The objective of this study was to evaluate the toxicity and safety of novel Mycobacterium tuberculosis fusion strain protein derivatives,referred to as B/R strain active proteins.In cellular experiments,RAW264.7 cells were treated with each vaccine preparation,and apoptosis rates were measured.In subsequent animal experiments,C57BL/6 mice were immunized via subcutaneous injection,and their survival and body weight changes were monitored and recorded at 2,4,8,12,and 16 weeks.The lungs and spleens were harvested to calculate organ coefficients,and pathological examinations were conducted.At the eighth week of immunization,the mice were infected with high concentrations of BCG,and pathological changes in the lungs and spleens were observed 4 weeks post-infection.The apoptosis rate at 6 hours was significantly higher in the experimental group than the PBS group(P<0.05).At 12 and 24 hours,the apoptosis rate in the experimental group remained higher than that in the PBS group,although this difference was not statistically significant.After immunization,mice in all four groups exhibited normal growth patterns,as indicated by stable body weight changes.At 4 and 12 weeks post-immunization,the lung coefficients in the protein group were significantly higher than those in the PBS group at the same time points.Additionally,the lung coefficients in the BCG group were significantly elevated across all time periods(P<0.05).The spleen coefficients in the protein and BCG groups were significantly higher than those in the PBS group at 2,4,8,12,and 16 weeks,whereas the ICD B/R group showed higher spleen coefficients than the PBS group only at week 8(P<0.05).Pathological examination revealed normal lung and spleen tissues in the PBS group.However,during the 2-8 weeks immunization period,lung and spleen tissues in all experimental groups exhibited varying degrees of damage,which gradually diminished by 12-16 weeks.Notably,no tuberculosis nodules were observed in any experimental group.After infection with high concentrations of BCG,no overt pathological changes were observed on the surfaces of the lungs and spleens in any group.Microscopic examination revealed less severe pathological changes in the lungs and spleens of mice in the experimental groups than the PBS group.Furthermore,no statistically significant differences were observed between the protein group and the BCG group.Our findings suggested that the B/R strain active proteins'toxicity and safety profiles were comparable to those of BCG,and showed immunoprotective effects.This study provides an experimental foundation for the development of a novel tuberculosis vaccine.

BACKGROUND:Allicin,the main active ingredient in garlic,has anti-inflammatory and antioxidant effects,but the role and mechanism of allicin in ameliorating rheumatoid arthritis remain unclear.OBJECTIVE:To analyze differentially expressed genes in rheumatoid arthritis using gene expression data and explore the regulatory role of alliin in rheumatoid arthritis.METHODS:Gene expression data for rheumatoid arthritis were collected from the GSE45291 and GSE93777 datasets,and differential expression analysis was then performed.Co-expression network analysis was conducted on the common differentially expressed genes identified in both datasets to identify the module genes most closely related to rheumatoid arthritis,followed by functional enrichment analysis.The SwissTargetPrediction database was used to predict the target genes of allicin in the differentially expressed genes.The area under the receiver operator characteristic curve(AUC)of the differentially expressed target genes was calculated.Human immortalized rheumatoid arthritis fibroblast-like synoviocytes were seeded onto pore plates and cultured for 24 hours,and the cells were then divided into five groups.The control group was not treated;methotrexate was added to the positive drug group;and 20,80,and 160 μg/mL allicin was added to the low-,medium-,and high-dose allicin groups,respectively.After 48 hours of treatment,cell activity was assessed using cell counting kit-8 assay and cell apoptosis was detected using TUNEL assay.Levels of oxidative stress and inflammatory factors were measured by ELISA.Changes in mitochondrial membrane potential were detected by JC-1 staining.The expression levels of target genes and related signaling pathways were detected by RT-qPCR and western blot.RESULTS AND CONCLUSION:A total of 6 487 common differentially expressed genes were identified in the GSE45291 and GSE93777 datasets,and 12 co-expression modules were obtained.The magenta module had the highest correlation with rheumatoid arthritis,with module genes primarily enriched in the phosphatidylinositol 3-kinase/protein kinase B signaling pathway.Database predictions revealed that seven differentially expressed genes were targeted by allicin,with epidermal growth factor receptor having the highest AUC value in the GSE45291 dataset and kynureninase having the highest AUC value in the GSE93777 dataset.Treatment of human immortalized rheumatoid arthritis fibroblast-like synoviocytes with allicin significantly inhibited cell activity,promoted cell apoptosis,decreased the levels of reactive oxygen species,malondialdehyde,interleukin-6,interleukin-1β,increased the expression of mitochondrial membrane potential,decreased the mRNA and protein expression of epidermal growth factor receptor,kynureninase,mitochondrial dynamin-related protein 1,elevated the mRNA and protein expression of mitochondrial fusion protein 2,and increased the protein expression of p-AKT and p-PI3K.To conclude,allicin plays the potential therapeutic effects on rheumatoid arthritis through the regulation of target genes such as epidermal growth factor receptor and kynureninase.

Objective:To compare the safety and efficacy of transarterial chemoembolization (TACE) combined with donafenib and programmed death protein 1 (PD-1) inhibitors and TACE combined with donafenib in the treatment of unresectable hepatocellular carcinoma (uHCC).Methods:Clinical data of 148 patients with uHCC treated at the First Affiliated Hospital of Zhengzhou University from December 2021 to December 2022 were retrospectively analyzed, including 127 males and 21 females, aged (56.6±9.9) years. Patients were divided into two groups: the TACE combined with donafenib and PD-1 inhibitors group (TACE+ DP, n=73) and TACE combined with single donafenib (TACE+ D, n=75). The overall survival (OS), progression-free survival (PFS), objective response rate (ORR), disease control rate (DCR), and the occurrence of treatment-related adverse events (TRAEs) of the two groups of patients were observed. Kaplan-Meier analysis was used for survival assessment, and the log-rank test was used for comparison. The related factors affecting the prognosis of patients were indentified and analyzed. Results:The median PFS of patients in the TACE+ D group and the TACE+ DP group were 7.2 months (95% CI: 5.7-8.3 months) and 10.5months (95% CI: 8.9-11.3 months), respectively. The median OS was 13.2 months (95% CI: 12.3-13.7 months) and 16.9 months (95% CI: 15.1-19.8 months), respectively. All these differences were statistically significant ( χ2=17.81, 26.92, respectively, both P<0.001). The ORR and DCR of TACE+ DP group were both higher than those in TACE+ D group [53.4% (39/73) vs 36.0% (27/75), χ2=4.55, P=0.031; and 90.4% (66/73) vs 77.3% (58/75), χ2=4.66, P=0.044]. No grade 4 or above adverse events occurred in either the TACE+ DP or the TACE+ D group. The most common treatment-related adverse events in TACE+ D and TACE+ DP group were hand-foot syndrome [46.7% (35/75) vs 49.3% (36/73)], hypertension [26.7% (20/75) vs 30.1% (22/73)], fatigue [22.7% (17/75) vs 24.7% (18/73)], diarrhea [26.7% (20/75) vs 28.8% (21/73)], and thrombocytopenia [25.3% (19/75) vs 28.8% (21/73)]. There was no significant difference in the incidence and severity of TRAEs between the groups ( χ2=0.08, P=0.774). TACE+ DP treatment was a favorable prognostic factor for PFS ( HR=0.33, 95% CI: 0.22-0.49, P<0.001) and OS ( HR=0.19, 95% CI: 0.11-0.33, P<0.001) of patients. Conclusion:Compared to TACE combined with donafenib, TACE combined with donafenib and PD-1 inhibitors, with good efficacy and safety, significantly improved the treatment response and survival in patients with uHCC.

The objective of this study was to evaluate the toxicity and safety of novel Mycobacterium tuberculosis fusion strain protein derivatives,referred to as B/R strain active proteins.In cellular experiments,RAW264.7 cells were treated with each vaccine preparation,and apoptosis rates were measured.In subsequent animal experiments,C57BL/6 mice were immunized via subcutaneous injection,and their survival and body weight changes were monitored and recorded at 2,4,8,12,and 16 weeks.The lungs and spleens were harvested to calculate organ coefficients,and pathological examinations were conducted.At the eighth week of immunization,the mice were infected with high concentrations of BCG,and pathological changes in the lungs and spleens were observed 4 weeks post-infection.The apoptosis rate at 6 hours was significantly higher in the experimental group than the PBS group(P<0.05).At 12 and 24 hours,the apoptosis rate in the experimental group remained higher than that in the PBS group,although this difference was not statistically significant.After immunization,mice in all four groups exhibited normal growth patterns,as indicated by stable body weight changes.At 4 and 12 weeks post-immunization,the lung coefficients in the protein group were significantly higher than those in the PBS group at the same time points.Additionally,the lung coefficients in the BCG group were significantly elevated across all time periods(P<0.05).The spleen coefficients in the protein and BCG groups were significantly higher than those in the PBS group at 2,4,8,12,and 16 weeks,whereas the ICD B/R group showed higher spleen coefficients than the PBS group only at week 8(P<0.05).Pathological examination revealed normal lung and spleen tissues in the PBS group.However,during the 2-8 weeks immunization period,lung and spleen tissues in all experimental groups exhibited varying degrees of damage,which gradually diminished by 12-16 weeks.Notably,no tuberculosis nodules were observed in any experimental group.After infection with high concentrations of BCG,no overt pathological changes were observed on the surfaces of the lungs and spleens in any group.Microscopic examination revealed less severe pathological changes in the lungs and spleens of mice in the experimental groups than the PBS group.Furthermore,no statistically significant differences were observed between the protein group and the BCG group.Our findings suggested that the B/R strain active proteins'toxicity and safety profiles were comparable to those of BCG,and showed immunoprotective effects.This study provides an experimental foundation for the development of a novel tuberculosis vaccine.

BACKGROUND:Allicin,the main active ingredient in garlic,has anti-inflammatory and antioxidant effects,but the role and mechanism of allicin in ameliorating rheumatoid arthritis remain unclear.OBJECTIVE:To analyze differentially expressed genes in rheumatoid arthritis using gene expression data and explore the regulatory role of alliin in rheumatoid arthritis.METHODS:Gene expression data for rheumatoid arthritis were collected from the GSE45291 and GSE93777 datasets,and differential expression analysis was then performed.Co-expression network analysis was conducted on the common differentially expressed genes identified in both datasets to identify the module genes most closely related to rheumatoid arthritis,followed by functional enrichment analysis.The SwissTargetPrediction database was used to predict the target genes of allicin in the differentially expressed genes.The area under the receiver operator characteristic curve(AUC)of the differentially expressed target genes was calculated.Human immortalized rheumatoid arthritis fibroblast-like synoviocytes were seeded onto pore plates and cultured for 24 hours,and the cells were then divided into five groups.The control group was not treated;methotrexate was added to the positive drug group;and 20,80,and 160 μg/mL allicin was added to the low-,medium-,and high-dose allicin groups,respectively.After 48 hours of treatment,cell activity was assessed using cell counting kit-8 assay and cell apoptosis was detected using TUNEL assay.Levels of oxidative stress and inflammatory factors were measured by ELISA.Changes in mitochondrial membrane potential were detected by JC-1 staining.The expression levels of target genes and related signaling pathways were detected by RT-qPCR and western blot.RESULTS AND CONCLUSION:A total of 6 487 common differentially expressed genes were identified in the GSE45291 and GSE93777 datasets,and 12 co-expression modules were obtained.The magenta module had the highest correlation with rheumatoid arthritis,with module genes primarily enriched in the phosphatidylinositol 3-kinase/protein kinase B signaling pathway.Database predictions revealed that seven differentially expressed genes were targeted by allicin,with epidermal growth factor receptor having the highest AUC value in the GSE45291 dataset and kynureninase having the highest AUC value in the GSE93777 dataset.Treatment of human immortalized rheumatoid arthritis fibroblast-like synoviocytes with allicin significantly inhibited cell activity,promoted cell apoptosis,decreased the levels of reactive oxygen species,malondialdehyde,interleukin-6,interleukin-1β,increased the expression of mitochondrial membrane potential,decreased the mRNA and protein expression of epidermal growth factor receptor,kynureninase,mitochondrial dynamin-related protein 1,elevated the mRNA and protein expression of mitochondrial fusion protein 2,and increased the protein expression of p-AKT and p-PI3K.To conclude,allicin plays the potential therapeutic effects on rheumatoid arthritis through the regulation of target genes such as epidermal growth factor receptor and kynureninase.

Objective:To compare the safety and efficacy of transarterial chemoembolization (TACE) combined with donafenib and programmed death protein 1 (PD-1) inhibitors and TACE combined with donafenib in the treatment of unresectable hepatocellular carcinoma (uHCC).Methods:Clinical data of 148 patients with uHCC treated at the First Affiliated Hospital of Zhengzhou University from December 2021 to December 2022 were retrospectively analyzed, including 127 males and 21 females, aged (56.6±9.9) years. Patients were divided into two groups: the TACE combined with donafenib and PD-1 inhibitors group (TACE+ DP, n=73) and TACE combined with single donafenib (TACE+ D, n=75). The overall survival (OS), progression-free survival (PFS), objective response rate (ORR), disease control rate (DCR), and the occurrence of treatment-related adverse events (TRAEs) of the two groups of patients were observed. Kaplan-Meier analysis was used for survival assessment, and the log-rank test was used for comparison. The related factors affecting the prognosis of patients were indentified and analyzed. Results:The median PFS of patients in the TACE+ D group and the TACE+ DP group were 7.2 months (95% CI: 5.7-8.3 months) and 10.5months (95% CI: 8.9-11.3 months), respectively. The median OS was 13.2 months (95% CI: 12.3-13.7 months) and 16.9 months (95% CI: 15.1-19.8 months), respectively. All these differences were statistically significant ( χ2=17.81, 26.92, respectively, both P<0.001). The ORR and DCR of TACE+ DP group were both higher than those in TACE+ D group [53.4% (39/73) vs 36.0% (27/75), χ2=4.55, P=0.031; and 90.4% (66/73) vs 77.3% (58/75), χ2=4.66, P=0.044]. No grade 4 or above adverse events occurred in either the TACE+ DP or the TACE+ D group. The most common treatment-related adverse events in TACE+ D and TACE+ DP group were hand-foot syndrome [46.7% (35/75) vs 49.3% (36/73)], hypertension [26.7% (20/75) vs 30.1% (22/73)], fatigue [22.7% (17/75) vs 24.7% (18/73)], diarrhea [26.7% (20/75) vs 28.8% (21/73)], and thrombocytopenia [25.3% (19/75) vs 28.8% (21/73)]. There was no significant difference in the incidence and severity of TRAEs between the groups ( χ2=0.08, P=0.774). TACE+ DP treatment was a favorable prognostic factor for PFS ( HR=0.33, 95% CI: 0.22-0.49, P<0.001) and OS ( HR=0.19, 95% CI: 0.11-0.33, P<0.001) of patients. Conclusion:Compared to TACE combined with donafenib, TACE combined with donafenib and PD-1 inhibitors, with good efficacy and safety, significantly improved the treatment response and survival in patients with uHCC.

Objective:To compare and analyze the changes of aggregation rate and routine parameters of platelets stored in temperature cycle,cold storage at 4 ℃ and oscillating storage at 22 ℃,so as to provide more experimental data for platelet preservation methods.Methods:Blood samples were collected at 5 time points on the 1st,2nd,3rd,4th and 6th day after platelet cycling preservation at temperature,cold storage at 4 ℃,and oscillating storage at 22 ℃.Platelet maximum aggregation rate(MAR)and routine parameters including platelet count(PLT),mean platelet volume(MPV),platelet distribution width(PDW)and platelet-larger cell ratio(P-LCR)were detected.Results:The platelet MAR of three groups showed a significant decrease trend with the preservation time,the fastest decrease was in the 22 ℃ group,the slowest was in the 4 ℃ group,and the temperature cycle group was between the two groups.On the 3rd day of preservation,the platelet MAR in 4 ℃ group was still in the normal range(MAR＞60％),while in temperature cycle group was about 50％,and in 22 ℃ group was the lowest.On the 4th day of preservation,platelet MAR in all the three groups was lower than 50％,and that in temperature cycle group was significantly lower than in 4 ℃ group but higher than in 22 ℃ group(both P＜0.05).On the 6th day of preservation,platelet MAR in the temperature cycle group was significantly lower than that in the 4 ℃ group(P＜0.05),but there was no statistically significant difference compared to 22 ℃ group(P＞0.05).PLT values in the three groups were all significantly decreased with the preservation time extension,and were significantly lower than those in the early stage of preservation within 6 days(all P＜0.05).PDW in temperature cycle group had no significant change within 6 days of preservation,but MPV and P-LCR were significantly increased.MPV,PDW and P-LCR all decreased significantly in 4 ℃ group within 6 days of preservation but increased in 22 ℃ group.Under the same storage days,PLT value of temperature cycle group had no significant difference with that of 4 ℃ group and 22 ℃ group,while MPV,PDW and P-LCR values were significantly higher than 4 ℃ group but lower than 22 ℃ group(all P＜0.05).Conclusion:The aggregation function and routine parameters changes of temperature circulating preserved platelets are between 4 and 22 ℃.

Studies have shown that a variety of diseases such as cardiovascular disease, renal disease and cancer are closely related to trimethylamine oxide (TMAO). Clinically, abnormal elevation of TMAO has been used as an evaluation index of atherosclerosis (AS) prior to imaging. In this study, we investigated the effects of lipid metabolism disorders as well as pharmacological interventions on urinary TMAO using a hyperlipidemic golden gopher model. The study used 48 Syrian golden hamster modeled with a high-fat diet for 2 weeks, and then ezetimibe, simvastatin, ezetimibe and simvastatin groups were administered for 4 consecutive weeks, as well as the clinical trial drug, IMM-H007, for pharmacological intervention. The animal experiment was conducted in accordance with the regulations of the Ethics Committee for Experimental Animal Management and Animal Welfare of Institute of Materia Medica, Chinese Academy of Medical Sciences (approval number: SCXK (Beijing) 2021-0011). Urine from rats was analyzed for 2D band selective heteronuclear single quantum coherence (2D bs-HSQC) at week 2 and 4 after drug administration. The results indicated that, in comparison to the control group, the high-fat diet significantly elevated urinary TMAO levels in the model group of hamsters after both 2 and 4 weeks of treatment (P < 0.05). Urinary TMAO levels were significantly reduced (P < 0.05) in the model group after 2 or 4 weeks of intervention with ezetimibe, simvastatin, combination therapy, and IMM-H007, showing a marked decrease even after 2 weeks of treatment. The detection of TMAO could precede the measurement of serum biochemical indicators, facilitating earlier efficacy assessment. This study evaluated the modulatory effects of clinical drugs and clinical trial drugs on TMAO, which provides useful information for clinical drug use and drug research. It also provides a means of TMAO detection based on 2D NMR technology, which is helpful for the clinical application of TMAO detection index.

Objective Little is known about the association between whole-blood nicotinamide adenine dinucleotide(NAD+)levels and nabothian cysts.This study aimed to assess the association between NAD+levels and nabothian cysts in healthy Chinese women. Methods Multivariate logistic regression analysis was performed to analyze the association between NAD+levels and nabothian cysts. Results The mean age was 43.0±11.5 years,and the mean level of NAD+was 31.3±5.3 μmol/L.Nabothian cysts occurred in 184(27.7%)participants,with single and multiple cysts in 100(15.0%)and 84(12.6%)participants,respectively.The total nabothian cyst prevalence gradually decreased from 37.4%to 21.6%from Q1 to Q4 of NAD+and the prevalence of single and multiple nabothian cysts also decreased across the NAD+quartiles.As compared with the highest NAD+quartile(≥34.4 μmol/L),the adjusted odds ratios with 95%confidence interval of the NAD+Q1 was 1.89(1.14-3.14)for total nabothian cysts.The risk of total and single nabothian cysts linearly decreased with increasing NAD+levels,while the risk of multiple nabothian cysts decreased more rapidly at NAD+levels of 28.0 to 35.0 μmol/L. Conclusion:Low NAD+levels were associated with an increased risk of total and multiple nabothian cysts.

This study aims to explore the anti-depression mechanism of Zuojin Pills based on the plasma constituents, network pharmacology, and experimental verification. UHPLC-TOF-MS was used for qualitative analysis of Zuojin Pills-containing serum. Targets of the plasma constituents and the disease were retrieved from PharmMapper and GeneCards. Then the protein-protein interaction(PPI) network was constructed and core targets were screened for GO term enrichment and KEGG pathway enrichment. Cytoscape 3.7.2 was employed construct the "compound-target-pathway" network and the targets and signaling pathways of Zuojin Pills against depression were predicted. CUMS-induced depression mouse model was established to verify the key targets. The results showed that a total of 21 constituents migrating to blood of Zuojin Pills were identified, which were mainly alkaloids. A total of 155 common targets of the constituents and the disease and 67 core targets were screened out. KEGG enrichment and PPI network analysis showed that Zuojin Pills may play a role in the treatment of depression through AMPK/SIRT1, NLRP3, insulin and other targets and pathways. Furthermore, the results of animal experiments showed that Zuojin Pills could significantly improve the depression behaviors of depression, reduce the levels of IL-1β, IL-6 and TNF-α in hippocampus and serum, activate AMPK/SIRT1 signaling, and reduce the protein expression of NLRP3. In conclusion, Zuojin Pills may play a role in the treatment of depression by activating AMPK/SIRT1 signaling pathway, and inhibiting NLRP3 activation and neuroinflammation in the hippocampus of mice.
Animals ; Mice ; Network Pharmacology ; AMP-Activated Protein Kinases ; Chromatography, High Pressure Liquid ; NLR Family, Pyrin Domain-Containing 3 Protein ; Sirtuin 1 ; Drugs, Chinese Herbal/pharmacology* ; Molecular Docking Simulation