Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.

Objective To investigate the origin of Biomphalaria straminea in China, so as to provide insights into assessment of schistosomiasis mansoni transmission risk and B. straminea control. Methods Guanlan River, Dasha River, Shenzhen Reservoir, upper and lower reaches of Kuiyong River, and Xinzhen River in Shenzhen, China, were selected as sampling sites. Ten Biomphalaria samples were collected from each site, and genomic DNA was extracted from Biomphalaria samples. DNA samples were obtained from 15 B. straminea sampled from 5 sampling sites in Minas Gerais State, Pará State, Federal District, Pernambuco State, and Sao Paulo State in Brazil, South America. Cytochrome c oxidase I (COI) and mitochondrial 16S ribosomal RNA (16S rRNA) genes were sampled using the above DNA templates, and the amplified products were sequenced. The COI and 16S rRNA gene sequences were downloaded from GenBank, and the sampling sites were acquired. All COI and 16S rRNA gene sequences were aligned and evolutionary trees of B. straminea were created based on COI and 16S rRNA gene sequences to identify the genetic similarity and evolutionary relationship between B. straminea samples from China and South America. Results A total of 60 COI gene sequences with a length of 529 bp and 3 haplotypes were obtained from B. straminea sampled from China. There were 165 COI gene sequences of B. straminea retrieved from GenBank, and following alignment with the above 60 gene sequences, a total of 33 haplotypes were obtained. Phylogenetic analysis showed that the three haplotypes of B. straminea from China were clustered into one clade, among which the haplotype China11 and three B. straminea samples from Brazil retrieved from GenBank belonged to the same haplotype. Geographical evolution analysis showed that the B. straminea samples from three sampling sites along eastern coasts of Brazil had the same haplotype with China11, and B. straminea samples from other two sampling sites were closely, genetically related to China11. A total of 60 16S rDNA gene sequences with approximately 322 bp in length were amplified from B. straminea in China, with 2 haplotypes identified. A total of 70 16S rDNA gene sequences of B. straminea were captured from GenBank. Phylogenetic analysis showed that Biomphalaria snails collected from China were clustered into a clade, and the haplotype China64 and the haplotype 229BS from Brazil shared the same haplotype. The 49 16S rDNA gene sequences of B. straminea from 25 sampling sites in southern Brazil, which were captured from GenBank, were included in the present analysis, and the B. straminea from 3 sampling sites shared the same haplotype with China64 in China. Geographical evolution analysis based on COI and 16S rRNA gene sequences showed that B. straminea sampled from eastern coastal areas of Brazil shared the same haplotypes in two gene fragment sequences with Biomphalaria snails collected from China. Conclusions The Biomphalaria snails in China are characterized as B. straminea, which have a low genetic diversity. The Biomphalaria snails in China have a high genetic similarity with B. straminea sampled from eastern coastal areas of Brazil, which may have originated from the eastern coastal areas of Brazil.

OBJECTIVE：To estab lish a me thod for simultaneous determination of morroniside ，loganin，echinacoside and acteoside in Huanshao capsules. METHODS ：HPLC method was adopted. The determination was performed on Zhongpuhong RD-C18 column with mobile phase consisted of acetonitrile- 0.1％ formic acid solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm （morroniside，loganin） and 330 nm （echinacoside，acteoside）. The column temperature was set at 35 ℃，and sample size was 10 μL. RESULTS：The linear range were 5.29-105.80 μg/mL for morroniside， 4.49-89.88 mg/L for loganin ，16.26-325.25 mg/L for echinacoside and 16.31-326.25 mg/L for acteoside ，r values were 0.999 9. RSDs of precision ，stability （24 h），reproducibility and durability tests were all lower than 2.0％ . The recoveries were 94.34％ -96.23％（RSD＝0.81％ ，n＝6），97.04％ -98.89％（RSD＝0.73％ ，n＝6），96.23％ -98.08％（RSD＝0.82％ ，n＝6）， 95.40％-98.47％（RSD＝1.23％，n＝6），respectively. The contents of above 4 components in 11 batches of Huanshao Capsules were 0.190-0.704，0.439-0.857，2.723-4.475 and 0.589-1.035 mg/g，respectively. CONCLUSIONS ：Established method is specific ， precise and can be used for content determination of 4 components in Huanshao capsules.

The burden of schistosomiasis remains a global public health problem,especially in sub-Saharan Africa despite progress in terms of morbidity control.Successful control efforts achieved by China in the last six decades came with considerable experience and lessons that could benefit schistosomiasis control programs in other endemic countries.China's role and commitment to global health cooperation has become increasingly important;this has created a platform for partnership with developing partners for the establishment of Forum on China-Africa health cooperation which prioritizes the pursuit of global elimination target for schistosomiasis and malaria,control of HIV/AIDS,and improved access to reproductive health care.Chinese government's commitment towards achieving schistosomiasis elimination in Africa prompted the establishment of Institution-based Network on China-Africa Cooperation for Schistosomiasis Elimination (INCAS),by the National Institute of Parasitic Diseases to promote schistosomiasis elimination in Africa.Schistosomiasis experts from six provincial institutions and counterparts from 10 African countries participated in the first workshop on China-Africa cooperation for Schistosomiasis Elimination in Africa at Lilongwe,Malawi,in 2015.Experts at the inaugural meeting shared experiences from their national schistosomiasis control programs,as well as identified areas for collaborative synergy targeting schistosomiasis elimination in Africa.The establishment of INCAS,which comprises of 28 member-institutions from China and Africa,was proposed at this meeting.We,therefore,provide information on INCAS activities,cooperation mechanism,as well as assess the strengths,weaknesses,opportunities,and threats as we target schistosomiasis elimination in Africa using the INCAS platform.

Objective To study the effect of enhanced MK gene expression in hepatic carcinoma cells.Methods The recombinant plasmid pIRES2-EGFP-MK was transfected into SMMC 7721 cells.The mRNA and protein expression levels of MK gene in these cells were determined by real-time PCR,Western blotting and flow cytometry.The intracellular DNR accumulation of these cells was measured by flow cytometry.To investigate the effect of MK gene mediated multidrug resistance,MTT assay was employed to determine the cellular sensitivity of different chemotherapeutic drugs in MK-overexpressed SMMC 7721 cells.Results The mRNA and protein expression levels of MK gene significantly increased after the recombinant plasmid pIRES2-EGFP-MK transfected into SMMC 7721 cells,suggesting that the recombinant plasmid pIRES2-EGFP-MK can enhance the transcription of MK effectively.The DNR accumulation of MK transfected cells decreased significantly (4.06 ± 0.88,P ＜ 0.05),and IC50 of MK transfected cells to ADM/5-FU increased significantly (15 ± 3,27 ± 4,P ＜ 0.05).Conclusions After the recombinant plasmid pIRES2-EGFP-MK transfected into hepatic carcinoma cells,expression of midkine increased,enhancing the resistance of hepatic carcinoma cells to chemotherapeutic drugs.

Objective To compare the diversity of mitochondrial genomes of Angiostrongylus cantonensis in the mainland of China. Methods According to the population genetic of A. cantonensis,seven female worms were selected to characterize the mi-tochondrial(MT)genomes. Twelve primer pairs based on known MT genome(GQ398121)were used for PCR. The target frag-ments were sequenced and aligned. The gene localization,genome structure,composition of nucleotide,distribution of variable sites,and phylogeny were analyzed by employing multiple softwares. Results Five distinct types were identified from seven com-plete MT genomes. They were similar in size and structure,i.e.,ranging 13 491-13 502 bp,including 12 protein-coding genes,2 ribosomal genes,22 tRNA genes,and 2 major non-coding regions. All the genes were localized at the same strand and had the same transcription direction. A total of 745 variable sites were identified,accounting for 5.5%. Among the variable sites,59 were deletion/insert mutations,105 transversions,and 581 transitions. The variable sites distributed evenly at the complete genome. Conclusion The study reveals the mutation profile in the whole MT genome of A. cantonensis and thus will facilitate the develop-ment of intraspecific differential diagnosis.

Objective To investigate the regulatory effects of IFN-γon Treg cells from HIV/AIDS patients receiving highly active antiretroviral therapy (HAART) for one year.Methods Thirty HIV/A1DS patients whose CD4+T cells were below 350/μ1 were recruited for HAART therapy.Blood samples were collected at the time points of 0,24,48 weeks after HAART.PBMCs were isolated and randomly divided into two culture groups.One group was cultured directly in medium and another group was co-cultured with IFN-γ (40 pg/ml).The supernatants and cells were separated after 5 days of culture for analysis.The concentrations of IL-12 and CD4+CD25+Foxp3 Treg cells were measured by ELISA and flow cytometry,respectively.Results The levels of IL-12 in the supernatants from the culture without IFN-γ at time points of 0,24,48 weeks after HAART were lower than those from the co-cultured group [(37.02±12.76) vs (41.79± 15.02),t=2.336,P=0.03; (41.76±17.01) vs (47.2±14.26),t=2.702,P=0.014; (48.01± 11.84) vs (53.44± 11.30),t =3.14,P =0.003].The percentages of CD4+ CD25 + Foxp3 Treg cells in CD4+ T cells from the direct-cultured group were higher than those from the co-cultured group at the three time points [(10.41±1.10)％ vs (2.40±1.11)％,t=13.89,P=0.000; (8.33±2.03)％ vs (1.99± 0.86)％,t=12.93,P=0.000; (5.65±1.55)％ vs (1.32±0.73)％,t=10.61,P=0.000].Moreover,the results within the same group at the time points of 0,24,48 weeks upon HAART were also significantly different.Conclusion With the interference of HAART,IL-12 levels were increased,while CD4+CD25+ Foxp3 Treg cells were decreased in patients with HIV/AIDS.IFN-γ plays an important role in this process.

OBJECTIVE: To investigate the effect of nasal instillation of vitamin D3 on patients with allergic rhinitis symptoms including nasal itching, sneezing, clear nasal discharge, and nasal congestion. METHOD: Thirty subjects with allergic rhinitis proved by skin prick test (SPT) and 30 subjects with deviated septum alone were recrui ted and administrated with 300 000 IU of vitamin D3 by nasal instillation weekly. Seven days after the intervention, the four major symptoms including nasal itching, sneezing, clear nasal discharge, and nasal congestion were evaluated by score. RESULT: After intranasal instillation of vitamin D3, the symptoms in allergic rhinitis group in cluding nasal itching, sneezing, nasal discharge and nasal congestion, and serum 25-hydroxyvitamin D level has statistical differences (P < 0.05). CONCLUSION Vitamin D3 could be well absorbed through nasal mucosa. It demonstrated to have significantly effect on serum 25-hydroxyvitamin D to improve the symptoms for patients with allergic rhinitis. Vitamin D3 may be a kind of adjuvant therapy for prevention and treatment of allergic rhinitis.
Administration, Intranasal ; Adult ; Cholecalciferol ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Rhinitis, Allergic ; drug therapy ; Young Adult

OBJECTIVE: To explore the distribution of allergen tested by skin prick test (SPT) in about 5,843 allergic rhinitis patients in south Shanghai. METHOD: SPT test was conducted in 5,843 allergic rhinitis patients who came to our clinic from January 2007 to August 2012. The result was analyzed by age, sex and year. RESULT: The top three allergens by percentage are dermatophagoides pteronyssinus, dermatophagoides culinae and fungus among 15 common allergens. Incidence rate between male and female in each year had statistical significance, both of which showed no increasing trend with year. Incidence rates among different age groups aging from 6 to 17 years' old had no statistically significant difference, but statistically significant difference among different age groups existed in other age groups. Incidence rate showed increasing trend with year in age group of 40-65, which was not observed in other groups. The incidence rate showed decreasing trend with age in male and female, while the incidence rate in male was always higher than female. CONCLUSION In south Shanghai, primary allergens causing allergic rhinitis are dermatophagoides pteronyssinus, dermatophagoides culinae and fungus. Statistically significant difference about allergic rhinitis existed in age and sex. SPT has important significance in diagnosis of allergens.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Allergens ; immunology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Rhinitis, Allergic ; diagnosis ; epidemiology ; immunology ; Skin Tests ; Young Adult

Objective To study chronic bronchitis rats oxidant/antioxidant related indicators Ho flavored Dingchuan Decoction.Methods 50 SD rats were adopted improved smoked method to make chronic bronchitis model.From the 20th day of the modeling,blank control group and model control group gavage were treated with saline.The positive control group were treated with Cough Mixture.The experimental groupwere dealed with Ho flavored Dingchuan Decoction high,medium and low-dose.20 day later,the cotent of SOD,CAT,MDA in rat serum,lung tissue and balf were observed.Results Compare to blank control group,the cotent of SOD[(188.11 ± 9.37)U/ml、(276.11 ±30.28) U/ml,(8.80 ± 1.49)U/ml],CAT[(9.40 ±2.04)U/ml,(26.57±4.26)U/ml,(1.58±0.26) U/ml] in rat serum,lung tissue of the experimental group significantly reduced (P ＜0.01) ; MDA [(5.64 ± 0.52) μmol/L,(6.32 ± 2.10) μmol/L,(28.08 ± 2.43) μ mol/L) increased (P ＜0.05 or P ＜ 0.01).Ho flavored Dingchuan Decoction can effectively reverse these changes,and showed a significant dose-dependent.The Ho flavored Dingchuantang,in middle and high-dose serum SOD [(253.05 ± 7.81) U/ml,(256.92 ± 5.37)U/ml],CAT [(14.86 ± 2.49)U/ml,(18.22 ± 2.65)U/ml] higher than those of blank control group.And the Ho flavored Dingchuantang,in middle and high-dose lung tissue SOD [(470.91 ± 11.46)U/mgprot,(482.91 ± 14.32)U/mgprot),CAT [(46.87 ± 3.75)U/mgprot,(55.49 ±3.33)U/mgprot] higher than those of blank control group.Content close to the normal level.Conclusion The Ho flavored Dingchuan Decoction has significantly reduce the oxidative stress in chronic bronchitis.