Objective To assess the risk of human Spirometra mansoni infections and investigate the knowledge, attitude and practice (KAP) towards sparganosis mansoni among residents in Henan Province, so as to provide insights into formulation of the sparganosis mansoni control measures. Methods Qinling Village in Fugou County of Zhoukou City, Bali Village in Yancheng District of Luohe City, Duzhai Village in Puyang County of Puyang City and Doushan Village in Luoshan County of Xinyang City were sampled as survey sites in Henan Province from July to August 2023, and more than 40 frogs were sampled from ponds or streams in each survey site for detection of Sparganum mansoni infections. At least 150 residents were sampled using a cluster sampling method from each survey site, and the sero-prevalence of anti-S. mansoni IgG antibody was estimated. In addition, a questionnaire survey was conducted on the KAP towards sparganosis mansoni among participants, and the proportion of eligible KAP, rate of correct KAP and KAP scores were calculated. Results A total 229 frogs were collected from 4 survey sites in 2023, and the overall prevalence of S. mansoni infection was 4.37% (10/229) in frogs, with 7.75% (10/129) prevalence in wild frogs and 0 in farm-bred frogs. A questionnaire survey was performed among 649 residents sampled from 4 survey sites, and 649 serum samples were collected. The seroprevalence of anti-S.mansoni IgG antibody was 0.15% (1/649) and the overall proportion of eligible KAP was 23.73% (154/649) among participants. There were age- (χ² = 30.905, P = 0.000), educational level- (χ² = 41.011, P = 0.000), and occupation-specific proportions of eligible KAP among participants (χ² = 10.721, P = 0.005), and the proportion of eligible KAP decreased with age (χ² _trend = 22.717, P = 0.000) and increased with education levels (χ² _trend = 40.025, P = 0.000). The rates of correct KAP towards sparganosis mansoni were 40.81% (2 119/5 192), 96.66% (1 882/1 947) and 63.81% (3 727/5 841) (χ² = 1 913.731, P = 0.000) among residents, respectively. The rates of correct KAP towards sparganosis mansoni varied significantly among survey sites (χ² = 136.872, 42.347 and 255.157; all P values= 0.000, with the highest rate of correct knowledge (51.94%, 748/1 440) and practices (75.86%, 1 229/1 620) in Yancheng District of Luohe City and the highest rate of correct attitudes in Puyang County of Puyang City (99.11%, 446/450) (all P values< 0.05). Conclusions There is still a high transmission risk of sparganosis mansoni in Henan Province, and the KAP towards sparganosis mansoni is required to be improved among residents.

Purpose/Significance To reveal secondary infection knowledge related to infectious diseases by mining public literature data,and to promote the research and construction of the secondary infection monitoring platform,so as to improve the prevention and control level of infectious diseases in China.Method/Process The literature based discovery method is firstly adopted to mine and ana-lyze the secondary diseases from large-scale biomedical literature data,taking viral hepatitis,human immunodeficiency virus(HIV)infection and tuberculosis infection as the examples.Result/Conclusion 3 kinds of secondary diseases including infectious diseases,non-infectious diseases and even tumors,are found from more than 36.8 million PubMed literature.The research results not only validate that this method provides a new approach for systematically and comprehensively reveal secondary infection knowledge related to infectious diseases,but also provide more powerful literature evidences for effective monitoring and early intervention of secondary diseases.

Objective To identify and verify the interacting protein of α-11 giardin, so as provide the experimental evidence for studies on the α-11 giardin function. Methods The yeast two-hybrid cDNA library of the Giardia lambia C2 strain and the bait plasmid of α-11 giardin were constructed. All proteins interacting with α-11 giardin were screened using the yeast two-hybrid system. α-11 giardin and all screened potential interacting protein genes were constructed into pBiFc-Vc-155 and pBiFc-Vn-173 plasmids, and co-transfected into the breast cancer cell line MDA-MB-231. The interactions between α-11 giardin and interacting proteins were verified using bimolecular fluorescence complementation (BiFC). Results The yeast two-hybrid G. lambia cDNA library which was quantified at 2.715 × 10⁷ colony-forming units (CFU) and the bait plasmid containing α-11 giardin gene without an autoactivation activity were constructed. Following two-round positive screening with the yeast two-hybrid system, two potential proteins interacting with α-11 giardin were screened, including eukaryotic translation initiation factor 5A (EIF5A), calmodulin-dependent protein kinase (CAMKL) and nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase (NADP-GDH), hypothetical protein 1 (GL50803_95880), hypothetical protein 2 (GL50803_87261) and a protein from Giardia canis virus. The α-11 giardin and EIF5A genes were transfected into the pBiFc-Vc-155 and pBiFc-Vn-173 plasmids using BiFC, and the recombinant plasmids pBiFc-Vc-155-α-11 and pBiFc-Vn-173-EIF5A were co-tranfected into MDA-MB-231 cells, which displayed green fluorescence under a microscope, indicating the interaction between α-11 giardin and EIF5A protein in cells. Conclusion The yeast two-hybrid cDNA library of the G. lambia C2 strain has been successfully constructed, and six potential protein interacting with α-11 giardin have been identified, including EIF5A that interacts with α-11 giardin in cells.

Objective:To construct the chimeric virus ChimZIKV of Japanese encephalitis virus and Zika virus and evaluate whether the chimeric virus can be developed into a candidate vaccine strain against Zika virus.Methods:The infectious clone of chimeric virus ChimZIKV was constructed by replacement the prM/E of Japanese encephalitis virus vaccine strain with that of Zika virus. RNA of chimeric virus ChimZIKV had been transcribed in vitro and was electro-transfected into BHK-21 cells to rescue chimeric virus. Afterward, the chimeric virus was identified by plaque assay, immunofluorescence and gene sequencing. The growth was shown by the growth curve. The neurobirulence, viremia, neutralizing antibody production and immune protective effect of chimeric virus were tested by animal experiments.Results:The result of restriction enzyme digestion showed that the infectious clone of chimeric virus was successfully constructed, and the immunofluorescence assay and sequencing showed that chimeric virus ChimZIKV was successfully rescued. The cell proliferation activity test showed that the cell proliferation activity of the chimeric virus infected group was higher than that of the infected group. The result of animal experiments showed that chimeric virus showed very low neurovirulence to mice and suckling mice, and the result of viremia showed that chimeric virus had no obvious viremia in mice. Plaque reduction neutralization test (PRNT) showed 100% positive conversion of neutralizing antibody in sera of immunized mice, and the highest neutralizing antibody titer (GMT=85) was seen in the third week. The immune protection experiment showed that the protection rate against JE/ZIKV(MR766) was 30%.Conclusions:This study showed that the chimeric virus ChimZIKV is immunogenic and with low neurovirulence, and it may be further developed as a candidate vaccine strain against Zika virus.

Objective: To explore the impact of MR T2WI high signal rim sign (T2-rim sign) on the immediate efficacy of high intensity focused ultrasound (HIFU) in treatment of uterine myoma. Methods: Data of 360 uterine myoma patients treated with HIFU were retrospectively analyzed. The ablation effect was judged based on the percentage of non-perfusion area (NPV) of myoma after HIFU treatment and the volume of myoma before treatment (NPV%). According to the exist of preoperative MR T2-rim sign, the patients were divided into T2-rim sign group (with T2-rim sign) and no T2-rim sign group (without T2-rim sign). Univariate and multivariate Logistic analysis were performed to evaluate the impact of NPV%. Results: Immediately after ablation, contrast-enhanced ultrasound showed a marked reduction in blood supply of myoma, with 60 cases NPV%<75% and 300 cases NPV%≥75%, and significant differences of myoma location, irradiation time, treatment intensity, MR T2WI signal intensity and with or without T2-rim sign were found between them (all P<0.05). NPV% was correlated with MR T2WI signal intensity, T2-rim sign, irradiation time and treatment intensity (all P<0.05). The irradiation time, treatment intensity and T2-rim sign were independent risk factors for NPV% (all P<0.05). Compared with no T2-rim sign group, the irradiation time and treatment intensity of T2-rim sign group all increased (both P<0.05) and NPV% decreased (P<0.05). Conclusion: T2-rim sign is one of the impact factors of immediate therapeutic responses of HIFU ablation for uterine myoma. Uterine myoma patients with T2-rim sign are able benefit from HIFU.

Objective:To explore the effect of D393E mutation on its intracerebral neurovirulence in mice after infectious clone of the chimeric Japanese encephalitis virus (JEV)/Zika virus (ZIKV) containing envelope protein D393E mutant was built and the virus was rescued.Methods:The full-length cDNA plasmid of JEV/ZIKV chimeric virus containing D393E mutation in the envelope protein was built by SOE PCR and gene cloning technology. The linearized fragment of the plasmid was used for in vitro transcription to obtain viral RNA, followed by electrotransfection into BHK-21 cells to rescue the mutant virus JEV/ZIKV(D393E). The plaque size and growth characteristics of JEV/ZIKV (D393E) and JEV/ZIKV were compared after BHK-21 cells were inoculated with the viruses. The intracerebral neurovirulence of the viruses was calculated after the viruses were inoculated in mice intracerebrally (i.c.).Results:Restriction enzyme digestion evaluation confirmed that the infectious clone of mutant virus was correctly built. The plaque assay demonstrated that the plaque diameter of JEV/ZIKV (D393E) was about 0.7±0.2 mm, which was smaller than that of JEV/ZIKV (1.3±0.2 mm). The LD 50 value of the mutant was 31.51 PFU when injected in mice via i. c. route, which was greater than 2.21 PFU of JEV/ZIKV. Conclusions:The D393E mutation of ZIKV envelope protein in JEV/ZIKA chimeric virus decreases the neurovirulence of the virus in mice.

Objective To investigate the cell origin of interleukin (IL)-22-secreting cell of mice infected with Trichinella spiralis (T.spiralis) at the early encapsulated stage.Methods Twelve Balb/c mice were divided into the infected group and the control group according to body weight by random number table.The infected mice were intragastrically administrated with 300 muscle larvae of T.spiralis,and the control mice were given the same amount of normal saline.The IL-22-secreting cell subsets in mouse splenic lymphocytes were detected by flow cytometry at the fourth week after infection.Results The proportion of IL-22-secreting cells in splenic lymphocytes of T.spiralis infected mice was increased when compared with control group [(0.88 ± 0.25)％ vs (0.28 ±0.17)％,t =-4.899,P ＜ 0.05].There was no significant difference between the proportion of CD3+IL-22+ cells and CD3-IL-22+ cells in the splenic lymphocytes of the infected group [(0.29 ± 0.17)％ vs (0.51 ± 0.17)％,t =-2.195,P ＞ 0.05],and the percentage of CD3-IL-22+ cells were similar between the infected group and the control group [(0.51 ± 0.17)％ vs (0.44 ± 0.22)％,t =-0.600,P ＞ 0.05].The proportion of CD3+IL-22+ cells in the infected group was significantly higher than that in the control group [(0.29 ± 0.17)％ vs (0.07 ± 0.06)％,t =-3.068,P ＜ 0.05],and the percentage of CD4+IL-22+ T cells and γδTCR+IL-22+ T cells were obviously increased in CD3+ lymphocytes [(1.28 ± 0.54)％ vs (0.16 ± 0.07)％,(0.33 ± 0.22)％ vs (0.02 ± 0.00)％,t =-4.997,-3.342,P ＜ 0.05].Conclusions The proportion of IL-22-secreting splenic lymphocytes is increased in mice infected with T.spiralis at the early encapsulated stage.The rise is caused by increased numbers of IL-22-secreting CD3 + lymphocytes,especially CD4+ T cells and γδT cells.

Objective To investigate the distribution characteristics and risk factors of intracranial and extracranial aterial lesions in Chinese patients with ischemic stroke . Methods In this multi‐center study ,2 310 continuously inpatients with ischemic stroke diagnosed in 20 stroke screening and prevention project base hospitals from June 2015 to M ay 2016 were enrolled . Carotid ultrasonography and transcranial color‐coded sonography or transcranial Doppler were performed in all patients to confirm the presence of cerebral artery stenosis or occlusion . According to the distribution of lesions ,the subjects were divided into 2 groups :the simple intracranial artery stenosis group and the simple extracranial artery stenosis group . T he difference of risk factors between the two groups was compared . Results Of the 2 310 patients with ischemic stroke ,1 516 ( 65 .6% ) had simple intracranial artery stenosis and 794 ( 34 .4% ) had simple extracranial artery stenosis . T he incidence of anterior circulation artery stenosis was higher in the group of intracranial artery stenosis than that in the extracranial artery stenosis group ( 68 .1% vs 48 .7% , P ＜0 .001) . Posterior circulation artery stenosis and combined anterior with posterior circulation artery stenosis were more common in patients with extracranial artery stenosis group than those in intracranial artery stenosis group ( 36 .4% vs 22 .1% ,14 .9% vs 9 .8% ;all P ＜0 .001) . Univariate analysis of risk factors for stroke showed that patients with intracranial arterial stenosis had a higher prevelence of hypertension , diabetes ,obesity ,and family history of stroke ,and their systolic blood pressure ,diastolic blood pressure , body mass index ( BM I) ,fasting blood‐glucose ,glycosylated hemoglobin ,triacylglycerol ,total cholesterol , and low‐density lipoprotein cholesterol were significantly higher than those in the extracranial arterial stenosis group ( all P ＜ 0 .05 ) . T he proportion of elderly ( ≥ 65 years old ) ,male and smokers in the extracranial arterial stenosis group was significantly higher than that in the intracranial arterial stenosis group ( all P ＜0 .05) . Multivariate logistic regression analysis showed that elderly ( ≥65 years old) ,male , and smoking history were independent risk factors for extracranial arterial stenosis ( OR＝ 2 .012 ,1 .637 , 1 .325 ,respectively ;all P ＜0 .05) . While hypertension ,diabetes ,less physical activity ,and high BM I levels were independent risk factors for simple intracranial arterial disease ( OR ＝ 1 .301 ,1 .252 ,1 .248 ,1 .030 , respectively ;all P ＜0 .05) . Conclusions There are significant differences in the distribution characteristics and risk factors of intracranial and extracranial aterial lesions in patients with ischemic stroke in China .

Objective To investigate the effect of long-term high-fat diet on cognitive function and hippocampus neurons ultrastructure in obese rats.Methods Forty SD rats were randomly assigned to a high fat diet (HFD) group and a common diet (CD) group.Meanwhile,HFD-induced obese rat model were established.The spatial learning and memory were measured by the Morris water maze,and the neurons ultrastructural changes in rat hippocampus CA1 region at the corresponding period were observed by transmission electron microscopy.Results The average weight of rats was 25％,28％,and 22％ higher in the HFD group than in the CD group at the 12,16,and 20 weeks,respectively;the Lee's indexes were 6％,4％,and 8％ higher;the average swimming latency were 52％,44％,and 40％ longer;the average swimming distance were 85％,45％,and 51％ longer;the average swimming speed were 57％,34％,and 18％ higher;the duration of staying in the target quadrant were 32％,54％,and 63％ shorter;and the average times of crossing the plate form were 30％,34％,and 34％ shorter,respectively (all P ＜0.001).In comparison of ultrastructure in hippocampus CA1 region of rats at corresponding time points,the amounts of degenerated and necrosis neurons,of the deformed and vacuolar mitochondria,and of the less rough endoplasmic reticulum were significantly more at 12,16,and 20 weeks in the HFD group than in the CD group.Conclusions Long-term HFD-induced obesity damages the structure of neurons in the hippocampus,impairs spatial learning and memory function,and accelerates cognitive aging in rats.

AIM:To observe and analyze the effect of cardiopulmonary bypass ( CPB) for 30 min on surface ultra-structure and mechanical properties of the erythrocyte membrane by atomic force microscopy (AFM).METHODS:Ten cases of elective patients in cardiac surgery were selected in the study and divided into control ( CON) group and CPB group.The central venous blood (2 mL) before surgery and 30 min after CPB was collected with heparin anticoagulation . The non-circular red blood cells were counted under a stand fluorescence microscope .AFM was used to examine the ultra-structure of the membrane surface and measure the force curve of the erythrocytes .RESULTS:The percentage of non-cir-cular red blood cells in CPB group showed no statistically significant differences as compared with CON group .AFM images showed that the significant differences of membrane surface concave and convex , evenness , particle distribution , the sur-face average roughness (Ra), the surface root mean square roughness (Rq) and cell membrane adhesion between CPB group and CON group were observed .However, the membrane deformation resilience and curve slope had no significant difference between the 2 groups.CONCLUSION:Cardiopulmonary bypass for 30 min changes the morphology and ultra-structure of the erythrocyte membrane surface , and increases the adhesion between cells .