1.Analysis of polymorphism of Pfhrp2 gene in Plasmodium falciparum from falciparum malaria patients in Yunnan Province
Ying DONG ; Aiming SUN ; Mengni CHEN ; Yanchun XU ; Xianghua MAO ; Yan DENG ; Henlin YANG
Chinese Journal of Schistosomiasis Control 2016;28(4):411-417
Objective To analyze the polymorphism of histidine rich protein 2(HRP II)gene in Plasmodium falciparum (Pfhrp2)from falciparum malaria patients in Yunnan Province,so as to lay the foundation for studying the defection of antigen genes of Plasmodium. Methods The filter paper blood samples and related information of falciparum malaria cases reported were obtained in Yunnan Province from August 2012 to September 2015. Under the guidance of the specific primers,the exon2 regions in Pfhrp2 gene in P. falciparum from DNA samples were amplified by PCR,and the PCR products were sequenced. The sequences of exon2 region in Pfhrp2 gene were blasted by comparing with the reference sequences AY816237,AY816240,and AY816301. Next,the polymorphism of the sequence in exon2 region of Pfhrp2 gene was analyzed by MEGA 5.04 software. The conserved sites and genetic distances between sequences were calculated by using the software as well,and the clustering tree was drawn according to the genetic distances between the amino acid sequences. Results A total of 218 bloods samples from the falciparum malaria cases in 15 prefectures of Yunnan Province were collected,and the sources of infection included Yun?nan,Africa and Myanmar. The PCR results showed that the exon2 regions in Pfhrp2 genes of 155 samples were positive by am?plification and their products were sequenced successfully. The sequence analysis showed that the length range of the amino acid residues of exon2 region in Pfhrp2 gene was from 115 aa to 298 aa,the average length was 239.7 aa. There was no statistically significance among the means of the amino acid residues of the isolates from Africa( 239.9 aa),Myanmar(239.5 aa)and Yun?nan(241.6 aa)(F=0.025,P>0.05). All the 155 amino acid sequences ended with type 12 repeat,98.1%(152/155)of them started with type 1 repeat and 1.9%(3/155)of them started with type 2. Type 2 presented most frequently repeat in all the se?quences and the average repeat times were 12.9. The homologous locus of the DNA sequences in exon2 regions of the 155 Pfhrp2 genes was 894 bp,among which the conservative sites accounted for 20.6%(186/894),and the variable sites for 78.2%(699/894). The genetic distances between the sequences of Africa isolates ranged from 0 to 0.741,and those of the Myanmar and Yun?nan isolates were 0-0.948 and 0-0.750,respectively. The cluster analysis showed that all the 155 sequences clustered into 3 cat?egories on genetic distances between amino acid sequences according to the size of the amino acid sequence length. At the same level,the sequences had approximate lengths and amino acid repeat types. Conclusion The sequence of exon2 region in Pfhrp2 gene of P. falciparum from falciparum malaria cases in Yunnan Province is highly polymorphic,the P. falciparum iso?lates are clustered mainly according to the size of the amino acid sequence of exon2 region in Pfhrp2 gene.
2.Geographical features of malaria in Yunnan Province
Xianghua MAO ; Chun WEI ; Yerong TANG ; Ying DONG ; Yan DENG ; Jian WANG ; Mengni CHEN ; Yanchun XU ; Xiaodong SUN
Chinese Journal of Schistosomiasis Control 2017;29(1):24-27
Objective To understand the geographical features of malaria in Yunnan Province,so as to provide the refer?ence for malaria elimination. Methods The data of malaria in Yunnan Province from 2012 to 2015 were collected and ana?lyzed. Results Totally 2 586 malaria cases were reported in Yunnan Province from 2012 to 2015,in which 274(10.60%) were local cases and 2 311(89.37%)were abroad imported,and one(0.03%)was domestic imported. The imported malaria cases and local cases were analyzed according to the sources and locations respectively,and the arithmetic means of the num?bers of imported and local cases were 96.29 and 10.96 respectively,the standard deviations of the numbers of imported and local cases were 421.18 and 19.12 respectively,and the difference of the means was not significant(Z=-0.326,P>0.10). Both the imported and local malaria cases could be clustered into five sections by the number of 5. The Herfendal?Hirshman indexes of the imported and local malaria cases were 8 121 and 1 598 respectively. Conclusions There is no significant difference of the distribution between the imported and local malaria cases,and they should be attaching equal importance. The non?uniform de?gree of imported cases is higher than that of the local cases,while both of them could be divided into five major clusters in the prevention and control work.
3.Production of hispidin polyphenols from medicinal mushroom Sanghuangporus vaninii in submerged cultures.
Wenwen YUAN ; Weihua YUAN ; Rong ZHOU ; Guoying LV ; Mengni SUN ; Yanxia ZHAO ; Weifa ZHENG
Chinese Herbal Medicines 2023;15(4):594-602
OBJECTIVE:
The medicinal mushroom Sanghuangporus vaninii produces pharmaceutically valuable hispidin polyphenols in natural habitats. However, due to the slow growth in nature, S. vaninii grown in the field (sclerotia) is not reliable for pharmaceutical purposes. Although higher biomass of fungal mycelia can be obtained in submerged cultures, the accumulation of hispidin polyphenols is rare.
METHODS:
In this study, the polyunsaturated fatty acids (PUFAs), linoleic acid (LA), linolenic acid (ALA), and methyl jasmonate (MeJa) were employed as the stimulant agents to coordinate the accumulation of biomass and hispidin polyphenols in its submerged cultures.
RESULTS:
The addition of LA and ALA promoted the mycelial accumulation, while the addition of MeJa inhibited the growth of S. vaninii concomitant with reduced total polyphenols. UPLC-Triple-TOF-MS analysis revealed an increased production of hispidin, phellinstatin, pinnilidine, and its derivatives upon the addition of LA and ALA, and hypholomine B and its isomer, 3,14'-bihispidinyl, and phelligridin E upon the addition of MeJa on day 13. Intriguingly, total polyphenols from the MeJa-supplementing cultures harbored a high capacity in scavenging free radicals. Chemical structural analysis showed that hispidin polyphenols had higher antioxidant activity due to more hispidin moieties induced by MeJa.
CONCLUSION
The supplement of PUFAs affects the synthesis and composition of hispidin polyphenols in S. vaninii. Our results provide a possibility to coordinate the production of hispidin polyphenols via submerged cultures of S. vaninii.