Objective To investigate the expression level of virulence-associated genes in promastigotes and amastigotes of different Leishmania spp.. Methods Total RNA was extracted from the promastigotes and amastigotes of Leishmania donovani, L. infantum, L. tropica, L. major and L. mexicana, and relevant strains. According to the reported gene sequences in GenBank, primers were designed in relation to the virulence-associated genes [GDP-mannose pyrophosphorylase (GDPMP), 3′a2rel-related protein (A2rel), beta-galactofuranosyl transferase (LPG1), lipophosphoglycan biosynthetic protein (LPG2), kinetoplast membrane protein 11 (KMP-11), cpc gene for cysteine proteinase (CPC), hydrophilic acylated surface protein (HASPB1), cathepsin L-like cysteine protease (CPB2), cathepsin L-like cysteine proteinase lmcpb2.8 (CPB2.8), Mr 100 000 heat shock protein (CLP b)], and control genes (alpha tubulin gene and GAPDH). Semi-quantitative RT-PCR was performed to detect expression level of these genes in promastigotes and amastigotes of different Leishmania spp. Results There was a significant difference in the expression profiles of the genes among the promastigotes and amastigotes of different Leishmania spp. The HASPB1 was detected in the amastigotes of all strains and promastigotes of L. donovani, the GDPMP, LPG1, LPG2, CPB2.8, CPB2, CPC, A2rel and CLP b were expressed in the promastigotes and/or amastigotes of the specific Leishmania spp, respectively. None of the stains carried the KMP-11 gene, whereas the amastigotes of L. donovani SC10 strain and L. major 5ASKH strain possessed CPC. Conclusion The expression profile of the virulence-associated genes shows species-specific and stage-specific differences.

Objective To explore the protein profile and identify developmentally regulated proteins of the promastigotes and axenic amastigotes with comparative proteomics technique.Methods The total proteins of promas-tigotes and axenic amastigotes of Leishmania donovani SC6 strain were separated by two-dimensional electrophoresis(2-DE) in a broad pH range(3-10) , and the gel was stained with Coomassie blue.The images were analyzed by PDQuest 1.0 software, and the major developmentally regulated proteins were identified by electrospray mass spectrometry.Results Approximately 700 protein spots were revealed in equivalent proteins of the promastigotes and axenic amastigotes separated by 2-DE, among which more than 90% protein spots showed equivalent quantity and distribution, with 6 proteins up-regulated and 3 proteins down-regulated in axenic amastigotes compared with promastigotes.Five of the 6 up-regulated proteins were with known function, respectively ascribed as Reiske iron-sulfur protein precursor, ?-tubulin, peroxidoxin 1, dihydrolipoamide acetyltransferase precursor, and mannose-1-phosphate guanyltransferase.Two of the 3 down-regulated proteins were identified as heat shock protein 70 and ?-tubulin.The functions of the developmentally regulated proteins were related to the carbohydrate/energy metabolism, stress response, or formation of cell membrane/cytoskeleton.Conclusion The findings demonstrate the differences in protein expression profiles between promastigotes and amastigotes.

Objective] By sequencing of SSU rRNA gene cloning from Xinjiang cutaneous leishmaniasis pathogen (XJCLP) to provide evidence for identification of the pathogen. [Methods] By PCR assay with primers R222 and R333, the specific fragment had been produced from SSU rRNA gene of XJCLP , L infantum, L tropica and cloned into pGEM ○[KG-6/7]R T Easy vector .The clones were sequenced by the Sanger dideoxy mediated chain termination method, analysis of SSU rRNA gene sequences from XJCLP, L tropica, L infantum with DNASIS. [Results] Sequence analysis showed that the specific fragment of SSU rRNAgene from XJCLP, L infantum,L tropica , were all 394 bp in length. There were 391 bases identical and three point mutations between the sequences of XJCLP and L tropica , the similarity being 99 2%; 390 bases identical and three point mutations and one insertion /deletion between the sequences of XJCLP and L infantum , the similarity being 99 0%. One insertion/deletion between the sequences of L tropica and L infantum , the similarity being 99 7%. The primary and secondary structures of SSU rRNA gene from XJCLP differed from those of L infantum and L tropica .A retrieval from GenBank confirmed that these 394 bp sequence are new gene sequences. [Conclusion]The primary and secondary structures of SSU rRNA gene from XJCLP, L infantum , L tropica were different. 394 bp sequence from SSU rRNA gene of XJCLP is a new gene sequence.

Objective:To analyze the clinical characteristics of patients with acute glyphosate herbicide poisoning and the differences in the severity of poisoning.Methods:A retrospective analysis was performed on patients with acute glyphosate herbicide poisoning admitted to the First Affiliated Hospital of Zhengzhou University from January 2014 to December 2020. The general information, exposure time, poisoning dose, poisoning cause, poisoning route, clinical manifestations, laboratory examination results during hospitalization, treatment measures, hospital stays and prognosis of the patients were collected. The patients were graded according to the poisoning severity scoring standard of Chinese Expert Consensus on Diagnosis and Treatment of Acute Poisoning in 2016. The highest severity score during hospitalization was used as the final grade. According to the final grade, asymptomatic and mild patients were included in the mild group, and moderate, severe and death patients were included in the severe group. The independent sample T test or Mann-Whitney U test was used for measurement data, and χ2 test or Fisher's exact test was used for counting data. The differences of general data and clinical data between the two groups were compared. Results:According to the inclusion and exclusion criteria, 83 patients with acute glyphosate herbicide poisoning were selected as the study subjects. All patients survived, mainly mild poisoning (56.6%), with a male to female ratio of 33∶50, and an average age of 39 years. The number of poisoning cases increased yearly (the highest in 2019), and most cases occurred in spring and summer. The main cause of poisoning was suicide (71.1%), direct oral administration (83.1%) was the primary route of poisoning, and the dominating clinical manifestations were digestive symptoms (71.1%). Laboratory tests showed increased white blood cell count (WBC), neutrophil percentage (NEUT %) and D-dimer, and decreased hemoglobin and potassium. Compared with the mild group, patients in the severe group were older [(51±17) years vs. (35±19) years], had a higher proportion of suicide and direct oral administration, a longer hospital stay [8.0 (4.8, 12.0) d vs. 3.0 (2.0, 5.5) d], a higher dose of poisoning [200.0 (50.0, 200.0) mL vs. 30.0 (11.3, 57.5) mL], and higher NEUT % within 24 h of admission [(83.4±10.4) vs. (73.2±12.8)]. The increase of WBC, NEUT %, aspartate aminotransferase, prothrombin time, D-dimer and the decrease of serum potassium were more common in the severe group than the mild group, with statistical significance (all P<0.05). Conclusions:The number of patients with acute glyphosate herbicide poisoning is increasing yearly. Generally, the condition is mild and the prognosis is satisfying. The severity is more serious in the middle-aged and elderly patients andthose with direct oral administration, high toxic dose, and high NEUT % within 24 h of admission. Severe poisoning is more likely to cause changes in laboratory indicators.