To study the immunoregulatory effects of lycorine on mice infected with Toxoplasma gondii(T.gondii),BALB/c mice were treated with 20 mg/kg lycorine solution after peritoneal in-fection with T.gondii RH strain.The serum and spleen of mice were collected at the 1st,3rd,5th and 7th day,respectively,and the spleen index of mice was calculated.The proportion of T lympho-cyte subtypes and NK cells were detected in mice by flow cytometry,and the changes of cytokine levels in mice were measured using ELISA kit,so as to evaluate the immunomodulatory effect of lycorine on mice infected with T.gondii.At the same time,the heart,liver,spleen,lungs,and kid-neys of mice were collected at the 7th day,and the pathological changes of the mouse organs were observed through pathological tissue sections,and the amount of the parasite in the liver,lung,kid-ney,and brain of the mice was detected by fluorescence quantitative PCR.The results showed that the survival time of the mice treated with lycorine was significantly extended,and the survival rate reached 80％.The spleen index of mice treated with lycorine was lower than that of the control group.Lycorine up-regulates the ratio of the CD4+T lymphocytes and NK cells in the spleen of mice infected with T.gondii,and helps to improve the ability of mice to resist T.gondii infection.Lycorine can up-regulate the levels of anti-inflammatory cytokine IL-4 and down-regulate the lev-els of pro-inflammatory factors IFN-γ,IL-1β,and IL-9 in serum of mice infected with T.gondii,which is conducive to reducing the damage of inflammatory response and enhancing the ability of anti-T.gondii infection.In addition,lycorine can alleviate the pathological damage of T.gondii to the liver,lungs,spleen,and kidneys of mice,and significantly reduce the amount of the parasite in the lung of mice.The results showed that lycorine could up-regulate the proportion of immune cells CD4+T lymphocytes and NK cells,inhibit the inflammatory response of mice infected with T.gondii,reduce the pathological damage of mice organs,and enhance the ability of mice to resist T.gondii infection,which is expected to become a novel anti-T.gondii drug.

To construct a safe and effective multi-epitope vaccine against the S1 protein of chicken infectious bronchitis virus(IBV).In this study,homologous and non-homologous dominant epitopes of IBV M41,T,QX and H120 virulent strain S1 proteins were screened by various online bioprediction software,respectively,and a new peptide W with high immunogenicity was construc-ted by connecting the screened B-cell and T-cell epitopes with a linker peptide.W was ligated to the truncated sequence of the four viral strains by T2A yietding to the eukaryotic expression vector pEGFP-N1,and it was identified by PCR and double digestion,the obtained recombinant plasmid was transfected into HEK293A cells and target protein expression was measured by Western blot.The constructed plasmid was injected intramuscularly twice to detect the antibody level,cytokine level,and peripheral blood T cell subsets were detected after two immunizations.The epitope pro-tein W was successfully constructed,which was structurally stable,antigenic,and soluble;the re-combinant plasmid pEGFP-WMQtH,pEGFP-W,and pEGFP-MQtH matched the expected size;anti-IBV IgG antibody levels in pEGFP-N1 was increased greatly compared to the PBS group.cyto-kines IL-2,and γ interferon(IFN-γ)were increased greatly(P＜0.05);peripheral blood CD4+/CD8a value(P＜0.05)was increased greatly.The W epitope protein was successfully constructed,which can effectively activate the humoral immunity and cellular immunity against four infectious bronchitis viruses(IBV),laying a foundation for the development of an effective vaccine against IB.

To construct a safe and effective multi-epitope vaccine against the S1 protein of chicken infectious bronchitis virus(IBV).In this study,homologous and non-homologous dominant epitopes of IBV M41,T,QX and H120 virulent strain S1 proteins were screened by various online bioprediction software,respectively,and a new peptide W with high immunogenicity was construc-ted by connecting the screened B-cell and T-cell epitopes with a linker peptide.W was ligated to the truncated sequence of the four viral strains by T2A yietding to the eukaryotic expression vector pEGFP-N1,and it was identified by PCR and double digestion,the obtained recombinant plasmid was transfected into HEK293A cells and target protein expression was measured by Western blot.The constructed plasmid was injected intramuscularly twice to detect the antibody level,cytokine level,and peripheral blood T cell subsets were detected after two immunizations.The epitope pro-tein W was successfully constructed,which was structurally stable,antigenic,and soluble;the re-combinant plasmid pEGFP-WMQtH,pEGFP-W,and pEGFP-MQtH matched the expected size;anti-IBV IgG antibody levels in pEGFP-N1 was increased greatly compared to the PBS group.cyto-kines IL-2,and γ interferon(IFN-γ)were increased greatly(P＜0.05);peripheral blood CD4+/CD8a value(P＜0.05)was increased greatly.The W epitope protein was successfully constructed,which can effectively activate the humoral immunity and cellular immunity against four infectious bronchitis viruses(IBV),laying a foundation for the development of an effective vaccine against IB.

To study the immunoregulatory effects of lycorine on mice infected with Toxoplasma gondii(T.gondii),BALB/c mice were treated with 20 mg/kg lycorine solution after peritoneal in-fection with T.gondii RH strain.The serum and spleen of mice were collected at the 1st,3rd,5th and 7th day,respectively,and the spleen index of mice was calculated.The proportion of T lympho-cyte subtypes and NK cells were detected in mice by flow cytometry,and the changes of cytokine levels in mice were measured using ELISA kit,so as to evaluate the immunomodulatory effect of lycorine on mice infected with T.gondii.At the same time,the heart,liver,spleen,lungs,and kid-neys of mice were collected at the 7th day,and the pathological changes of the mouse organs were observed through pathological tissue sections,and the amount of the parasite in the liver,lung,kid-ney,and brain of the mice was detected by fluorescence quantitative PCR.The results showed that the survival time of the mice treated with lycorine was significantly extended,and the survival rate reached 80％.The spleen index of mice treated with lycorine was lower than that of the control group.Lycorine up-regulates the ratio of the CD4+T lymphocytes and NK cells in the spleen of mice infected with T.gondii,and helps to improve the ability of mice to resist T.gondii infection.Lycorine can up-regulate the levels of anti-inflammatory cytokine IL-4 and down-regulate the lev-els of pro-inflammatory factors IFN-γ,IL-1β,and IL-9 in serum of mice infected with T.gondii,which is conducive to reducing the damage of inflammatory response and enhancing the ability of anti-T.gondii infection.In addition,lycorine can alleviate the pathological damage of T.gondii to the liver,lungs,spleen,and kidneys of mice,and significantly reduce the amount of the parasite in the lung of mice.The results showed that lycorine could up-regulate the proportion of immune cells CD4+T lymphocytes and NK cells,inhibit the inflammatory response of mice infected with T.gondii,reduce the pathological damage of mice organs,and enhance the ability of mice to resist T.gondii infection,which is expected to become a novel anti-T.gondii drug.

To construct a recombinant attenuated Salmonella strain carrying the p57 and BLID genes and to evaluate its inhibitory effect on breast cancer cells and apoptosis induction in vitro,the recombinant attenuated Salmonella LH430/pEGFP-p57-BLID was constructed.RT-PCR and Western blot were used to detect the transcription and expression of the recombinant attenuated Salmonella in cancer cells.The CCK-8 method was used to assess the safety of the recombinant at-tenuated Salmonella and its impact on the activity of breast cancer cells.The cell scratch assay was used to analyze the influence of the recombinant attenuated Salmonella on the migratory ability of cancer cells.Flow cytometry and Western blot were used to analyze the effect of the recombinant attenuated Salmonella on the apoptosis of MDA-MB-231 cells.The bands of target genes carried by the recombinant attenuated Salmonella could be amplified by PCR,and these target genes could be transcribed and expressed in MDA-MB-231 cells.The recombinant attenuated Salmonella showed good growth characteristics and genetic stability.Compared with the PBS group,recombi-nant attenuated Salmonella LH430/pEGFP-p57-BLID significantly reduced(P＜0.01)the activity of MDA-MB-231 cells and significantly inhibited(P＜0.01)the migration of MDA-MB-231 cells.Recombinant attenuated Salmonella LH430/pEGFP-p57-BLID can obviously increase the number of apoptosis in MDA-MB-231 cells,significantly upregulate(P＜0.01)the expression of apoptotic protein Bax,and significantly downregulate(P＜0.01)the expression of apoptotic protein Bcl-2.The results showed that the recombinant attenuated Salmonella had an inhibitory effect on breast cancer MDA-MB-231 cells and could induce apoptosis,which laid a foundation for the subsequent study of anti-tumor in vivo.

To construct a recombinant attenuated Salmonella strain carrying the p57 and BLID genes and to evaluate its inhibitory effect on breast cancer cells and apoptosis induction in vitro,the recombinant attenuated Salmonella LH430/pEGFP-p57-BLID was constructed.RT-PCR and Western blot were used to detect the transcription and expression of the recombinant attenuated Salmonella in cancer cells.The CCK-8 method was used to assess the safety of the recombinant at-tenuated Salmonella and its impact on the activity of breast cancer cells.The cell scratch assay was used to analyze the influence of the recombinant attenuated Salmonella on the migratory ability of cancer cells.Flow cytometry and Western blot were used to analyze the effect of the recombinant attenuated Salmonella on the apoptosis of MDA-MB-231 cells.The bands of target genes carried by the recombinant attenuated Salmonella could be amplified by PCR,and these target genes could be transcribed and expressed in MDA-MB-231 cells.The recombinant attenuated Salmonella showed good growth characteristics and genetic stability.Compared with the PBS group,recombi-nant attenuated Salmonella LH430/pEGFP-p57-BLID significantly reduced(P＜0.01)the activity of MDA-MB-231 cells and significantly inhibited(P＜0.01)the migration of MDA-MB-231 cells.Recombinant attenuated Salmonella LH430/pEGFP-p57-BLID can obviously increase the number of apoptosis in MDA-MB-231 cells,significantly upregulate(P＜0.01)the expression of apoptotic protein Bax,and significantly downregulate(P＜0.01)the expression of apoptotic protein Bcl-2.The results showed that the recombinant attenuated Salmonella had an inhibitory effect on breast cancer MDA-MB-231 cells and could induce apoptosis,which laid a foundation for the subsequent study of anti-tumor in vivo.

{L-End}Objective To analyze the spectrum characteristics of noise hazards in the metal products industry. {L-End}Methods A total of six metal product industries were selected as research subjects using stratified sampling method. The noise intensity (A/C-weighted) and noise spectrum (Z-weighted) of workplaces and job positions were detected. The characteristics of the noise spectrum of each job position were analyzed using the difference between equivalent continuous C-weighted sound pressure level (L_Ceq)-equivalent continuous A-weighted sound pressure level (L_Aeq), the dominant frequency of the noise spectrum, and cluster analysis methods. {L-End}Results The workplace noise and job position noise of 11 main noise positions in the metal products industry were exceeded national standard, with incidence of 69.2% and 78.1%, respectively. The average of normalization of equivalent continuous A-weighted sound pressure level to a normal 40 hours working week exceeded the national standard in 100.0% of hydraulic workers, welders, and ultrasonic cleaning workers. The result of spectrum analysis showed that the noise in the metal product industry was mainly broadband. The noise of the collision welder position was classified as low-frequency broadband noise, while the noise of the painter position was classified as low-frequency narrowband noise. The noise spectrum characteristics of other positions were similar and classified into one category, all of which were broadband noise. Among them, numerical control lathe workers, welders, threaders, machinists, and cutters were exposed to high-frequency broadband noise, while press workers were exposed to mid-frequency broadband noise, and grinders, hydraulic workers, and ultrasonic cleaning workers were exposed to low-frequency broadband noise. The detection rate of binaural high-frequency hearing threshold improvement among workers was 61.7%, and there was no statistical correlation between the detection rate of binaural high-frequency hearing threshold improvement and the noise level intensity of each spectrum. {L-End}Conclusion The level of noise hazards in the metal products industry is severe. Effective engineering control measures and hearing protection measures should be implemented based on the spectral characteristics of noises.

Objective:To investigate the clinical and electrophysiological characteristics of facial onset sensory motor neuronopathy (FOSMN) syndrome.Methods:Ten patients diagnosed with FOSMN syndrome in Peking Union Medical College Hospital from January 2012 to December 2022 were included. The clinical and electrophysiological characteristics of patients were analyzed and summarized, and the genetic testing was also performed in these patients.Results:The age of onset was (56.6±6.5) years, and the longest survival duration of disease was 10 years. All patients had numbness around the face and mouth as the first symptom and abnormal blink reflex. A total of 52 sensory nerve conduction nerves were detected, among which 2 median nerves and 2 μlnar nerves showed decreased amplitude of sensory nerve action potential. Needle electromyography showed neurogenic lesions, with both progressive and chronic denervation. Whole exome sequencing identified the heterozygous variant c.272A>C in the exon 4 of the SOD1 gene resulting in the amino acid change p.Asp90Ala in 1 patient. In all patients, the disease progressed relentlessly and eventually led to involvement of respiratory muscle. Conclusion:FOSMN syndrome is characterized by abnormal blink reflex and sometimes abnormal sensory nerve conduction may be shown on electrophysiologic testing.

Objective: To evaluate the value of cerebrospinal fluid markers expecially total-tau protein (T-tau), phosphorylated-tau protein (P-tau) in diagnosis and differentiation of sporadic Creutzfeldt-Jakob disease (sCJD). Methods: sCJD (according to 2009 Brain criteria, 2018 Neurology amended criteria), Alzheimer's disease (AD; the National Institute on Aging at National Institutes of Health and the Alzheimer's Association revised guidelines 2011 criteria) and other patients without cognitive impairment, matched for sex and age, in the Department of Neurology, Peking Union Medical College Hospital from 2018 to 2019 were enrolled. Twelve sCJD patients, 49 AD patients and 14 normal controls were enrolled. Cerebrospinal fluid (CSF) specimens were collected through gravity dropping directly, and further stored in -80 ℃ and disposed according to widely used standards. The levels of T-tau and P-tau were measured by ELISA. The data on electroencephalogram and neuroimaging findings of sCJD patients were recorded. Moreover, specimens of sCJD patients were sent to the Chinese Center for Disease Control and Prevention to test 14-3-3 protein and PRNP genotype. Results: Using Mann-Whitney U test, T-tau concentrations were found higher in patients with sCJD (1 211(448, 2 227) pg/ml) than in AD patients (549(314, 1 078) pg/ml; U=178, P=0.034 9), and both groups had higher T-tau than the control group (127(79, 192) pg/ml; U=20, 73, P<0.01). The level of P-tau was significantly increased in AD patients (72(58,109) pg/ml) compared to the control group (27(15, 42) pg/ml; U=82, P<0.01), but not in sCJD patients (32(24, 47) pg/ml). The T-tau/P-tau ratio was higher in sCJD patients (29.77(20.01, 54.53)) than in AD patients (7.45(4.79, 10.43); U=87, P<0.01). Twelve sCJD patients had cotical hyperintensity on diffusion weighted imaging and five had periodic three-phase waves on electroencephalogram. Nine sCJD patients, whose CSF samples were tested in the Chinese Center for Disease Control and Prevention, carried an M/M genotype at codon 129 and E/E at codon 219. Conclusion The CSF tau level and T-tau/P-tau ratio are significantly increased in sCJD, which may promote the diagnosis and differentiation of sCJD in routine clinical setting.

Identifying and comparing the chemical constituents of wild silkworm cocoon and silkworm cocoon is of great significance for understanding the domestication of silkworm. In this study, we used high temperature and high pressure and methanol-water system to extract cocoon chemical constituents. We used UHPLC-MS to identify and compare cocoon chemical constituents of wild silkworm and domestic silkworm Dazao and Haoyue strains. The cocoon metabolic fingerprints of wild silkworm and domestic silkworm Dazao and Haoyue strains were obtained by using the UHPLC-MS in the positive ion mode and negative ion mode. By annotation, we found that cocoon chemical compounds with high abundances contained amino acids, flavonoids, alkaloids, terpenes, organic acids, and lignans. PLS-DA showed that the cocoon components were significantly different among the wild silkworm and two domestic silkworm strains Dazao and Haoyue. Proline, leucine/isoleucine and phenylalanine showed significantly higher abundances in the cocoon of domestic silkworm Dazao strain than in those of wild silkworm and domestic silkworm Haoyue strain. The flavonoid secondary metabolites are abundant in the Dazao cocoon, including quercetin, isoquercetin, quercetin 3-O-sophoroside, quercetin-3-O-α-L-rhamnoside, quercetin-3-O- rutinoside, and kaempferol. The other secondary metabolites, alkaloids, terpenes and lignans, showed higher abundances in the wild silkworm cocoon than in the domestic silkworm cocoon, including neurine, candicine, pilocarpidine, artemisiifolin, eupassopin, and eudesobovatol. By exposing cocoons to UV light and observing the green fluorescence of flavonoids, we found that Dazao cocoon had the most flavonoids, and Haoyue cocoon had least flavonoids and wild silkworm cocoon had mediate flavonoids. Alkaloids and organic acids are good anti-insect and antimicrobial agents, which have high abundance in the wild silkworm cocoon and could enhance the defense ability of wild silkworm cocoon. Flavonoids are abundant in the cocoon of domestic silkworm Dazao strain, which the main factors are leading to the yellow-green cocoon of Dazao.
Animals ; Bombyx ; Chromatography, High Pressure Liquid ; Flavonoids ; Mass Spectrometry