified.Determination of the minimum inhibitory concentration(MIC)of eight antibiotics(tylvalosin,tiamulin,tilmicosin,lincomycin,enrofloxacin,chlortetracycline,doxycycline and tylosin)for isolates showed that tylvalosin,doxycycline and tiamulin were effective against 52 clinical isolates(MIC values≤0.0625-0.25μg/mL,≤0.0625-1μg/mL,and 0.25-2μg/mL,respectively).Tilmicosin,enrofloxacin and lincomycin had high MICo values(>32μg/mL).An artificial M.synoviae infection model was established in chickens for evaluation of the short-term therapeutic effect of these antibiotics.After 5 days of medication,doxycycline(200 mg/L)showed a superior ability to inhibit M.synoviae compared with other groups,as did tylvalosin(200 mg/L).Furthermore,the therapeutic efficacy of tylvalosin(0.4μg/mL)on intra-embryo-injected M.synoviae was higher than that of tiamulin at the same dose.A combination of MiC values determined in vitro and therapeutic effects observed in vivo revealed that tylvalosin and doxycycline had the best therapeutic effects.Tylvalosin also showed better inhibitory effects on the vertical transmis-sion of M.synoviae than tiamulin.

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.

Advances in the Detection of Mycoplasma hyopneumoniae by Polymerase Chain Reaction (PCR) Technology

Mycoplasma hyopneumoniae is an important pathogen causing Mycoplasmal pneumonia of swine, which generally causes secondary infections and mixed infections, thus seriously threats the development of swine industry and resulting in huge economic losses. Using PCR technology has very important significance to the correct diagnosis of Mycoplasmal pneumonia at the early stage. In this paper, specific target genes of Mycoplasma hyopneumoniae, methods for clinical sample collection, key technical factors of DNA sample processing method, and the research progress, main advantages and disadvantages, and application of general PCR technology, multiple PCR technology, nested-PCR technology, real-time fluorescence quantitative PCR technology, gene chip detection technology and loop-mediated isothermal amplification in detection of Mycoplasma hyopneumoniae were summarized, which provided convenience for the effective diagnosis and prevention of Mycoplasmal pneumonia of swine.

Antibody Detection on Mycoplasma capricolum subsp. Capripneumoniae in Qinghai Province

[Objective] The aim was to investigate the prevalence of Mycoplasma capricolum subsp. Capripneumoniae in Qinghai Province. [Method] By using indirect hemagglutination test kit for detecting Mycoplasma capricolum subsp. Capripneumoniae,208 goat serums were detected. [Result] The positive rate of goat sera was 16.3%,and the positive rate of sera from different regions ranged from 6.7% to 24.3%. [Conclusion] The infection rate of Mycoplasma capricolum subsp. Capripneumoniae was high in Qinghai Province,so it is necessary to strengthen the prevention and control of this disease.

Establishment of a Collection and Detection Technology of Mycoplasma hyopneumoniae in Aerosol

ObjectiveThis study was to establish a simple method for collecting and detecting Mycoplasma hyopneumoniae （Mhp） in aerosol. MethodBased on the mechanisms of liquid impinger and filtration sampler, a double concentration aerosol sampler was designed for collecting Mhp aerosol. Firstly, the collection was performed in a closed environment full of artificial aerosol of Mhp. Secondly, collection efficiency was detected by real-time PCR. Thereafter, the clinical feasibility of the designed equipment was tested by collecting aerosol samples in different pig herds. In one assay, the samples were collected at different times from one pig house challenged with Mhp. In another assay, the samples was collected from the delivery room, nursery and fattenning house of a MPS outbreak farm as well as a Mhp infection positive pig farm without obvious clinical symptoms. All the aerosol samples were then detected by real-time PCR or nested PCR. ResultThe collection efficiency of the designed bioaerosol sampler was （37.04±6.43） %, Mhp could be detected 7 d after intratracheal challenge with pneumonic lung homogenate suspension. Aerosol samples of 11 pig houses from the two Mhp positive pig farms with or without clinical symptoms all showed a positive result of PCR, the positivity rate was 100%. ConclusionA high sensitive collecting and detecting technology of aerosol was successfully established, which can be applied to clinical detection of Mhp in aerosol.

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 paraffin embedded 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 (chi(2) = 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 (chi(2)=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.

Secretory Expression of Mycoplasma hyopneu-moniae P97R1 Gene in Pichia pastoris and Primary Application of the Expression Product

[Objective] This study aimed to investigate the secretory expression of P97R1 gene of Mycoplasma hyopneumoniae（Mhp） in Pichia pastoris expression system and the primary application; of the expression product. [Method] A pair of specific primers was designed to conduct PCR according to the Mhp P97R1 gene sequence in Genbank, and the amplified P97R1 gene was cloned into the pPICZa-A yeast expression vector to construct the secretory recombinant expression vector pPICZa-A-P97R1. The plasmid pPICZa-A-p97R1 linearized by Sac I was transformed into P. pastoris GSl15 by electroporation. Positive transformant identified by PCR was incubated to express P97R1 protein after methanol induction. And the expression product was identified using SDS-PAGE and Western-blotting anal.wsis. [Result] P97R1 protein was successfully expressed in the P. pastoris system, with a secre- tory amount of 499μg/ml, and revealed good reactogenicity. Meanwhile, an indirect ELISA method was established with P97R1 protein after the optimization of each reaction factor, which showed good specificity and repeatability according to repeated tests. [Conclusion] This study provides bases for developing the ELISA Kit for anti- body detection and genetically engineered vaccine to Mhp.

In vitro Antimicrobial Activity of Golden Buckwheat Extract and Therapeutic Effect of its Preparation on Chicken Mycoplasma Infection

[Objective] This study aimed to identify the in vitro antibacterial activity of golden buckwheat extract and investigate the therapeutic effect of its preparation on mycoplasma infection. [Method] Through measuring the minimum inhibitory concentra-tion, the in vitro antibacterial activity of golden buckwheat water extract was deter-mined; meanwhile, the therapeutic effect of golden buckwheat oral solution on my-coplasma infection was determined by artificial y infecting chickens with Mycoplasma gal isepticum culture. [Results] The golden buckwheat water extract had obvious in-hibitory effects against Pseudomonas aeruginosa and Escherichia coli, and a certain inhibitory effect on Salmonel a and Staphylococcus aureus; administration of golden buckwheat oral solution at the dose of 0.5%-1.0% continuously for 5 d had a good therapeutic effect against mycoplasma infection. [Conclusion] The study provides sci-entific bases for further study on the antibacterial activity of golden buckwheat and its application.

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。