[Cloning, Expression and Immunodiagnostic Evaluation of the Fasciola gigantica Thioredoxin Peroxidase].

OBJECTIVE: To immunoscreen the gene encoding thioredoxin peroxidase (TPx) from a cDNA library made from adult Fasciola gigantica worms, clone and express the gene, and evaluate the immunodiagnostic value of TPx recombinant protein. METHODS: The A ZAP cDNA library was immunoscreened with pooled serum of fascioliasis gigantica patients. The obtained positive clones were sequenced and analyzed by multiple sequence alignment. The full-length (rFgTPx) and N-termianal truncated (rFgTPx_nt) sequence of FgTPx was subcloned into prokaryotic plasmid pET28a(+) with a non-fusion expression technique, respectively. The recombinant proteins of rFgTPx and rFgTPx_nt were purified by His-bind affinity column (Ni-NTA). rFgTPx and rFgTPx_nt were used in indirect ELISA to test the antibody response of the serum samples. Sera of 27 fascioliasis gigantica patients, 15 patients with schistosomaisis japonica, 15 clonorchiasis sinensis patients, and 32 healthy donors were tested by using the recombinant protein based ELISA. RESULTS: The TPx recombinant proteins were obtained through expression, purification and renaturation, the relative molecular mass of rFgTPx and rFgTPx_nt were Mr 30,000 and Mr 26,000, respectively. The total diagnostic coincidence rate, sensitivity and specificity of rFgTPx_nt-based ELISA was 87.6% (78/89), 66.7% (18/27), and 96.8% (60/62), respectively. The cross reaction with Schistosoma japonicum and Clonorchis sinensis was 0 and 1/15 for rFgTPx_nt, respectively. Before and after treatment, A450 value of the serum samples from fascioliasis patients was 0.233 ± 0.088 and 0.129 ± 0.072, respectively (t = 4.27, P < 0.01). CONCLUSION: The gene encoding TPx is expressed in the prokaryotic expression system. The recombinant protein shows proper sensitivity and high specificity for the serodiagnosis of Fasciola gigantica infection.

[Epidemiological investigation of fascioliasis and analysis of a chronic human fascioliasis case in Binchuan County, Yunnan Province].

From February to March 2014, six natural villages in Zhoucheng Town, Binchuan County of Yunan Province, were randomly selected by cluster sampling. Serum anti-fascioliasis IgG was detected by ELISA. The sero-positive individuals were further tested for Fasciola infection using sediment detection with nylon bag (260 meshes) and Kato-Katz method. Among 1207 sampled persons, the sero-positive rate was 3.0% (36/1207). The rate in males and females was 2.3% (12/530) and 3.6% (24/677) (u=1.46, P>0.05). The sero-positive rate in Zhoucheng Village and Baizhuang Village was 4.0% (24/616) and 2.0% (12/591), respectively (u=2.07, P<0.05). The positive rate of stool examination in serum-positive persons was 6.5% (2/31). One stool-egg-positive patients was the case in 2012 outbreak, and the eggs were stale. The other patient was newly infected, and further clinical diagnosis indicated that it was a case of chronic fascioliasis.

Cross talk between Cu excess and Fe deficiency in the roots of rice.

Copper (Cu) and iron (Fe) share similar characteristics and participate as coenzymes in several physiological processes. Both Cu excess and Fe deficiency result in chlorosis, however, the crosstalk between the two is not clear in rice. In this study, we performed transcriptome analysis for Cu excess and Fe deficiency in rice. Some WRKY family members (such as WRKY26) and some bHLH family members (such as late flowering) were selected as novel potential transcription factors involved in the regulation of Cu detoxification and Fe utilization, respectively. These genes were induced under corresponding stress conditions. Many Fe uptake-related genes were induced by Cu excess, while Cu detoxification-related genes were not induced by Fe deficiency. Meanwhile, some genes, such as metallothionein 3a, gibberellin 3beta-dioxygenase 2 and WRKY11, were induced by Cu excess but repressed by Fe deficiency. Concisely, our results highlight the crosstalk between Cu excess and Fe deficiency in rice. Cu excess caused Fe deficiency response, while Fe deficiency did not lead to Cu toxicity response. Metallothionein 3a might be responsible for Cu toxicity-induced chlorosis in rice. The crosstalk between Cu excess and Fe deficiency might be regulated by gibberellic acid.

Mechanism of anti-hyperuricemia of isobavachin based on network pharmacology and molecular docking.

BACKGROUND: Hyperuricemia is a more popular metabolic disease caused by a disorder of purine metabolism. Our previous study firstly screened out a natural product Isobavachin as anti-hyperuricemia targeted hURAT1 from a Chinese medicine Haitongpi (Cortex Erythrinae). In view of Isobavachin's diverse pharmacological activities, similar to the Tranilast (as another hURAT1 inhibitor), our study focused on its potential targets and molecular mechanisms of Isobavachin anti-hyperuricemia based on network pharmacology and molecular docking. METHODS: First of all, the putative target genes of compounds were screen out based on the public databases with different methods, such as SwissTargetPerdiction, PharmMapper and TargetNet,etc. Then the compound-pathways were obtained by the compounds' targets gene from David database for Gene Ontology (GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enrichment analysis. The cross pathways of compound-pathways and the diseases pathways of hyperuricemia from Comparative Toxicogenomics Database were be considered as the compound-disease pathways. Next, based on the compound-disease pathways and the PPI network, the core targets were identified based on the retrieved disease-genes. Finally, the compound-target-pathway-disease network was constructed by Cytoscape and the mechanism of isobavachin anti-hyperuricemia was discussed based on the network analysis. RESULTS: Our study demonstrated that there were five pathways involved in Isobavachin against hyperuricemia, including Drug metabolism-other enzymes, Metabolic pathways, Bile secretion, Renin-angiotensin system and Renin secretion. Among the proteins involved in these pathways, HPRT1, REN and ABCG2 were identified as the core targets associated with hyperuricemia, which regulated the five pathways mentioned above. It is quite different from that of Tranilast, which involved in the same pathways except Bile secretion instead of purine metabolism. CONCLUSION: This study revealed Isobavachin could regulate the pathways including Drug metabolism-other enzymes, Metabolic pathways, Bile secretion, Renin-angiotensin system, Renin secretion by core targets HPRT1, REN and ABCG2, in the treatment of hyperuricemia effect. Among them, the Bile secretion regulated by ABCG2 probably would be a novel pathway. Our work provided a theoretical basis for the pharmacological study of Isobavachin in lowering uric acid and further basic research.

Transcripts related with ammonium use and effects of gibberellin on expressions of the transcripts responding to ammonium in two invasive and native Xanthium species.

Soil nitrogen forms are important for exotic plant invasions. However, little effort has been made to study the molecular mechanisms underlying the utilization of different N forms in co-occurring invasive and native plants. The invasive plant Xanthium strumarium prefers nitrate relative to ammonium, and mainly invades nitrate-dominated environments, while it co-occurring native congener X. sibiricum prefers ammonium. Here, we addressed the genetic bases for the interspecific difference in ammonium use and the effects of gibberellin (GA). Twenty-six transcripts related with GA biosynthesis and ammonium utilization were induced by ammonium in X. sibiricum, while only ten in X. strumarium and none for ammonium uptake. XsiAMT1.1a, XsiGLN1.1 and XsiGLT1b may be crucial for the strong ability to absorb and assimilate ammonium in X. sibiricum. All tested transcripts were significantly up-regulated by GA1 and GA4 in X. sibiricum. XsiGA3OX1a, which was also induced by ammonium, may be involved in this regulation. Consistently, glutamine synthetase activity increased significantly with increasing ammonium-N/nitrate-N ratio for X. sibiricum, while decreased for X. strumarium. Our study is the first to determine the molecular mechanisms with which invasive and native plants use ammonium differently, contributing to understanding the invasion mechanisms of X. strumarium and its invasion habitat selection.

Fabrication of cellulose derivative coated spherical covalent organic frameworks as chiral stationary phases for high-performance liquid chromatographic enantioseparation.

Porous spherical silica-based chiral stationary phases (CSPs) have been commercially used in the field of chiral separation, however, the scope of their application is, to some extent, limited by the instability of silica towards mobile phase containing strong base or acid. As such, developing new matrix-based CSPs is one of the effective strategies to overcome this bottleneck in studies of chiral separation materials. In this work, we have demonstrated that stable spherical covalent organic frameworks (SCOFs) can be utilized as matrixes for the fabrication of new CSPs for the first time. Specifically, a porous imine-linked SCOF with good crystallinity, large surface area, and high chemical stability is synthesized at room temperature. Then, cellulose-tris (3,5-dimethylphenylcarbamate) (CDMPC), a typical cellulose derivative, is selected as a potential chiral selector and coated onto the robust SCOFs, giving rise to the fabrication of new CDMPC@SCOF CSPs. The as-synthesized stable SCOF-based CSPs are exploited for high-performance liquid chromatographic (HPLC) enantioseparation, showing high resolution abilities for the separation of racemic compounds such as metalaxyl, 1-(1-naphthalenyl)ethanol, epoxiconazol, trans-stilbene oxide, and so on. Moreover, the prepared SCOF-based CSPs exhibit more superior acid and base stability than those of the silica-based CSPs. Our work not only uncovers the great potential of SCOFs as matrixes for constructing novel CSPs, but also expands the application of COFs in the field of enantiomeric separation under harsh base and acid conditions.

Joint effect of pre-operative anemia and perioperative blood transfusion on outcomes of colon-cancer patients undergoing colectomy.

BACKGROUND: Both pre-operative anemia and perioperative (intra- and/or post-operative) blood transfusion have been reported to increase post-operative complications in patients with colon cancer undergoing colectomy. However, their joint effect has not been investigated. The purpose of this study was to evaluate the joint effect of pre-operative anemia and perioperative blood transfusion on the post-operative outcome of colon-cancer patients after colectomy. METHODS: We identified patients from the American College of Surgeons National Surgical Quality Improvement Program (NSQIP) database 2006-2016 who underwent colectomy for colon cancer. Multivariate logistic regression analysis was employed to assess the independent and joint effects of anemia and blood transfusion on patient outcomes. RESULTS: A total of 35,863 patients-18,936 (52.8%) with left-side colon cancer (LCC) and 16,927 (47.2%) with right-side colon cancer (RCC)-were identified. RCC patients were more likely to have mild anemia (62.7%) and severe anemia (2.9%) than LCC patients (40.2% mild anemia and 1.4% severe anemia). A total of 2,661 (7.4%) of all patients (1,079 [5.7%] with LCC and 1,582 [9.3%] with RCC) received a perioperative blood transfusion. Overall, the occurrence rates of complications were comparable between LCC and RCC patients (odds ratio [OR] = 1.01; 95% confidence interval [CI] = 0.95-1.07; P = 0.750). There were significant joint effects of anemia and transfusion on complications and the 30-day death rate (P for interaction: 0.010). Patients without anemia who received a transfusion had a higher risk of any complications (LCC, OR = 3.51; 95% CI = 2.55-4.85; P < 0.001; RCC, OR = 3.74; 95% CI = 2.50-5.59; P < 0.001), minor complications (LCC, OR = 2.54; 95% CI = 1.63-3.97; P < 0.001; RCC, OR = 2.27; 95% CI = 1.24-4.15; P = 0.008), and major complications (LCC, OR = 5.31; 95% CI = 3.68-7.64; P < 0.001; RCC, OR = 5.64; 95% CI = 3.61-8.79; P < 0.001), and had an increased 30-day death rate (LCC, OR = 6.97; 95% CI = 3.07-15.80; P < 0.001; RCC, OR = 4.91; 95% CI = 1.88-12.85; P = 0.001) than patients without anemia who did not receive a transfusion. CONCLUSIONS: Pre-operative anemia and perioperative transfusion are associated with an increased risk of post-operative complications and increased death rate in colon-cancer patients undergoing colectomy.

[Research of water contact and labor practice of residents in plateau mountain schistosomiasis endemic areas in Yunnan Province].

OBJECTIVE: To understand the status of water contact and labor practice of residents in plateau mountain schisto- somiasis endemic areas in Yunnan Province, so as to provide the evidence for formulating the strategy of prevention and control of schistosomiasis in the next step of work. METHODS: A total of 412 residents aged 6-65 years old in 2 villages were randomly sampled and investigated with questionnaires for their water contacts and labor practices in plateau valley schistosomiasis endem- ic areas. Then the schistosome infection status of the residents was surveyed with the indirect hemagglutination assay (IHA). RE- SULTS: Among the 412 residents investigated, the rate of water contact was 88.35%, and the main causes of water contact were watering, and swimming and playing. The percentages of residents who had schistosomiasis history were 1.16%, 0.00%, 30.80%, 3.85%, and 0 in them who had swimming and playing water-contacts, bathing, watering, washing hands, and others, respectively (χ2 = 38.96, P < 0.01). The positives of IHA in the above-mentioned residents were 18.60%, 0.00%, 37.60%, 23.08%, and 0, respectively (χ2 = 12.61, P < 0.05). CONCLUSIONS: The productive infested water contact is the main way of schistosome infection. Therefore, the changes of labor practices and water contact of the residents are very important for the ef- fective prevention and control of schistosomiasis transmission.

[Research of freely grazing and wild feces behaviors in schistosomiasis endemic areas of plateau mountain area in Yunnan Province].

OBJECTIVE: To understand the situation of the freely grazing and wild feces behaviors of residents in plateau mountain area of schistosomiasis endemic areas in Yunnan Province. METHODS: Two villages of Xidian and Moguang in Heqing County, Yunnan Province were selected as the study area and the questionnaire surveys were performed to the randomly selected villagers aged 6 to 65 years with the sampling ratio of 30%. Then the respondents were tested for the infection of schistosomiasis by indirect hemagglutination assay (IHA). RESULTS: Totally 412 residents were surveyed. In all the responds, the ratios of "captive breed", "freely grazing "unknown" and "no response" were 55.34%, 4.85%, 26.94% and 12.86% respectively; and the ratios of "no grazing", "less than 5/week" and "no less than 5/week" were 75.49%, 16.02% and 8.50% respectively; and the ratios of "no wild feces", "less than 5/week", "no less than 5/week" and "no answer" were 68.45%, 27.67%, 2.91% and 0.97% respectively. CONCLUSIONS: Freely grazing and wild feces behaviors of residents in schistosomiasis endemic area of Heqing County are widespread, could heavily affect the control of schistosomiasis in Heqing County. The further work is to strengthen the management of human and animal feces and grazing and consolidate the results of the prevention and control of schistosomiasis and finally achieve the aim of the transmission interruption.

[Epidemiological investigation on fascioliasis and its risk factors in population of Binchuan County, Yunnan Province].

OBJECTIVE: To understand the infection status of Fasciola and related risk factors in population of Binchuan County, Yunnan Province. METHODS: In February and March, 2014, 6 natural villages from 2 administrative villages in Zhoucheng Town were selected as investigated sites, and the permanent residents aged ≥ 5 years were examined by ELISA for the serum antibody detection, and then the positive ones were examined by Kato-Katz technique and the hatching method for Fasciola egg detection. Meanwhile, the fascioliasis cases that checked out by the above methods were further surveyed, and the residents who with the same gender, similar age and negative results in the blood examination were selected as the controls, then the risk factors were explored through a case control study. RESULTS: Totally 1 207 people were included in the blood examination, and 36 positives were found with a positive rate of 2.98%. Though the differences between the positive rates of the serum antibody in people with different genders and age were not statistically significant (χ2 = 1.686, 7.106, both P > 0.05) , those between the rates of people in different administrative villages and natural villages were statistically significant (χ2 = 4.103, 13.181, both P < 0.05). Two cases were determined as positives in fecal examinations with a positive rate of 6.45% (2/31), among which, 1 case was newly discovered. The case control study showed that OR values of eating out- purchased cold dish (4.30), raw mint (1.22) , raw leek (4.30) and drinking unboiled-water (3.75) were relatively higher, but all of them were not statistically significant (χ2 = 0.524, 0.046, 0.524, 1.571, all P > 0.05). CONCLUSIONS: There is still sporadic Fasciola infection in Binchuan County. Eating raw vegetables and drinking unboiled water which are contaminated by Fasciola metacercariae may be the infection routes of local people, and to find out the exact entrance way is the key points for the infection control.

[Effect of comprehensive schistosomiasis control measures based on infection source control in mountainous areas of Yunnan Province].

OBJECTIVE: To evaluate the effect of comprehensive schistosomiasis control measures based on infection source control in plateau mountain areas of Yunnan Province. METHODS: From 2006 to 2004, four administrative villages were selected as test areas from plateau canyon and plateau basin endemic areas in Jindun Town, Heqing County, two villages each type, and the comprehensive control measures were implemented, including the examination and treatment of schistosomiasis, Oncomelania hupensis snail survey and control, health education, improving drinking water and lavatories, banning grazing, constructing sanitary pen of livestock, replacing cattle with machine, etc. The schistosome infection state and snail status in 2006 were treated as the baseline information, and the effect of the comprehensive measures were evaluated. Results The infection rate of human in plateau canyon areas decreased from 4.94% in 2006 to 0.06% in 2014, and that of livestock decreased from 1.11% to 0. In plateau basin areas, there was only 1 case of schistosomiasis found in Xiaolian Village in 2007, and no any other cases found in the other years, the infection rates of livestock dropped from 7.38% to 0. Compared with 2006, the snail areas in the two type areas decreased by 74.89% and 75.30%, respectively, meanwhile, the percentage of snail area, the occurrence rate of frames with snails, as well as the average density of living snails also decreased, and no infected snails were found since 2008. Xidian and Xinzhuang villages in plateau canyon area reached the criteria of schistosomiasis transmission controlled in 2009, and Xiao-lian and Kangfu villages in plateau basin reached the criteria of transmission interrupted in 2014. CONCLUSIONS: The comprehensive schistosomiasis control measures based on infection source control can effectively control the endemic situation of schistosomiasis in plateau areas of Yunnan Province. In the future, we should pay an equal attention to the infection sources control and snail control to consolidate and amplify the achievement of schistosomiasis control.

An Outbreak of Human Fascioliasis gigantica in Southwest China.

Fascioliasis is a common parasitic disease in livestock in China. However, human fascioliasis is rarely reported in the country. Here we describe an outbreak of human fascioliasis in Yunnan province. We reviewed the complete clinical records of 29 patients and performed an epidemiological investigation on the general human population and animals in the outbreak locality. Our findings support an outbreak due to Fasciola gigantica with a peak in late November, 2011. The most common symptoms were remittent fever, epigastric tenderness, and hepatalgia. Eosinophilia and tunnel-like lesions in ultrasound imaging in the liver were also commonly seen. Significant improvement of patients' condition was achieved by administration of triclabendazole®. Fasciola spp. were discovered in local cattle (28.6%) and goats (26.0%). Molecular evidence showed a coexistence of F. gigantica and F. hepatica. However, all eggs seen in humans were confirmed to be F. gigantica. Herb (Houttuynia cordata) was most likely the source of infections. Our findings indicate that human fascioliasis is a neglected disease in China. The distribution of triclabendazole®, the only efficacious drug against human fascioliasis, should be promoted.

[Effect of comprehensive control and management of schistosomiasis at plateaus regions in Yunnan Province].

OBJECTIVE: To evaluate the effect of comprehensive control and management of schistosomiasis at plateaus regions in Yunnan Province. METHODS: Xiaolian and Kangfu villages at plateaus regions where schistosomiasis was endemic were selected as study areas from 2006 to 2011, the schistosomiasis comprehensive control measures were conducted, and these measures included the health education, chemotherapy, stool and water management, grazing forbidden, Oncomelania snail control, etc. combined with the infrastructure of farmland water conservancy, agricultural comprehensive development, adjustment of industry structure, returning farmland to forest, ditch hardening, and aquaculture. RESULTS: At Xiaolian Village, no schistosomiasis patient and livestock were found from 2008; and at Kangfu Village, no schistosomiasis patient was detected from 2006 and no schistosomiasis domestic animal was found from 2008. In 2011, the snail area, percent of frames with living snails, the number of snails, average concentration of living snails, and the highest concentration of living snails descended by 69.99%, 81.86%, 88.86%, 89.71%, 57.95% at Xiaolian Village, and descended by 27.65%, 1.11%, 94.71%, 92.16%, 88.00% at Kangfu Village, respectively, compared with those in 2006. The ratios of infected snail area to snail area were 68.91% in 2006 and 69.13% in 2007 at Xiaolian Village, and 61.73% in 2006 and 43.24% in 2007 at Kangfu Village. There were no infected snails from 2008 in the two villages. CONCLUSION: The comprehensive control and management measures can effectively control the schistosomiasis prevalence at plateaus regions.

[Investigation on outbreak of angiostrongyliasis cantonensis due to consumption of snail food in Dali City].

OBJECTIVE: To investigate the potential cause of an outbreak of eosinophilic meningitis in Dali City in March, 2011. METHODS: A retrospective investigation was performed among 18 consumers who had meal together. The clinical documents of inpatients were analyzed and the diagnoses were checked. The outpatients were investigated through a case survey. Pomacea snails on sale were examined for the infection status of Angiostrongylus cantonensis. RESULTS: Of 18 persons, 16 consumed Pomacea snail food and 9 were infected with an infection rate of 56.3%. The two persons who did not consumed snail food were not infected. Serum samples of seven patients were examined for antibody to Angiostrongylus cantonensis in peripheral blood and 3 were proved positive (42.9%). The snails consumed by the patients came from the market, and the surveillance data showed that the infection rate of Angiostrongylus cantonensis of Pomacea snails was 7.3%. CONCLUSION: According to the epidemiological survey, clinical manifestations, and laboratory examinations, the outbreak of eosinophilic meningitis is due to the consumption of Pomacea snails infected with Angiostrongylus cantonensis.