Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.

Objective To understand how warfarin inhibition is hindered by the vitamin K epoxide reductase(VKORC1)resistant mutations.Methods An electrophoretic mobility assay was conducted based on the principle that binding with warfarin resulted in changes in VKORC1 electrophoretic mobility.The activity of VKORC1 and the half maximal inhibitory concentration(ICs0)of warfarin inhibiting VKORC1 were detected by ELISA.These experiments evaluated the binding ability of VKORC1 resistant mutants to warfarin.Results With warfarin bound,VKORC1 was protected from N-ethylmaleimide(NEM)modification and showed increased electrophoretic mobility,which was dependent on an unmodified cysteine on the flexible transmembrane helix 1(TM1)of VKORC1.In-creasing the warfarin concentration could shift more VKORC1 towards the species with the fast mobility.Compared to the wildtype VKORC1,the fast mobility fraction became less or disappeared in warfarin-resistant mutants,indi-cating weakened binding of warfarin.In addition,VKORC1 mutants with the weaker electrophoretic mobility shift indicated the stronger warfarin resistant.Conclusion Weakened warfarin binding is the primary cause of warfarin resistance.

Objective:To analyze the risk factors of recurrent wheezing in children with bronchiolitis and to construct a predictive model.Methods:Prospective research methods was used. One hundred and eighty children with bronchiolitis who were treated in Hefei Eighth People's Hospital from February 2017 to February 2019 were selected as the study subjects, and the included children were separated into a modeling group (126 cases) and a validation group (54 cases) according to 7∶3. The children were followed up for 3 years, and then the modeling group was divided into wheezing group (48 cases) and no wheezing group (78 cases) according to whether the children had recurrent wheezing. The Hosmer-Lemeshow fitting curve and receiver operating characteristic (ROC) curve were drawn to evaluate the validity and accuracy of the constructed prediction model.Results:Multivariate Logistic regression analysis showed that artificial feeding ( OR = 8.838, 95% CI 2.601 to 30.027), family history of allergies ( OR = 6.709, 95% CI 1.825 to 24.665), underlying diseases ( OR = 8.114, 95% CI 1.638 to 40.184), and higher IgE level ( OR = 1.020, 95% CI 1.012 to 1.029) were the independent risk factors for recurrent wheezing in children with bronchiolitis ( P<0.05). The area under the curve of the modeling group was 0.917 (95% CI 0.855 to 0.959), and the sensitivity and specificity were 83.33% and 85.90%, respectively; the area under the curve of the validation group was 0.911 (95% CI 0.847 to 0.954), and the sensitivity and specificity were 89.58% and 79.49%, respectively. Conclusions:Artificial feeding, family history of allergies, underlying diseases, and higher IgE level are the independent risk factors for recurrent wheezing in children with bronchiolitis. The constructed prediction model has good accuracy and validity, and can be used as an effective tool for clinical prediction of recurrent wheezing in children with bronchiolitis.

Pulmonary blast injury has become the main type of trauma in modern warfare, characterized by externally mild injuries but internally severe injuries, rapid disease progression, and a high rate of early death. The injury is complicated in clinical practice, often with multiple and compound injuries. Currently, there is a lack of effective protective materials, accurate injury detection instrument and portable monitoring and transportation equipment, standardized clinical treatment guidelines in various medical centers, and evidence-based guidelines at home and abroad, resulting in a high mortality in clinlcal practice. Therefore, the Trauma Branch of Chinese Medical Association and the Editorial Committee of Chinese Journal of Trauma organized military and civilian experts in related fields such as thoracic surgery and traumatic surgery to jointly develop the Clinical treatment guideline for pulmonary blast injury ( version 2023) by combining evidence for effectiveness and clinical first-line treatment experience. This guideline provided 16 recommended opinions surrounding definition, characteristics, pre-hospital diagnosis and treatment, and in-hospital treatment of pulmonary blast injury, hoping to provide a basis for the clinical treatment in hospitals at different levels.

Objective: To investigate the expression pattern，underlying function and clinical significance of Guanylate-binding protein 1 ( GBP1) in pulmonary tuberculosis ( pTB) ． Methods: Immunohistochemical staining was applied to detect the expression of GBP1 in pTB specimensand control samples． Combined with Gene Expression Omnibus ( GEO) datasets ，including GSE83456 and GSE34608，receiver operating characteristic ( ROC) curve was depicted to assess the diagnostic value of GBP1 in pTB．Then，the correlation between GBP1 and related regulatory factors was analyzed by protein-protein interaction network ( PPI) ; Finally，the potential molecular mechanism of GBP1 in pTB was probed by Gene Set Enrichment Analysis( GSEA) . Results: Compared with the control group，GBP1 was significantly overexpressed in human pTB samples，including lung tissue and blood．The positive rate of GBP1 protein in pTB was 73. 9% . ROC curve analysis revealed that GBP1 might have important value in early diagnosis of pTB．GSEA analysis suggested that the hyper-expression of GBP1 was closely related to the host inflammatory response，IFN-γ/ α signaling pathway and TNF-α/ IL-6 signal transduction． Conclusion GBP1 is highly expressed in pTB tissues and is involved in the process of inflammatory response and host anti-tuberculosis infection ; GBP1 may be used as an early diagnostic marker or therapeutic target for pTB.

Preclinical studies have shown that metformin, an activator of AMP-activated protein kinase (AMPK) , can inhibit the growth of lymphoma cells without affecting the function of normal lymphocytes, promote the apoptosis of lymphoma cells and improve the tumor immune environment. Clinical studies have shown that metformin can improve the efficacy of chemotherapy and prognosis of non-Hodgkin lymphoma (NHL) . Through the influence on glucose metabolism, it can improve blood glucose level and reduce the incidence of steroid diabetes. The main molecular mechanisms of metformin against NHL include activating AMPK in tumor cells, inhibiting mammalian target of rapamycin signal, inhibiting key cholesterol synthesis, improving glucose metabolism, enhancing highly cytotoxic T lymphocyte (CTL) activity and so on. Current research shows that metformin may become a new strategy for the treatment of NHL.

Selenium (Se) is an essential trace element for organisms. Se deficiency will cause diseases such as Keshan disease and Kashin-Beck in human being, and huge loss to animal husbandry. Currently available Se supplements have such problems as low Se content, poor bioavailability, and poor safety. Chlorella pyrenoidosa can produce bioavailable and safe organic Se under suitable conditions, which is thus a promising Se supplement. Therefore, in this study, we tried to improve the Se tolerance and accumulation of C. pyrenoidosa by directional adaptation. To be specific, we gradually increased the concentration of Na₂SeO₃ in medium to domesticate C. pyrenoidosa and optimized the adapting time and concentration gradient of Na₂SeO₃ during the adaptation. The results showed that the adapted C. pyrenoidosa was more tolerant to Se and had stronger Se enrichment ability. In 5 L fermenter, the adapted strains could tolerate 40 mg/L Na₂SeO₃ and the synthesis rate of organic Se was 175.6% higher. Then, Se addition method in the 5 L fermenter was optimized. The result demonstrated that addition of Na₂SeO₃ at 40 mg/L during heterotrophic culture achieved the final dry weight of C. pyrenoidosa cells at 106.4 g/L, content of organic Se at 1 227 mg/kg, and synthesis rate of organic Se at 1.36 mg/(L·h). Compared with the reported highest cell density of 75 g/L and the highest organic Se content of 560 mg/kg, the corresponding figures in this study were 41.9% and 119.1% higher, respectively. In conclusion, directional adaptation can remarkably improve the Se tolerance and enrichment of C. pyrenoidosa.
Animals ; Humans ; Selenium/pharmacology* ; Chlorella ; Heterotrophic Processes

Objective:To analyze the genotype distribution of acute hepatitis B virus in China.Methods:A total of six hundred and twenty acute Hepatitis B cases reported to China Information System for Diseases Control and Prevention from 2015 to 2017 were selected. First, the full-length HBV genome was obtained by nested PCR amplification. In addition, the HBV genotype was determined by constructing a phylogeny tree. Finally, using primarydata, HBV genotype distribution was analyzed.Results:A total of 519 (83.71%, 519/620) sequences were obtained genotype of 620 acute hepatitis B cases, including A (0.19%, 1/519), B (27.17%, 141/519), C (62.04%, 322/519), D (9.06%, 47/519), I (0.77%, 4/519) and C/D (0.77%, 4/519); B2(95.03%, 134/141) and C2 (72.67%, 234/322) were the two major subgenotypes. Genotypes were distributed differently in seven regions of China. The proportion of genotype C appeared higher in Northeast China (94.55%, 52/55), North China (93.85%, 61/65), East China (78.87%, 56/71), and South China (58.14%, 50/86). The proportion of genotype B was higher in Central China (58.07%, 36/62) and Southwest China (52.94%, 45/85), the proportion of genotype D was the highest in Northwest China (48.42%, 46/95). A total of 515 cases were classified as serotypes, including 'adr' (57.48%, 296/515), 'adw' (30.87%, 159/515), 'ayr' (0.19%, 1/515), and 'ayw' (11.46%, 59/515). Genotype B was dominated by 'adw' serotype (92.14%, 129/140), genotype C was dominated by 'adr' serotype (91.88%, 294/320),all genotype D were 'ayw' serotype. The genotype of acute hepatitis B was correlated with serotype, 'adw' was dominant in genotype B, 'adr' was dominant in genotype C and 'ayw' was dominant in genotype D.In different gender and age group, there was no statistical significance ingenotype distribution ( P>0.05). Conclusions:The genotype of acute hepatitis B in China from 2015 to 2017 was mainly B, C, and D; genotype C was dominant in the Northeast China,North China, East China and South China; B and C were common in Central and Southwest China, and genotype B was dominant. Genotype D was primarily distributed in Northwest China. The genotype of acute hepatitis B was correlated with serotype, 'adw' was dominant in genotype B, 'adr' was dominant in genotype C and 'ayw' was dominant in genotype D. There was no difference in the distribution of acute hepatitis B genotypes among different genders and age groups.

γ-glutamyl carboxylase (GGCX), also known as vitamin K-dependent glutamyl carboxylase, catalyzes the posttranslational modification of specific glutamate residues in vitamin K-dependent proteins (VKDPs), and participates multiple biological functions including blood coagulation, bone metabolism, vascular calcification, and cell proliferation. It has been reported originally that GGCX pathogenic variation causes blood coagulation deficiency, which is called as vitamin K-dependent coagulation factor deficiency 1 (VKCFD1). Recently, it has been found that GGCX gene variation results in multiple clinical phenotypes, including dermatological, ophthalmological, skeletal or cardiac abnormalities. Among them, dermatological phenotype is the most common, which is known as pseudoxanthoma elasticum-like syndrome. This paper has reviewed the GGCX pathogenic variation associated phenotypes, in order to increase the recognition of GGCX-related genetic diseases and to help its diagnosis and treatment.

Objectives:To understand the awareness rate and its influencing factors of their HBV infection status among HBsAg-positive persons aged 15-69 years in China.Methods:A cross-sectional design was used to conduct a questionnaire survey on the awareness of their infection status among HBsAg-positive persons aged 15-69 years who were identified in the 2020 national hepatitis B seroepidemiology survey. The awareness rate of the whole respondent and respondents with different characteristics were described, and the differences were compared with the χ2 test. The logistic regression model was used to analyze the factors influencing the awareness rate. Results:The overall awareness rate among the respondents was 43.10% (1 828/4 241). The awareness rate was lower in males than in females (41.30% vs. 44.65%). The awareness rate was lower in the 60-69-years-old age group than in other age groups (30.38% vs. 36.77%-57.58%). The awareness rate was lower in rural areas than in urban areas (39.43% vs. 47.32%). The awareness rate was lower in regions with a per capita gross domestic product (GDP) below RMB 54 000 than in regions with a per capita GDP of RMB 54 000 and above (36.81% vs. 41.61%-50.30%). The awareness rate was lower in respondents without other liver diseases than with other liver diseases (41.52% vs. 60.68%). The awareness rate was lower in respondents without a family history of hepatitis B-related disease or unknown family history than with a family history (43.58% vs. 68.26%; 24.71% vs. 68.26%). Multivariate logistic regression analysis showed that male [odds ratio ( OR)=0.841, 95% confidence interval ( CI): 0.734-0.964], high school and below [primary school and below, junior middle school, high school/technical secondary school, OR (95% CI): 0.247 (0.190-0.321), 0.451 (0.352-0.577), 0.634 (0.486-0.827)], rural areas ( OR=0.822, 95% CI: 0.715-0.945) and regions with a per capita GDP below RMB 80 000 [54 000-80 000, OR (95% CI): 0.810 (0.688-0.954), below RMB 54 000, OR (95% CI): 0.793 (0.669-0.941)] were the negative factors influencing the awareness rate. While 30-39-years-old ( OR=2.089, 95% CI: 1.626-2.683) and 40-49-years-old ( OR=1.590, 95% CI: 1.250-2.023) age groups, with other liver diseases ( OR=2.244, 95% CI: 1.754-2.871) and family history related to hepatitis B ( OR=2.688, 95% CI: 2.242-3.223) were the positive factors influencing the awareness rate. Conclusion:The overall awareness rate of their infection status among HBsAg-positive persons aged 15-69 years is 43.10% in China. Health promotion and coverage expansion on HBV screening should be further strengthened to achieve the proposed World Health Organization's target of 90% HBV infection diagnosis rate by 2030.