Background In recent years, regional high-temperature weather in summer occurs frequently in China. Heat stroke is the most representative meteorological disease caused by high temperature. In order to improve monitoring, early warning, prevention, and control of heat stroke, it is of great significance to understand the epidemiological characteristics of heat stroke and the associated impact of heatwave. Objective To understand the epidemiological characteristics of heat stroke cases in Jinan City, and to explore the effects of heatwave exposure on heat stroke. Methods Case reports of heat stroke and daily data of meteorological factors in Jinan City from 2017 to 2022 were collected. We described the temporal, population, and regional distribution characteristics of heat stroke cases in Jinan City, and used a time-stratified case-crossover design combined with conditional logistic regression model to explore the effects of heatwave exposure on heat stroke under 12 heatwave definitions (different combinations of intensity and duration). The cut-off percentiles used for heatwave definitions were the 90th (P₉₀), 95th (P₉₅), 97.5th (P_97.5), and 99th (P₉₉) percentiles of daily mean temperature; the durations were ≥ 2 d, ≥ 3 d, and ≥ 4 d, respectively. P_i(k), where i is temperature threshold, and k is duration. For example, the definition of a heatwave was notated as P₉₀(2), indicating that the daily mean temperature is ≥ P₉₀ and lasts for ≥ 2 d. Alternatively, lag01 denotes the cumulative lag effect with a 1 d lag, and so on. Results A total of 1394 cases of heat stroke were reported in Jinan City from 2017 to 2022, including 581 mild cases and 813 severe cases, and 85 deaths were reported, with a cumulative fatality rate of 6.10%. The cases of heat stroke reported each year during the study period were concentrated from June to August and peaked in July (665 cases, 47.70%). The sex ratio of males to females in heat stroke cases was 2.02:1. A high incidence of heat stroke was in 50-89 years, with a smaller peak occurring in the age group of 50-59 years and a larger peak in the age group of 70-79 years, respectively. The high-incidence areas of heat stroke were distributed in the western part of Jinan City where city centers situated (Tianqiao District, 274 cases, 19.66%; Huaiyin District, 223 cases, 16.00%) and in the surrounding rural areas (Pingyin County, 254 cases, 18.22%). The effect of heatwave exposure on heat stroke was statistically significant during the study period. The largest effect estimates for the effect on heat stroke occurred under the heatwave definitions of P₉₉(2), P_97.5(3), and P_97.5(4) at lag04, lag03, and lag04, where corresponding OR (95%CI) values were 9.27 (4.71, 14.24), 8.95 (6.17, 12.98), and 8.22 (4.91, 13.78), respectively. The exposure-response curve showed that the risk of heat stroke tended to increase with the increase of average daily temperature. Conclusion July is the key period for the occurrence of heat stroke among Jinan City residents, while male cases are predominant, more serious cases, age concentration in the 50-89 years. The occurrence of heatwave can further increase the risk of heat stroke with a significant lag effect.

Objective:To study the levels and variation of equilibrium factor in indoor environment.Methods:A one-year continuous measurement of radon concentration and equilibrium equivalent radon concentration was carried out in an indoor office building of Nanning city. The effective data acquisition rates of radon gas and radon progeny were 99.9% and 86.7%, respectively.Results:The annual average activity concentration and equilibrium equivalent radon concentration in indoor environment were (50.9±20.7)and (15.5±10.1)Bq/m 3, both of which had the same diurnal and seasonal variation. The average annual value of equilibrium factor was 0.30±0.12, showing no obvious diurnal variation. The distribution of monthly mean value of equilibrium factor showed a similar trend to that of radon and radon progeny. The highest and the lowest value appeared in November and June, respectively, with 0.47±0.24 and 0.19±0.06. Conclusions:Due to the large variation range of monthly mean value of equilibrium factor in indoor environment, when annual effective dose of radon exposure was estimated based on radon gas concentration, attention should be paid to choose the quantity value of equilibrium factor and the uncertainty caused by the change of equilibrium factor should be considered.

The potency of an improved recombinant multi-epitope vaccine against FMDV type Asia1 was evaluated in this study .A multi-epitope gene based on FMDV type Asia1 was designed and a recombinant expression plasmid (pRE-oIgG) was constructed .The proteins ,RE-oIgG and 3D were expressed in E .coli cells and purified with Ni-NTA agarose resin by affinity chromatography .The proteins ,RE-oIgG ,3D and RE-oIgG plus 3D ,were emulsified in an oil adjuvant ISA 206 .Twenty-five female guinea pigs were randomly divided into five groups and intramuscularly vaccinated for with RE-oIgG ,3D ,RE-oIgG plus 3D ,an inactivated FMDV vaccine (type Asia1) ,and PBS .All animals were vaccinated for two times .Anti-FMDV specific an-tibodies ,neutralization antibodies ,protection potency ,and lymphoproliferation assay were detected by ELISA ,virus neutrali-zation assay ,challenge test ,and flow cytometry ,respectively .Results showed that RE-oIgG plus 3D elicited significant high-level anti-FMDV specific antibodies compared to RE-oIgG alone (P<0 .05) .All the vaccinated animals induced higher level lymphoproliferation responses in vitro except PBS .Both 3D alone and PBS produced the negligible neutralizing antibodies and anti-FMDV specific antibodies .RE-oIgG plus FMDV 3D not only elicited high levels of anti-FMDV neutralizing antibodies ,but also induced significant lymphoproliferation responses .More importantly ,RE-oIgG plus 3D conferred complete protection to guinea pigs against challenge with 1 000 GPID50 .Interestingly ,two of five vaccinated animals with 3D alone were full protected against challenge ,and other three animals significantly showed a delay of 2-3 days in the onset of clinical signs .Therefore ,we considered that RE-oIgG plus 3D induces strong humoral and cellular immune responses ,which may be used for control and prevention of FMD in the future .

To identify linear epitopes on the non-structural protein 3AB of foot-and-mouth disease virus (FMDV),BABL/c mice were immunized with the 3AB protein and splenocytes of BALB/c mice were fused with myeloma Sp2/0 cells.Two hybridoma monoclonal antibodies (mAbs) cell lines against the 3AB protein of foot-and-mouth disease virus (FMDV) were obtained,named C6 and E7 respectively.The microneutralization titer was 1∶1024 for mAb C6,and 1∶512 for E7.Both mAbs contain kappa light chains,and were of subclass IgG2b.In order to define the mAbs binding epitopes,the reactivity of these mAbs against FMDV were examined by indirect ELISA.The results showed that both mAbs can react with FMDV,but had no cross-reactivity with Swine Vesicular Disease (SVD) antigens.The titers in abdomen liquor were 1∶5×106 for C6 and 1∶2×106 for E7.In conclusion,the mAbs obtained from this study are specific for the detection of FMDV,can be used for etiological and immunological researches on FMDV,and have potential use in diagnosis and future vaccine designs.

To develop a sensitive and specific ELISA for detection of antibodies to the nonstructural protein of FMDV. We cloned and expressed FMDV nonstructural protein 3AB in Escherichia coli expression system. The recombinant protein 3AB was purified with Ni-NTA HisBind Resins and characterized by Western blotting. An indirect ELISA based on purified protein 3AB as a coating antigen was established. The specificity and sensitivity of this assay were evaluated by comparison with a commercial 3ABC-ELISA kit in detecion of serum samples. The results showed that the recombinant protein 3AB was expressed as a formation of inclusion bodies in Escherichia coli. The purified protein could specificially react with FMDV infection antibodies in Western blotting assay, but no reaction with the immune antibodies induced with vaccine. Two assays were no significant differences in specificity and sensitivity for detection of field samples (P>0.05). Therefore, we speculated that the recombinant protein 3AB is a promising molecular marker, which may effectively differentiate FMD-infected from vaccinated animals in a herd.
Animals ; Antibodies, Viral ; analysis ; Antigens, Viral ; biosynthesis ; genetics ; immunology ; Cattle ; Cattle Diseases ; diagnosis ; immunology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Foot-and-Mouth Disease ; diagnosis ; immunology ; Foot-and-Mouth Disease Virus ; chemistry ; genetics ; isolation & purification ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Viral Nonstructural Proteins ; biosynthesis ; genetics ; immunology

Caspases are an evolutionarily conserved family of aspartate-specific cystein-dependent proteases with essential functions in apoptosis and normally exist in ceils as inactive proenzymes.In addition to the inflammatory caspases,the initiator and effector caspases have been shown to have an important role in regulating the immune response,but are involved in different ways.We give a brief introduction on the benefit of apoptosis on the clearance of invasive pathogens,and the caspase functions involved in the immune response.Then we construct a working model of caspases during pathogen invasion.A detailed description of the three modes is given in the discussion.These three modes are regulated by different inhibitors,and there may be a novel way to treat intracellular pathogen and autoimmune diseases based on the specific inhibitors.

To develop a safe and efficient recombinant subunit vaccine to foot-and-mouth disease virus(FMDV)type Asia 1 in sheep,a tandem repeated multiple-epitope gene consisting of residues 137-160 and 197-211 of the VP1 gene of FMDV was designed and artificially synthesized.The biologically functional molecule,the ovine IgG heavy constant region(oIgG)as a protein carrier was introduced for design of the multiple-epitope recombinant vaccine and recombinant expression plasmids pET-30a-RE and pET-30a-RE-oIgG were successfully constructed.The recombinant proteins,RE and RE-oIgG,were expressed as a formation of inclusion bodies in E.coli.The immune potential of this vaccine regime in guinea pigs and sheep was evaluated.The results showed that IgG could significantly enhance the immune potential of antigenic epitopes.The recombinant protein RE-oIgG could not only elicit the high levels of neutralizing antibodies and lymphocytes proliferation responses in the vaccinated guinea pigs,but confer complete protection in guinea pigs against virus challenge.Although the recombinant protein RE could not confer protection in the vaccinated animals,it could delay the appearance of the clinical signs and reduce the severity of disease.Inspiringly,the titers of anti-FMDV neutralizing antibodies elicited in sheep vaccinated with RE-oIgG was significantly higher than that for the RE vaccination.Therefore,we speculated that this vaccine formulation may be a promising strategy for designing a novel vaccine against FMDV in the future.

In order to develop an anti-FMDV A Type monoclonal antibo by (mAb),BABL/c mice were immunized with FMDV A type.Monoclonal antibodies (mAbs) 7B11 and 8H4 against Foot-and-mouth disease virus (FMDV) serotype A were produced by fusing SP2/O myeloma cells with splenocyte from the mouse immunized with A/AV88.The microneutralization titer of the mAbs 7B11 and 8H4 were 1024 and 512,respectively.Both mAbs contain kappa light chains,the mAbs were IgG1.In order to define the mAbs binding epitopes,the reactivity of these mAbs against A Type FMDV,were examined using indirect ELISA,the result showed that both mAbs reacted with A Type FMDV.These mAbs may be used for further vaccine studies,diagnostic methods,prophylaxis,etiological and immunological research on FMDV.Characterization of these ncindicated that prepared anti-FMDV A mAbs had no cross-reactivity with Swine Vesicular Disease (SVD) or FMDV O,Asial and C Type antigens.Their titers in abdomen liquor were 1:5×106 and 1:2×106,respectively.7B11 was found to be of subtype IgG1,8H4 was classified as IgG2b subtype.The mAbs prepared in this study,are specific for detection of FMDV serotype A,and is potentially useful for pen-side diagnosis.

In this study,the coding region of type O FMDV capsid protein VP1 and a series of codon optimized DNA sequences coding for VP1 amino acid residues 141-160(epitopel),tandem repeat 200-213(epitope2(+2))and the combination of two epitopes(epitope1-2)was genetically cloned into the prokaryotic expression vector pPROExHTb and pGEX4T-1,respectively.VP1 and the fused epitopes GST-E1,GST-E2(+2)and GST-E1-2 were successfully solubly expressed in the cytoplasm of Escherichia coli and Western blot analysis demonstrated they retained antigenicity.Indirect VP1-ELISA and epitope ELISAs were subsequently developed to screen a panel of 80 field pig sera using LPB-ELISA as a standard test.For VP1-ELISA and all the epitope ELISAs,there were clear distinctions between the FMDV-positive and the FMDV-negative samples.Cross-reactions with pig sera positive to the viruses of swine vesicular disease virus that produce clinically indistinguishable syndromes in pigs or guinea pig antisera to FMDV strains of type A,C and Asial did not occur.The relative sensitivity and specificity for the GST-E1 ELISA,GST-E2(+2),GST-E1-2 ELISA and VP1-ELISA in comparison with LPB-ELISA were 93.3% and 85.0%,95.0% and 90%,100% and 81.8%,96.6% and 80.9% respectively.This study shows the potential use of the aforementioned epitopes as alternatives to the complex antigens used in current detection for antibody to FMDV structural proteins.

VP1 is a major antigenic protein of foot-and-mouth disease virus(FMDV), which induces the immune response against FMDV infection, and contains several epitopes of the virus. We designed and chemically synthesized a DNA fragment which encoding a tandem repeat protein of 136-160aa and 198-211aa of a strain of type Asia I FMDV, and cloned the gene of heavy chain constant region of sheep IgG. By using the BamH I, EcoR I and Xho I sites, both genes were cloned into pPROExHTb vector in turn to form a recombinant plasmid pPRO-FshIgG A chimeric protein, named FshIgG, was obtained after transforming the pPRO-FshIgG into Escherichia coli BL21 (DE3) host cell and induced by IPTG. Inoculation with 100 microg FsIgG induced strong neutralizing antibody response in guinea pigs, and FshIgG inoculated guinea pigs were also protected against 200 ID50 FMDV challenge. Our study indicated that the heavy chain constant region of sheep IgG can act as the carrier protein for FMDV peptide epitopes, and FshIgG is a potential multiepitope peptide vaccine candidate to prevent FMDV infection.
Animals ; Antibodies, Viral ; blood ; Capsid Proteins ; biosynthesis ; genetics ; Epitopes ; immunology ; Escherichia coli ; genetics ; metabolism ; Foot-and-Mouth Disease Virus ; genetics ; immunology ; Genetic Vectors ; genetics ; Guinea Pigs ; Immunization ; Immunoglobulin G ; genetics ; Immunoglobulin Heavy Chains ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Sheep ; Viral Vaccines ; genetics ; immunology