ObjectiveTo investigate the role of DEAD-box helicase 3 X-linked （DDX3X） in immune-mediated liver injury （ILI）， and to clarify its mechanism by regulating endoplasmic reticulum stress （ERS）-dependent apoptotic pathway and its association with the clinical progression of hepatitis B. MethodsMice were given injection of concanavalin A （ConA） via the caudal vein to establish a model of ILI， PBS （control group） and different concentrations of ConA were injected into the tail vein of hepatocyte-specific DDX3X-knockout mice （DDX3X^ΔHep and DDX3X-flox mice （DDX3X^fl/fl）， respectively.. The log-rank survival analysis， measurement of the serum levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）， and HE staining of liver tissue were performed to assess liver injury， and qRT-PCR and Western Blot were used to measure the mRNA and protein expression levels of glucose-regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein homologous protein （CHOP）， and DDX3X in liver tissue. Intraperitoneal injection of 4-phenylbutyric acid （4-PBA， 100 mg/kg） was performed to inhibit ERS. Serum samples （n=30） and liver tissue samples （n=6） were collected from healthy controls， chronic hepatitis B （CHB） patients， and hepatitis B virus-associated liver failure （HBV-LF） patients； ELISA was used to measure the serum level of DDX3X， and qRT-PCR/Western Blot was used to analyze the expression of targets in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group of mice， the expression of DDX3X in the liver of mice induced by ConA was significantly increased after liver injury （P<0.05）， and hepatocyte-specific DDX3X knockout increased the 72-hour survival rate of mice by 55% （compared with 20% in the DDX3X^fl/fl group）， with significant reductions in the serum levels of ALT and AST （P<0.000 1） and the expression levels of the ERS markers GRP78 and CHOP （P<0.05）. After ERS was inhibited by 4-PBA， there was alleviation of liver injury （with reductions in ALT and AST， P <0.001） and a reduction in DDX3X expression （P<0.01）. The analysis of clinical samples showed that the mRNA and protein expression levels of liver DDX3X in CHB patients and HBV-LF patients were significantly higher than those in healthy controls （all P<0.01）， and there was a significant increase in the serum level of DDX3X in HBV-LF patients （P<0.000 1）. ConclusionDDX3X exacerbates ILI by regulating the ERS-dependent apoptotic pathway （GRP78/CHOP）， and its expression is associated with the progression of hepatitis B. Therefore， it can be used as a potential therapeutic target.

Objective To investigate the expression profile, prognostic value, gene co-expression network, and immunomodulatory role of BRF1 in a pan-cancer context, and to explore its biological functions and molecular regulatory mechanisms in esophageal squamous cell carcinoma (ESCC). Methods The pan-cancer dataset from The Cancer Genome Atlas (TCGA) was utilized to analyze the differential expression of BRF1 in tumor versus normal tissues, its association with patient survival, pathway enrichment for co-expressed genes, and immune features (including immune checkpoints, cytokines, and immune cell infiltration). The expression profile of BRF1 in ESCC was validated using the Gene Expression Omnibus (GEO) database. In vitro, BRF1 was knocked down in ESCC cells using siRNA. Cell proliferation and migration were assessed by MTT and Transwell assays, respectively. The expression levels of proliferation- and migration-related proteins were detected by Western blotting. The correlation between BRF1 and ferroptosis was analyzed using TCGA data. Results BRF1 was significantly upregulated in over 20 types of cancer, and its high expression was associated with poor prognosis in patients with adrenocortical carcinoma and prostate adenocarcinoma. BRF1 was found to positively regulate the T-cell-mediated cell death pathway in esophageal adenocarcinoma and was associated with the circadian rhythm regulation pathway in pancreatic adenocarcinoma. The correlation of BRF1 with immune checkpoints, cytokine networks, and immune cell infiltration was found to be cancer type-specific. In vitro experiments demonstrated that knocking down BRF1 significantly inhibited the proliferation of ESCC cells, accompanied by the downregulation of the proliferation marker PCNA. Cell migration was also significantly impaired, with decreased expression of Vimentin and MMPs and increased expression of E-cadherin. Furthermore, the expression of BRF1 was positively correlated with that of ferroptosis-antagonizing genes, such as GPX4, HSPA5, and SLC7A11. Conclusion BRF1 plays complex roles in pan-cancer, participating in the regulation of tumorigenesis, progression, and immune infiltration. BRF1 promotes the proliferation and migration of ESCC cells, a mechanism potentially associated with the regulation of ferroptosis resistance. These findings suggest that BRF1 could be a potential therapeutic target for ESCC.

Objective: To investigate the characteristics of allele frequencies for 9 red blood cell (RBC) blood group systems in the Hui ethnic voluntary blood donor population of Suzhou using real-time fluorescence PCR technology, so as to provide technical support for establishing a RBC blood group genetic database. Methods: PCR-TaqMan technology was employed to perform genotyping detection for 9 RBC blood group systems using 144 samples from Hui voluntary blood donors in Suzhou, collected between October 2023 and August 2024. Results: Blood group allele frequencies among Suzhou Hui voluntary blood donors were distributed as follows: MNS system (M=0.566 0, N=0.434 0; S=0.079 9, s=0.920 1); Lutheran system (Lu =0.003 5, Lu =0.996 5; Au =0.895 8, Au =0.104 2); Kell system (K=0.000 0, k=1.000 0; Kp =0.003 5, Kp =0.996 5; JS =0.000 0, JS =1.000 0); Duffy system (Fy =0.899 3, Fy =0.100 7); Kidd system (JK =0.451 4, JK =0.548 6); Diego system (Di =0.041 7, Di =0.958 3); Yt system (Yt =0.996 5, Yt =0.003 5); Dombrock system (Do =0.128 5, Do =0.871 5); Colton system (Co =1.000 0, Co =0.000 0). The PCR-TaqMan-based RBC blood group genotyping technology successfully completed testing for all samples. Conclusion: The MNS, Lutheran, Duffy, Kidd, Diego, and Dombrock blood group systems in the Suzhou Hui population exhibited polymorphic distribution patterns, whereas the Colton system was monomorphic. Standardized application of PCR-TaqMan technology facilitates the establishment of an RBC blood group genetic database.

Objective To understand and grasp the status quo of resistance of Culex pipiens pallens to four commonly used insecticides in Hefei City, and to provide a scientific basis for the chemical control of mosquito larvae. Methods From June to July 2023, Cx. pipiens pallens larvae were collected from 9 counties (cities and districts) in Hefei City. The LC50 of late third-instar to early fourth-instar larvae of Cx. pipiens pallens to commonly used insecticides was determined by larval immersion method (sensitive baseline method). Results Cx.pipiens pallens larvae in Hefei City exhibited different degrees of resistance to four insecticides: permethrin, beta-cypermethrin, temephos, and propoxur. The relative resistance coefficients to permethrin and beta-cypermethrin were 26.96 and 21.17, respectively, indicating the moderate resistance level. The relative resistance coefficients to propoxur were 6.70, indicating a low resistance level. The relative resistance coefficient to temephos was 2.43, indicating a sensitivity level. Culex pipiens pallens against pyrethroids such as 0.25% permethrin, 0.025% deltamethrin and 0.025% cypermethrin in 1 h knockout rate and 24 h mortality rates were 3.25% (4/123) and 46.34% (57/123), 3.60% (5/139) and 35.97% (50/139), 3.85% (6/156) and 40.38% (63/156), respectively. For 5% malathion and 0.1% propoxur, the 1 h knockdown rate and 24 h mortality rate were 97.69% (127/130) and 99.23% (129/130), 94.48% (137/145) and 100.00% (145/145), respectively. It showed resistance to 0.25% permethrin, 0.025% deltamethrin and 0.025% cypermethrin, and sensitivity to 5% malathion and 0.1% propoxur. Conclusions Culex pipiens pallens in Hefei City have developed varying degrees of resistance to parathyroid and carbamate insecticides. In the control of mosquito vectors, it is essential to strengthen the scientific and rational use of chemical control in combination with environmental and physical control measures to form an integrated control strategy. This approach will improve the control efficiency while delaying the occurrence and development of insecticide resistance.

[Objective] To investigate the antigen and gene frequency distribution of the MNS blood group system in the Han population of voluntary blood donors in Suzhou, and to explore the polymorphism of rare MNS blood group genes, in order to improve the construction of the local rare blood group database. [Methods] A total of 8 034 whole blood samples were randomly collected from Han blood donors at our station from October 2023 to June 2024. The MNS blood group phenotypes were identified using serological methods. Gene frequencies were analyzed and compared with those of ethnic populations in other regions. Rare MNS phenotype samples were subjected to gene sequencing. [Results] The distribution of MNS blood group system phenotypes within the population was as follows: the MM, NN, and MN phenotypes accounted for 23.00%, 27.12%, and 49.88% respectively; the SS, ss, and Ss phenotypes accounted for 0.30%, 90.99%, and 8.70% respectively. The gene frequencies of M, N, S, and s were 0.4794, 0.5206, 0.0465, and 0.9534 respectively. Chi-squared tests confirmed adherence to Hardy-Weinberg equilibrium with P-values of 0.997 and 0.349, showing statistical significance compared to some other regional ethnic populations (P<0.05). Additionally, one rare serological phenotype, S-s-, with a frequency of 0.01%, was identified. [Conclusion] The MNS blood group system in the Han population of voluntary blood donors in Suzhou exhibits polymorphism and regional distribution characteristics. Gene frequencies differ from those observed in other regions of China. It is essential to enhance the establishment of a rare blood type database in Suzhou to provide data support for precise clinical transfusion.

Objective: To classify the ABO blood group phenotypes of 5 cases from a family, and to explore the molecular mechanism for reduced B antigen expression in ABO blood group system. Methods: Serological identification of the ABO blood group was performed using microcolumn gel assay and saline tube method. The soluble antigens in saliva were detected by the agglutination inhibition assay. The full-length sequences and upstream promoter regions of ABO gene were sequenced for genotyping using PacBio SMRT sequencing technology. Results: The results of serological tests indicated the expression of B antigen decreased in 3 out of 5 blood samples. A mixed-field agglutination was observed with anti-B antibody. B antigen was not detected in all 5 saliva samples. The ABO genotype for all samples were ABO B.01/ABO O.01.02, and a novel mutation c. 28+5875C>T within the DNA-binding region of RUNX1 in +5.8-kb site were found in the B allele for 3 samples with reduced expression of B antigen. Conclusion: Results of serological and genetic analyses classify the 3 cases with reduced B antigen expression as B phenotype. The novel mutation c. 28+5875C>T of RUNX1 could be the key reason for reduced B antigen expression in 3 cases with B phenotype.

Objective To observe the difference of iron deposition in brain deep nuclei of systemic lupus erythematosus(SLE)patients with different disease activity levels using quantitative susceptibility mapping(QSM).Methods Fifty-four SLE patients were retrospectively enrolled and divided into moderate to severe activity group(score＞9,n=25)and mild activity group(score≤9,n=29)according to SLE disease activity index(SLEDAI)score.Patients' general clinical data,as well as the mean QSM of bilateral head of caudate nuclei,putamina,globi pallidi,red nuclei,substantiae nigrae,dentate nuclei and thalami were compared between groups.Pearson or Spearman coefficients were performed to analyze correlations of the mean QSM of nuclei being significant different between groups and conventional clinical indicators.Then receiver operating characteristic(ROC)curve was drawn,and the area under the curve(AUC)was calculated to evaluate the efficacy of the mean QSM for distinguishing moderate to severe and mild activity SLE.Results Compared with mild activity group,moderate to severe activity group had higher serum ferritin,erythrocyte sedimentation rate(ESR),C reactive protein(CRP),anti-double stranded DNA(anti-dsDNA)antibody levels,also higher proportion of positive anti-ribosomal P protein(anti-P)antibodies,but lower complement C3 and C4 levels(all P＜0.05).The mean QSM of bilateral putamina and thalami were significantly higher in moderate to severe activity group than in mild activity group(both P＜0.05).The mean QSM of bilateral putamina was positively correlated with SLEDAI scores,ferritin levels and positivity of anti-P antibodies in SLE patients(with r or rs of 0.447,0.526 and 0.473,respectively,and all P＜0.05).The AUC for distinguishing moderate to severe SLE and mild activity SLE based on the mean bilateral putamina QSM was 0.810.Conclusion There were significant differences of iron deposition in deep brain nuclei of SLE patients with moderate to severe and mild activity.The mean QSM of bilateral putamina could be used to distinguish SLE with moderate to severe activity and mild activity.

Objective To investigate the short-term efficacy of arthroscopic total internal suture combined with platelet-rich plasma(PRP)in the treatment of lateral meniscal popliteal hiatus area injuries.Methods Forty-eight patients diagnosed with lateral meniscal popliteal hiatus area injuries in our hospital from January 2020 to December 2022 were selected and divided into Fast-Fix total internal suture group(FF group,n=23)and Fast-Fix total internal suture combined with PRP group(PRP group,n=25)according to treatment methods.The positive rate of McMurray test at 6 and 12 months after surgery,the preoperative and postoperative visual analogue scale(VAS)of pain,the International Knee Documentation Committee(IKDC)scores,the Lysholm Knee Function Scores,and the Knee Society Scores(KSS)were compared between the two groups.Results No adverse events such as vascular and nerve injury,fever and infection occurred in both groups.There was no significant difference between the two groups in the positive rates of McMurray test at 6 and 12 months(χ2=0.880,P=0.348;Fisher's exact test,P=0.479).In the PRP group,25 cases were followed up for11-14 months,with an average of(12.2±0.8)months;in the FF group,23 cases were followed up for11-14 months,with an average of(12.8±0.8)months.The VAS scores at 6 months and 12 months after surgery were significantly lower than those before surgery in both groups(all P=0.000).The VAS scores of the PRP group at 6 and 12 months after surgery were significantly lower than those of the FF group(P<0.05).Both groups showed significant improvements in IKDC,Lysholm,and KSS scores at 6 and 12 months after surgery compared to preoperative levels(all P=0.000).The IKDC,Lysholm,and KSS scores of the PRP group at 6 and 12 months after surgery were significantly higher than those of the FF group(P<0.05).Conclusion Compared to Fast-Fix total internal suture alone,PRP joint cavity adjuvant therapy based on Fast-Fix total internal suture surgery could reduce the postoperative pain of patients,and promote the functional recovery of the knee joint,which is more recommended in clinical practice.

Objective:To compare the clinical outcomes of APTT-HTO and Biplanar-high tibial osteotomy (Biplanar-HTO) in treating medial compartment knee osteoarthritis.Methods:A non-randomized controlled trial was conducted. Twenty-eight patients with medial compartment knee osteoarthritis who underwent HTO at the Affiliated Hospital of Guizhou Medical University from August 2021 to January 2022 were enrolled. Based on the patients' surgical preference, they were assigned to either the APTT-HTO group ( n=15) or the Biplanar-HTO group ( n=13), followed up for 12 months postoperatively. Postoperative pain Visual Analog Scale (VAS) scores, Knee Society Score (KSS), changes in patellar height (Caton-Deschamps Index, CDI), and posterior tibial slope (PTS) were compared between the two groups. Results:The APTT-HTO group demonstrated a significantly shorter operative time (64.13±4.85 min) compared to the Biplanar-HTO group (81.54±6.09 min) ( P<0.05). No significant differences were observed in intraoperative correction (APTT-HTO: 12.19°±4.85°; Biplanar-HTO: 11.23°±3.02°) or postoperative drainage volume (APTT-HTO: 47.00±13.79 ml; Biplanar-HTO: 47.00±11.17 ml) ( P>0.05). At 12-month follow-up (APTT-HTO: 13.93±2.05 months; Biplanar-HTO: 14.08±2.14 months; no dropouts), the APTT-HTO group showed no significant changes in PTS (9.32°±2.04° vs. preoperative 8.82°±1.89°) or CDI (0.95±0.11 vs. 0.98±0.11) ( P>0.05), while the Biplanar-HTO group exhibited increased PTS (13.27°±1.99° vs. 8.86°±1.99°) and decreased CDI (0.64±0.10 vs. 0.97±0.16) ( P<0.05). The differences in PTS and CDI between the APTT HTO group and the Biplanar HTO group at 12 months after surgery were statistically significant ( P<0.05). Both groups achieved significant clinical improvements: in APTT-HTO, VAS decreased (preopreation 4.80±1.01 to postopreation 1.06±0.88), KSS knee scores increased (47.67±12.03 to 87.93±4.38), and KSS function scores improved (48.00±4.93 to 67.00±5.91); in Biplanar-HTO, VAS reduced (5.08±1.12 to 1.85±1.14), KSS knee scores rose (46.85±11.48 to 85.85±5.11), and KSS function scores enhanced (46.92±5.60 to 66.92±5.22) ( P<0.05 for all). Complications included soft tissue irritation (2 cases per group), with Biplanar-HTO additionally reporting deep vein thrombosis (1 case), hinge fracture (1 case), and patella baja (3 cases). Conclusions:Both APTT-HTO and Biplanar-HTO effectively treat medial compartment knee osteoarthritis. However, APTT-HTO outperforms Biplanar-HTO in preventing postoperative patella infera and minimizing alterations in PTS.

Objective:To construct a method for hepatitis B virus (HBV) DNA detection based on recombinase-mediated isothermal amplification (RAA)-clustered regularly interspaced short palindromic repeats and their associated protein 13a (CRISPR-Cas13a).Methods:Through the alignment and screening of HBV DNA sequences, a positive plasmid was constructed, and recombinase-aided amplification (RAA) primers and CRISPR RNA (crRNA) were designed. A method for detecting HBV DNA based on the RAA-CRISPR-Cas13a system was developed, and the specificity and sensitivity were evaluated. Utilizing the CRISPR-Cas13a system, 70 clinical samples from HBV DNA-positive patients with various viral loads collected at Beijing You′an Hospital from 2019 to 2021 were analyzed. The detection results were further compared with those results using real-time quantitative polymerase chain reaction (qPCR).Results:The optimal RAA amplification primers and crRNA were first screened using the RAA-CRISPR-Cas13a method, with the sensitivities for detecting HBV DNA standards and for clinical samples at 1 IU/ml and<10 IU/ml, respectively, demonstrating specificity for HBV DNA detection. Compared with qPCR (the gold standard), the detection consistency between the two methods was 100% (70/70).Conclusion:This study established a method for detecting HBV DNA by integrating recombinase-aided amplification (RAA) technology with CRISPR/Cas13a technology.