Objective To establish a multiplex nucleic acid assay for rapid detection of Echinococcus multilocularis, E. granulosus and E. shiquicus based on the recombinase-aided isothermal amplification assay (RAA) and to preliminarily assess its diagnostic efficiency. Methods The mitochondrial genomic sequences of E. multilocularis (GenBank accession number: NC_000928), E. granulosus (GenBank accession number: NC_044548) and E. shiquicus (GenBank accession number: NC_009460) were used as target sequences, and three pairs of primers were designed based on the RAA primer design principle and synthesized for the subsequent multiple RAA amplification. The genomic DNA of E. multilocularis, E. granulosus and E. shiquicus at different concentrations and the recombinant plasmids containing the target gene at various concentrations were amplified to evaluate the diagnostic sensitivity of the multiplex RAA assay, and the genomic DNA of E. multilocularis, E. granulosus, E. shiquicus, Taenia multiceps, T. saginata, T. asiatica, Dipylidium caninum, T. hydatigena, Toxocara canis, Fasciola hepatica, T. pisiformis, Mesocestoides lineatus and Cryptosporidiumn canis was detected using the multiplex RAA assay to evaluate its specificity. In addition, the reaction condition of the multiplex RAA assay was optimized, and was then employed to detect the tissues with echinococcosis lesions, simulated canine fecal samples and field captured fox fecal samples to examine its application values. Results The multiplex RAA assay was effective to specifically amplify the mitochondrial gene fragments of E. multilocularis, E. granulosus and E. shiquicus within 40 min at 39 °C, with sequence lengths of 540, 430 bp and 200 bp, respectively. This multiplex RAA assay showed the minimum detection limits of 2.0, 2.5 pg/μL and 3.1 pg/μL for detection of the genomic DNA of E. multilocularis, E. granulosus and E. shiquicus, and presented the minimum detection limit of 200 copies/μL for detection of the recombinant plasmids containing E. multilocularis, E. granulosus and E. shiquicus target genes. This multiplex RAA assay was effective to simultaneously detect single and multiple infections with E. multilocularis, E. granulosus and E. shiquicus, but failed to amplify the genomic DNA of T. multiceps, T. saginata, T. asiatica, D. caninum, T. hydatigena, T. canis, F. hepatica, T. pisiformis, M. lineatus and C. canis. In addition, the optimized multiplex RAA assay was effective to detect all positive samples from the tissue samples with echinococcosis lesions, simulated canine fecal samples and field captured fox fecal samples, which was fully consistent with the detection of the single PCR assay. Conclusion A sensitive and specific multiplex nucleic acid assay for rapid detection of E. multilocularis, E. granulosus and E. shiquicus has been successfully established.

Objective To understand the current status and changing tendency of human hookworm infections in Anhui Province. Methods According to the unified national survey scheme, a total of 48 survey sites were sampled from 16 counties (cities) in 4 ecological regions of Anhui Province using a stratified cluster random sampling method from 2014 to 2015. The hookworm eggs were detected in the fecal samples from permanent residents at ages of over one year living in the survey sites using a modified Kato-Katz thick smear method, and the subjects’health knowledge and behaviors were investigated using questionnaire survey. Results A total of 12 300 persons were examined in the 48 survey sites from 4 ecological regions of Anhui Province between 2014 and 2015, and 259 subjects were identified with hookworm infections, with a mean prevalence of 2.11%. Among the four ecological regions, the North China Plain had the highest prevalence of human hookworm infections (3.02%) and in all survey sites, Linquan County had the highest prevalence (7.03%). Ancylostoma duodenale was the predominant hookworm species identified (62.16%), and 65.64% had mild infections. The prevalence of human hookworm infections was significantly greater in women than in men (χ² = 4.16, P < 0.05), and showed a tendency towards a rise with ages (χ²_trend = 113.36, P < 0.01). In addition, the prevalence of human hookworm infections varied in occupations (χ² = 159.41, P < 0.01) and education levels (χ² = 34.95, P < 0.01). Questionnaire survey showed low prevalence of human hookworm infections in subjects knowing the question“how hookworm infection occurs”and denying“using fresh stools for fertilization”(χ² = 15.05, P < 0.01; χ² = 4.19, P < 0.05). Conclusions The prevalence of human hookworm infections has greatly decreased in Anhui Province; however, the prevalence remains relatively high in some regions and populations. The North China Plain should be regarded as the key area for hookworm disease prevention and control, and housewives and populations with advanced ages and low educational levels are key targeted populations in Anhui Province.

The purpose of this paper is to study the use rules of drugs for lung diseases in internal medicine department of Xin'an Wang's family, discuss the compatibility of common drugs for lung diseases, guide clinical application, and inherit Xin'an medicine. By retrospective study on lung diseases cases in Wang's internal medicine works, the lung diseases and use frequency of common drugs treated by Wang's medicine were counted, and the systematic clustering and association rule analysis of common drugs were conducted by using SPSS Statistic 20 and SPSS Modeler 18.0, respectively. The results showed that asthma, cold and cough were the main lung diseases treated by Wang's medicine, and the commonly used medicines included antitussive and antiasthmatic drugs, spleen-invigorating and dampness-removing drugs, and expectorants. The medicine taste was mainly bitter, pungent and sweet, with cold and warm properties in a balanced way, without severely cold or hot herbs, mainly attributing to the lung and stomach meridians. In clustering analysis, 10 drug combinations were obtained; association analysis showed that two, three, four association rules respectively had 11, 21, and 10 groups, and each drug group had 11, 16, and 5 items. Core combinations: Poria, Armeniacae Semen Amarum, Asteris Radix et Rhizome, Coicis Semen, Farfarae Flos, Dendrobii Caulis, Perilla Frutescens, Stemonae Radix, Citri Reticulatae Pericarpium, Cynanchi Stauntonii Rhizome et Radix, Meretricis Concha Cyclinae Concha, Belamcandae Rhizoma, and Pinelliae Rhizome. Xin'an Wang's medicine paid attention to the lung nature when treating lung diseases. Lung is a delicate organ, not resistant to coldness or heat, so severely cold or hot herbs shall not be used, and the clear and light drugs with functions of dispersing lung Qi, clearing phlegm evil, strengthening spleen, eliminating phlegm, and relieving cough and asthma are often used. Lung deficiency is a kind of deficiency of Qi and Yin, so both Qi and Yin shall be regulated. Deficiency of Yin would burn the lung and make the lung collaterals blocked. In this case, the lung collaterals shall be dredged for hemostasis. Long time of lung deficiency would hinder the distribution of body fluid, and lung shall be regulated to dissipate phlegm.
Data Mining ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lung Diseases ; drug therapy ; Medicine, Chinese Traditional ; Meridians ; Retrospective Studies

Coenurosis is an important zoonotic helminthic disease caused by the larval stage of the tapeworm Taenia multiceps. This parasite typically infects the brain of the intermediate hosts, including sheep, goat, cattle and even humans. We report a case of T. multiceps infection in a yak confirmed by clinical symptoms, morphological characteristics, and molecular and phylogenetic analyses. The coenurus was thin-walled, whitish, and spherical in shape with a diameter of 10 cm. The parasite species was identified as T. multiceps by PCR amplification and sequencing of the 18S rRNA, cox1 and nad1 genes. Three gene sequences all showed high homology (all above 97%) with the reference sequences from different hosts. Moreover, phylogenetic reconstructions with the 3 published Taenia gene sequences confirmed that the Qinghai yak isolate was closely related to T. multiceps. Although there are advanced diagnosis and treatment methods for coenurosis, early infection is difficult to diagnose. Importantly, the findings of yak infection case should not be ignored due to its zoonotic potential.
Animals ; Brain ; Cattle ; Cestoda ; China ; Diagnosis ; Goats ; Helminths ; Humans ; Parasites ; Polymerase Chain Reaction ; Sheep ; Taenia

OBJECTIVETo investigate the relationship between lipid accumulation and autofluorescence intensity of injury site after spinal cord injury (SCI), and explore whether CuSO⁴ can eliminate autofluorescence in the injury site after SCI.

METHODSThirty six Wild Type mice at age of 8 to 12 weeks (weight 18 to 24 g) were randomly divided into normal control group (4) and SCI group (32). Respectively, 8 mice of SCI group were sacrificed randomly at 1 week, 2 weeks, 4 weeks and 8 weeks after injury. Frozen sections of spinal cord tissue with injury site at the center were made to observe autofluorescence under green channel of fluorescence microscope (Specimens of normal control group were taken from the same segment of the spinal cord, and fixed with 4% paraformaldehyde solution). Oil Red O staining was applied to visualize the lipid accumulation in the injury site, and correlation between lipid accumulation and autofluorescence intensity was analyzed. Furthermore, sections were incubated with CuSO⁴ buffer to eliminate autofluorescence, and CuSO⁴ concentration and incubation time was optimized.

RESULTSNo obvious autofluorescence or lipid staining was found in normal spinal cord tissue sections. By contrast, autofluorescence appeared in the injury site of spinal cord sections, and the intensity increased with passing time after injury. Oil Red O staining showed that lipid accumulated in the injury site with passing time after injury as well, and the correlation between lipid accumulation and autofluorescence intensity was positive. After incubation with CuSO⁴ buffer, the autofluorescence in the injury site was significantly reduced, especially after optimizing CuSO⁴ concentration and incubation time.

CONCLUSIONSLipid accumulation may play an important role to determine the autofluorescence intensity of injury site after SCI, and the autofluorescence intensity can be used as a simple index for evaluating lipid peroxidation damage. Optimized method of using CuSO⁴ can significantly reduce the autofluorescence in the injury site after SCI, which will be beneficial to the application of immunofluorescence staining technique in the study of SCI.

OBJECTIVETo study the mechanism of interleukin 7/interleukin 7 receptor (IL-7/IL-7R) in promoting cell proliferation and inducing lymphangiogenesis of non-small cell lung cancer (NSCLC) in vivo and in vitro.

METHODSImmunohistochemical study for IL-7, IL-7R, cyclin D1 and vascular endothelial growth factor-D (VEGF-D) was carried out in NSCLC tissues from 95 patients. The relationship between IL-7/IL-7R expression and various parameters was analyzed. The mechanism of IL-7/IL-7R in promoting cell proliferation and inducing lymphangiogenesis was studied by methylthiazolyldiphenyl-tetrazolium bromide, fluorescence-activated cell sorting, reverse transcriptase-PCR, Western blot, co-immunoprecipitation, chromatin immunoprecipitation and nude mice experiments with xenograft tumors.

RESULTSIL-7 (63.2%, 60/95), IL-7R (61.1%, 58/95), cyclin D1 (52.6%, 50/95) and VEGF-D (58.9%, 56/95) showed that high level of expression in NSCLC. IL-7/IL-7R over-expression correlated with cyclin D1 expression (P < 0.01, P < 0.01), VEGF-D expression (P < 0.01, P < 0.01), increased lymphovascular density (P = 0.005, P = 0.013), advanced clinical stage (P = 0.008, P = 0.005) and presence of lymph node metastasis (P < 0.01, P < 0.01). IL-7/IL-7R could promote proliferation of A549 cell, increase cyclin D1 and VEGF-D expression, and enhance c-Fos/c-Jun expression and phosphorylation, resulting in formation of heterodimer. Furthermore, IL-7/IL-7R could induce binding of c-Fos/c-Jun to cyclin D1/VEGF-D promoters and regulate their transcription. IL-7/IL-7R could also promote proliferation and lymphangiogenesis of lung cancer xenograft tumors.

CONCLUSIONSIL-7/IL-7R promotes c-Fos/c-Jun expression and activity in NSCLC. This further facilitates cyclin D1 expression and accelerates proliferation of cells and VEGF-D-induced lymphovascular formation.

Animals ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Female ; Humans ; Interleukin-7 ; metabolism ; physiology ; Lung Neoplasms ; metabolism ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Staging ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-fos ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Receptors, Interleukin-7 ; metabolism ; physiology ; Vascular Endothelial Growth Factor D ; metabolism

OBJECTIVETo observe the exposure levels of extremely low frequency electromagnetic fields in workplaces and to analyze the effects of extremely low frequency electromagnetic radiation on cardiovascular system of occupationally exposed people.

METHODIntensity of electromagnetic fields in two workplaces (control and exposure groups) was detected with EFA-300 frequency electromagnetic field strength tester, and intensity of the noise was detected with AWA5610D integral sound level. The information of health physical indicators of 188 controls and 642 occupationally exposed workers was collected. Data were analyzed by SPSS17.0 statistic software.

RESULTSThe intensity of electric fields and the magnetic fields in exposure groups was significantly higher than that in control group (P < 0.05), but there was no significant difference of noise between two workplaces (P > 0.05). The results of physical examination showed that the abnormal rates of HCY, ALT, AST, GGT, ECG in the exposure group were significantly higher than those in control group (P < 0.05). There were no differences of sex, age, height, weight between two groups (P > 0.05).

CONCLUSIONExposure to extremely low frequency electromagnetic radiation may have some effects on the cardiovascular system of workers.

Adult ; Cardiovascular System ; radiation effects ; Case-Control Studies ; Electromagnetic Fields ; adverse effects ; Electromagnetic Radiation ; Humans ; Male ; Occupational Exposure ; adverse effects ; Young Adult

Objective To investigate the genetic stability of non-replicating recombinant adenovirus which used Ad41 as vector and could express VP6 gene of group A rotavirus during continous passage,in order to develop the vaccine of rotavirus.Methods The recombinant adenovirus rvAd41-VP6 (o) was prepared by our laboratory early,it then was continuously propagated on 293TE7 cells for 14 passages.After that samples of the infected cells were collected at every 2 passages for the detection of the integration of the VP6 gene by PCR,and the expression of the target protein was detected by Western Blot analysis.Results Analysis by PCR revealed that,there was stable integration of specific VP6 gene in the rvAd41-VP6 (o),Western Blot analysis confirmed that rvAd41-VP6 (o) could stably expressed the group-specific antigen structural protein VP6 (o),and it had preferable genetic stability.Conclusion The recombinant adenovirus rvAd41-VP6(o) which could stably express the VP6(o) gene had favorable biological property in vitro,and it has provided a basis for further research of animal immunization.

Objective To study the quality of life and its influential factors among HIV positive population. Methods Quality of life was evaluated among 2608 HIV positive population by WHOQOL (Chinese Version) to calculate the total and four domains' scores of quality of life. Multiple linear regression model was used to analyze the relationship between the factors and the scores of the four domains and the total score of quality of life. Results Physical,psychological,social,environmental and the total scores of the HIV positive population were 12.96±1.94,11.79± 1.19,13.79±2.44,12.40±1.93 and 51.02±6.03,respectively. Females had a higher scores than males in the four domains and the total score of quality of life. Through a multiple linear regression model,we found that the influential factors would include gender,age,occupation,family conflict and appetite etc. Conclusion People living with HIV had higher scores in social domain,but lower scores in psychological domain,suggesting that psychological intervention should he strengthened.