Rapid isolation and determination of bisphenol A in complicated matrices by magnetic molecularly imprinted electrochemical sensing.

Because of its widespread distribution in the environment, bisphenol A (BPA) has become a global concern as an endocrine disruptor and a threat to human health through the food chain. Thus an efficient determination method is urgently needed for monitoring the levels of BPA. Herein, a novel electrochemical technique for the detection of BPA was performed by synchronous extraction and pre-concentration of BPA onto magnetic molecularly imprinted polymer (BMMIP), with subsequent readout on a magneto-actuated glassy carbon electrode (MGCE) by differential pulse voltammetry. Compared to the current methods of BPA determination, this BMMIP-based electrochemical sensor (BMMIPs@MGCE) not only simplifies the sample handling procedures substantially, without filtration, centrifugation, or other complex operations, but also can be easily renewed by a controllable magnetic field. As a sensor component, the core-shell BMMIPs exhibited excellent binding capacity (Qe = 82.5 mg g-1), short adsorption equilibrium time (30 s), and outstanding selectivity (k' = 7.239) towards BPA, as well as stability and recyclability. Importantly, the BMMIPs@MGCE sensor was successfully applied for the on-site monitoring and rapid detection of BPA in complicated real-world specimens, with good recoveries (81.31-119.77%) and a low limit of detection (0.133 µmol L-1). Therefore, the stable and low-cost BMMIPs@MGCE sensor provides a new approach for the rapid determination of BPA in the field of environmental control and food safety. Graphical abstract.

En bloc right hemicolectomy with pancreatoduodenectomy for right-sided colon cancer invading duodenum.

BACKGROUND: En bloc right hemicolectomy with pancreatoduodenectomy (RHCPD) is the optimum treatment to achieve the adequate margin of resection (R0) for locally advanced right-sided colon cancer with duodenal invasion. Information regarding the indications and outcomes of this procedure is limited. METHOD: In this retrospective study, 2269 patients with right colon cancer underwent radical right colectomy between October 2010 and May 2019, in which 19 patients underwent RHCPD for LARCC were identified. The overall survival (OS), disease-free survival (DFS), operative mortality, postsurgical complications, gene mutational analysis, and prognostic factors were evaluated. Survival was estimated using Kaplan-Meir method. RESULTS: Of these 19 patients who underwent LARCC, the OS was 88%, 66%, and 58% at 1, 3, and 5 years. The DFS was 72%, 56%, and 56% at 1, 3, and 5 years. The median operative time was 320 min (range: 222-410 min), and the median operative blood loss was 268 mL (range: 100-600 mL). The OS was significantly better among patients with well-differentiated tumor, N0 stage, and high microsatellite instability (MSI) and in patients who received adjuvant chemotherapy. The major postoperative complications occurred in 8 patients (42%), with pancreatic fistula (PF) being the most common. On the basis of the univariate analysis, poorly differentiated tumor, regional lymph node dissemination, MSI status, and no perioperative chemotherapy were the significant predictors of poor survival (P < 0.05). CONCLUSIONS: This study suggests that RHCPD is feasible and can achieve complete tumor clearance with favorable outcome, particularly in patients with lymph node-negative status.

Visceral Leishmaniasis in China: an Endemic Disease under Control.

Visceral leishmaniasis (VL) caused by Leishmania spp. is an important vector-borne and largely zoonotic disease. In China, three epidemiological types of VL have been described: anthroponotic VL (AVL), mountain-type zoonotic VL (MT-ZVL), and desert-type ZVL (DT-ZVL). These are transmitted by four different sand fly species: Phlebotomus chinensis, P. longiductus, P. wui, and P. alexandri. In 1951, a detailed survey of VL showed that it was rampant in the vast rural areas west, northwest, and north of the Yangtze River. Control programs were designed and implemented stringently by the government at all administrative levels, resulting in elimination of the disease from most areas of endemicity, except the western and northwestern regions. The control programs consisted of (i) diagnosis and chemotherapy of patients, (ii) identification, isolation, and disposal of infected dogs, and (iii) residual insecticide indoor spraying for vector control. The success of the control programs is attributable to massive and effective mobilization of the general public and health workers to the cause. Nationally, the annual incidence is now very low, i.e., only 0.03/100,000 according to the available 2011 official record. The overwhelming majority of cases are reported from sites of endemicity in the western and northwestern regions. Here, we describe in some depth and breadth the current status of epidemiology, diagnosis, treatment, and prevention of the disease, with particular reference to the control programs. Pertinent information has been assembled from scattered literature of the past decades in different languages that are not readily accessible to the scientific community. The information provided constitutes an integral part of our knowledge on leishmaniasis in the global context and will be of special value to those interested in control programs.

[Analysis of Antibodies with Superior Antigen-recognition in Serum from Patients Infected by Necator americanus].

Objective: To evaluate the reactivity of adult hookworm antigens to serum from patients with hookworm disease, and analyze in the serum class- or subclass-specific antibodies that show superior antigen recognition. Methods: Sera from healthy participants, patients infected by Necator americanus and those with other parasitic infections were processed for ELISA, which used raw antigens extracted from adult worms of Necator americanus as the coating antigen, and different classes or subclasses of anti-human antibody labeled with HRP as the secondary antibody. The sensitivity and specificity of the assay with various secondary antibodies were compared. Results: The ELISA using IgM, IgD，IgE, IgG, IgG1, IgG2, IgG3, or IgG4 as the secondary antibody showed a sensitivity of 41.84%, 2.04%, 1.02%, 92.93%, 19.39%, 25.51%, 17.35%, and 88.78%, respectively; specificity of 77.61% 97.01%, 92.54%, 79.10%, 95.52%, 92.53%, 92.53%, and 92.53%, respectively; and diagnostic efficiency of 56.36%, 40.61%, 38.18%, 87.88%, 50.30%, 52.7%, 47.88%, and 90.30%, respectively. The sensitivity when using IgG4 and IgG as the secondary antibody had far exceeded that when using IgM, IgD, IgE, and other three subclasses of IgG (P<0.05). There was no difference in sensitivity between tests using IgG4 and IgG (χ2=1.61, P＞0.05). However, the test using IgG4 revealed significantly higher specificity than that using IgG (χ2=4.97, P＜0.05). Conclusion: Use of IgG4 as the enzyme-linked secondary antibody shows advantages in overall diagnostic efficiency over other classes/subclasses in ELISA.

[Molecular networks and mechanisms of epithelial-mesenchymal transition regulated by miRNAs in the malignant melanoma cell line].

Melanoma is a malignant cutaneous cancer of high metastasis and lethal rates. Epithelial-mesenchymal transition (EMT) plays an important role in the embryonic developmental process that is often activated during tumorigenesis and metastasis. In this study, we integrated of mRNA and miRNA transcriptome sequencing data of melanocyte and melanoma cell lines to identify genes involved in the process of tumor EMT in the first place, and uncovered 11 miRNAs including miR-130a-3p, miR-130b-3p, miR-125a-5p, miR-30a-3p, miR-195-5p, miR-345-5p, miR-509-3-5p, miR-374a-5p, miR-509-5p, miR-148a-3p and miR-330-3p, negatively related with EMT genes using the Mirsystem software. Bioinformatics analysis with target genes of these miRNAs revealed two networks closely related with cellular development and cell-to-cell interactions, as well as multiple signaling pathways participating in EMT. Validation of the 11 miRNAs with molecular biology experiments demonstrated that four miRNAs regulated oncogenes in melanomas, including miR-195-5p, miR-130a-3p, miR-509-5p, and miR-509-3-5p. Our study integrates two kinds of omics data to screen for EMT-related miRNAs, providing a new research idea in the precision genomics of cancer research.

Differential diagnosis of cystic and alveolar echinococcosis using an immunochromatographic test based on the detection of specific antibodies.

Human cystic and alveolar echinococcoses are zoonotic diseases caused by the larval stages of Echinococcus granulosus and Echinococcus multilocularis, respectively. As the diseases are co-endemic in many areas of the world, a simple and rapid test for the differential diagnosis of cystic echinococcosis (CE) and alveolar echinocoocosis (AE) is needed. Here, we describe the development of an immunochromatographic test (ICT) using crude hydatid cyst fluid and a recombinant 18-kDa protein (rEm18) as antigens for the detection of E. granulosus and E. multilocularis antibodies in serum samples. The ICT was evaluated with serum samples from 195 echinococcosis patients from different endemic areas in northwestern China. These included 144 from CE patients, 51 from AE patients, 67 from patients with other parasitic diseases, 13 from patients with serous hepatic cysts, and 60 from healthy individuals. The sensitivity and specificity of the ICT for CE were 91.0 and 96.9% and for AE were 98.0 and 99.3% with diagnostic efficiencies of 94.1 and 99.1%, respectively. No significant differences and high degrees of agreement were found between the ICT and an enzyme-linked immunosorbent assay for both CE and AE. Five serum samples from cysticercosis patients and one serum sample from a healthy control were found positive for CE with the ICT. These findings indicate that this test allows for discrimination between both forms of human echinococcosis. In conclusion, the ICT developed in this study is a promising tool for the simultaneous detection and discrimination of CE and AE. This test will be useful for serodiagnosis of CE and AE in clinical settings and screening programs.

[Rapid risk assessment on the import of American trypanosomiasis to China].

American trypanosomiasis, as one of the "neglected tropical diseases", is a zoonosis induced by Trypanosoma cruzi. It is endemic in 18 countries in the Central and South America, especially in rural areas. A rapid risk assessment was carried out to analyze the potential threat of imported cases to China, which would provide information to policy makers in health authorities.

[Research progress on the diagnosis technology of canine visceral leishmaniasis].

Canine visceral leishmaniasis is caused by Leishmania infantum. Infected dogs, either symptomatic or asymptomatic, are considered as the major reservoirs for zoonotic visceral leishmaniasis. Accurate and rapid detection of canine leishmanial infection is crucial for control of human visceral leishmaniasis due to its role in the transmission of the infection to vectors. Various techniques based on parasitology, immunology and molecular biology have been studied and evaluated for detecting canine leishmanial infection. This article reviews the progress in techniques and methods for its diagnosis.

[Establishment and evaluation of colloid gold labeled immunochromatographic strip test for rapid diagnosis of alveolar echinococcosis].

OBJECTIVE: To establish and evaluate a colloid gold immunochromatographic strip test for the diagnosis of alveolar echinococcosis. METHODS: Total RNA was prepared from Echinococcus multilocularis protoscoleces collected from Xinjiang Uygur Autonomous Region. Em18 gene was obtained by reverse transcription-polymerase chain reaction (RT-PCR). The PCR product was sequenced and cloned into pGEX-3X vector. The recombinant plasmid was expressed and induced by isopropyl-beta-D-thiogalactopyranoside (IPTG) to obtain recombinant protein. The anti-human IgG monoclonal antibodies was conjugated with colloid gold as detecting reagent; the recombinant Em18 antigen and goat anti-mouse IgG were immobilized on nitrocellulose in proper position. The prepared immunochromatographic strip was evaluated using serum samples from patients with alveolar echinococcosis (56), cystic echinococcosis (87), cysticercosis (30), schistosomiasis japonica (10), toxoplasmosis (10) and healthy subjects (50) . Comparison between the immunochromatographic strip test and ELISA was made by kappa statistics. RESULTS: Sensitivity detected by the immunochromatographic strip test was 92.9% (52/56). The cross-reactivity to cystic echinococcosis and cysticercosis was 9.2% (8/87) and 3.3% (1/30), respectively. There was no cross reactivity with schistosomiasis japonica and toxoplasmosis. 4 samples out of 50 healthy people showed false positive reaction. The overall specificity was 93.0 (174/187). Sensitivity and specificity both showed no statistical difference between immunochromatographic strip test and ELISA. High degree of agreement was observed between the strip test and ELISA (kappa = 0.98). CONCLUSION: The developed immunochromatographic strip test using recombinant Em18 antigen as coated antigen is a sensitive, specific, simple and rapid assay for diagnosing alveolar echinococcosis.

[Detection and species identification of two imported cases of cutaneous leishmaniasis].

OBJECTIVE: To diagnose and identify pathogen of two suspected cases of cutaneous leishmaniasis. METHODS: Two cases of dermatosis with several major ulcers on the skin were examined, who worked and returned from Algeria (case 1) and Saudi Arabia (case 2), respectively. The stained smears of skin tissue from lesions were observed by microscope. Extravasate from lesions was cultured in NNN medium to search protozoan parasites, which were obtained by centrifugation. Two pairs of species-specific primers, ITS1-ITS2 and K13A-K13B, were used to amplify inter-nal transcribed spacer of rDNA and kinetoplast DNA, respectively. The products were sequenced and analyzed by Blast. RESULTS: There were Leishmania amastigotes in the tissue smear of case 2, while none in that of case 1. Promastigotes were found in culture medium of both cases. The PCR products of ITS1-ITS2 and K13A-K13B from 2 cases were about 330 bp and 120 bp with respective homology of 100% and 96% to corresponding sequences of Leishmania major. The accession numbers of 4 sequences were JF831924-JF831927. CONCLUSION: Two cases of dermatosis are diagnosed as imported cutaneous leishmaniasis and the pathogen is L. major.

Label-Free Quantitative Proteomic Analysis of Three Strains of Viscerotropic Leishmania Isolated from Patients with Different Epidemiological Types of Visceral Leishmaniasis in China.

BACKGROUND: There are three epidemiological types of visceral leishmaniasis in China, which are caused by Leishmania strains belonging to the L. donovani complex. The mechanisms underlying their differences in the population affected, disease latency, and animal host, etc., remain unclear. We investigated the protein abundance differences among Leishmania strains isolated from three types of visceral leishmaniasis endemic areas in China. METHODS: Promastigotes of the three Leishmania strains were cultured to the log phase and harvested. The protein tryptic digests were analyzed with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), followed by label-free quantitative analysis. The MS experiment was performed on a Q Exactive mass spectrometer. Raw spectra were quantitatively analyzed with the MaxQuant software (ver 1.3.0.5) and matched with the reference database. Differentially expressed proteins were analyzed using the bioinformatics method. The MS analysis was repeated three times for each sample. RESULTS: A total of 5012 proteins were identified across the KS-2, JIASHI-5 and SC6 strains in at least 2 of the three samples replicate. Of them, 1758 were identified to be differentially expressed at least between 2 strains, including 349 with known names. These differentially expressed proteins with known names are involved in biological functions such as energy and lipid metabolic process, nucleotide acid metabolic process, amino acid metabolic process, response to stress, cell membrane/cytoskeleton, cell cycle and proliferation, biological adhesion and proteolysis, localization and transport, regulation of the biological process, and signal transduction. CONCLUSION: The differentially expressed proteins and their related biological functions may shed light on the pathogenicity of Leishmania and targets for the development of vaccines and medicines.

Contributions of the National Institute of Parasitic Diseases to the control of visceral leishmaniasis in China.

Visceral leishmaniasis (VL) caused by Leishmania spp. is an important vector-borne disease prevalent in China. VL was rampant in the vast area of China north of the Yangtze River before the founding of the People's Republic of China in 1949. As a result of strenuous interventions, the disease was basically eliminated in most of the former epidemic areas in 1958-60. At present, only sporadic cases occur in the western regions of China. In the process, National Institute of Parasitic Diseases at China CDC and the Chinese Center for Tropical Diseases Research (NIPD-CTDR) have achieved great impact in controlling the diseases as well as in research on Leishmania spp. This review summarized the contribution of experts from NIPD-CTDR to the control and elimination of VL in various aspects, such as understanding the epidemiological features of VL, confirmation of VL vectors and their distribution, development of control tools including diagnostics and insecticides, monitoring and evaluation supported by information management, technical supports to the control programmes, as well as analysis of the challenges faced. At the same time, it puts forward constructive suggestions for the ultimate interruption of VL transmission in China.

[Public health situation and needs assessment after Wenchuan earthquake in Anxian, Sichuan province].

OBJECTIVE: To explore the public health situation and needs in Anxian after Wenchuan earthquake so as to make an effective strategy for disease control and prevention. METHODS: 69 concentrated settlements with 100 residents were investigated. Probability proportion to size was adopted for sampling of 2200 residents from 687 scattered households (about 440 000 scattered residents). The content of this survey included drinking water, food hygiene, environment sanitation, planning immunity and medical health service, disease surveillance and so on. SPSS 16.0 was used for data analysis, and statistical interpretation was used to describe the results. RESULTS: 90.9% (31/66) resettled residents in Anxian lived in tents, 7.6% (5/66) lived in the movable-plate house, 93.3% (621/666) scattered households lived in tents and 71.9% (446/621) of them lived in tents which were built by residents themselves; the rate of drinking water disinfection in resettlement sites and scattered households were 97.1% (66/68) and 94.6% (650/687); 12.8% scattered residents had mouldy or food; 50% of resettlement sites raised animals; 43.6% (17/39) medical station didn't have bacterin inoculation service; 66.7% (10/15) lacked sufficient disinfection equipment; register rate was 50.0% (33/66) and report rate of symptoms and infectious diseases was 56.1% (37/66). CONCLUSION: There was still some risk of enteric and vector-borne diseases in Anxian, therefore, some tailored measures should be very important.

[Mental health assessment among scattered residents after Wenchuan earthquake in Anxian, Sichuan province].

OBJECTIVE: To explore the mental health status of residents scattered living in Anxian after Wenchuan earthquake so as to provide scientific basis for further mental health intervention. METHODS: A face to face interview was conducted among the scattered residents with designed questionnaire, which had three parts of the physical and emotional reaction, the relax methods and the social care and supports expected. Two-stage probability proportional to size (PPS) sample method was performed to sample 2184 from 0.44 million scattered residents in Anxian. On the basis of statistical description, mental health of different characteristics groups was compared. RESULTS: Three main symptoms of posttraumatic stress disorders in 2184 residents (11.23+/-3.44) were higher than the 103 fire victims in Hunan in 2003 (10.06+/-3.26), three factor scores of SCL-90 (5.76+/-1.74) were higher than normal in 1998 repair mode (n=23 891) (4.72+/-1.44), and the statistical difference was observed (t=10.77, P<0.05; t=706.04, P<0.05). Comparing the mental health of different groups, some significant differences were found by age, gender and education background. CONCLUSION: The earthquake disaster brought prevalent physical and emotional reaction. Elderly people, female, junior students need mental intervention immediately. Therefore, strengthen the mental education and assistance (especially in high risk groups) would be of more significance.

[Field evaluation of gold-immunochromatographic assay for diagnosis of vivax malaria].

OBJECTIVE: To evaluate a gold-immunochromatographic assay (GICA) for malaria diagnosis in an endemic area of vivax malaria. METHODS: Blood samples were collected from febrile patients in 5 township-hospitals of Mengcheng County, Anhui Province, between September and October 2008. The samples were examined by GICA and microscopy under double blind condition and the results were compared. RESULTS: Among 292 blood samples, 181 were found P. vivax-positive by microscopy, and 163 were positive by GICA. Altogether, the coincidence of the two methods stood for 92.8% (271/292), including 108 negatives and 163 positives. 21 samples with discrepancy covered 18 microscopy positive but GICA negative, 3 microscopy negative but GICA positive. The GICA positive rate in patients with a parasitaemia of > 1,000 parasites/microl, 100-1,000 parasites/microl, and < 100 parasites/microl was 93.5% (115/123), 86.0% (43/50), and 62.5% (5/8), respectively. CONCLUSION: GICA is a useful diagnosis method for endemic area of vivax malaria.

[Preliminary evaluation on the transmission potential of visceral leishmaniasis after earthquake in Wenchuan].

Data of visceral leishmaniasis cases since 2005 were collected through the National Infectious Disease Monitoring System. Number of reported cases in 2005, 2006, 2007 and January to June in 2008 was 59, 49, 77 and 30 in Sichuan Province, and 92, 106, 162 and 83 respectively in Gansu Province. With an increase of the number of stray dogs and susceptible human population, damage of the medical services including diagnosis and treatment capacity after the earthquake, there might be a strengthened transmission potential and possible spread of the disease.

Field evaluation of an immunochromatographic test for diagnosis of cystic and alveolar echinococcosis.

BACKGROUND: The larval stages of the tapeworms Echinocoocus granulosus and Echinococcus multilocularis are the causative agents of human cystic echinococcosis (CE) and human alveolar echinococcosis (AE), respectively. Both CE and AE are chronic diseases characterised by long asymptomatic periods of many years. However, early diagnosis of the disease is important if treatment and management of echinococcosis patients are to be successful. METHODS: A previously developed rapid diagnostic test (RDT) for the differential detection of CE and AE was evaluated under field conditions with finger prick blood samples taken from 1502 people living in the Ganzi Tibetan Autonomous Prefecture, China, a region with a high prevalence for both forms of human echinococcosis. The results were compared with simultaneously obtained abdominal ultrasonographic scans of the individuals. RESULTS: Using the ultrasonography as the gold standard, sensitivity and specificity, and the diagnostic accuracy of the RDT were determined to be greater than 94% for both CE and AE. For CE cases, high detection rates (95.6-98.8%) were found with patients having active cysts while lower detection rates (40.0-68.8%) were obtained with patients having transient or inactive cysts. In contrast, detection rates in AE patients were independent of the lesion type. The positive likelihood ratio of the RDT for CE and AE was greater than 20 and thus fairly high, indicating that a patient with a positive test result has a high probability of having echinococcosis. CONCLUSIONS: The results suggest that our previously developed RDT is suitable as a screening tool for the early detection of human echinococcosis in endemic areas.

[Establishment and evaluation of colloid gold labeled immunochromatographic strip test for rapid diagnosis of malaria].

OBJECTIVE: To establish and evaluate a gold immunochromatographic strip test for detection and differentiation of Plasmodium vivax and P. falciparum. METHODS: The monoclonal antibodies, F4H12, G4C9 and D8F7, were conjugated with colloid gold as detecting reagent; monoclonal antibody B2G10 (against P. vivax/ P. falciparum) and D6A7 (only against P. falciparum) were immobilized on nitrocellulose in proper position. Blood samples from 107 febrile patients from endemic area of malaria and 17 patients with visceral leishmaniasis were used for evaluating the specificity. Blood samples of malaria patients (110 with P. vivax and 54 with P. falciparum) were used for evaluating the sensitivity. RESULTS: 5 samples out of 107 febrile patients and 17 patients with visceral leishmaniasis showed false positive reaction with a specificity of 96.0% (119/124), all the 17 samples from patients with visceral leishmaniasis were negative. 164 blood samples of malaria patients showed a sensitivity of 92.3% (153/164), 92.7% (102/110)and 94.4% (51/ 54) for patients infected with P. vivax or P. falciparum, respectively. CONCLUSION: The immunochromatographic strip test based on antigen-capturing is a sensitive, specific, simple and rapid assay for malaria diagnosis.

[Asymptomatic Leishmania infection in human population of Wenxian County, Gansu Province].

OBJECTIVE: To analyze the status of Leishmania infantum asymptomatic infection in human population of a Kala-azar endemic area in Wenxian County, Gansu Province, and to evaluate the tests used. METHODS: Blood samples were tested by PCR using two pairs of primers, RV1-RV2 and K13A-K13B, for detecting Leishmania-specific DNA. ELISA and rK39-dipstick were used to detect Leishmania-specific antibodies. RESULTS: The positive rate of PCR, ELISA and rK39-dipstick was 30.9%(83/269). 24.2%(65/269) and 0 (0/269) respectively. CONCLUSION: The prevalence of asymptomatic infection of L. infantum in humans is high in the area. PCR test based on RV1-RV2 and K13A-K13B primer pairs is a sensitive and specific method for detecting the asymptomatic infection.

[Study on PCR method for detecting the asymptomatic infection of Leishmania infantum].

OBJECTIVE: To establish PCR method for the detection of the asymptomatic infection of Leishmania infantum. METHODS: Six primer pairs were selected for detecting Chinese strain of L. infantum by optimizing conditions which affect amplification. Their sensitivity and specificity were compared by using DNAs extracted from human blood seeded with cultured L. infantum promastigotes (MHOM/CN/86/GS) as template. Blood samples of the inhabitants without symptoms of visceral leishmaniasis in the endemic area were analyzed with two selected primer pairs with good sensitivity and specificity. RESULTS: The specificity of all six primer pairs reached 100%, and the sensitivity varied among the primer pairs. The primer pairs RV1-RV2 (0.1 parasite/ml blood) and K13A-K13B (1 parasite/ml blood) were most sensitive. Leishmania DNA was detected in 33% (33/100) and 30% (30/100) human blood samples by RV1-RV2 and K13A-K13B primer pairs respectively. CONCLUSION: This study suggests that RV1-RV2 and K13A-K13B primer pairs are suitable in detecting the asymptomatic infection of L. infantum, and the prevalence of the asymptomatic infection is high in human population in the endemic area.