Purpose: This study assessed the performance of the ultrasound-guided attenuation parameter (UGAP) in diagnosing and grading hepatic steatosis in patients with metabolic dysfunctionassociated steatotic liver disease (MASLD). Magnetic resonance imaging proton density fat fraction (MRI-PDFF) served as the reference standard. Methods: Patients with hepatic steatosis were enrolled in this prospective study and underwent UGAP measurements. MRI-PDFF values of ≥5%, ≥15%, and ≥25% were used as references for the diagnosis of steatosis grades ≥S1, ≥S2, and S3, respectively. Spearman correlation coefficients and area under the receiver operating characteristic curves (AUCs) were calculated. Results: Between July 2023 and June 2024, the study included 88 patients (median age, 40 years; interquartile range [IQR], 36 to 46 years), of whom 54.5% (48/88) were men and 45.5% (40/88) were women. Steatosis grades exhibited the following distribution: 22.7% (20/88) had S0, 50.0% (44/88) had S1, 21.6% (19/88) had S2, and 5.7% (5/88) had S3. The success rate for UGAP measurements was 100%. The median UGAP value was 0.74 dB/cm/MHz (IQR, 0.65 to 0.82 dB/ cm/MHz), and UGAP values were positively correlated with MRI-PDFF (r=0.77, P<0.001). The AUCs of UGAP for the diagnoses of ≥S1, ≥S2, and S3 steatosis were 0.91, 0.90, and 0.88, respectively. In the subgroup analysis, 98.4% (60/61) of patients had valid controlled attenuation parameter (CAP) values. UGAP measurements were positively correlated with CAP values (r=0.65, P<0.001). Conclusion Using MRI-PDFF as the reference standard, UGAP demonstrates good diagnostic performance in the detection and grading of hepatic steatosis in patients with MASLD.

Purpose: This study assessed the performance of the ultrasound-guided attenuation parameter (UGAP) in diagnosing and grading hepatic steatosis in patients with metabolic dysfunctionassociated steatotic liver disease (MASLD). Magnetic resonance imaging proton density fat fraction (MRI-PDFF) served as the reference standard. Methods: Patients with hepatic steatosis were enrolled in this prospective study and underwent UGAP measurements. MRI-PDFF values of ≥5%, ≥15%, and ≥25% were used as references for the diagnosis of steatosis grades ≥S1, ≥S2, and S3, respectively. Spearman correlation coefficients and area under the receiver operating characteristic curves (AUCs) were calculated. Results: Between July 2023 and June 2024, the study included 88 patients (median age, 40 years; interquartile range [IQR], 36 to 46 years), of whom 54.5% (48/88) were men and 45.5% (40/88) were women. Steatosis grades exhibited the following distribution: 22.7% (20/88) had S0, 50.0% (44/88) had S1, 21.6% (19/88) had S2, and 5.7% (5/88) had S3. The success rate for UGAP measurements was 100%. The median UGAP value was 0.74 dB/cm/MHz (IQR, 0.65 to 0.82 dB/ cm/MHz), and UGAP values were positively correlated with MRI-PDFF (r=0.77, P<0.001). The AUCs of UGAP for the diagnoses of ≥S1, ≥S2, and S3 steatosis were 0.91, 0.90, and 0.88, respectively. In the subgroup analysis, 98.4% (60/61) of patients had valid controlled attenuation parameter (CAP) values. UGAP measurements were positively correlated with CAP values (r=0.65, P<0.001). Conclusion Using MRI-PDFF as the reference standard, UGAP demonstrates good diagnostic performance in the detection and grading of hepatic steatosis in patients with MASLD.

Objective To investigate the effects of ischemia time and storage periods on RNA quality in fresh-frozen breast cancer(BC)and esophageal cancer(EC)tissue samples in order to establish evidence-based protocols for biobank sample management.Methods The tumor(T)and paired normal(N)tissue samples from 6 cases of BC and 6 cases of EC were collected and cryopreserved in Biobank,Shantou Central Hospital.Mirror paraffin-embedded tissues were simultaneously prepared into sections for morphological analysis.The samples were divided into two groups of<15 min and 15-30 min according to ischemia time,and RNA quality was analyzed at 4 storage periods of 8-10 months(T1),14-16 months(T2),26-28 months(T3)and 38-40 months(T4).Results In 96 analyzed samples,93.8%(90/96)exhibited high quality(RIN≥6),with 89.6%(43/48)in BC and 97.9%(47/48)in EC.Significant differences in RIN were observed between BC group and EC group(8.050 vs.8.600,P=0.009).In EC group,RIN value was significantly negatively correlated with RNA yield(P<0.001).Moreover,RIN values of tumor-normal pairs exhibited markedly significant differences(7.550 vs.9.000,P<0.001).In contrast,no significant difference was detected in BC group(8.200 vs.7.700,P=0.348).Statistical analysis showed that RIN value was positively correlated with 28S/18S(P<0.001),but had no correlation with tumor content(P=0.676)and necrotic content(P=0.055).Neither ischemia time(<15 min vs.15-30 min:8.200 vs.8.300,P=0.932)nor storage periods(T1-T4:8.400,7.700,8.450,8.600,P=0.163)compromised RNA quality.Conclusion Organ origin and tissue type could influence RNA quality of fresh-frozen tissue samples.However,limited ischemia time(≤30 min)and long-term storage period(38-40 months)do not adversely affect RNA quality in fresh-frozen breast cancer and esophageal cancer tissue samples.

Aim To detect the non-coding RNA(ncRNA)expression profile of neuroglioma cells via whole transcriptome sequencing,establish the ceRNA network and reveal the molecular mechanism of ncRNA participating in the inhibition of neuroglioma cell prolif-eration by melatonin.Methods Neuroglioma cells were intervened with by 0,2,4,6 and 8 mmol·L-1 melatonin for 24,48 and 72 h,and the inhibitory effect of melatonin on cell proliferation was detected via CCK-8;after the intervention of 0 and 4 mmol·L-1 melatonin to U251 cells for 24 h,differentially ex-pressed miRNA(DEmiRNA),lncRNA(DElncRNA)and mRNA(DEmRNA)were detected through whole transcriptome sequencing,along with GO and KEGG enrichment analysis of DEmRNA;the ceRNA network was constructed,and the key gene expression of ceR-NA was verified through qRT-PCR.Results Melato-nin exerts a time-dose-dependent inhibitory effect on the proliferation of neuroglioma cells;a total of 5049 DEmRNA,635 DElncRNA and 146 DEmiRNA in 0 and 4 mmol·L-1 melatonin groups were screened out via whole transcriptome sequencing;DEmRNAs were mainly enriched in cancer-related signaling pathways,such as ferroptosis,mTOR signaling pathway,FoxO signaling pathway and cell cycle;the ceRNA network included 4 lncRNAs,3 miRNAs and 48 mRNAs.As verified through real-time PCR,the expressions of hsa-miR-129-5p,hsa-miR-362-5p,LINC00707 and SLC16A1-AS1 of U251 cells were consistent with the sequencing results,and the gene expression of U87 cells was basically consistent with the sequencing re-sults.Conclusions Melatonin affects cancer-related signaling pathways through the differential expression of ncRNA so as to inhibit the proliferation of U251 cells;the ceRNA network composed of LINC00707,SLC16A1-AS1,hsa-miR-129-5p and hsa-miR-362-5p may take a part in the molecular mechanism of melato-nin in inhibiting neuroglioma cell proliferation.

Objective To investigate the protective effect of astaxanthin(ASTA)on gouty chondrocyte injury induced by monosodium urate crystal(MSU)and its mechanism.Methods The gout cell model by sodium urate crystals was established.C-28I2 cells were randomly divided into blank group(conventional culture),model group(200 μg·mL-1MSU),experimental-L group(200 μg·mL-1 MSU+20 μmol·mL-1 ASTA),experimental-H group(200 μg·mL-1 MSU+40 μmol·L-1 ASTA),experimental-H+Vector group(transfected with Vector+200 μg·mL-1 MSU+40 μmol·L-1 ASTA),experimental-H+NLRP3 group(transfected with NLRP3 plasmid+200 ig·mL-1 MSU+40 μmol·L-1 ASTA).Cell counting kit-8(CCK-8)assay was used to detect the cell proliferation rate;Western blot assay was used to detect the expression of related proteins;the levels of Hyp and GAG were detected by enzyme-linked immunosorbent assay(ELISA).Results The cell proliferation rate of blank group,model group,experimental-L group and experimental-H group were(100.00±5.40)％,(67.41±4.52)％,(72.69±5.05)％and(81.47±7.73)％,respectively.The above indicators showed statistically significant differences between the model group and the blank group,between the experimental-L,experimental-H groups and the model group(all P＜0.05).Nucleotide binding oligomeric domain-like receptor protein 3(NLRP3)protein expression levels in blank group,model group,experimental-L group,experimental-H group,experimental-H+Vector group and experimental-H+NLRP3 group were 0.44±0.04,0.86±0.06,0.72±0.10,0.52±0.03,0.51±0.03 and 1.13±0.10,respectively;the expression levels of cleaved Caspase-1(Cl-caspase-1)protein were 0.33±0.05,0.73±0.08,0.61±0.07,0.42±0.04,0.39±0.04 and 0.70±0.06,respectively;the mature interleukin(m-IL)-1 β protein expression levels were 0.26±0.03,0.91±0.05,0.70±0.11,0.57±0.08,0.58±0.06 and 0.79±0.08,respectively;the Hyp levels were(1.95±0.22),(3.33±0.25),(2.53±0.18),(2.22±0.21),(2.24±0.20)and(3.25±0.38)μg·mL-1,respectively;the GAG levels were(2.30±0.20),(3.71±0.26),(3.29±0.33),(2.90±0.16),(2.97±0.24)and(3.50±0.34)ng·mL-1,respectively.The above indicators showed statistically significant differences between the model group and the blank group,between the experimental-L,experimental-H groups and the model group,between the experimental-H+NLRP3 group and the experimental-H+Vector group(all P＜0.05).Conclusion Astaxanthin can regulate NLRP3 to play an anti-inflammatory role and has a protective effect on MSU-induced gouty cartilage injury.

Aim To investigate the effect of tetrameth-ylpyrazine(TMP)on neuroinflammation after cerebral ischemia and hypoxia via mannose-binding lectin(MBL).Methods Patients diagnosed with ischaemic stroke at Shanxi Provincial People's Hospital were in-cluded in the study,and their clinicopathological data,as well as blood and urine samples,were collected with the consent of the patients and their families.Using these biological samples,differential proteins and tar-gets were identified by proteomic analysis and subse-quently verified with animal experiments.The mice were divided into the sham,dMCAO,and TMP(10,20,40 mg·kg-1)treatment groups.After seven days of drug administration,the modified neurological sever-ity score(mNSS)was used to assess the neurological function.TTC staining was used to detect the volume of cerebral infarction.Motor function was evaluated be-haviourally,and ELISA was used to detect MASP1,sC5b-9,TNF-α,IL-6,and IL-1β.Western blot was used to determine the expression of relevant proteins,such as MBL2,MASP2,and C3.Results Compared with the sham group,the dMCAO group exhibited in-creased neurological impairment,which was signifi-cantly ameliorated by TMP treatment.The expression levels of MBL2,C3 and MASP2 were elevated in the dMCAO group and were reduced following TMP treat-ment.Additionally,the dMCAO group showed elevat-ed expression of inflammatory factors IL-1 β,IL-6 and TNF-α,which were then suppressed by TMP treat-ment.Conclusion TMP inhibits the inflammatory re-sponse after ischemia and hypoxia by regulating MBL,thus attenuating brain injury.

Streptococcus equinus is a zoonotic disease that can cause illness in various animals under specific environmental condi-tions.No reports have described isolation of this bacterium from the liver in affected sheep.This study successfully isolated and identi-fied a strain of Streptococcus equinus through bacterial isolation and culture,Gram staining,drug sensitivity testing,mouse sensitivity testing,bacterial biochemical testing,and whole genome sequencing.The strain was found to have pathogenicity toward Kunming white mice,and to be sensitive to four antibiotics(penicillin,ampicillin,ceftiofur sodium,and ceftriaxone sodium)but resistant to four antibiotics(streptomycin,amoxicillin,tetracycline,and gentamicin).On the basis of drug sensitivity testing,targeted treatment of the affected sheep flock with ceftiofur sodium effectively controlled the disease within 2 days,and no new cases occurred.This study provides a reference for biological characterization of ovine Streptococcus equinus;public health;and the investigation of disease pre-vention,control,and epidemiology.

To analyze the ethnic differences in the genotype distribution of thalassemia between the Han and Li ethnic groups in the Qiongnan region (southern Hainan). A cross-sectional study employing a stratified multistage sampling method was conducted from January 2019 to December 2023. A total of 4 493 high-risk individuals (2 734 Han and 1 759 Li) from southern Hainan (including Sanya, Ledong, Baoting, Lingshui, and other counties) underwent thalassemia genetic testing. The genotype distribution was statistically analyzed. Inter-group comparisons were performed using χ2 test or Fisher′s exact test. The results showed an overall thalassemia positivity rate of 66.70% (2 997/4 493), with carrier, intermediate and major thalassemia rates of 62.01% (2 786/4 493), 3.98% (179/4 493) and 0.71% (32/4 493), respectively. The positivity rates for thalassemia were 87.83% (1 545/1 759) in the Li ethnic group and 53.11% (1 452/2 734) in the Han ethnic group. Among them, the Li ethnic group exhibited significantly higher positivity rates for α-thalassemia (71.12% vs. 40.64%, χ2=398.90, P<0.001) and α/β-compound thalassemia (13.36% vs. 3.33%, χ2=160.06, P<0.001) compared to the Han ethnic group, whereas the Han ethnic group had a higher β-thalassemia rate (9.14% vs. 3.35%, χ2=56.03, P<0.001). Both ethnic groups shared common α-thalassemia alleles (-α 3.7 and -α 4.2), but the -- SEA allele proportion was significantly higher in Han (21.33% vs. 4.34%, χ2=231.45, P<0.001). Six rare -α 21.9 mutations (0.26%) were exclusively identified in the Li ethnic group, whereas none were found in Han. For β-thalassemia, the β CD41-42 allele was predominant in Li (96.60% vs. 71.01%, χ2=77.24, P<0.001), whereas other alleles (β IVS-II-654, β CD71-72, β CD17, and β -28) were more prevalent in Han (11.01%, 6.96%, 4.64%, and 3.19% vs. 1.54%, 0.00%, 0.31%, and 0.62%, respectively),all P<0.05. In conclusion, distinct ethnic disparities in thalassemia genotype distribution are observed in southern Hainan. The Li ethnic group is predominantly characterized by α-thalassemia and α/β-compound genotypes with a predominant β CD41-42 mutation. In contrast, the Han ethnic group displays higher -- SEA proportion and heterogeneous β-thalassemia genotypes.

To analyze the ethnic differences in the genotype distribution of thalassemia between the Han and Li ethnic groups in the Qiongnan region (southern Hainan). A cross-sectional study employing a stratified multistage sampling method was conducted from January 2019 to December 2023. A total of 4 493 high-risk individuals (2 734 Han and 1 759 Li) from southern Hainan (including Sanya, Ledong, Baoting, Lingshui, and other counties) underwent thalassemia genetic testing. The genotype distribution was statistically analyzed. Inter-group comparisons were performed using χ2 test or Fisher′s exact test. The results showed an overall thalassemia positivity rate of 66.70% (2 997/4 493), with carrier, intermediate and major thalassemia rates of 62.01% (2 786/4 493), 3.98% (179/4 493) and 0.71% (32/4 493), respectively. The positivity rates for thalassemia were 87.83% (1 545/1 759) in the Li ethnic group and 53.11% (1 452/2 734) in the Han ethnic group. Among them, the Li ethnic group exhibited significantly higher positivity rates for α-thalassemia (71.12% vs. 40.64%, χ2=398.90, P<0.001) and α/β-compound thalassemia (13.36% vs. 3.33%, χ2=160.06, P<0.001) compared to the Han ethnic group, whereas the Han ethnic group had a higher β-thalassemia rate (9.14% vs. 3.35%, χ2=56.03, P<0.001). Both ethnic groups shared common α-thalassemia alleles (-α 3.7 and -α 4.2), but the -- SEA allele proportion was significantly higher in Han (21.33% vs. 4.34%, χ2=231.45, P<0.001). Six rare -α 21.9 mutations (0.26%) were exclusively identified in the Li ethnic group, whereas none were found in Han. For β-thalassemia, the β CD41-42 allele was predominant in Li (96.60% vs. 71.01%, χ2=77.24, P<0.001), whereas other alleles (β IVS-II-654, β CD71-72, β CD17, and β -28) were more prevalent in Han (11.01%, 6.96%, 4.64%, and 3.19% vs. 1.54%, 0.00%, 0.31%, and 0.62%, respectively),all P<0.05. In conclusion, distinct ethnic disparities in thalassemia genotype distribution are observed in southern Hainan. The Li ethnic group is predominantly characterized by α-thalassemia and α/β-compound genotypes with a predominant β CD41-42 mutation. In contrast, the Han ethnic group displays higher -- SEA proportion and heterogeneous β-thalassemia genotypes.

Objective To investigate the protective effect of astaxanthin(ASTA)on gouty chondrocyte injury induced by monosodium urate crystal(MSU)and its mechanism.Methods The gout cell model by sodium urate crystals was established.C-28I2 cells were randomly divided into blank group(conventional culture),model group(200 μg·mL-1MSU),experimental-L group(200 μg·mL-1 MSU+20 μmol·mL-1 ASTA),experimental-H group(200 μg·mL-1 MSU+40 μmol·L-1 ASTA),experimental-H+Vector group(transfected with Vector+200 μg·mL-1 MSU+40 μmol·L-1 ASTA),experimental-H+NLRP3 group(transfected with NLRP3 plasmid+200 ig·mL-1 MSU+40 μmol·L-1 ASTA).Cell counting kit-8(CCK-8)assay was used to detect the cell proliferation rate;Western blot assay was used to detect the expression of related proteins;the levels of Hyp and GAG were detected by enzyme-linked immunosorbent assay(ELISA).Results The cell proliferation rate of blank group,model group,experimental-L group and experimental-H group were(100.00±5.40)％,(67.41±4.52)％,(72.69±5.05)％and(81.47±7.73)％,respectively.The above indicators showed statistically significant differences between the model group and the blank group,between the experimental-L,experimental-H groups and the model group(all P＜0.05).Nucleotide binding oligomeric domain-like receptor protein 3(NLRP3)protein expression levels in blank group,model group,experimental-L group,experimental-H group,experimental-H+Vector group and experimental-H+NLRP3 group were 0.44±0.04,0.86±0.06,0.72±0.10,0.52±0.03,0.51±0.03 and 1.13±0.10,respectively;the expression levels of cleaved Caspase-1(Cl-caspase-1)protein were 0.33±0.05,0.73±0.08,0.61±0.07,0.42±0.04,0.39±0.04 and 0.70±0.06,respectively;the mature interleukin(m-IL)-1 β protein expression levels were 0.26±0.03,0.91±0.05,0.70±0.11,0.57±0.08,0.58±0.06 and 0.79±0.08,respectively;the Hyp levels were(1.95±0.22),(3.33±0.25),(2.53±0.18),(2.22±0.21),(2.24±0.20)and(3.25±0.38)μg·mL-1,respectively;the GAG levels were(2.30±0.20),(3.71±0.26),(3.29±0.33),(2.90±0.16),(2.97±0.24)and(3.50±0.34)ng·mL-1,respectively.The above indicators showed statistically significant differences between the model group and the blank group,between the experimental-L,experimental-H groups and the model group,between the experimental-H+NLRP3 group and the experimental-H+Vector group(all P＜0.05).Conclusion Astaxanthin can regulate NLRP3 to play an anti-inflammatory role and has a protective effect on MSU-induced gouty cartilage injury.