吉林省孢子丝菌病的特点及致病菌毒力的研究
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摘要
申克孢子丝菌(Sporothrix schenckii)是一种条件致病菌,广泛存在于自然界中。孢子丝菌病(sporotrichosis)是由申克孢子丝菌感染引起的一种慢性肉芽肿性疾病。临床上孢子丝菌病分为固定型、淋巴管型和皮肤播散型,在吉林省则以固定型比较常见。申克孢子丝菌感染发病的机制不仅与机体免疫状态有关,而且与不同菌株的致病毒力有关。但有关吉林省孢子丝菌病的特点及致病菌毒力尚未进行深入研究。
本研究通过流行病学调查、患者及实验小鼠血清NO含量测定、不同型别菌株基因型鉴定及致病真菌超微结构观察,分析申克孢子丝菌的致病特点,旨在探讨吉林省孢子丝菌病发病的原因及不同型别菌株的毒力。
研究结果表明,吉林省为孢子丝菌病高发区,冬、春季多发;女性发病多于男性,农民为易感人群,从2004年起发病率明显增加。面部、双手、四肢等暴露部位多见;临床表现固定型多于淋巴管型,皮肤播散型少见。各型孢子丝菌病患者治疗后的血清NO含量均较治疗前明显降低;淋巴管型患者血清NO含量明显高于固定型,固定型和淋巴管型均明显高于正常对照组;在实验小鼠免疫状态和菌株培养时间相同的情况下,血清NO含量与菌株的致病力呈正相关;同一菌株致病力的大小与培养时间有关,培养7 d菌株的致病力高于培养14 d的菌株;吉林省申克孢子丝菌线粒体DNA(mtDNA)分型以4型为主。扫描电镜观察显示,三型菌株孢子表面结构存在差异。
本研究主要创新点是,在省级范围内进行大规模流行病学调查;孢子丝菌病患者治疗后血清NO含量降低,淋巴管型患者血清NO含量明显高于固定型;感染小鼠血清NO含量与菌株致病力呈正相关;培养7 d菌株的毒力高于培养14 d的菌株;吉林省申克孢子丝菌线粒体DNA分型具有一定特点;不同型别菌株超微结构上存在差异。
Sporotrichosis, caused by Sporothrix schenckii infection, is a chronic histomycosis. The Sporothrix Shenckii is widespread in nature, they can be contaminated in soil and Fumu, Bacillus subtilis, reed, corn stalks and other plant corruption. This fungi not only violates the skin, but can cause harm to visceral organs. People who have minor injury are easily infected. Farmers, masonry, paper mill workers, mine workers and gardeners are the susceptible population.
At present, it is much seen in the northern area of China, and Jilin Province has high incidence rate. Now, with the irregular application of broad-spectrum antibiotics and widely use of corticosteroids and immunosuppressive drugs in the field of medicine, some skin lesion lack of typical syndrome is easily misdiagnosed or missed. Outside the Skin tissue, it is difficult to get early diagnosis, delaying treatment time often has serious consequences. Thus the early diagnosis of it is critical importance to its treatment and prognosis.
To further understanding the epidemiological characteristics of the disease and its treatment and prevention, this research selects 2863 cases with out-patient and in-patient treatment from Dermatology of Jilin Province Now, only 2863 cases belong to Jilin province would be retrospectively analysed and made conclusion. Research will be carried out with classification and mutual-control study. All materials are analysed and prossed with statistical methods.
Sporothrix infection can cause skin, lymphatics ,skin tissues and other organs involved. The clinical symptoms are mainly associated with the host's immune status, also with strain virulence and infection pathway. In recent years, studies have shown that the immune mechanism of Sporothrix infection is mainly related to cellular immunity by activation of T cell and macrophages, while NO is the main product released from macrophages. Study shows that NO are defensive factor against Sporothrix . it is ofen seen that the NO values develop dynamic change in a lot of infectious diseases. So the changes in NO concentration can indirectly reflect the post-infection immune mechanism progress status.
The classification of fungi is a huge system engineering . The traditional fungi toxicitology depends on the morphological , physiobiochemistry and antigen construct feature. However , since there are so many species of fungi and obvious individual polymorphisms that it can not reflect the entity characteristics of fungi. Our research intends to identify the strains of different types of Sporothrix Schenckii and to find a simple and rapid method to identification of Sporothrix Schenckii, which includes the identification of mitochondria DNA types, ribosomal gene ITS/NTS fragments, PCR and DNA consequence information in GENEBANK.
The morphological characters of different types of Sporothrix Schenckii can not be distinguished by optical microscope. Otherwise, we can explain the relationship between morphology and different clinical types by ultrastructures.
In the recent thirty years, with the rapid development of molecular biology, fungal taxonomy has entered the new era from phenotype to genetype.
The main results of this study are as follows:
1.Jilin Province was a high incidence area of sporotrichosis, especially in the areas of Jiutai, Nongan and Tongyu. Female was more than male, particularly for the woman workers. It was the most easily infective mass from 40 to 60, especially the farmers. Exposed parts such as face ,hands and limbs were more seen. The incidence was higher in winter and spring, which obviously increased the occurrence trend from 2004. The fixed type sporochosis was more than lymphatic vessel-type, and cutaneous disseminated type was rarely seen. It was the main method for prevention by promoting the recycle mechanism of corn stalks and reed, while adding iodine for areas lack of iodine.
2.The NO contents were much different in thr three types of sporotrichosis: lymphatic vessel-type was higher than that in fixed type, and fixed type was more than control group. The NO contents for all types before treatment were higher than that after treatment. Thus serum NO levels could be used as a biochemical indicator for mornitoring the infection of sporotrichosis, which had a positive correlation with the virulence of Sporothrix schenckii with the same immunological status and cultivating time of mice. So the virulence of Sporothrix schenckii with the same immunological status could be evaluated. The virulence had a correlation with cultivating time for the same strain, and the virulence of the strain cultivated for 7days was higher than that of the strain cultivated for 14 doys.
3.The strains of three types of sporotrichosis could not be recognized with routine fungal morphological methods. The mRNA of strains of Sporothrix schenckii in Jilin Province consists of type 4, 20 and 24, and nearly 90% of which was type 4. The ITS zone of Sporothrix schenckii was highly conservative, which of types was completely the same. So the recognization and fast diagnosis in genes by measuring ITS zone could made. Meanwhile, the NTS zone of cutaneous disseminated type was the same with the other two types.
4.There were some difference on the surface of spores for the three types by using scanning electron microscope, and nebular adhesion on the spore surface of lymphatic vessel-type and cutaneous disseminated type was found, which causes the increased virulence. The constructions of spores and hyphae were the same for three types by using transmission electron microscope. So the virulence of Sporothrix schenckii was depended on the materials on the surface of cells.
The creative points of the present study are: it is the first epidemiological study of sporotrichosis in Jilin Provence and so far, no report had been published for the measuring of NO between before and after treatment and serum NO of immunosuppressed mice for 4w. Meanwhile, no report has been published for the research of mitochondria DNA type or the difference of ultrastructure among all types.
本研究通过流行病学调查、患者及实验小鼠血清NO含量测定、不同型别菌株基因型鉴定及致病真菌超微结构观察,分析申克孢子丝菌的致病特点,旨在探讨吉林省孢子丝菌病发病的原因及不同型别菌株的毒力。
研究结果表明,吉林省为孢子丝菌病高发区,冬、春季多发;女性发病多于男性,农民为易感人群,从2004年起发病率明显增加。面部、双手、四肢等暴露部位多见;临床表现固定型多于淋巴管型,皮肤播散型少见。各型孢子丝菌病患者治疗后的血清NO含量均较治疗前明显降低;淋巴管型患者血清NO含量明显高于固定型,固定型和淋巴管型均明显高于正常对照组;在实验小鼠免疫状态和菌株培养时间相同的情况下,血清NO含量与菌株的致病力呈正相关;同一菌株致病力的大小与培养时间有关,培养7 d菌株的致病力高于培养14 d的菌株;吉林省申克孢子丝菌线粒体DNA(mtDNA)分型以4型为主。扫描电镜观察显示,三型菌株孢子表面结构存在差异。
本研究主要创新点是,在省级范围内进行大规模流行病学调查;孢子丝菌病患者治疗后血清NO含量降低,淋巴管型患者血清NO含量明显高于固定型;感染小鼠血清NO含量与菌株致病力呈正相关;培养7 d菌株的毒力高于培养14 d的菌株;吉林省申克孢子丝菌线粒体DNA分型具有一定特点;不同型别菌株超微结构上存在差异。
Sporotrichosis, caused by Sporothrix schenckii infection, is a chronic histomycosis. The Sporothrix Shenckii is widespread in nature, they can be contaminated in soil and Fumu, Bacillus subtilis, reed, corn stalks and other plant corruption. This fungi not only violates the skin, but can cause harm to visceral organs. People who have minor injury are easily infected. Farmers, masonry, paper mill workers, mine workers and gardeners are the susceptible population.
At present, it is much seen in the northern area of China, and Jilin Province has high incidence rate. Now, with the irregular application of broad-spectrum antibiotics and widely use of corticosteroids and immunosuppressive drugs in the field of medicine, some skin lesion lack of typical syndrome is easily misdiagnosed or missed. Outside the Skin tissue, it is difficult to get early diagnosis, delaying treatment time often has serious consequences. Thus the early diagnosis of it is critical importance to its treatment and prognosis.
To further understanding the epidemiological characteristics of the disease and its treatment and prevention, this research selects 2863 cases with out-patient and in-patient treatment from Dermatology of Jilin Province Now, only 2863 cases belong to Jilin province would be retrospectively analysed and made conclusion. Research will be carried out with classification and mutual-control study. All materials are analysed and prossed with statistical methods.
Sporothrix infection can cause skin, lymphatics ,skin tissues and other organs involved. The clinical symptoms are mainly associated with the host's immune status, also with strain virulence and infection pathway. In recent years, studies have shown that the immune mechanism of Sporothrix infection is mainly related to cellular immunity by activation of T cell and macrophages, while NO is the main product released from macrophages. Study shows that NO are defensive factor against Sporothrix . it is ofen seen that the NO values develop dynamic change in a lot of infectious diseases. So the changes in NO concentration can indirectly reflect the post-infection immune mechanism progress status.
The classification of fungi is a huge system engineering . The traditional fungi toxicitology depends on the morphological , physiobiochemistry and antigen construct feature. However , since there are so many species of fungi and obvious individual polymorphisms that it can not reflect the entity characteristics of fungi. Our research intends to identify the strains of different types of Sporothrix Schenckii and to find a simple and rapid method to identification of Sporothrix Schenckii, which includes the identification of mitochondria DNA types, ribosomal gene ITS/NTS fragments, PCR and DNA consequence information in GENEBANK.
The morphological characters of different types of Sporothrix Schenckii can not be distinguished by optical microscope. Otherwise, we can explain the relationship between morphology and different clinical types by ultrastructures.
In the recent thirty years, with the rapid development of molecular biology, fungal taxonomy has entered the new era from phenotype to genetype.
The main results of this study are as follows:
1.Jilin Province was a high incidence area of sporotrichosis, especially in the areas of Jiutai, Nongan and Tongyu. Female was more than male, particularly for the woman workers. It was the most easily infective mass from 40 to 60, especially the farmers. Exposed parts such as face ,hands and limbs were more seen. The incidence was higher in winter and spring, which obviously increased the occurrence trend from 2004. The fixed type sporochosis was more than lymphatic vessel-type, and cutaneous disseminated type was rarely seen. It was the main method for prevention by promoting the recycle mechanism of corn stalks and reed, while adding iodine for areas lack of iodine.
2.The NO contents were much different in thr three types of sporotrichosis: lymphatic vessel-type was higher than that in fixed type, and fixed type was more than control group. The NO contents for all types before treatment were higher than that after treatment. Thus serum NO levels could be used as a biochemical indicator for mornitoring the infection of sporotrichosis, which had a positive correlation with the virulence of Sporothrix schenckii with the same immunological status and cultivating time of mice. So the virulence of Sporothrix schenckii with the same immunological status could be evaluated. The virulence had a correlation with cultivating time for the same strain, and the virulence of the strain cultivated for 7days was higher than that of the strain cultivated for 14 doys.
3.The strains of three types of sporotrichosis could not be recognized with routine fungal morphological methods. The mRNA of strains of Sporothrix schenckii in Jilin Province consists of type 4, 20 and 24, and nearly 90% of which was type 4. The ITS zone of Sporothrix schenckii was highly conservative, which of types was completely the same. So the recognization and fast diagnosis in genes by measuring ITS zone could made. Meanwhile, the NTS zone of cutaneous disseminated type was the same with the other two types.
4.There were some difference on the surface of spores for the three types by using scanning electron microscope, and nebular adhesion on the spore surface of lymphatic vessel-type and cutaneous disseminated type was found, which causes the increased virulence. The constructions of spores and hyphae were the same for three types by using transmission electron microscope. So the virulence of Sporothrix schenckii was depended on the materials on the surface of cells.
The creative points of the present study are: it is the first epidemiological study of sporotrichosis in Jilin Provence and so far, no report had been published for the measuring of NO between before and after treatment and serum NO of immunosuppressed mice for 4w. Meanwhile, no report has been published for the research of mitochondria DNA type or the difference of ultrastructure among all types.
引文
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