Objective: To study the chemical constituents from the cane of Pueraria lobata. Methods: The compounds were isolated by column chromatography over silica gel and purified by Sephadex LH-20 column chromatography and preparative TLC. The structures were elucidated by means of physicochemical properties and spectroscopic analyses. Results: Eleven compounds were separated and identified as: dehydrovomifoliol (1), blumenol A (2), 3β-hydroxy-5α,6α- epoxy-7-megastigmen-9-one (3), liquiritigenin (4), garbanzol (5), ferulaldehyde (6), 2,4-dihydroxybenzaldehyde (7), 4-hydroxy-2-methoxybenzaldehyde (8), (-)-tuberosin (9), coumestrol (10), and β-sitosterol (11). Conclusion: Compounds 1-3 are sesquiterpenoids, which are firstly isolated from the plants of Pueraria DC., and compounds 4-8 are obtained from the plants of Pueraria DC. for the first time.

The relationships between serum endothelin-1,calcitonin gene-related peptide and hypoxic-ischemic brain injury(HIBD) has been widely importance.Their respective functions,mutual relations and their relationships with HIBD has become a hot research.Understanding of their biological characteristics,mechanism and regulation of gene expression characteristics,is conducive to more in depth study on its pathogenic characteristics,provide a theoretical basis for clinical diagnosis and treatment.

Objective To produce prokaryotic recombinant protein glyceraldehyde-3-phosphate dehydrogenase of Clonorchis sinensis (CsGAPDH), analyze its enzyme activity and immunological function. Methods The recombinant CsGAPDH was purified according to the protocol of GST?Bind~TM kit and was digested with thrombin proteinase and eluted with wash buffer. The BALB/c mice were inoculated with the purified protein. The antisera collected from the mice were used to detect the titres of IgG antibodies by ELISA, and Western blotting was used to identify the specificity of the antisera with the purified CsGAPDH. S-P immunohistochemistry method was used to confirm the expression and distribution of CsGAPDH in adult Clonorchis sinensis with the polyclonal antibodies from immunized BALB/c mice. The CsGAPDH catalytic activity was evaluated employing the conventional substrate glyceraldehydes-3-phosphate (3-GAP). Results SDS-PAGE showed a single purified protein band. Gel scanning analysis revealed that the protein purity of CsGAPDH was 90%. ELISA analysis showed an increased IgG value. S-P immunohistochemistry analysis demonstrated that the recombinant plasmid pGEX-4T-1-GAPDH expressed and distributed in muscle cell membrane of immune mice. Western blotting result suggested that CsGAPDH protein contained essential epitopes with high antigenic activities. This protein CsGAPDH could catalyzed 3-GAP with enzymatic active unit of 2 872 U min~-1ml~-1. Conclusion The recombinant protein CsGAPDH shows a proper enzymatic activity and immunogenicity.

Objective To construct prokaryotic recombinant plasmids of transcriptional co-activator (TC) gene of Clonorchis sinensis, express and purify the recombinant protein and analyze its biological function. Methods A pair of primers was designed according to the known sequence of TC gene. The TC gene fragment was amplified by PCR. After purification and digestion with BamHⅠ and SalⅠ , the TC gene was connected to the prokaryotic expression vectors, pGEX-4T-1 and pET30a(+). By cloning target gene into these vectors, pGEX-4T-1 and pET30a(+), prokaryotic recombinant plasmids of TC gene were constructed and transferred into E.coli BL21. The positive expressed recombinants were detected by SDS-PAGE and Western blotting. Immobilized metal (Ni 2+ ) chelation affinity chromatography was used to purify His-TC produced by the expression of the recombinant protein pET30a(+)-TC. Results The recombinant plasmids, pGEX-4T-1-TC and pET30a(+)-TC, were constructed successfully. SDS-PAGE testified that the molecular weight of the recombinant protein was correct. Western blot analysis of GST-TC recombinant protein testified that the recombinant protein could be recognized by immunized rabbit serum, which means the protein is GST-immune active and the clone can express recombinant Clonorchis sinensis antigen. After affinity chromatography of the pET-TC protein, there was only one protein band with expected size on the SDS-PAGE gel. Conclusion The TC gene was screened from cDNA library of adult Clonorchis sinensis, cloned, expressed and purified. The purified protein of TC gene will be of importance for further research on the biological function of the gene.

Background It is well known that tissue factor (TF) is expressed in tumor cells and neovascular endothelial cells of tumor.It plays an important role in the formation of new blood vessels as well as the growth and metastasis of tumor.However,whether TF is expressed or not in choroidal melanoma(CM)is unclear.Objective This study is to investigate the expression of TF in a choroidal melanoma cell line and human choroidal melanoma.Methods The expression of TF was studied in the optimal choroidal melanoma-1 (OCM-1) cell line and ten specimens from CM patients using immunhistochemistry.Ten normal eye specimens from donators were used as controls.Results The TF protein was mainly expressed in the cytoplasm.It is over-expressed in OCM-1 cells with positive rate of 85.33±5.47%.Hyper-expression of TF also was found in human choroidal melanoma with a positive rate of 41.60±14.17%.The integrated optical density (IOD) of positive cells in CM was 33853.67±16445.30,and only 5.65±4.26% of positive cells was found in normal human choroidal tissue.The IOD of TF expression in normal human choroidal tissue was 426.43±316.62.Conclusion The overexpression of TF in CM cells may be a new immunotherapy target for CM treatment.

Objective To observe the effect of lipopolysaccharide (LPS)-mediated infection during pregnancy on the expression of mineralocorticoid receptor (MR) and density of dendritic spines in CA1 region of the dorsal hippocampus of rat offspring,so as to explore the mechanisms for learning and memory injury of rat offspring which were infected during prenatal period,then to provide scientific experimental evidence for the prevention of prenatal infection-induced delayed neuropsychiatric sequelae which contributed to learning and memory dysfunction.Methods Ten-week-old female Sprague-Dawley rats (n =30) were matched with male rats (1 ∶ 1).Pregnant rats were randomly divided into a control group (n =10) and an experimental group (n =20).The pregnant rats in experimental group were treated with LPS (66 μg/kg,intraperitoneally),and the pregnant rats in control group were intraperitoneally injected with same volume of saline on gestational day 10.On postnatal day 48,Morris water maze was used to estimate the ability of learning and memory;the brain tissues of offspring were taken and paraffin sections were stained with hematoxylin eosin (HE) for histological observation of CA1 region of the dorsal hippocampus;frozen sections were treated with indirect immunofluorescence to observe the expression of MR in CA1 region of the dorsal hippocampus;Golgi-Cox method was used to observe the density of dendritic spines of CA1 region.Results In Morris water maze test,from the third day the time of escape latency in experimental group [the 3rd day:(42.603 ± 9.837) s;the 4th day:(30.222 ± 9.789) s;the 5th day:(28.808 ± 12.526) s] was significantly higher than that of the control group [the 3rd day:(28.078±14.088) s;the 4th day:(20.692±13.099) s;the 5th day:(14.632 ±11.624) s] (the 3rd day:t =-3.274,P＜0.01;the 4th day:t =-2.257,P ＜0.05;the 5th day:t =-3.213,P＜0.01);the swimming time in the target quadrant [(14.660 ± 7.337) times] and the number of crossing platform [(0.933 ± 0.704) times] in experi mental group were significantly decreased compared with those of the control group [time:(23.820 ± 6.321) s;num bers:(2.000 ± 0.756) times] (t =3.663,4.000,all P ＜ 0.01).Hematoxylin eosin staining showed that the nerve cells of the hippocampus in the control group distributed in order,nucleuses were round or oval,nucleoli were obvious,and chromatins were homogeneous;but the cells in the experimental group distributed in disorder and pathological changes were detected,such as cellular swelling,necrosis and obvious nuclear pyknosis.By immunofluorescence staining,the average optical density (AOD) of MR in CA1 region decreased significantly in the experimental group (0.067 ± 0.017) compared with that of the control group (0.083 ± 0.009) (t =2.644,P ＜ 0.05).In Golgi-Cox method,the density of dendritic spines in experimental group [(7.705 ± 0.791)/10 μm] was below that of the control group [(9.655 ± 1.391)/10 μm] (t =3.852,P ＜ 0.01).Conclusions LPS-mediated infection during pregnancy might lead to hippocampus-dependent learning and memory dysfunction which might be associated with the reduced expression of MR and the low density of dendritic spines in CA1 region of the dorsal hippocampus.

Objective To explore the risk factors and protective factors for stomach cancer in Ganyu county.Methods Data of 319 pairs of new stomach cancer cases and health controls(1:1 matched)were studied by conditional Logistic regression method on the influential factors.Results The main risk factors for stomach cancer were the habits of eating hot foods(OR= 2.21,95%CI:1.57～3.12),the history of upper gastrointestinal inflammation(4.26,2.59～7.03),family history of cancer(2.29, 1.37～3.82);The main protective factors were often eating raw garlic(OR=0.64,95%CI:0.42～0.96),regular drinking tea(0.37, 0.25～0.54).Conclusion Unhealthy dietary habits,the history of upper gastrointestinal inflammation and family history of cancer are related to stomach cancer,while eating raw garlic and regular drinking tea may be effective in preventing the occurrence of stomach cancer.The results could be applied in stomach cancer prevention activities in Ganyu county.

Objective: To investigate the level of health literacy among residents in Karamay City, Xinjiang Uygur Autonomous Region, so as to provide insights into formulating health promotion strategies. Methods: Based on the Health Literacy Surveillance Program in Karamay City, demographic information and health literacy data were collected from residents aged 15 to 69 years in Karamay City in 2022, and levels of health literacy was analyzed. Factors affecting health literacy were identified using a multivariable logistic regression model. Results: A total of 1 473 respondents were included, with a mean age of (39.62±15.09) years. There were 751 men (50.98%) and 722 women (49.02%). The level of health literacy was 28.31%, and the prevalence rates of basic healthy knowledge and attitudes, healthy lifestyles and behaviors and basic health-related skills were 39.31%, 27.36% and 30.75%, respectively. The prevalence rates of safety and first aid, scientific health outlook, health information, chronic disease prevention, infectious disease prevention and basic medical care were 58.93%, 49.29%, 48.88%, 33.67%, 29.74% and 27.77%, respectively. Multivariable logistic regression analysis identified age (45 to 54 years OR=2.329, 95%CI: 1.005-5.398), ethnicity (ethnic minority, OR=0.459, 95%CI: 0.341-0.617), educational level (primary school and below, OR=0.213, 95%CI: 0.102-0.444; junior high school, OR=0.302, 95%CI: 0.193-0.471; high school, OR=0.579, 95%CI: 0.428-0.784), and annual household income (less than 10 000 Yuan, OR=0.352, 95%CI: 0.199-0.624; 10 000 to 30 000 Yuan, OR=0.447, 95%CI: 0.279-0.717; 30 000 to 50 000 Yuan, OR=0.598, 95%CI: 0.407-0.880) as factors affecting health literacy among residents in Karamay City. Conclusion The level of health literacy was 28.31% among residents in Karamay City in 2022. Health education and promotion should be carried out targeting residents of different ages, ethnicities, educational levels and annual household income, in order to further enhance the level of health literacy among residents in Karamay City.

This article aimed to study the antigenicity of nucleocapsid proteins (NPs) in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents. NPs of six pathogenic phleboviruses were expressed and purified using a prokaryotic expression system and rabbits were immunized with individual recombinant NPs. Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses, and the sera titer was determined by indirect ELISA. Furthermore, sera from SFTS patients were also detected by each recombinant NP as a coating antigen using indirect ELISA. The cross-reactions and the sera titer were subsequently determined. Both the concentration and purity of recombinant NPs of six pathogenic phleboviruses met the standards for immunization and detection. The results of indirect ELISA and Western blotting showed that each anti-phlebovirus NP rabbit immune serum had potential serological cross-reactivity with the other five virus NP antigens. Furthermore, the sera from SFTS patients also had cross-reactivity with the other five NP antigens to a certain extent. Our preliminary study evaluated the antigenicity and immune reactivity of six pathogenic phleboviruses NPs and laid the foundation for the development of diagnostic reagents.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Cross Reactions ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Phlebotomus Fever ; diagnosis ; immunology ; virology ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

High altitude polycythemia (HAPC) had become one of the main common chronic diseases, which had seriously threatened the health of Plateau people. In the Tibetan medicine classic bookSi-Bu Yi-Dian, there were recordings on HAPC treatment methods and medications, which had the unique advantages of identified therapeutic effect with little side effect. This article analyzed Tibetan medicine in the prevention and treatment of HAPC from aspects such as etiology and pathogenesis, clinical treatment advantages and modern innovation study. Questions were also raised on lacking of standardization on HAPC clinical effectiveness, as well as Tibetan medicine compound material basis and action mechanisms were unclear. It was proposed that based on the inheritance of Tibetan medicine theoretical basis and clinical therapeutic effect, the Tibetan medicine original thinking should be combined with modern science and technology, in order to strengthen the analysis of ancient literature collection in HAPC treatment and data mining in medication experiences. The clinical treatment standards and medication plan should be standardized. Methods of systems biology, such as metabolomics, can be used in the further study of the scientific connotation of HAPC treatment by Tibetan medicine.