Carbon budget of diversified cropping systems in southwestern China: Revealing key crop categories and influencing factors under different classifications.

Cropping systems are considered the largest source of agricultural GHG emissions. Identifying key categories and factors affecting cropping systems is essential for reducing these emissions. Most studies have focused on the carbon budget of cropping systems from the perspective of a single crop or crop category. Comprehensive studies quantifying the carbon budget of diversified cropping systems, including farmland and garden crops, are still limited. This study aims to fill this gap by quantifying the carbon budget of diversified cropping systems, clarifying their carbon attributes, and identifying key crop categories and influencing factors within different classifications of the system. This study analyzed the carbon budget of a diversified cropping system consisting of 19 crops in Yunnan Province, southwestern China, using a crop-based net greenhouse gas balance methodology based on the "cradle-to-farm" life cycle idea. Crops were categorized into three levels of categories to assess the potential impact of categorization within the cropping system on its carbon balance. Results showed that Yunnan's diversified cropping system is a significant carbon sink, with net sequestration of 33.1 Mt CO2 eq, total emissions of 37.4 Mt CO2 eq, and total sequestration of 70.5 Mt CO2 eq. Cereals, vegetables, and hobby crops were the main contributors to carbon emissions, accounting for 41.61%, 21.87%, and 15.37%, respectively. Cereal crops also made the largest contribution to carbon sequestration at 53.18%. Bananas had the highest emissions per unit area (11.45 t CO2 eq ha-1), while walnuts had the highest sequestration (20.64 t CO2 eq ha-1). In addition, this study highlights effective strategies to reduce greenhouse gas emissions, such as reducing nitrogen fertilizer use, minimizing reactive nitrogen losses, and controlling methane emissions from rice fields. By elucidating the impact of carbon dynamics and crop categories, this study provides insights for sustainable agricultural practices and policies.

Genetic polymorphism analysis and forensic application evaluation of 57 insertion/deletion polymorphisms from Yi ethnic group in Yunnan.

BACKGROUND: As a new kind of diallelic genetic marker, insertion/deletion (InDel) polymorphisms have recently been used in forensic science. However, there are relatively few studies on the forensic evaluation of InDel genetic polymorphisms from different populations. AIM: The aim of the present work is to assess the genetic polymorphism and forensic applicability of 57 InDels from the Yi ethnic group and explore the genetic background of this group. SUBJECTS AND METHODS: A total sample of 122 unrelated individuals of Yi group from the Yunnan province were genotyped by the AGCU indel 60 Kit. Multiplex population genetic analyses on the same 57 InDels were carried out among the Yunnan Yi group and 29 reference populations. RESULTS: The average allele frequency of these loci in the Yi ethnic group was 0.485. Heterozygosity, polymorphism information content, and the power of discrimination were 0.477, 0.362, and 0.612, respectively. The combined power of discrimination and the combined power of exclusion reached to 0.99999999999999999669 and 0.999962965, respectively. The results showed that 57 InDels polymorphisms have high genetic polymorphisms in the Yi ethnic group. CONCLUSIONS: The 57 InDels could be used for forensic individual identification, paternity testing, and intercontinental population discrimination, with the potential for use in biogeographic ancestry inference.

Cystoderma yongpingense sp. nov. (Squamanitaceae, Agaricales) a new species from southwestern China.

The present study introduces a novel fungus, Cystoderma yongpingense, which was identified in the southwestern region of China. The new species is characterized by a pileus that ranges in color from light orange-red to orange-red; the pileus has a wrinkled surface and is accompanied by a persistent annulus that is membranous and floccose-scaly. Above the annulus, the color transitions from white to yellowish brown. This proposal is substantiated through analyses encompassing both morphological characteristics and phylogenetic relationships. The phylogenetic position of the newly discovered species has been further corroborated through comprehensive maximum likelihood and Bayesian sequence analyses of the ITS + nrLSU DNA regions. Additionally, the technical description of C. yongpingense is enhanced by detailed illustrations and comparative studies with species that are closely related.

Molecular evidence for potential transovarial transmission of Jingmen tick virus in Haemaphysalis longicornis fed on cattle from Yunnan Province, China.

Jingmen tick virus (JMTV) is a novel tick-borne virus first identified from Jingmen city, Hubei Province of China in 2010. It has been proved that JMTV can cause human diseases and is widely distributed both inside and outside of China. However, the survival mode and transmission characteristics of JMTV still need further research, particularly in terms of transovarial transmission. In this study, an investigation was conducted to explore the presence of JMTV from engorged female ticks to their offspring. All engorged female adult ticks were collected from domestic cattle and allowed to lay eggs in appropriate humidity and temperature conditions. Maternal ticks, eggs and larvae were screened for JMTV RNA through real-time polymerase chain reaction (RT-PCR) and nested PCR methods. The results revealed the positive rate of 10.53% (10/95) in engorged ticks, 9.09% (2/22) in eggs and 8% (4/50) in larvae pools, respectively. Phylogenetic analysis confirmed that sequences from eggs and larvae had closer relationship with those isolates from maternal engorged ticks with more than 99.7% homology and JMTV manifested with evolutional conservatism. Our study has identified for the first time that JMTV could be transmitted from mother generation to offspring of Haemaphysalis Longicornis. Nonetheless, the efficiency of transovarial transmission in JMTV and the significance of ticks as amplification hosts still need to be further illustrated.

Characteristics of molecular markers associated with chloroquine resistance in Plasmodium vivax strains from vivax malaria cases in Yunnan Province, China.

BACKGROUND: Chloroquine (CQ) has been the preferred clinical treatment for vivax malaria in Yunnan Province since 1958, with over 300,000 patients. This study aimed to help make trend predictions regarding variations the in anti-malarial drug susceptibility of Plasmodium vivax distributed in Yunnan Province and effectively implement monitoring measures on the efficacy of anti-malarial drugs for vivax malaria. METHODS: Blood samples collected from patients with mono-P. vivax infections were employed in this study based on the principle of cluster sampling. The whole gene of P. vivax multidrug resistance 1 protein gene (pvmdr1) was amplified by nested-PCR techniques and the PCR amplification produce were sequenced by Sanger bidirectional sequencing. The mutant loci and haplotypes of coding DNA sequence (CDS) were identified by comparison with the reference sequence (NC_009915.1) of the P. vivax Sal I isolate. Parameters such as Ka/Ks ratio were calculated using MEGA 5.04 software. RESULTS: A total of 753 blood samples from patients infected with mono-P. vivax were collected, of which 624 blood samples yielded the full gene sequence (4392 bp) of the pvmdr1 gene, with 283, 140, 119, and 82 sequences from 2014, 2020, 2021 and 2022, respectively. A total of 52 single nucleotide polymorphic (SNP) loci were detected for the 624 CDSs, of which 92.3% (48/52), 34.6% (18/52), 42.3% (22/52), and 36.5% (19/52) SNPs were detected in 2014, 2020, 2021 and 2022, respectively. All of 624 CDSs were defined for a total of 105 mutant haplotypes, with CDSs of 2014, 2020, 2021, and 2022 containing 88, 15, 21, and 13 haplotypes, respectively. Of the 105 haplotypes, the threefold mutant haplotype (Hap_87) was the starting point for stepwise evolution, and the most drastic tenfold mutations were Hap_14 and Hap_78, and the fivefold, sixfold, sevenfold, and eightfold mutations. CONCLUSIONS: In the majority of vivax malaria cases in Yunnan Province, most of them were infected with strains carrying demonstrating highly mutated in pvmdr1 genes. However, the dominant mutation strains types varied from year to year, which warrants further exploration in order to confirm the correlation between with phenotypic changes in P. vivax strains and their susceptibility to anti-malarial drugs such as chloroquine.

Genetic polymorphisms and phylogenetic characteristics of Tibeto-Burman-speaking Lahu population from southwest China based on 41 Y-STR loci.

BACKGROUND: Male sex-linked Y-chromosome short tandem repeats (Y-STRs) have been widely used in forensic cases and population genetics research. At present, the forensic-related Y-STR data in the Chinese Lahu population are still poorly understood. AIM: To enrich the available Y-STR data of this Chinese minority population and investigate its phylogenetic relationships with other reported populations. SUBJECTS AND METHODS: The genetic polymorphisms of 41 Y-STR loci were analysed in 299 unrelated healthy Lahu male individuals from Southwest China. Phylogenetic analyses were performed by multidimensional scaling analysis and neighbor-joining phylogenetic tree construction. RESULTS: A total of 379 alleles were observed at the 41 Y-STR loci. The allele frequencies ranged from 0.0033 to 0.9666. The genetic diversity values ranged from 0.0653 to 0.9072. A total of 254 different haplotypes of the 41 Y-STR loci were observed in 299 individuals. The values of haplotype diversity, haplotype match probability, and discrimination capacity were 0.9987, 0.0047, and 0.8495, respectively. The phylogenetic analysis indicated that the Tibeto-Burman-speaking Lahu population showed a close genetic relationship with the Yunnan Yi population. CONCLUSIONS: The haplotype data of the present study can enrich the forensic databases of this Chinese minority population and will be useful for population genetics and forensic DNA application.

Molecular phylogeny and morphology reveal a new wood-rotting fungal species, Sistotrema yunnanense sp. nov. from the Yunnan-Guizhou Plateau.

Wood-rotting fungi are important components of woody plant ecosystems and play an active role in the decomposition and turnover of nutrients from wood, and are among the major groups of Basidiomycota. In this study, a new species of wood-rotting fungus, Sistotrema yunnanense, was proposed based on morphological characteristics and molecular evidence. It is characterized by resupinate basidiomata, a monomitic hyphal system having generative hyphae with clamp connections, suburniform to urniform basidia, and short-cylindrical to oblong ellipsoid basidiospores (4.5-6.5 × 3-4 µm). Phylogenetic analyses performed using the large subunit nuc rDNA indicated that S. yunnanense was nested within the genus Sistotrema s.l. of the family Hydnaceae, within the order Cantharellales.

Identification and genome characterization of novel parechovirus sequences from Hipposideros armiger in China.

BACKGROUND: Bats were identified as a natural reservoir of emerging and re-emerging infectious pathogens threatening human health and life. METHODS: This study collected 21 fecal samples of Hipposideros armiger in Mengla County of Xishuangbanna Prefecture Yunnan Province to combine one pool for viral metagenomic sequencing. RESULTS: Two nearly complete genomes of parechoviruses, BPeV11 and BPeV20, were sequenced. Genome analysis revealed that BPeV11 and BPeV20 follow a 3-3-4 genome layout: 5' UTR-VP0-VP3-VP1-2A-2B-2C-3A-3B-3C-3D-3' UTR. The prevalence of BPev11 and BPev20 by Nested-PCR showed that 1 of 21 fecal samples was positive. Based on amino acid identity comparison and phylogenetic analysis of P1, 2C, and 3D, BPeV11 and BPeV20 were closely related to but distinct from FPeVs. CONCLUSION: It was probably proposed to be a novel species in the genus Parechovirus of the family Picornaviridae. The isolation of BPev11 and BPev20 from H. armiger in China is the first complete genome of parechovirus isolations from bat feces of the genus Hipposideros.

Genetic polymorphism investigation of 16 X-STR loci in the Bai ethnic minority in Yunnan Province, Southwest China.

To investigate the genetic variation and forensic efficiency of 16 X-chromosomal short tandem repeat (X-STR) loci (DX6795, DXS9902, DXS8378, HPRTB, GATA165B12, DXS7132, DXS7424, DXS6807, DXS6803, GATA172D05, DXS6800, DXS10134, GATA31E08, DXS10159, DXS6789, and DXS6810) in the Bai minority, we calculated allele frequencies, forensic parameters, and haplotype frequencies in 424 (202 males and 222 females) unrelated, healthy Bai individuals from Dali Bai Autonomous Prefecture in Yunnan Province, China. We observed a total of 132 alleles; 5-19 alleles were detected in each locus, and the corresponding allele frequencies ranged from 0.0016 to 0.7589. All of the loci detected were highly polymorphic in the Bai population in Yunnan Province, except DXS6800. The values for the combined power of discrimination in females (PDf) and males (PDm) were 0.999999999999996 and 0.999999997487061, respectively. According to a phylogenetic tree, neighboring populations and different nationalities in the same area appeared to have relatively close evolutionary relationships. This study provides and complements X-chromosome genetic polymorphism data for the Bai people in Yunnan Province, Southwest China, and enriches the available reference materials for this Chinese minority population.

Evaluation of different factors on metal leaching from nickel tailings using generalized additive model (GAM).

Compared with sulfide tailings, the oxidation and transformation of certain substances in oxidized tailings into more soluble forms may affect the bioaccumulation and biomagnification properties and enhance the risk of toxic effects in the ecosystem. This study aimed to apply the generalized additive model (GAM) to evaluate factors affecting heavy metal leaching from nickel (Ni) tailings. We created an orthogonal experiment table (L18(37)) to evenly distribute the different combinations of factor values. The Ni tailings were immersed in solutions with different combinations of factor values for 16 d, and samples were taken on days 1, 2, 4, 7, 11, and 16 to measure the pH and heavy metal concentration of the leachate. The GAM was used to fit the concentration of heavy metals of the leachate and the initial factors of the leaching solution. The results showed that the pH and Cr concentration of the leachate increased with time and stabilized after 1 d (pH of approximately 7), while the Mn, Ni, and Tl concentrations gradually decreased and stabilized after peaking on the first day. An analysis of the GAM results showed that the Cr concentration was highly sensitive to the solid-liquid ratio (F = 127.8) and tailing particle size (F = 10.7). The Cr concentration of the leachate was significantly higher when exposed to a high solid-liquid ratio or a fine particle size, whereas the Mn, Ni, and Tl concentrations were highly sensitive to the KCl concentration and solid-liquid ratio (F = 77.4, 146.9, and 315.9 respectively). The GAM identified interactions between key factors, which have complex and strong effects on the leaching of tailings and the migration of heavy metals, either promotional or antagonistic. The prediction of the minimum Cr leaching concentration shows that GAM can be used to determine the conditions associated with minimum leaching concentrations of heavy metals and to effectively predict the metal concentrations of leachate. As such, the results of this study can be applied to the management of nickel tailings.

Genetic diversity of 15 STR loci in Yunnan Va ethnic minority and the phylogenetic relationships with 26 other populations.

BACKGROUND: The Va (also called "Wa") people are an ethnic minority living mainly in the southwest of Yunnan Province. AIM: This study was conducted to obtain the genetic information and forensic statistical parameters of 15 autosomal short tandem repeat (STR) loci included in the AmpFlSTR®Identifiler™ kit (Applied Biosystems, Foster City, CA) in the Yunnan Va population, with a view to enriching the genetic databases of the Chinese Va population. SUBJECTS AND METHODS: A total of 508 unrelated Chinese Va individuals were genotyped with this 15 STR kit, the genetic polymorphisms and associated forensic parameters were calculated. The genetic relationships between Chinese Va and 26 other Chinese populations were also evaluated. RESULTS: All of the STR loci reached the Hardy-Weinberg equilibrium after Bonferroni correction. A total of 159 alleles were observed with allele frequencies ranging from 0.000984 to 0.606299. The combined discrimination power (CDP) and the cumulative probability of excluding (CPE) of the 15 STR loci were 0.999 999 999 999 999 988 126 and 0.999 995 734, respectively. Our results indicated that the geographically adjacent or ethnically close populations showed a higher genetic affinity. CONCLUSIONS: The results of this study will enrich the forensic databases of the Chinese Va population and could be applied in forensic analysis.

Undocumented Migrants Reintroducing COVID-19, Yunnan Province, China.

To limit the spread of severe acute respiratory syndrome coronavirus 2, the government of China has been monitoring infected travelers and minimizing cold-chain contamination. However, other factors might contribute to recurring outbreaks. We analyze the role of undocumented migrants as potential transmitters of severe acute respiratory syndrome coronavirus 2 in China.

First report of the prevalence and genetic characterization of Giardia duodenalis and Cryptosporidium spp. in Yunling cattle in Yunnan Province, southwestern China.

Yunling cattle is an unique cattle breed distributed in Yunnan Province, southwestern China. It is yet to know whether Yunling cattle are infected with Giardia duodenalis and Cryptosporidium spp.. The objectives of the present study were to investigate the prevalence and characterize the assemblages of G. duodenalis and species of Cryptosporidium spp. in Yunling cattle in Yunnan province. The overall prevalence of G. duodenalis and Cryptosporidium spp. were 10.49% (41/391) and 0.77% (3/391), respectively. The age was considered as the risk factor for Yunling cattle infection with G. duodenalis (χ2 = 8.082, OR = 2.56, P = 0.004). Two assemblages of G. duodenalis, assemblage A (n = 1) and assemblage E (n = 40), were identified by amplification of the ß-giardin (bg) and glutamate dehydrogenase (gdh) gene loci using the nested PCR methods. Furthermore, Cryptosporidium andersoni (n = 1) and Cryptosporidium ryanae (n = 2) were detected by nested PCR targeting the small subunit (SSU) rRNA gene. This is the first report of G. duodenalis and Cryptosporidium spp. in Yunling cattle in China, which provided baseline date for further studies of the prevalence, genetic identity, and public health potential of these parasites in Yunling cattle.

Prediction of the CYP2D6 enzymatic activity based on investigating of the CYP2D6 genotypes around the vivax malaria patients in Yunnan Province, China.

BACKGROUND: In recent years, the incidence rate of vivax malaria recurrence still had 3.1% in Yunnan Province population after eradication therapy using primaquine (PQ). In order to understand the specific failure reasons for preventing vivax malaria relapses, a preliminary exploration on the CYP2D6 enzyme activity was carried out in the vivax malaria patients in Yunnan Province population by analysing mutational polymorphism in the coding region of CYP2D6 gene. METHODS: Blood samples were collected from vivax malaria patients with suspected relapse (SR) and non-relapsed (NR) malaria in Yunnan Province. The DNA fragments containing 9 exons regions of human CYP2D6 gene were amplified by performing PCR and sequenced. The sequencing results were aligned by using DNAStar 11.0 to obtain the coding DNA sequence (CDS) of CYP2D6 gene. DnaSP 6.11.01 software was used to identify mutant polymorphisms and haplotypes of the CDS chain. The waterfall function of GenVisR package in R was utilized to visualize the mutational landscape. The alleles of CYP2D6 gene were identified according to the criteria prescribed by Human Cytochrome P450 (CYP) Allele Nomenclature Committee Database and the CYP2D6 enzyme activity was predicted based on diploid genotype. RESULTS: A total of 320 maternal CDS chains, including 63 from SR group and 257 from NR group, were obtained. Twelve mutant loci, including c.31 (rs769259), c.100 (rs1065852), c.271 (rs28371703), c.281 (rs28371704), c.294 (rs28371705), c.297 (rs200269944), c.336 (rs1081003), c.408 (rs1058164), c.505 (rs5030865), c.801 (rs28371718), c.886 (rs16947), and c.1,457 (rs1135840) were observed on the 640 CDS chains (including 320 maternal and 320 paternal chains). The high-frequency mutation at rs1135840 (0.703) and low-frequency mutation, such as rs28371703, were detected only in the SR group. The frequency of mutant rs1058164 and rs1135840 were significantly increased in the SR group ([Formula: see text]= 4.468, 5.889, P < 0.05), as opposed to the NR group. Of the 23 haplotypes (from Hap_1 to Hap_23), the nomenclatures of 11 allelic forms could be found: Hap_3 was non-mutant, Hap_2 accounted for the highest frequency (36.9%, 236/640), and Hap_9 had the most complex sequence structure, containing 7 loci mutations. Allele *10 was the most frequent among these genotypes (0.423). Among the allele *10 standard named genotypes, *1/*10, *1/*1 and *2/*10 were significantly more frequent in the NR group ([Formula: see text]= 3.911, P < 0.05) and all showed uncompromised enzyme activity; the impaired genotype *10/*39 was more frequent in the SR group ([Formula: see text]= 10.050, P < 0.05), and genotype *4/*4was detected only in the SR group. CONCLUSION: In the patients receiving PQ dosage in Yunnan Province population, both rs1135840 single nucleotide polymorphism and *10 allele form was common in the CYP2D6 gene. Low-frequency mutation sites, such as rs28371703, were only presented in patients with vivax malaria relapse.

A thematic analysis of experiences of HIV risks among female sex workers in the Yunnan-Vietnam Chinese border region.

BACKGROUND: The Yunnan province is located near the "Golden Triangle" border region between China, Myanmar and Thailand, which has the highest HIV/AIDS prevalence in China. Female sex workers (FSWs) in the Yunnan province are highly vulnerable to HIV infection. The objective of this study was to examine the experiences of FSWs in the Yunnan to better understand the risk of infection and the potential for transmission of HIV. METHODS: Semi-structured interviews were conducted between May 2018 and June 2018 with 20 FSWs recruited in Hekou County, Yunnan Province, China. Thematic analysis was conducted to identify themes that highlighted increased exposure of FSWs to the risk of HIV infection and transmission. RESULTS: The findings showed that FSWs' primary source for HIV information was gynecologists, with few visiting the local HIV charity Red Ribbon. FSWs reported infrequent visits for check-ups with some seeing a gynecologist once a year. FSWs felt that the onus was on them to prevent STI/HIV infection by using a condom during sex, regardless of their ability to negotiate use. FSWs were also reluctant to see a gynecologists for treatment. Instead, they resorted to douching as a way of preventing HIV/STIs and treating vaginal health problems, such as leucorrhea. Most FSWs worked without the influence of alcohol and drugs. A small number of FSWs reported heroin addiction and injecting drug use. CONCLUSION: The findings suggest a need for innovative HIV prevention strategies among FSWs and their clients in the Chinese border region. Governmental agencies should continue to implement practical strategies in terms of HIV prevention education and condom use through tailored interventions that are localized. Such strategies should include localized tailored interventions that dispel myths about douching as a method of HIV/STI prevention and incorporate a mobile outreach approach, similar to the 'roadside restaurant' outreach that has been successful in rural China.

The Pattern of Vitamin D Levels in Children 0-4 Years of Age in Yunnan Province.

OBJECTIVE: Vitamin D is an essential nutrient that regulates the activity of calcium and bone hormones throughout life; however, vitamin D levels in children, which is the most crucial period during human development, has not been established. METHODS: As the first descriptive study of serum vitamin D levels in children in Yunnan Province, we determined the serum vitamin D levels in children 0‒4 years of age who underwent physical examinations at Kunming Children's Hospital, and the association between the serum vitamin D level and the calcium, phosphorus and alkaline levels. RESULTS: Vitamin D levels in children were highest in the summer months and lowest in the winter months. Vitamin D deficiency was more common in girls than boys. A social-economic effect was shown, as evidenced by the significantly higher serum vitamin D levels in children from the top five cities compared with the lower-ranked cities. Moreover, we also demonstrated a significant correlation between vitamin D and serum calcium levels. CONCLUSION: Our study suggested that sex and age affected the vitamin D levels of children, and a reasonable reference range in children 0-4 years of age in Yunnan Province was determined.

Cadmium and Lead Pollution Characteristics of Soils, Vegetables and Human Hair Around an Open-cast Lead-zinc Mine.

Atmospheric deposition of cadmium (Cd) and lead (Pb) was investigated together with the accumulation, distribution and health risks from potentially toxic metals in soils, vegetables and human hair at a mining area in southwest China. Annual atmospheric deposition of Cd and Pb were 41.1 and 192 g ha- 1, respectively, and consisted mainly of dry deposition. Agricultural soils experienced high levels of metal pollution around the mine, with 66.4% and 57.3 % of vegetable samples grown on these polluted fields exceeding maximum permissible Cd and Pb concentrations, particularly the leafy vegetables. Residents living near the mining area had high Cd (0.75 mg kg- 1) and Pb (6.87 mg kg- 1) concentrations in their hair, and the maximum values occurred in occupationally exposed individuals. Long-term mining activities have resulted in high health risks to the local population due to Cd and Pb deposition and accumulation from the atmosphere, soils and vegetables.

Analysis of initial laboratory diagnosis of malaria and its accuracy compared with re-testing from 2013 to 2018 in Yunnan Province, China.

BACKGROUND: According to China's Malaria Eradication Action Plan, malaria cases diagnosed and reported by health authorities at the county level must be further re-confirmed by provincial laboratories. The Yunnan Province Malaria Diagnostic Reference Laboratory (YPMDRL) began the synchronous implementation of microscopic examinations and nested polymerase chain reaction (nested-PCR) testing to re-test the malaria cases initially diagnosed by county-level laboratories and to evaluate the consistency of Plasmodium species identified between by YPMDRL and by the county-level laboratories from 2013 to 2018 in Yunnan Province. METHODS: Data on malaria initial diagnosis completed by county-level laboratories in Yunnan Province were collected weekly from the "China Disease Prevention and Control Information System" from 2013 to 2018. The YPMDRL performed Plasmodium microscopic examination and 18S rRNA gene nested-PCR testing on every malaria case managed by the China Disease Prevention and Control Information System. The re-testing detection results were fed back to the initial diagnosis and reporting unit for revision of malaria case types. RESULTS: A total of 2,869 malaria cases were diagnosed and reported by county-level laboratories in Yunnan Province from 2013 to 2018. The re-testing rate was 95.6% (2,742/2,869), and the re-testing rate increased from 2013 to 2018. Among the re-tested 2,742 cases, 96.7% (2651/2742), 2.2% (59/2742), and 1.1% (32/2742) were doubly examined by microscopy and by nested-PCR, only by microscopy, and only by nested-PCR, respectively. The total Plasmodium species accuracy rate at county-level laboratories was 92.6% (2,543/2,742) reference to the diagnosis by YPMDRL. Among the inconsistent 199 cases, they were identified as including 103 negative cases, 45 falciparum malaria cases, 30 vivax malaria cases, 11 ovale malaria cases, and 10 malariae malaria cases by YPMDRL. From 2013 to 2018, the revised and registered malaria cases by the China Disease Prevention and Control Information System in Yunnan Province was 2,747 cases, including 2,305 vivax malaria cases, 421 falciparum malaria cases, 11 ovale malaria cases, and 10 malariae malaria cases. CONCLUSIONS: The double re-testing strategy by microscopy and by gene testing increases the accuracy of diagnoses malaria in Yunnan Province, and gene testing can reliably differentiate Plasmodium species. The re-testing results provided by YPMDRL are the authoritative basis for revising malaria kind in Yunnan Province.

Polymorphism analysis of propeller domain of k13 gene in Plasmodium ovale curtisi and Plasmodium ovale wallikeri isolates original infection from Myanmar and Africa in Yunnan Province, China.

BACKGROUND: Eighteen imported ovale malaria cases imported from Myanmar and various African countries have been reported in Yunnan Province, China from 2013 to 2018. All of them have been confirmed by morphological examination and 18S small subunit ribosomal RNA gene (18S rRNA) based PCR in YNRL. Nevertheless, the subtypes of Plasmodium ovale could not be identified based on 18S rRNA gene test, thus posing challenges on its accurate diagnosis. To help establish a more sensitive and specific method for the detection of P. ovale genes, this study performs sequence analysis on k13-propeller polymorphisms in P. ovale. METHODS: Dried blood spots (DBS) from ovale malaria cases were collected from January 2013 to December 2018, and the infection sources were confirmed according to epidemiological investigation. DNA was extracted, and the coding region (from 206th aa to 725th aa) in k13 gene propeller domain was amplified using nested PCR. Subsequently, the amplified products were sequenced and compared with reference sequence to obtain CDS. The haplotypes and mutation loci of the CDS were analysed, and the spatial structure of the amino acid peptide chain of k13 gene propeller domain was predicted by SWISS-MODEL. RESULTS: The coding region from 224th aa to 725th aa of k13 gene from P. ovale in 83.3% of collected samples (15/18) were amplified. Three haplotypes were observed in 15 samples, and the values of Ka/Ks, nucleic acid diversity index (π) and expected heterozygosity (He) were 3.784, 0.0095, and 0.4250. Curtisi haplotype, Wallikeri haplotype, and mutant type accounted for 73.3% (11/15), 20.0% (3/15), and 6.7% (1/15). The predominant haplotypes of P. ovale curtisi were determined in all five Myanmar isolates. Of the ten African isolates, six were identified as P. o. curtisi, three were P. o. wallikeri and one was mutant type. Base substitutions between the sequences of P. o. curtisi and P. o. wallikeri were determined at 38 loci, such as c.711. Moreover, the A > T base substitution at c.1428 was a nonsynonymous mutation, resulting in amino acid variation of T476S in the 476th position. Compared with sequence of P. o. wallikeri, the double nonsynonymous mutations of G > A and A > T at the sites of c.1186 and c.1428 leads to the variations of D396N and T476S for the 396th and 476th amino acids positions. For P. o. curtisi and P. o. wallikeri, the peptide chains in the coding region from 224th aa to 725th aa of k13 gene merely formed a monomeric spatial model, whereas the double-variant peptide chains of D396N and T476S formed homodimeric spatial model. CONCLUSION: The propeller domain of k13 gene in the P. ovale isolates imported into Yunnan Province from Myanmar and Africa showed high differentiation. The sequences of Myanmar-imported isolates belong to P. o. curtisi, while the sequences of African isolates showed the sympatric distribution from P. o. curtisi, P. o. wallikeri and mutant isolates. The CDS with a double base substitution formed a dimeric spatial model to encode the peptide chain, which is completely different from the monomeric spatial structure to encode the peptide chain from P. o. curtisi and P. o. wallikeri.

[Analysis of EGFR mutation and clinical features of lung cancer in Yunnan].

Objective: To analyze the EGFR mutation profile of lung cancer patients in Yunnan, and to provide evidence for clinical personalized treatment. Methods: Demographic and clinical data of 2 967 lung cancer patients undergoing EGFR identification were collected and analyzed from January 2014 to August 2019 in Yunnan Cancer Hospital. Results: The proportion of EGFR mutation in 2 967 patients with lung cancer was 46.2%. Univariate analysis showed that the proportion of EGFR mutation in women was higher than that in men (P<0.001) and displayed a downward trend with age (P=0.03). The mutation rate of ethnic minorities was higher than Han (P=0.012). Mutation rate in patients without smoking history was higher than those with smoking history (P<0.001), and patients without drinking history was higher than patients with drinking history (P<0.001). Mutation rate in patients without family history of lung cancer was higher than those with family history (P=0.008). The mutation rate of adenocarcinoma was higher than other pathological types (P<0.001). The mutation rate was different among stages, and it was higher in early patients than that in advanced patients (P<0.001). The mutation rate of tissue specimens was higher than those of cytology and peripheral blood samples (P<0.001). The mutation rate of Xuanwei area was lower than that in non-Xuanwei area (P<0.001). Multivariate analysis showed that gender (P<0.001), age (P=0.036), smoking history (P<0.001), pathological type (P<0.001), specimen type (P<0.001), and whether or not Xuanwei area (P<0.001) were the independent factors of EGFR mutation.The EGFR mutation was more common in female, non-smokers, adenocarcinoma, non-Xuanwei area, tissue specimen and young lung cancer patients.The mutation types of EGFR in 1 370 cases mainly included 19-Del and L858R. The predominant mutation of EGFR in Xuanwei area was L858R, while in non-Xuanwei area was 19-Del.The mutation rates of G719X, G719X+ L861Q, G719X+ S768I, and S768I in Xuanwei were higher while the mutation rates of 19-Del, L858R, and 20-ins were lower than non-Xuanwei area (P<0.05). The 19-Del mutation rate of ethnic minorities is higher than that of Han (P<0.001). The combined mutation rate of G719X, L861Q in Han was higher than that of ethnic minorities (P=0.005). Conclusions: The EGFR mutation rate in lung cancer patients in Yunnan is similar to Asian and Chinese, and higher in female, non-smokers, adenocarcinomas, young and non-Xuanwei area patients. The most common types of EGFR mutation in Yunnan are 19-Del and L858R. The predominant mutation of EGFR in Xuanwei area is L858R, while in non-Xuanwei area is 19-Del. The mutation rates of G719X, G719X+ L861Q, G719X+ S768I and S768I are higher in Xuanwei patients than those in non-Xuanwei patients. The combined mutation rate of G719X and L861Q in Han nationality is higher than that of ethnic minorities.