Molecular isolation and identification of Mycobacterium avium subsp.hominissuis in Didelphis virginiana from Hidalgo,Mexico

Objective:To isolate and identify the exact species of the genus Mycobacterium from Didelphis(D.)virginiana,and the direct implications of this bacterium to public health and veterinary medicine.Methods:Thirty-one D.virginiana were captured and necropsied in Hidalgo,Mexico.Tissue samples were collected to culture mycobacteria present and examine individual specimens’histopathology.Mycobacterium identification was obtained through the application of amplification and sequencing of 16S rDNA techniques.Results:Three strains were isolated and identified as Mycobacterium(M.)avium subsp.hominissuis by utilizing M.avium complexspecific primers.Granulomatous lesions were observed in the subpleural zone(granuloma gradeⅡ)and bronchial(granuloma gradeⅠ)of the lungs of D.virginiana with positive isolation.Conclusions:Three strains of M.avium subsp.hominissuis,from lung tissue samples of D.virginiana were identified.This subspecies of M.avium has important implications in public health and veterinary medicine.

High prevalence of viable Mycobacterium avium subspecies paratuberculosis in Crohn's disease

AIM:To examine the detection rate of viable Mycobacterium avium subspecies paratuberculosis(MAP) in patients with inflammatory bowel disease [Crohn's disease(CD) and ulcerative colitis(UC)].METHODS:Thirty patients with CD(15 with at least one NOD2/CARD15 mutation),29 with UC,and 10 with no inflammatory bowel disease(IBD).were tested for MAP by polymerase chain reaction(specific IS900 fragment) and blood culture.RESULTS:MAP DNA was detected in all original blood samples and 8-wk blood cultures(CD,UC and non-IBD).Positive MAP DNA status was confirmed by dot blot assays.All 69 cultures were negative by acid-fast Ziehl-Neelsen staining.Viable MAP,in spheroplast form,was isolated from the 18-mo blood cultures of all 30 CD patients,one UC patient,and none of the non-IBD controls.No association was found between positive MAP cultures and use of immunosuppressive drugs or CDassociated single nucleotide polymorphisms.CONCLUSION:MAP is widely present in our area and MAP DNA can be recovered from the blood of CD,UC and non-IBD patients.However,MAP spheroplasts were only found in CD patients.

Mycobacterium avium subspecies paratuberculosis and its relationship with Crohn's disease

The hypothesis postulating that Mycobacterium avium paratuberculosis(MAP) is the cause of Crohn's disease(CD) has been circulating for many years.Advances in molecular techniques,such as polymerase chain reaction and culture methods,have enabled researchers to demonstrate that there is an association between MAP and CD.Recently,genome-wide association studies have identified novel susceptibility genes for CD,which are critical for generation of an adaptive immune response that is protective against intracellular pathogens,including M.tuberculosis infection.However,the role of MAP as a cause of CD suffered a setback with the report that administration of antimycobacterial therapy failed to lead to a sustained response in CD patients.Accordingly,this review sought neither to confirm nor refute this,but instead to survey recent literature on the role of MAP in CD.

Relationship between Crohn’s disease, infection with Mycobacterium avium subspecies paratuberculosis and SLC11A1 gene polymorphisms in Sardinian patients

AIM: To study the association between Crohn’s disease (CD), Mycobacterium avium subspecies paratubercu- losis (MAP), and genetic factors by examining the role of natural resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms (now SLC11A1) in Sar dinian patients with CD and controls. METHODS: Thirty-seven CD patients and 34 controls with no inflammatory bowel disease (IBD) were recruited at the University of Sassari after giving written con sent. Six SCL11A1 polymorphisms previously reported to be the most significantly associated with IBD were searched. M. paratuberculosis was identified by IS900 PCR and sequencing. Logistic regression was used to cal culate odds ratios (OR) for the associations among CD presence of MAP, and 6 loci described above. RESULTS: For the first time, a strong association was observed between polymorphisms at NRAMP1 locus 823C/T and CD. While CD was strongly associated withboth NRAMP1 and MAP, NRAMP1 polymorphisms and MAP themselves were not correlated. CONCLUSION: Combined with previous work on the NOD2/CARD15 gene, it is clear that the interplay of ge- netic, infectious, and immunologic factors in the etiology of CD is complex.

On deaf ears, Mycobacterium avium paratuberculosis in pathogenesis Crohn's and other diseases

The historic suggestion that Mycobacterium avium subsp. paratuberculosis(Map) might be a zoonotic pathogen was based on the apparent similarity of lesions in the intestine of patients with Crohn's disease(CD) with those present in cattle infected with Map, the etiological agent of Johne's disease. Reluctance to fully explore this possibility has been attributed to the difficulty in demonstrating the presence of Map in tissues from patients with CD. Advances in technology have resolved this problem and revealed the presence of Map in a significant proportion of patients with CD and other diseases. The seminal finding from recent investigations, however, is the detection of Map in healthy individuals with no clinical signs of disease. The latter observation indicates all humans are susceptible to infection with Map and lends support to the thesis that Map is zoonotic, with a latent stage of infection similar to tuberculosis, where infection leads to the development of an immune response that controls but does not eliminate the pathogen. This clarifies one of the reasons why it has been so difficult to document that Map is zoonotic and associated with the pathogenesis of CD and other diseases. As discussed in the present review, a better understanding of the immune response to Map is needed to determine how infection is usually kept under immune control during the latent stage of infection and elucidate the triggering events that lead to disease progression in the natural host and pathogenesis of CD and immune related diseases in humans.

The relative frequency of Mycobacterium tuberculosis and Mycobacterium avium infections in HIV positive patients,Ahvaz,Iran

Objective:To estimate the prevalence of Mycobacterium tuberculosis(M.tuberculosis) and Mycobacterium avium(M.avium) infections in HIV -positive patients suspected to have puimonar) and extrapulmonary mycobacterial co-infection using PCR technique.Methods: Totally 50 samples comprising sputum,pleural fluid and CSF taken from HIV positive patients suspected to have mycobacterial infection,were processed.The demographic information and results of acid fast staining and culture were recorded for each patient.The PCR for detecting of M.tuberculosis comprised of specific primers targeting IS6110 gene sequence.For detecting of M.avium,PCR with primers that amplifies the mig gene were used.Results:From 50 samples processed,45 were sputum(90%),3 pleural fluid(6%) and 2 CSK(4%).In total.8(16%) were culture positive,7 had positive acid fast staining 114%) and 13 samples(26%)were positive using PCR technique.All the positive samples were sputum and belonged to patients with pulmonary infection.01 these.9 were positive for M.tuberculosis(69.2%) and 4 were identified as 1/.avium (30.8%).which 2 out of 13 positive samples showed mixed infections by both mycobacteria. Conclusions:The PCR shows the highest detection rate(26%) of mycobacteria compared with culture and acid last staining.The majority of infections were with M.tuberculosis(18%) and this shows the importance of this mycobacterial co-infection in HIV positive patients in the region of study.

Mycobacterium avium subspecies paratuberculosis infects and multiplies in enteric glial cells

AIM: To establish the role of enteric glial cells during infection with Mycobacterium avium subspecies paratuberculosis (MAP) in Crohn's disease. METHODS: In order to establish the role of enteric glial cells during infection with M. avium subspecies paratuberculosis (MAP) in Crohn's disease, Map adhesion experiments on enteric glial cells were performed as well as expression analysis of Map sigma factors during infection. RESULTS: In this study, for the first time, we found a high affinity of MAP to enteric glial cells and we analyzed the expression of MAP sigma factors under different conditions of growth. CONCLUSION: The fact that Map showed a high affinity to the glial cells raises concerns about the complicated etiology of the Crohn's disease. Elucidation of the mechanisms whereby inflammation alters enteric neural control of gut functions may lead to novel treatments for Crohn's disease.

Mycobacterium avium subspecies paratuberculosis in the causation of Crohn's disease

Mycobacterium avium subspecies paratuberculosis(MAP),originally called Johne’s bacillus was firstdescribed from Germany in 1895 as the cause of achronic inflammatory disease of the intestine in acow.As the 20th century progressed,clinical andsub-clinical MAP infection in farm animals inWestern Europe appeared to become moreprevalent.Among the early reviews available

Mycobacterium avium subspecies paratuberculosis in the etiology of Crohn's disease, cause or epiphenomenon?

The origin of inflammatory bowel disease is unknown. Attempts have been made to isolate a microorganism that could explain the onset of inflammation, but no pathological agent has ever been identified. Johne's disease is a granulomatous chronic enteritis of cattle and sheep caused by Mycobacterium avium subspecies paratuberculosis(MAP) and shows some analogies with Crohn's disease(CD). Several studies have tried to clarify if MAP has a role in the etiology of CD. The present article provides an overview of the evidence in favor and against the "MAP-hypothesis", analyzing the methods commonly adopted to detect MAP and the role of antimycobacterial therapy in patients with inflammatory bowel disease. Studies were identified through the electronic database, MEDLINE, and were selected based on their relevance to the objective of the review. The presence of MAP was investigated using multiple diagnostic methods for MAP detection and in different tissue samples from patients affected by CD or ulcerative colitis and in healthy controls. On the basis of their studies, several authors support a close relationship between MAP and CD. Although increasing evidence of MAP detection in CD patients is unquestionable, a clear etiological link still needs to be proven.

Pulmonary <i>Mycobacterium avium </i>Complex Disease Requiring Differentiation from Recurrence of Lung Cancer during the Follow-Up Period for Lung Cancer

Case 1 was a 49-year-old woman who visited with a dry cough. She had an underlying disease of lung adenocarcinoma and received cancer immunotherapy because of an ALK-positive response and several cancer chemotherapies. The clinical effect was a complete response. Chest CT was performed because of continuous dry cough, and a new tumor shadow was recognized in the lingula portion of the left upper lobe. We performed CT-guided lung biopsy and could aspirate pus-fluid. The culture test for acid-fast bacilli was positive and the causative microorganism was identified as Mycobacterium avium by the DDH method. The final diagnosis was pulmonary abscess due to M. avium. Treatment using combined chemotherapy including CAM was performed and a good clinical response was obtained. Case 2 was a 67-year-old man who had a past history of surgical resection of lung adenocarcinoma eight and two years ago and received several cancer chemotherapies and radiation therapy. Because a new nodular shadow appeared in the right middle lobe one year ago and showed strong positivity on PET/CT, surgical resection was performed with the suspected recurrence of lung cancer. Subsequently, the histological diagnosis was epithelioid granuloma and a culture test of acid-fast bacilli was positive, with the identification of Mycobacterium intracellulare by the DDH method. Combined chemotherapy was not performed because the lesion was completely resected. Afterwards, a new nodular shadow appeared in the left lower lobe again and bronchoscopy was performed. Because M. intracellulare was isolated from the local specimen, we diagnosed the patient with recurrence of pulmonary MAC disease and combined chemotherapy including CAM was performed for one year. Finally, the nodular lesion disappeared. It is difficult to differentiate pulmonary MAC disease from lung cancer. Therefore, careful follow-up of patients with lung cancer while keeping in mind the possible complication of pulmonary MAC disease is necessary.

Mycobacterium avium Paratuberculosis (MAP) and Cytomegalovirus (CMV) Are Frequently Detected in the Saliva of Patients Recently Diagnosed with Crohn Disease (CD) Whereas Oral Propionibacterium Acnes (PA) or Methylacetate (MA) in Their Breath Is Rare

We investigated the presence of MAP, CMV, or Epstein-Barr virus (EBV) in the saliva of patients presenting with periodontal disease, gingivitis or oral aphthosis. We also measured methylacetate in their breath and looked for the presence of PA on their tongue. All patients were prospectively enrolled into a two-year cohort study. We compared the group presenting with CD versus the group without CD. 167 patients were enrolled. 24 patients presented with CD. CMV and MAP were found concomitantly in 67% of CD patients (17 patients) whereas only 1 patient (<0.1%;p < 0.001) has these infectious agents in the comparative group. PA was rare in CD (25% versus 70.6%;p < 0.001). MA was low, especially with an empty stomach (0.59 ppm +/? 1.45 versus 5.14 ppm +/? 8.97;p < 0.001). It was concluded that the association MAP + CMV was frequently found in oral lesions of CD at an early stage. Detection of PA, MAP and CMV, as well as breath test could be easily performed and may participate to the early diagnosis of CD.

Concurrent pulmonary Mycobacterium avium complex (MAC) infection and active Hürthle cell thyroid carcinoma: is there a connection?

We present two cases of pulmonary MAC infection in women with Hürthle cell thyroid carcinoma. Both cases were asymptomatic octogenarian women with active Hürthle cell thyroid carcinoma and prolonged periods of hypothyroidism prior to diagnosis of pulmonary MAC. Mycobacterium avium complex has never been reported in association with any type of thyroid cancer, specifically Hürthle cell carcinoma. A review of the literature and possible associations between the two are discussed in this article.

The 19 kDa Protein from <i>Mycobacterium avium subspecies paratuberculosis</i>Is a Glycolipoprotein

This study characterizes the 19 kDa protein expressed by Mycobacterium avium subspecies paratuberculosis (MAP) as a glycolipoprotein, providing the foundation for future experiments regarding its antigenicity and role in disease pathogenicity. We have previously shown that a 4.8 kb insert from MAP will produce a 16 kDa recombinant protein when expressed in Escherichia coli and 19 kDa recombinant protein when expressed in M. smegmatis (smeg19K). The difference of 3 kDa in size of these expressed proteins may be related to post translational modifications that occur in Mycobacterium species. We hypothesized that smeg19K is a glycolipoprotein since BLAST analysis revealed approximately 76% amino acid identity between the MAP 19 kDa protein and a known lipoglycoprotein, the 19 kDa protein of M. tuberculosis. This prediction was confirmed by the following positive staining of smeg19K with Sudan Black 4B, a postelectrophoresis dye used to stain for lipids. Smeg19K has also stained positively for glycosylation with the lectin concavalin A, a highly specific stain for mannose residues. As expected, treatment with tunicamycin (an antibiotic known to inhibit N-glycosylation) and treatment with deglycosylation assay (non-specific for mannose), showed no reduction in size of 19 kDa glycolipoprotein.

牛副结核病的病原学和防控研究进展

牛副结核病是一种由副结核分枝杆菌引起的主要影响反刍动物的慢性消耗性疾病,也是一种潜在的人兽共患病。该病可导致腹泻、渐进性消瘦,甚至死亡,增加生产成本,是影响全球奶牛业最严重的传染病之一。牛副结核病难以诊断,病原分离周期长,我国相关研究报道较少且存在误区。因为潜伏期长,患病动物在感染早期体液免疫水平低,排菌量低,导致细菌分离培养和抗体检测都很难做出诊断;即使最灵敏的实时荧光定量聚合酶链式反应(qPCR)检测方法也只能在部分感染动物的粪便样品中检出副结核分枝杆菌;皮肤变态反应试验和γ干扰素释放试验等方法易受其他结核分枝杆菌的影响,也难以普及。本文旨在提炼性的阐述牛副结核病的病原学研究进展,总结其流行病学和防控策略等信息,以期为该病的预防和控制工作提供参考。

副结核分枝杆菌免疫原蛋白的筛选及免疫保护效果评价

【背景】副结核病(paratuberculosis,PTB)是由副结核分枝杆菌(Mycobacterium avium subsp.paratuberculosis,MAP)主要引起反刍动物的一种慢性消耗性传染病。患病动物表现为肉芽肿性肠炎和肠道的功能失调、顽固性腹泻、渐进性消瘦、营养不良、贫血、嗜睡甚至死亡。PTB给畜牧业造成巨大的经济损失,并且严重威胁公共卫生安全。由于目前临床上对于PTB的检测和控制手段不够完善,现有的PTB疫苗保护效果不佳,并且干扰牛结核病的诊断,因此亟需研发免疫原性强、保护效果佳的疫苗以用于PTB的防控。【目的】通过筛选MAP免疫原蛋白,并对其免疫保护效果进行评价,为PTB的防控提供数据支持。【方法】根据MAP的p22、map1272c、map3531c、map3783、map3701c以及map3527这6个基因,分别构建5种重组质粒,在获得5种重组蛋白基础上,联合MONTANIDE ISA 61 VG佐剂皮下多点注射免疫小鼠,通过IFN-γELISPOT试验筛选出最佳免疫原;随后将最佳免疫原和已报道的66NC融合蛋白混合,经皮下多点注射免疫小鼠,在二免3周后用1×10^(8)CFU的MAP K-10菌株腹腔感染小鼠。通过IFN-γELISPOT试验,抗体监测和细胞因子检测,分析感染后小鼠的体重、肝脏病理学和组织病理学变化及其荷菌数差异,综合评价候选亚单位疫苗的免疫原性和免疫保护效果。【结果】以MAP p22、map1272c、map3531c、map3783以及map3701c为基础成功构建和表达了5种融合蛋白58F、62F、69F、46F和52F,其中免疫58F后产生的IFN-γ水平最高,是更具潜力的候选免疫原,且以融合蛋白组合66NC+58F诱导持久高滴度的IgG、IgM、IgG1和IgG2a水平,诱导特异性的IFN-γ、TNF-α和IL-17A的释放;在保护效果评价中,融合蛋白组合66NC+58F抵抗MAP感染造成的体重下降,显著减轻肝脏的病理损伤,降低MAP在肝脏中的定植。【结论】融合蛋白组合66NC+58F诱导小鼠产生Th1和Th17型免疫反应,对MAP感染有着一定免疫保护作用,是PTB重要的候选亚单位疫苗。

副结核分支杆菌实时荧光定量PCR检测方法的建立及应用

为加强副结核分支杆菌(MAP)的检测、监测与防控,建立MAP荧光定量PCR(qPCR)检测方法,根据国内MAP流行菌株特异性插入序列IS900设计1对特异性引物和探针,构建重组阳性质粒用作建立qPCR的模板,优化反应体系和条件,验证该方法的特异性、敏感性和重复性。结果显示,该方法最低检测限为5拷贝/μL;重复试验中批内和批间变异系数均小于4%;与牛传染性鼻气管炎病毒、牛副流感病毒3型、牛轮状病毒、牛病毒性腹泻病毒、牛支原体、肺炎克雷伯菌和曼氏杆菌等病原检测无交叉反应,能特异性检出MAP。应用建立的qPCR开展了内蒙古自治区6个市的MAP临床样品检测和流行情况分析,结果显示MAP平均阳性率为0.85%(6/704)。结果表明,成功建立了MAP的实时荧光定量PCR检测方法,可用于临床中MAP的监测和调查,为MAP疾病诊断与监控提供了快捷的方法。

艾滋病合并鸟分枝杆菌感染患者临床特征初步分析

目的分析艾滋病合并鸟分枝杆菌感染患者的临床特征并与艾滋病合并其他非结核分枝杆菌(nontuberculous mycobacteria,NTM)感染患者对比,为临床诊断与治疗鸟分枝杆菌病提供实验室数据支撑。方法回顾性分析2016年4月—2022年6月云南省传染病医院确诊的43例艾滋病合并鸟分枝杆菌感染患者(鸟分枝杆菌感染组)及15例艾滋病合并其他类型NTM感染患者(其他种类NTM感染组)的临床资料,包括人口学特征、临床症状、影像学特征、实验室检测指标、治疗方案及预后情况等。结果鸟分枝杆菌感染组主要临床症状为发热、咳嗽咳痰、乏力并伴体质量下降,占60.47%,其他种类NTM感染组则以发热、咳嗽咳痰、乏力为主(86.67%)。2组患者均以肺部条索状、片状、结节状影伴淋巴结肿大为主,分别占51.16%和53.33%。鸟分枝杆菌感染组不同CD4^(+)T细胞计数分组实验室指标整体无统计学差异,但其中CD4^(+)T细胞101~200个/μL组与CD4^(+)T细胞<50个/μL组患者白细胞计数与白细胞介素-6水平相比,差异均有统计学意义(H=10.316,H=16.130;P均<0.05)。其他种类NTM感染组患者与鸟分枝杆菌感染组患者相比,整体各项实验室指标更接近正常值范围,不同CD4^(+)T细胞计数与实验室指标间差异无统计学意义(P均>0.05)。58例患者中35例患者经治疗后好转出院,20例患者行抗鸟分枝杆菌治疗后病情依然严重但放弃治疗,3例患者死亡。鸟分枝杆菌感染组患者与其他种类NTM感染组患者预后相比,差异无统计学意义(P=0.890)。结论结合艾滋病合并感染NTM患者人口学特征、CD4^(+)T水平、胸部影像学特征以及临床症状,可辅助临床快速识别和诊断鸟分枝杆菌病,为患者的临床治疗及预后提供数据支撑。

Specific Antigens to Distinguish <i>M. tuberculosis</i>from <i>M. avium</i>

To distinguish Mycobacterium tuberculosis from Mycobacterium avium, specific M. tuberculosis antigens had been studied for improving the early differential diagnosis effect of tuberculosis caused by different Mycobacterium. The rabbit anti-M. avium sera and anti-M. tuberculosis sera were analyzed for antibody-based reactivity by matrix-assisted laser desorption-ionization mass spectrometry (MALDI-TOF Mass) against M. tuberculosis proteins. The immunoreactive spots, which were attributed to the proteins HspX, GroES and CFP-10, were mostly located at 10 - 60 kDa and PI 4 - 6, subsequently Western blotting result proved that HspX and CFP-10 were specific to M. tuberculosis and ELISA testing result of 30 M. avium positive sera showed that GroES were cross-reactive to M. avium. Lastly, positive and negative tuberculosis reference sera and based on the mechanism of indirect ELISA, the specificity and the sensitivity of the methods targeting the antibodies HspX, GroES or CFP-10 were evaluated at 37% and 26%, 12% and 97%, 81% and 98%, respectively. The combination of these three antibody detection methods allowed to reached a specificity of 42%, and of 39% without taken into account of the method targeting the GroES antibody. Using proteomics approach, we found three M. tuberculosis specific antigens showed good potential in tuberculosis diagnosis, providing basic study for serodiagnosis of tuberculosis.

禽结核菌素生产用菌种的制备与鉴定

为制备禽结核菌素国家标准品,研究禽结核菌素生产用菌株禽分枝杆菌的菌种特性,对1992年冻干保存的3株菌CVCC68201、CVCC68202和CVCC68203进行复苏,采用SPF鸡复壮,分离培养后冻干了新批次菌种,对菌种形态、生化特性、培养特性、毒力和抗原性进行了鉴定。结果表明,制备的菌种纯粹,形态、生化和培养特性符合禽分枝杆菌的生物学特征,免疫原性符合的兽用生物制品质量标准的规定。本研究为禽分枝杆菌的鉴定提供参考,也为制造禽结核菌素的工艺改进奠定基础。

获得性免疫缺陷综合征合并分枝杆菌感染患者的免疫学特征与耐药性分析

目的:分析获得性免疫缺陷综合征(acquired immunodeficiency syndrome,AIDS)患者合并结核分枝杆菌(Mycobacterium tuberculosis,MTB)或非结核分枝杆菌(non-tuberculous mycobacteria,NTM)感染的免疫学特征及其对常用抗结核药物的耐药特点,为临床AIDS患者并发MTB或NTM感染的诊断和治疗提供参考。方法:选取2017年1月—2022年12月苏州市第五人民医院收治的44例AIDS合并MTB感染患者(设为AIDS/MTB感染组)和44例AIDS合并NTM感染患者(设为AIDS/NTB感染组)作为研究对象,采集患者的年龄、性别、免疫学检查结果,以及MTB和NTB对常用抗结核药物的药敏试验结果等信息,分析AIDS患者合并MTB或NTM感染的免疫学特征与耐药特点。结果:AIDS/MTB感染组和AIDS/NTB感染组患者的性别均以男性为主(93.18%vs97.73%),而年龄上均主要集中在30~<50岁(56.82%vs 59.09%),但2组患者在性别和年龄上的分布情况经比较其差异无统计学意义(P>0.05);AIDS/NTB感染组患者的T-SPOT检查均为阴性,AIDS/MTB感染组患者T-SPOT检查的阳性率较高;AIDS/MTB感染组患者的CD4^(+)T细胞绝对值明显高于AIDS/NTB感染组(108.4±76.7 vs36.5±22.1,P<0.05);AIDS/NTB感染组患者中检出的主要病原菌为鸟分枝杆菌和堪萨斯分枝杆菌,其对链霉素、异烟肼、利福平、乙胺丁醇、对氨基水杨酸、阿米卡星的耐药率均大于50.00%(远高于MTB对其的耐药率),仅对丙硫异烟胺的耐药率较低(<5.00%)。结论:AIDS患者并发MTB或NTM感染好发于30~<50岁的男性,T-SPOT检查和CD4^(+)T细胞水平可以较好地对MTB感染和NTM感染进行鉴别诊断;此外,NTM感染的病原菌对多数抗结核药物耐药性较强,应尽量选用其他敏感的抗菌药物进行治疗,以保证对患者的治疗效果。