Objective: Scutellarin in Shuganning injection has potential allergenicity and is prone to causing anaphylactoid reactions. There is an urgent need to establish a simple and rapid chromatographic quantitative method to achieve the limit control of scutellarin in Shuganning injection.
Methods: For scutellarin, HPLC method was used in the quantitative analysis using a Shim-pack GIS C₁₈ (2.1 mm×150 mm, 5 μm) column. The mobile phase was a blend of 0.1% trifluoroacetic acid in water and acetonitrile, and gradient elution progress was applied for the whole process. The flow rate was 0.3 mL·min^-1 and the column temperature was maintained at 37 ℃. The detecting wavelength was 320 nm.
Results: The linear regression equation was y=51 456x-27 352, the correlation coefficient r=0.999 8, namely the calibration curve was linear in the range of 1.02-102.00 μg·mL^-1; the average recovery was 103.76%; the detection and quantification limit for determining scutellarin was 0.16 μg·mL^-1 and 0.57 μg·mL^-1, respectively; the injection sample was stable at room temperature for 48 h.
Conclusion: This method is easy to operate, reproducible and has high accuracy, thus can be used for quality control of Shuganning injection.

Objective To investigate the effect of ergothioneine(EGT)on tissue metabolism in middle-aged and aged mice.Methods Nine-month-old middle-aged and aged C57BL/6J mice were selected to be gavaged with EGT aqueous solution at the dose and frequency of 35 mg EGT per kg body weight every two days.The control group was gavaged with the same amount of water.Both groups were fed with EGT-free diet.After 7 weeks,the liver,kidney,and small intestine of mice were collected,and untargeted metabolomics analysis was performed using ultra-per-formance liquid chromatography-mass spectrometry(UHPLC-MS).The differential metabolites were selected for KEGG metabolic pathway enrichment analysis.Results Compared with the control group,the abundances of me-tabolites related to redox balance in liver,kidney and small intestine of middle-aged and aged mice treated with EGT did not change significantly.However,taurine and hypotaurine metabolism in liver(P＜0.01),purine metabo-lism in kidney(P＜0.01),and cysteine and methionine metabolism in small intestine(P＜0.001)were affected.Conclusions Non-redox-related metabolic changes in the liver,kidney and small intestine of middle-aged and aged mice by ergothioheine.

OBJECTIVE: To explore the genotype-phenotype relationship in a child with Opitz G/BBB syndrome (OS) with mild clinical phenotype. METHODS: A child with motor developmental delay as the initial symptom admitted to Xi'an Children's Hospital on June 10, 2021 was selected for this study. Clinical data were collected, and peripheral blood samples were obtained from the child and his mother. Whole exome sequencing (WES) was performed to identify genetic variant in the child. Candidate variant were verified by Sanger sequencing to assess inheritance patterns and pathogenicity. Real-time fluorescence quantitative PCR (RT-qPCR) and Western blot (WB) analyses were conducted to evaluate the effects of the variant on mRNA and protein expression, respectively, using recombinant expression plasmids generated in vitro. This study was approved by the Medical Ethics Committee of Xi'an Children's Hospital (Ethics No. 20240045). RESULTS: The child, a 9-month-and-7-day-old boy, presented with a low nasal bridge, hypertelorism, and difficulty sitting independently. Echocardiography revealed an atrial septal defect. WES identified a homozygous variant in the MIDI gene, c.1483C>T (p.R495X), which was confirmed by Sanger sequencing and found to be inherited from the mother.Recombinant expression plasmids were successfully constructed. RT-qPCR analysis showed that the variant significantly reduced MIDI gene mRNA expression, while WB results indicated that the variant led to the production of a truncated protein. CONCLUSION The mild clinical phenotype of OS in this child may be attributed to the mRNA degradation escape mechanism induced by the nonsense variant c.1483C>T (p.R495X) in the MIDI gene. These findings provide valuable diagnostic insights for this pedigree and contribute to the understanding of the genotype-phenotype correlation in OS.
Humans ; Male ; Infant ; Transcription Factors/metabolism* ; Microtubule Proteins/genetics* ; Craniosynostoses/genetics* ; Hypospadias/genetics* ; Codon, Nonsense/genetics* ; RNA, Messenger/metabolism* ; Female ; RNA Stability/genetics* ; Phenotype ; Nuclear Proteins/genetics* ; Ubiquitin-Protein Ligases ; Esophagus/abnormalities* ; Hypertelorism

Objective:To investigate the relationship between thyroid hormone sensitivity indicators and the risk of mild cognitive impairment (MCI) in patients with type 2 diabetes mellitus (T2DM) who have normal thyroid function.Methods:A retrospective study design was used to gather data from 100 patients with T2DM who had normal thyroid function and were treated at Shangluo Central Hospital between June 2022 and January 2024. The Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment (MoCA) were used for evaluation. Patients with an MMSE score of ≤ 26 or a MoCA score of ≤ 26 were included in the MCI group ( n = 22), whereas those with an MMSE score of > 26 and a MoCA score of > 26 were included in the non-MCI group ( n = 78). Multivariate logistic regression analysis was performed to identify factors influencing MCI in patients with T2DM. Additionally, receiver operating characteristic curve analysis was conducted to assess the predictive value of thyroid hormone sensitivity indicators for MCI in this patient population. Results:In the MCI group, 68.18% of patients had an education level of high school or below. The level of glycosylated hemoglobin in this group was (8.45 ± 1.95)%, insulin resistance index was (3.34 ± 0.25), the level of triglycerides was (2.59 ± 0.19) mmol/L, the level of uric acid was (354.76 ± 46.16) μmol/L, thyroid-stimulating hormone index was (3.09 ± 0.26), and thyroid feedback quantile-based index was [0.2 (0.0, 0.5)]. All of these values were significantly higher than those in the non-MCI group: [43.59%, (7.63 ± 1.61)%, (3.05 ± 0.17), (2.09 ± 0.15) mmol/L, (311.74 ± 67.15) μmol/L, (2.87 ± 0.13), 0.2 (-0.1, 0.4), χ2 = 4.15, t = 2.01, 6.32, 12.99, 3.47, 5.47, Z = -2.54, all P < 0.05]. In the MCI group, the level of free triiodothyronine was (4.32 ± 0.21) pmol/L, and thyrotroph T4 resistance index (TT4RI) was (34.54 ± 4.30), both of which were significantly lower than those in the non-MCI group [(4.61 ± 0.36) pmol/L, (37.15 ± 5.55), t = 2.58, 2.04, both P < 0.05]. There were no significant differences in gender, age, body mass index, alcohol consumption, smoking, history of cerebral infarction, hypertension, duration of diabetes, use of only metformin, fasting blood glucose, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, total cholesterol, alanine aminotransferase, aspartate aminotransferase, free thyroxine, and thyroid-stimulating hormone between the two groups (all P > 0.05). Logistic regression analysis indicated that education level, triglyceride, uric acid, free triiodothyronine, thyroid-stimulating hormone index, and TT4RI were risk factors for MCI (all P < 0.05). The predictive value of the combined thyroid hormone sensitivity indicators for MCI in patients with T2DM was high, with an area under the receiver operating characteristic curve of 0.778 [95% CI(0.676, 0.880), P < 0.001], a sensitivity of 0.538, a specificity of 0.909, and a maximum Youden index of 0.447. Conclusions:In patients with T2DM who have normal thyroid function, the thyroid hormone sensitivity indicators (TT4RI and thyroid feedback quantile-based index) exhibited a negative correlation with MCI, whereas TSHI demonstrated a positive correlation with MCI. The combined use of these thyroid hormone sensitivity indicators is valuable for predicting MCI and offers significant guidance for the early intervention of cognitive impairment in patients with T2DM.

Objective To investigate the relationship between the expression levels of mir-155/Th17 and mir-23b-3p/Tfh in peripheral blood and the prognostic value of secondary atopic dermatitis(AD)in AIDS patients.Methods In this study,80 AIDS patients were treated in our hospital from February 2021 to December 2023.The subjects were divided into AD group and non-AD group according to whether they had secondary AD.The levels of mir-155,Th17,mir-23b-3p and Tfh in peripheral blood of AD group and non-AD group were compared.Spearman method was used to analyze the relationship between levels of mir-155,Th17,mir-23b-3p and Tfh in peripheral blood and secondary AD in AIDS patients.At the same time,according to the prognosis of patients,the subjects were divided into good prognosis group and poor prognosis group,and the risk factors affecting the prognosis of patients were screened by Logistic regression analysis.Receiver operating characteristic(ROC)curve was drawn to analyze the predictive value of peripheral blood mir-155,Th17,mir-23b-3p and Tfh levels on the prognosis of AIDS patients.Results Compared with non-AD group,the levels of miR-155,Th17 and Tfh in peripheral blood of AD group were higher(P＜0.05),and the levels of miR-23b-3p were lower(P＜0.05).The level of miR-155,Th17 and Tfh in peripheral blood were positively correlated with secondary AD of AIDS(r=0.487,0.492,0.495,P＜0.05),and the level of miR-23b-3p in peripheral blood was negatively correlated with secondary AD of AIDS(r=-0.503,P＜0.05).The levels of HIV load(high dose),lymphoma,PLT,NLR,CRP,miR-155,Th17 and Tfh in the poor prognosis group were higher than those in the good prognosis group(P＜0.05),while the levels of albumin and miR-23b-3p in the poor prognosis group were lower than those in the good prognosis group(P＜0.05).Logistic regression analysis showed that lymphoma,PLT,NLR,low albumin,CRP,miR-155,Th17,Tfh and low miR-23b-3p were all risk factors for poor prognosis of AIDS patients(P＜0.05).ROC curve results showed that the AUC(95％CI)and sensitivity of combined detection of peripheral blood miR-155,Th17,Tfh and miR-23b-3p in predicting poor prognosis of AIDS patients were 0.883(0.805～0.992)and 96.70％,respectively.All of them were higher than the above peripheral blood indexes alone(P＜0.05).Conclusion The abnormal expression of miR-155,Th17,Tfh,and miR-23b-3p in peripheral blood is closely related to secondary AD in AIDS patients,and is a risk factor affecting the poor prognosis of AIDS patients.The combined detection of these indicators has a high predictive value for the poor prognosis of AIDS patients.

Objective To investigate the relationship between the expression levels of mir-155/Th17 and mir-23b-3p/Tfh in peripheral blood and the prognostic value of secondary atopic dermatitis(AD)in AIDS patients.Methods In this study,80 AIDS patients were treated in our hospital from February 2021 to December 2023.The subjects were divided into AD group and non-AD group according to whether they had secondary AD.The levels of mir-155,Th17,mir-23b-3p and Tfh in peripheral blood of AD group and non-AD group were compared.Spearman method was used to analyze the relationship between levels of mir-155,Th17,mir-23b-3p and Tfh in peripheral blood and secondary AD in AIDS patients.At the same time,according to the prognosis of patients,the subjects were divided into good prognosis group and poor prognosis group,and the risk factors affecting the prognosis of patients were screened by Logistic regression analysis.Receiver operating characteristic(ROC)curve was drawn to analyze the predictive value of peripheral blood mir-155,Th17,mir-23b-3p and Tfh levels on the prognosis of AIDS patients.Results Compared with non-AD group,the levels of miR-155,Th17 and Tfh in peripheral blood of AD group were higher(P＜0.05),and the levels of miR-23b-3p were lower(P＜0.05).The level of miR-155,Th17 and Tfh in peripheral blood were positively correlated with secondary AD of AIDS(r=0.487,0.492,0.495,P＜0.05),and the level of miR-23b-3p in peripheral blood was negatively correlated with secondary AD of AIDS(r=-0.503,P＜0.05).The levels of HIV load(high dose),lymphoma,PLT,NLR,CRP,miR-155,Th17 and Tfh in the poor prognosis group were higher than those in the good prognosis group(P＜0.05),while the levels of albumin and miR-23b-3p in the poor prognosis group were lower than those in the good prognosis group(P＜0.05).Logistic regression analysis showed that lymphoma,PLT,NLR,low albumin,CRP,miR-155,Th17,Tfh and low miR-23b-3p were all risk factors for poor prognosis of AIDS patients(P＜0.05).ROC curve results showed that the AUC(95％CI)and sensitivity of combined detection of peripheral blood miR-155,Th17,Tfh and miR-23b-3p in predicting poor prognosis of AIDS patients were 0.883(0.805～0.992)and 96.70％,respectively.All of them were higher than the above peripheral blood indexes alone(P＜0.05).Conclusion The abnormal expression of miR-155,Th17,Tfh,and miR-23b-3p in peripheral blood is closely related to secondary AD in AIDS patients,and is a risk factor affecting the poor prognosis of AIDS patients.The combined detection of these indicators has a high predictive value for the poor prognosis of AIDS patients.

Objective:To explore the genotype-phenotype relationship in a child with Opitz G/BBB syndrome (OS) with mild clinical phenotype.Methods:A child with motor developmental delay as the initial symptom admitted to Xi ′an Children′s Hospital on June 10, 2021 was selected for this study. Clinical data were collected, and peripheral blood samples were obtained from the child and his mother. Whole exome sequencing (WES) was performed to identify genetic variant in the child. Candidate variant were verified by Sanger sequencing to assess inheritance patterns and pathogenicity. Real-time fluorescence quantitative PCR (RT-qPCR) and Western blot (WB) analyses were conducted to evaluate the effects of the variant on mRNA and protein expression, respectively, using recombinant expression plasmids generated in vitro. This study was approved by the Medical Ethics Committee of Xi′an Children′s Hospital (Ethics No. 20240045).Results:① The child, a 9-month-and-7-day-old boy, presented with a low nasal bridge, hypertelorism, and difficulty sitting independently. Echocardiography revealed an atrial septal defect. ② WES identified a homozygous variant in the MIDI gene, c. 1483C>T (p.R495X), which was confirmed by Sanger sequencing and found to be inherited from the mother.③ Recombinant expression plasmids were successfully constructed. RT-qPCR analysis showed that the variant significantly reduced MIDI gene mRNA expression, while WB results indicated that the variant led to the production of a truncated protein. Conclusion:The mild clinical phenotype of OS in this child may be attributed to the mRNA degradation escape mechanism induced by the nonsense variant c. 1483C>T(p.R495X) in the MIDI gene. These findings provide valuable diagnostic insights for this pedigree and contribute to the understanding of the genotype-phenotype correlation in OS.

Objective:To explore the impact of the stimulator of interferon genes(STING)-interferon regulatory factor 3(IRF3)pathway on the senescence of rapid pacing HL-1 myocytes.Methods:HL-1 cells were divided into five groups: the control group(HL-1 cells without any treatment), pacing group(HL-1 cells paced for 48 hours), STING siRNA group(HL-1 cells paced for 48 hours and transfected with STING siRNA), NC siRNA group(HL-1 cells paced for 48 hours and transfected with NC siRNA), and H151 inhibitor group(HL-1 cells paced for 48 hours with the addition of 1 μmol/L STING inhibitor H151). Mitochondrial membrane potential was assessed in control and pacing group cells, and mitochondrial MitoTracker and TFAM co-localization staining was performed on these cells.Cellular senescence was evaluated using β-galactosidase staining in each group, and the positive rate of cellular senescence was observed and calculated.Western blotting was employed to detect the expression levels of STING, IRF3, P-IRF3, P16, P21, and P53 proteins in all groups.Immunofluorescence was utilized to examine the expression distribution of STING and P21 across the various groups.ELISA was performed to measure the concentrations of interleukin(IL)-1β, IL-6, and IL-8 in the cell supernatants from each group as part of the senescence-associated secretory phenotype(SASP).Results:Compared with the control group, the ratio of mitochondrial JC-1 multimer to monomer was significantly decreased in the pacing group( t=16.42, P<0.05), the co-localization of mitochondrial MitoTracker and TFAM in the cells was significantly weakened, the proportion of cells with positive cellular senescence-associated β-galactosidase staining significantly increased in the pacing group, the expression levels of STING, P-IRF3/IRF3, P16, P21, and P53 proteins were significantly elevated in the pacing group, and the concentrations of IL-1β, IL-6, and IL-8 in the cell supernatants were markedly increased.Compared with the pacing group, the proportion of cells with positive cellular senescence-associated β-galactosidase staining decreased in the STING siRNA group and H151 inhibitor group ( F= 18.13, P<0.05), the expression levels of STING, P-IRF3/IRF3, P16, P21, and P53 were reduced in the STING siRNA group and H151 inhibitor group ( F=16.84, 26.56, 74.70, 31.80, 31.23, all P<0.05), and the concentrations of IL-1β, IL-6, and IL-8 in the cell supernatants decreased( F=197.80、1 339.00、1 308.00, all P<0.001). Conclusions:Rapid pacing of HL-1 cells can promote mtDNA release into the cytoplasm, activate the STING-IRF3 pathway, accelerate cellular senescence, and enhance the secretion of SASP.Inhibiting the expression of STING can delay the senescence induced by the rapid pacing of HL-1 cells and reduce SASP secretion.

Objective:To analyze the conventional ultrasound（CUS）and contrast-enhanced ultrasound（CEUS）features of hepatic lymphoma，and to investigate the value of CEUS in the diagnosis of hepatic lymphoma.Methods:The images of 39 patients（39 lesions）with hepatic lymphoma pathologically confirmed by surgery and puncture from March 2012 to July 2024 at Zhongshan Hospital，Fudan University were retrospectively analyzed. Evaluations of CUS included the echogenicity，morphology，color Doppler flow imaging（CDFI）situation，evaluations of CEUS included enhancement type，enhancement degree compared to the peripheral normal liver parenchyma and time to enhancement.Results:In the 39 lesions，hypoechoic lesions were detected in 31（79.49%，31/39）patients on CUS. CDFI detected linear or branched color flow signals inside the lesions in 21（53.85%，21/39）lesions，and peripheral color flow signals around the lesions in 3 lesions，while arterial flow signals were detected in 16 of them，with a resistance index of 0.50～0.77（0.67 ± 0.02）. In addition，signs of non-compacted normal blood vessels passing through the lesions were detected in 5 lesions. After injection of contrast medium，39 lesions showed different degrees of enhancement，mainly showed entirety homogeneous enhancement，during the arterial phase of CEUS，34（87.18%，34/39）lesions showed fast enhancement，and when the enhancement reached the peak，26（66.67%，26/39）lesions revealed hyper-enhancement，showing “fast progression”. There were 38（97.44%，38/39）lesions in the portal and delayed phases showed “fast forward”.Conclusions:CUS and CEUS can provide some value in the diagnosis and differential diagnosis of hepatic lymphoma.

Objective:To explore the differences between clinical-pathological-ultrasound features in hepatocellular carcinoma（HCC）with negative and positive des-gamma-carboxy prothrombin（DCP）.Methods:A retrospective analysis was conducted on 649 patients with pathologically confirmed HCC at Zhongshan Hospital，Fudan University from April 2020 to May 2024. Patients were stratified into DCP-negative（177 cases，<40 mAU/ml）and DCP-positive（472 cases，≥40 mAU/ml）groups. Clinical data，pathological features，and ultrasound findings were collected. Conventional ultrasound and contrast-enhanced ultrasound（CEUS）imaging characteristics were analyzed and compared between the two groups，and the correlation between ultrasound features and pathological characteristics were analyzed.Results:The DCP-negative group exhibited a lower incidence of microvascular invasion（10.17% vs. 34.75%， P<0.001）and smaller median tumor diameter（23 mm vs. 40 mm， P<0.001）. Heterogeneous internal echogenicity was less frequent in DCP-negative tumors［48.59%（86/177） vs. 74.58%（352/472）， P<0.001］. CEUS revealed higher rates of arterial-phase iso-enhancement（6.78% vs. 1.69%）and absence of washout（13.56% vs. 4.45%）in DCP-negative HCC（both P<0.001）. CEUS LI-RADS classification showed fewer LR-5 lesions［50.85%（90/177） vs. 59.53%（281/472）］in DCP-negative group（ P<0.001）. Conclusions:HCC with different DCP states has different clinical-pathological-ultrasound features. DCP-negative HCCs are more likely to show atypical enhancement patterns characteristic of HCC.