A novel lectin(termed PML)was purified from fruiting bodies of the edible mushroom Phellodon melaleucus(division Basidiomycota)by ion exchange,hydrophobic interaction,and gel filtration chromatographies,with overall t...A novel lectin(termed PML)was purified from fruiting bodies of the edible mushroom Phellodon melaleucus(division Basidiomycota)by ion exchange,hydrophobic interaction,and gel filtration chromatographies,with overall titer recovery~60%and 20-fold purification.PML displayed hemagglutination activity 13319 units/mg toward rabbit erythrocytes.SDS-PAGE and gel filtration analyses revealed that PML is a homodimeric lectin with a molecular weight of 28.8 kDa.PML hemagglutination activity was not inhibited by various simple sugars or their derivatives,but was enhanced by cations Ca^(2+),Mg^(2+),Zn^(2+),and Cu^(2+).The activity was stable in pH range 6–9 and in the temperature range 20–60°C.Circular dichroism(CD)spectroscopic analysis showed that PML was composed primarily ofβ-sheets with lowα-helix content.In a B16 melanoma mouse model,PML treatment significantly inhibited tumor growth,and increased cytokine IL-10 content.Our findings suggest that PML is a potential anticancer therapeutic agent.展开更多
Severe climate change and global warming may impact significantly on vector-borne disease including Japanese encephalitis (JE) infection in human and animals. Thus, veterinary authority requires large quantity of diag...Severe climate change and global warming may impact significantly on vector-borne disease including Japanese encephalitis (JE) infection in human and animals. Thus, veterinary authority requires large quantity of diagnostic tools to survey vector-borne diseases. New producing method having a relation with JE antigen is needed to substitute conventional sucrose-acetone extraction method using suckling mouse. So, we developed new manufacturing method using polyethylene glycol (PEG) precipitation. Japanese encephalitis virus (JEV) was propagated in roller bottle containing Vero cell and inactivated with two kinds of inactivating reagents. Viability of the supernatant of bulk containing antigen was checked using Vero cell after inactivation. The supernatant did not show hemagglutination (HA) activity with goose erythrocytes. The antigen inactivated by binary ethylenimine (BEI) and concentrated by PEG precipitation method was found to be 2048 HA, but the antigen inactivated by 0.3% formaldehyde solution and concentrated by PEG precipitation method did not show HA titer. The antigen prepared from mice brain using sucrose-acetone extraction method showed 256 HA titer. This BEI inactivation method does not evoke animal welfare problem and can replace the conventional method that required biological hazardous reagents and suckling mice in preparing HA antigen. This new BEI inactivation method was safe in producing HA antigen against JEV in laboratory and can reduce environmental contamination of acetone.展开更多
[ Objective] To study the correlation between HI titer and protection against IBV challenge in SPF layers. [ Method ] SPF layers were randomly divided into four groups, namely group A1, A2, B1 and B2. The group A1 was...[ Objective] To study the correlation between HI titer and protection against IBV challenge in SPF layers. [ Method ] SPF layers were randomly divided into four groups, namely group A1, A2, B1 and B2. The group A1 was immunized with H120 live vaccine. The group A2 was first immunized with H120 live vaccine and later boosted with ND-IB-EDS trivalent inactivated vaccine. The group B1 was used as unimmunized chal- lenge control. The group B2 was kept as unimmunized unchallenged control. The blood samples were taken prior and post-vaccination at intervals and HI tests were conducted. At the laying peak, the group A1, A2 and B1 were challenged with IBV M4t virulent strain. The clinical features and egg production of layers were monitored and recorded. [Result] After 30 d post vaccination with H120 live vaccine, the HI titer reached 4.45 log2; after 30 days boosting with ND-IB-EDS trivalent inactivated vaccine, the HI titer reached to 7.35 log2. Before challenge, HI antibody titer in group A1, A2, B1 and B2 were respectively 4.24 log2, 7.40 Iog2, 2.10 log2 and 2.10 log2. After challenge, chickens in unimmunized challenge control group B1 showed respiratory symptoms, egg production dropped by 30.9%, and they produced more soft-shelled, no-shelled or abnormal eggs. In the group A1, some chickens had light respiratory symptoms and egg production dropped by 11.7%. In the group A2, the egg production of all chickens was as normal as the group B2. [ Conclusion] When the HI titer was over 6 log2, challenge by virulent virus had no impact on egg produc- tion; when the HI titer was 5 log2, 4 log2 and less 3 log2, egg production dropped by 6.0%, 11.3% and 29.6%, respectively. Thus, the HI anti- body level in chickens has close correlation with protection against IBV challenge.展开更多
Paramyxovirus Tianjin strain, a new genotype of Sendai virus, was isolated from the lungs of common cotton-eared marmoset that died of severe respiratory infection in the marmoset colonies. The 19.28% IgM positive rat...Paramyxovirus Tianjin strain, a new genotype of Sendai virus, was isolated from the lungs of common cotton-eared marmoset that died of severe respiratory infection in the marmoset colonies. The 19.28% IgM positive rate in the young children with acute respiratory tract infection suggested a close relationship between Tianjin strain and humans. Hemagglutinin-neuraminidase (HN) is its major transmembrane glycoprotein responsible for viral attachment, penetration and release. To clear the relationship between HN structure and function of paramyxovirus Tianjin strain, rHN1, rHN2 and rHN3 overlapping the ectodomain of HN protein were expressed. Their antigenicity and hemaglutination activity, as well as cross reactivity to standard antisera against influenza virus type A, type B were analyzed. The results indicated expressed rHNs have the natural antigenicity. The segment rHN2 possesses more linear epitopes exposed on the surface of the native HN protein than found in segments rHN3 and rHN1. The hemagglutination activity of segment rHN3 is higher than that of segments rHN2 and rHN1, and partially dependent on the three-dimensional conformation of HN3 protein. Cross-reactivity between rHNs and standard antisera against influenza virus type A, type B suggested that rHNs might not be the best alternative as specific antigens to detect virus in clinical serum specimens.展开更多
Objective: To investigate the effect of acupuncture on high hemagglutination state, blood sugar raising hormone and immunocyte factor levels in type II diabetes patients. Methods: A total of 120 inpatients and outpati...Objective: To investigate the effect of acupuncture on high hemagglutination state, blood sugar raising hormone and immunocyte factor levels in type II diabetes patients. Methods: A total of 120 inpatients and outpatients were randomly divided into acupuncture plus medication group (n=52) and medication group (n=50). In addition, 18 type II diabetes patients formed acupuncture group for comparing their therapeutic effects. Main acupoints used were Pishu (BL 20), Geshu (BL 17), Yishu, Shenshu (BL 23), Zusanli (ST 36), Sanyinjiao (SP 6), etc.. combined with other acupoints according to different sydroms. These acupoints were stimulated by manipulaing the filiform needles with uniform reinforcing and reducing method for 15 min and then stimulated electrically for 15 min with an electroacupuncture therapeutic apparatus. Western medicines used were Glipizide, Dimethyldiguanide Hydrochloride, etc.. The treatment was given once daily, with 10 sessions being a therapeutic course, 2～3 courses altogether. Indexes of external thrombosis length (ETL), platelet agglutination rate (PAgR), fibrinogen (FG), activated partial thromboplastin time (APTT), fasting blood glucose (FBG), prothrombin time(PT), adrenocoticortropic hormone (ACTH), cortisol (CS), growth hormone (GH), glucagon (GL), tumor necrosis factor alpha (TNF α), interleukin 6 (IL 6), insulin (INS) and C peptide (C P) were determined using radioimmunoassay. Results: After 2～3 courses of treatment, both acupuncture group and medication plus acupuncture group could significantly improve high hemagglutination state, lower blood sugar raising hormone level, regulate immunocyte factor level and raise the sensitivity of insulin, which were apparently superior to those of medication group (P<0.05～0.01). Conclusion: Acupuncture therapy can effectively regulate plasma blood sugar raising hormone, immunocyte factor levels, increase the sensitivity of insulin to target cells, resist blood coagulation and improve microcirculation.展开更多
Dear Editor,By targeting the programmed cell death 1(PD-1)pathway with monoclonal antibodies(mAbs),immune checkpoint therapy(ICT)has achieved unprecedented clinical success in the treatment of multiple tumors.1,2 Canc...Dear Editor,By targeting the programmed cell death 1(PD-1)pathway with monoclonal antibodies(mAbs),immune checkpoint therapy(ICT)has achieved unprecedented clinical success in the treatment of multiple tumors.1,2 Cancer cells evade the host immune system via both the tolerance of T cells and functional suppression of innate immune cells.3 CD47 provides a“do not eat me”signal by binding to signal regulatory protein alpha(SIRPα)to prevent innate immune cells from attacking host cells.4 Recently,macrophages were found to restore antitumor reactivity by blocking the interaction between upregulated CD47 on tumor cells and SIRPαon innate immune cells.5 However,it remains unknown whether there are blocking“hotspots”on CD47 for mAb-based anti-CD47 therapy or additional blocking hotspot regions within CD47 for therapeutic mAb development.Although it is overexpressed on tumor cells,CD47 is also expressed in many normal cells,including red blood cells and platelets.6 Some validated CD47-blocking mAbs under clinical investigation induce hemagglutination and anemia.7 Thus,designing an engineered CD47-blocking antibody to exert a therapeutic effect with limited hemagglutination is needed.展开更多
基金supported by grants from the China Agriculture Research System(CARS-20-01A)。
文摘A novel lectin(termed PML)was purified from fruiting bodies of the edible mushroom Phellodon melaleucus(division Basidiomycota)by ion exchange,hydrophobic interaction,and gel filtration chromatographies,with overall titer recovery~60%and 20-fold purification.PML displayed hemagglutination activity 13319 units/mg toward rabbit erythrocytes.SDS-PAGE and gel filtration analyses revealed that PML is a homodimeric lectin with a molecular weight of 28.8 kDa.PML hemagglutination activity was not inhibited by various simple sugars or their derivatives,but was enhanced by cations Ca^(2+),Mg^(2+),Zn^(2+),and Cu^(2+).The activity was stable in pH range 6–9 and in the temperature range 20–60°C.Circular dichroism(CD)spectroscopic analysis showed that PML was composed primarily ofβ-sheets with lowα-helix content.In a B16 melanoma mouse model,PML treatment significantly inhibited tumor growth,and increased cytokine IL-10 content.Our findings suggest that PML is a potential anticancer therapeutic agent.
文摘Severe climate change and global warming may impact significantly on vector-borne disease including Japanese encephalitis (JE) infection in human and animals. Thus, veterinary authority requires large quantity of diagnostic tools to survey vector-borne diseases. New producing method having a relation with JE antigen is needed to substitute conventional sucrose-acetone extraction method using suckling mouse. So, we developed new manufacturing method using polyethylene glycol (PEG) precipitation. Japanese encephalitis virus (JEV) was propagated in roller bottle containing Vero cell and inactivated with two kinds of inactivating reagents. Viability of the supernatant of bulk containing antigen was checked using Vero cell after inactivation. The supernatant did not show hemagglutination (HA) activity with goose erythrocytes. The antigen inactivated by binary ethylenimine (BEI) and concentrated by PEG precipitation method was found to be 2048 HA, but the antigen inactivated by 0.3% formaldehyde solution and concentrated by PEG precipitation method did not show HA titer. The antigen prepared from mice brain using sucrose-acetone extraction method showed 256 HA titer. This BEI inactivation method does not evoke animal welfare problem and can replace the conventional method that required biological hazardous reagents and suckling mice in preparing HA antigen. This new BEI inactivation method was safe in producing HA antigen against JEV in laboratory and can reduce environmental contamination of acetone.
文摘[ Objective] To study the correlation between HI titer and protection against IBV challenge in SPF layers. [ Method ] SPF layers were randomly divided into four groups, namely group A1, A2, B1 and B2. The group A1 was immunized with H120 live vaccine. The group A2 was first immunized with H120 live vaccine and later boosted with ND-IB-EDS trivalent inactivated vaccine. The group B1 was used as unimmunized chal- lenge control. The group B2 was kept as unimmunized unchallenged control. The blood samples were taken prior and post-vaccination at intervals and HI tests were conducted. At the laying peak, the group A1, A2 and B1 were challenged with IBV M4t virulent strain. The clinical features and egg production of layers were monitored and recorded. [Result] After 30 d post vaccination with H120 live vaccine, the HI titer reached 4.45 log2; after 30 days boosting with ND-IB-EDS trivalent inactivated vaccine, the HI titer reached to 7.35 log2. Before challenge, HI antibody titer in group A1, A2, B1 and B2 were respectively 4.24 log2, 7.40 Iog2, 2.10 log2 and 2.10 log2. After challenge, chickens in unimmunized challenge control group B1 showed respiratory symptoms, egg production dropped by 30.9%, and they produced more soft-shelled, no-shelled or abnormal eggs. In the group A1, some chickens had light respiratory symptoms and egg production dropped by 11.7%. In the group A2, the egg production of all chickens was as normal as the group B2. [ Conclusion] When the HI titer was over 6 log2, challenge by virulent virus had no impact on egg produc- tion; when the HI titer was 5 log2, 4 log2 and less 3 log2, egg production dropped by 6.0%, 11.3% and 29.6%, respectively. Thus, the HI anti- body level in chickens has close correlation with protection against IBV challenge.
基金National natural science foundation item (30471530)
文摘Paramyxovirus Tianjin strain, a new genotype of Sendai virus, was isolated from the lungs of common cotton-eared marmoset that died of severe respiratory infection in the marmoset colonies. The 19.28% IgM positive rate in the young children with acute respiratory tract infection suggested a close relationship between Tianjin strain and humans. Hemagglutinin-neuraminidase (HN) is its major transmembrane glycoprotein responsible for viral attachment, penetration and release. To clear the relationship between HN structure and function of paramyxovirus Tianjin strain, rHN1, rHN2 and rHN3 overlapping the ectodomain of HN protein were expressed. Their antigenicity and hemaglutination activity, as well as cross reactivity to standard antisera against influenza virus type A, type B were analyzed. The results indicated expressed rHNs have the natural antigenicity. The segment rHN2 possesses more linear epitopes exposed on the surface of the native HN protein than found in segments rHN3 and rHN1. The hemagglutination activity of segment rHN3 is higher than that of segments rHN2 and rHN1, and partially dependent on the three-dimensional conformation of HN3 protein. Cross-reactivity between rHNs and standard antisera against influenza virus type A, type B suggested that rHNs might not be the best alternative as specific antigens to detect virus in clinical serum specimens.
文摘Objective: To investigate the effect of acupuncture on high hemagglutination state, blood sugar raising hormone and immunocyte factor levels in type II diabetes patients. Methods: A total of 120 inpatients and outpatients were randomly divided into acupuncture plus medication group (n=52) and medication group (n=50). In addition, 18 type II diabetes patients formed acupuncture group for comparing their therapeutic effects. Main acupoints used were Pishu (BL 20), Geshu (BL 17), Yishu, Shenshu (BL 23), Zusanli (ST 36), Sanyinjiao (SP 6), etc.. combined with other acupoints according to different sydroms. These acupoints were stimulated by manipulaing the filiform needles with uniform reinforcing and reducing method for 15 min and then stimulated electrically for 15 min with an electroacupuncture therapeutic apparatus. Western medicines used were Glipizide, Dimethyldiguanide Hydrochloride, etc.. The treatment was given once daily, with 10 sessions being a therapeutic course, 2～3 courses altogether. Indexes of external thrombosis length (ETL), platelet agglutination rate (PAgR), fibrinogen (FG), activated partial thromboplastin time (APTT), fasting blood glucose (FBG), prothrombin time(PT), adrenocoticortropic hormone (ACTH), cortisol (CS), growth hormone (GH), glucagon (GL), tumor necrosis factor alpha (TNF α), interleukin 6 (IL 6), insulin (INS) and C peptide (C P) were determined using radioimmunoassay. Results: After 2～3 courses of treatment, both acupuncture group and medication plus acupuncture group could significantly improve high hemagglutination state, lower blood sugar raising hormone level, regulate immunocyte factor level and raise the sensitivity of insulin, which were apparently superior to those of medication group (P<0.05～0.01). Conclusion: Acupuncture therapy can effectively regulate plasma blood sugar raising hormone, immunocyte factor levels, increase the sensitivity of insulin to target cells, resist blood coagulation and improve microcirculation.
基金supported by the China National Grand S&T Special Project(2018ZX10302302)the Strategic Priority Research Program of the Chinese Academy of Sciences(CAS)(XDA12020358)G.F.G.and J.Y.are supported by the NSFC Innovative Research Group(Grant No.81621091).
文摘Dear Editor,By targeting the programmed cell death 1(PD-1)pathway with monoclonal antibodies(mAbs),immune checkpoint therapy(ICT)has achieved unprecedented clinical success in the treatment of multiple tumors.1,2 Cancer cells evade the host immune system via both the tolerance of T cells and functional suppression of innate immune cells.3 CD47 provides a“do not eat me”signal by binding to signal regulatory protein alpha(SIRPα)to prevent innate immune cells from attacking host cells.4 Recently,macrophages were found to restore antitumor reactivity by blocking the interaction between upregulated CD47 on tumor cells and SIRPαon innate immune cells.5 However,it remains unknown whether there are blocking“hotspots”on CD47 for mAb-based anti-CD47 therapy or additional blocking hotspot regions within CD47 for therapeutic mAb development.Although it is overexpressed on tumor cells,CD47 is also expressed in many normal cells,including red blood cells and platelets.6 Some validated CD47-blocking mAbs under clinical investigation induce hemagglutination and anemia.7 Thus,designing an engineered CD47-blocking antibody to exert a therapeutic effect with limited hemagglutination is needed.
