Objective The study aims to investigate the immunological functions of the nucleocapsid (N) protein of the novel coronavirus Omicron (BA.1, BA.2) and evaluate the differences among different N proteins of mutant strains in immunogenicity. Methods By aligning sequences, the mutation sites of the Omicron (BA.1, BA.2) N protein relative to prototype strain of the novel coronavirus (Wuhan-Hu-1) were determined. The pET-28a-N-Wuhan-Hu-1 plasmid was used as template to construct pET-28a-BA.1/BA.2-N through single point mutation or homologous recombination. The three kinds of N protein were expressed in prokaryotic system, purified through Ni-NTA affinity chromatography, and then immunized into mice. The titer and reactivity of the polyclonal antibody, as well as the expression level of IL-1β and IFN-γ in mouse spleen cells, were detected using indirect ELISA and Western blot assay. Results The constructed prokaryotic expression plasmids were successfully used to express the Wuhan-Hu-1 N, BA.1 N, and BA.2 N proteins in E.coli BL21(DE3) at 37 DegreesCelsius for 4 hours. The indirect ELISA test showed that the titers of polyclonal antibody prepared by three N proteins were all 1:51 200. All three N proteins can increase the expression of IFN-γ and IL-1β cytokines, but the effect of Omicron N protein in activing two cytokines was more obvious than that of Wuhan-Hu-1 N protein. Conclusion The study obtained three new coronavirus N proteins and polyclonal antibodies, and confirmed that mutations in the amino acid sites of the N protein can affect its immunogenicity. This provides a basis for developing rapid diagnostic methods targeting N protein of different novel coronavirus variants.
Animals ; Mice ; SARS-CoV-2/genetics* ; Coronavirus Nucleocapsid Proteins/immunology* ; Nucleocapsid Proteins/isolation & purification* ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Mice, Inbred BALB C ; Interferon-gamma/metabolism* ; Interleukin-1beta/metabolism* ; Female ; Escherichia coli/metabolism* ; Mutation ; Humans

Objective To evaluate the performance of the automated magnetic beads extraction method for extraction of serum fat-soluble vitamins by liquid chromatography-tandem mass spectrometry(LC-MS/MS).Methods A total of 120 clinical residual serum samples were collected automated magnetic beads extraction method was applied to extract serum fat-soluble vitamins A,D2,D3,E and K.The linearity,limit of quantification,precision,accuracy,carryover and matrix effect of fat-soluble vitamins A,D2,D3,E and K were detected by LC-MS/MS.The consistency of the method and conventional extraction method was compared.Results The linear correlation coefficients for extracting fat-soluble vitamins A,D2,D3,E and K by automated magnetic beads extraction method were all greater than 0.99,and the quantitative limits for these five fat-soluble vitamins were 5,0.25,0.25,125 and 0.025 ng/ml.The intra-batch precision and inter-batch precision were 0.66%～4.83%and 0.15%～3.70%,respectively,and the average spike recovery rate was 87.05%～111.11%.The matrix effect was 95.43%～99.07%,and the difference between the mean value of the cyclic injection results of high-low value samples and the mean value of the cyclic injection results of low-low value samples was less than 3s of the mean value of the cyclic injection results of low-low value samples.The statistical analysis showed that the correlation between the results of the automated magnetic beads extraction method and the conventional extraction method was good(r＞0.99),and there was no significant bias in the detection results of the two methods.Conclusion The automated magnetic beads extraction method for extraction of fat-soluble vitamins had good detection performance,which may be expected to improve sample flux and analysis efficiency.

Chronic chloropropene poisoning is a disease mainly caused by peripheral nerve damage due to close contact with chloropropene in industrial production, its clinical manifestations include varying degrees of sensory, motor, or tendon reflex disorders in the distal limbs, and neuromyography can show neurogenic damage. This article analyzed and summarized the clinical characteristics, diagnosis and treatment methods of three patients with occupational chronic chloropropene poisoning, in order to enhance the clinical understanding of occupational chronic chloropropene poisoning and provide reference for clinical diagnosis and treatment.

Objective To explore the impact of C1q/tumour necrosis factor-related protein 6(CTRP6)on doxorubicin(DOX)-induced apoptosis and oxidative stress injury of cardiomyocytes,along with its associated mechanism of action.Methods The H9C2 car-diomyocyte injury model was established by dividing the subjects into different groups.These groups included the control group(NS group),the DOX(1μmol/L)group,and the DOX(1μmol/L)group supplemented with CTRP6 at concentration gradients of 0.5 μg/ml,1μg/ml,3μg/ml,and 5μg/ml,respectively.After 24hours of culture,the survival rate of the cardiomyocytes was measured,the results showed that the highest increase in cardiomyocyte survival rate was observed at a CTRP6 concentration of 3 μg/ml,which was selected as the optimal concentration.Subsequently,the experimental groups of H9C2 cardiomyocytes consisted of the control group(NS group),the CTRP6 group,the DOX(1μmol/L)group,and the DOX(1μmol/L)group supplemented with CTRP6(3μg/ml)group.The impact of DOX stimulation on the transcriptional and translational levels of CTRP6 was assessed using fluorescence real-time quantitative polymer-ase chain reaction(RT-qPCR)and Western blot assay.Apoptosis levels were measured using Tunel assay,while enzyme-linked im-munosorbent assay(ELISA)kits were was used to detect the activity of cellular total glutathione peroxidase(GSH-Px)and superoxide dismutase(SOD),as well as the levels of malondialdehyde(MDA).Western blot assay was used to detect the any alterations in the lev-els of AdipoR1 protein and the protein expression of the protein kinase B/glycogen synthase kinase-3 β(Akt/GSK-3 β)signaling path-way.Results Compared with the control group(NS group),the expression levels of CTRP6 protein and mRNA were significantly de-creased in the DOX group(46.26％and 67.74％reduction,respectively,P＜0.05).In contrast,compared with DOX group,the DOX+CTRP6group showed a significant increase in CTRP6 protein expression levels(P＜0.05).Among the different concentrations of CTRP6 added to the DOX+CTRP6group,the survival rate of cardiomyocytes in DOX+3μg/ml CTRP6 treatment group was significantly increased by 26.48％(P＜0.05).Tunel staining revealed a decrease in apoptosis and an upregulation of Bcl-2 expression.In addi-tion,GSH-Px and SOD activities increased by 20.49％and 36.89％respectively,while MDA levels were significantly inhibited(de-creased by 47.09％,P＜0.05).The levels of AdipoR1 protein was significantly increased(P＜0.05).Furthermore,there was a signifi-cant elevation observed in the phosphorylation levels of the Akt/GSK-3β signaling pathway proteins.Conclusion CTRP6 ameliorates DOX-induced apoptosis and oxidative stress injury in cardiomyocytes,possibly through the protective effect of the Akt/GSK-3β signa-ling pathway.

Chronic chloropropene poisoning is a disease mainly caused by peripheral nerve damage due to close contact with chloropropene in industrial production, its clinical manifestations include varying degrees of sensory, motor, or tendon reflex disorders in the distal limbs, and neuromyography can show neurogenic damage. This article analyzed and summarized the clinical characteristics, diagnosis and treatment methods of three patients with occupational chronic chloropropene poisoning, in order to enhance the clinical understanding of occupational chronic chloropropene poisoning and provide reference for clinical diagnosis and treatment.

ObjectiveTo explore the clinical efficacy of Huanglian Jiedutang as an adjunctive treatment for acute cerebral infarction complicated with gastric motility disorder. MethodSixty patients with acute cerebral infarction complicated with gastric motility disorder with fire toxin syndrome were randomly divided into a western medicine control group （control group） and a traditional Chinese medicine （TCM） combined treatment group （observation group）， with 30 cases in each group. The control group received basic treatment for cerebral infarction and relevant western medical symptomatic treatment based on the patients' gastrointestinal symptoms. The observation group received Huanglian Jiedutang in addition to the treatment provided to the control group. The treatment course was 7 days. Neurological deficit scores and gastrointestinal dysfunction scores were assessed in both groups before treatment and on the 4^th and 7^th days of treatment. Gastrointestinal electrographic parameters， serum citrulline （CIT）， and motilin （MTL） levels were measured in both groups before treatment and on the 7^th day of treatment. Clinical efficacy was compared between the two groups. ResultCompared with the baseline in both groups， the neurological deficit scores and gastrointestinal dysfunction scores were significantly reduced on the 4th and 7^th days of treatment （P<0.05）. The reductions in these scores were more significant on the 7^th day compared with those on the 4^th day of treatment （P<0.05）. On the 4^th and 7^th days of treatment， the observation group showed a significantly greater reduction in neurological deficit scores and gastrointestinal dysfunction scores compared with the control group （P<0.05）. On the 7th day of treatment， compared with the baseline， both groups showed a significant increase in gastric antral and gastric body electric wave amplitudes as well as serum CIT and MTL levels （P<0.05）， and there were no statistically significant differences in the frequency of gastric antral and gastric body electric waves. On the 7th day of treatment， compared with the control group， the observation group had a significant increase in gastric antral and gastric body electric wave amplitudes as well as serum CIT and MTL levels （P<0.05）， and there were no statistically significant differences in the frequency of gastric antral and gastric body electric waves. After 7 days of treatment， the total effective rate in the observation group was 90.00% （27/30）， higher than 76.67% （23/30） in the control group， but the difference was not statistically significant. ConclusionAdjunctive treatment with Huanglian Jiedutang can effectively improve the symptoms of neurological function impairment and gastrointestinal dysfunction in patients with acute cerebral infarction complicated with gastric motility disorder， increase gastric antral and gastric body electric wave amplitudes， improve gastric motility disorder， and increase serum CIT and MTL levels， thereby improving the imbalanced secretion function of the gastrointestinal tract.

Background::Given that seizures may be triggered by vaccination, this study aimed to evaluate the risk and correlative factors of seizures in patients with epilepsy (PWE) after being vaccinated against coronavirus disease 2019 (COVID-19).Methods::This study retrospectively enrolled PWE who were vaccinated against COVID-19 in the epilepsy centers of 11 hospitals in China. We divided the PWE into two groups as follows: (1) patients who developed seizures within 14 days of vaccination were assigned to the SAV (with seizures after vaccination) group; (2) patients who were seizure-free within 14 days of vaccination were assigned to the SFAV (seizure-free after vaccination) group. To identify potential risk factors for seizure reccurence, the binary logistic regression analysis was performed. Besides, 67 PWE who had not been vaccinated were also included for elucidating the effects of vaccination on seizures recurrence, and binary logistic regression analysis was performed to determine whether vaccination would affect the recurrence rate of PWE who had drug reduction or withdrawal.Results::The study included a total of 407 patients; of which, 48 (11.8%) developed seizures within 14 days after vaccination (SAV group), whereas 359 (88.2%) remained seizure-free (SFAV group). The binary logistic regression analysis revealed that duration of seizure freedom ( P < 0.001) and withdrawal from anti-seizure medications (ASMs) or reduction in their dosage during the peri-vaccination period were significantly associated with the recurrence of seizures (odds ratio= 7.384, 95% confidence interval = 1.732–31.488, P = 0.007). In addition, 32 of 33 patients (97.0%) who were seizure-free for more than three months before vaccination and had a normal electroencephalogram before vaccination did not have any seizures within 14 days of vaccination. A total of 92 (22.6%) patients experienced non-epileptic adverse reactions after vaccination. Binary logistic regression analysis results showed that vaccine did not significantly affect the recurrence rate of PWE who had the behavior of ASMs dose reduction or withdrawal ( P = 0.143). Conclusions::PWE need protection from the COVID-19 vaccine. PWE who are seizure-free for >3 months before vaccination should be vaccinated. Whether the remaining PWE should be vaccinated depends on the local prevalence of COVID-19. Finally, PWE should avoid discontinuing ASMs or reducing their dosage during the peri-vaccination period.

This paper reported 1 case of poisoning caused by stramonium. Cases of Datura poisoning have been reported nationwide, Its effect on the central nervous system of patients is characterized by first excitation and then inhibition, clinical manifestations include decreased gland secretion, dilated pupils, and tachycardia, etc. Its poisoning mechanism is anticholinergic effect, the effect on Peripheral nervous system is to inhibit Parasympathetic nervous system. Hemoperfusion combined with neostigmine anticholinergic therapy at the early stage of poisoning can effectively improve the clinical symptoms of patients in a short time.

This paper reported 1 case of poisoning caused by stramonium. Cases of Datura poisoning have been reported nationwide, Its effect on the central nervous system of patients is characterized by first excitation and then inhibition, clinical manifestations include decreased gland secretion, dilated pupils, and tachycardia, etc. Its poisoning mechanism is anticholinergic effect, the effect on Peripheral nervous system is to inhibit Parasympathetic nervous system. Hemoperfusion combined with neostigmine anticholinergic therapy at the early stage of poisoning can effectively improve the clinical symptoms of patients in a short time.

【Objective】 To understand the structural characteristics of lapsed apheresis donors in our blood center and provide guidance for further improving recruitment and retention strategies by retrospectively analyzing the characteristics of lapsed apheresis donors in our center in recent years. 【Methods】 The apheresis donation and lapsed apheresis donors in Zhejiang Blood Center from 2016 to 2020 were statistically analyzed, and the general information of lapsed donors, including gender, blood type, age, education level and occupation composition, were compared and analyzed. The lapse of novel and long-term donors with different frequency were retrospectively analyzed. The SPSS 19.0 statistical software package was used for data analysis. 【Results】 In 2020, the total lapse of apheresis donors decreased by 16.6% as compared with 2016(P<0.05). The lapse rate of donors with blood groups A, B, O and AB was 1∶1∶1∶1, higher in female donors(59.0%) than males(50.0%), and dominated by age group of 18-35(66.3%). With the increase of age, the lapse rate decreased. Donors lapsed were mainly with college or above educational background(60.8%), with high proportion in students and the staff. In 2020, the lapse of novel apheresis donors decreased by 34.1% as compared with 2016, but the average lapse rate of novel apheresis donors was still as high as 70.5%. The average lapse rate of blood donors with different frequency was 52.5%, and the lapse rate decreased significantly with the increase of apheresis donation frequency. The average lapse rate of novel blood donors with whole blood donation experience was lower than those without (56.1% vs 82.9%). 【Conclusion】 Multiple measures for recruitment and retention have effectively reduced the lapse of apheresis donors. However, apheresis donors who are 18-35 years old, with college degree or above, students and staff were the main groups from lapsing. In addition, low-frequency and novel apheresis donors without whole blood donation experience were more likely to be lapsing. Targeted and personalized blood donation services should be further provided, and the management of " full cycle of blood donation" should be strengthened to reduce the lapsing, so as to retain more blood donors.