OBJECTIVE To investigate the effects and mechanism of Yigan huayu formula in alleviating liver fibrosis in mice. METHODS Mice were randomly divided into blank group （normal saline）， model group （normal saline）， Yigan huayu formula low- and high-dose groups （28.98， 57.96 g/kg， calculated by crude drug）， with 8 mice in each group. Except for the blank group， the liver fibrosis model was induced by intraperitoneal injection of 15%CCl 4 -olive oil solution. From the third week， the mice received the medicine/normal saline intragastrically， once a day， for 4 consecutive weeks. After the last medication， liver indexes were calculated， the activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum， as well as the hydroxyproline （HYP） content in liver tissue， were measured. Liver histopathology was evaluated. Differentially expressed proteins （DEPs） in liver tissue were analyzed based on proteomics， followed by bioinfo rmatics analysis. The expressions of core DEPs were validated using Western blot （WB） and immunohistochemistry （IHC） methods. RESULTS Compared with the blank group， the model group showed significantly elevated liver indexes， serum activities of ALT and AST， and hepatic HYP content （ P ＜0.05）， along with obvious pathological damage and collagen deposition. Compared with the model group， the above indexes of mice in the Yigan huayu formula high-dose group were decreased significantly （ P ＜0.05）， with marked improvement in liver pathological damage and collagen deposition. Proteomics identified 210 DEPs between the model group and Yigan huayu formula high-dose group. DEPs were significantly enriched in extracellular matrix （ECM）-receptor interaction and lipid metabolism pathways. WB and IHC confirmed that Yigan huayu formula could significantly inhibit the abnormally elevated expressions of collagen type Ⅳ alpha1 chain （COL4A1）， secreted protein acidic and rich in cysteine （SPARC）， vitronectin （VTN） and laminin subunit alpha5 （LAMA5） in liver tissue of mice （ P ＜0.05）. CONCLUSIONS Yigan huayu formula may exert anti-hepatic fibrosis effects by inhibiting the expressions of proteins such as COL4A1， LAMA5， SPARC， and VTN， thereby blocking the ECM-receptor interaction signaling pathway， and subsequently suppressing excessive ECM deposition and basement membrane remodeling.

Objective: To accurately determine ABO blood groups through serological and molecular biological tests in patients whose forward typing was group O but who showed weak antibodies on one side of reverse typing, thereby ensuring precise transfusion. Methods: A total of 160 cases from 2023 to 2025 at Tianjin Blood Center, initially screened as O type with weak antibodies or missing antibodies on one side of reverse typing, were collected. Serological tests absorption-elution tests, and molecular biological tests were performed to accurately identify their ABO blood groups and ensure transfusion safety. Results: Among the 160 patients with forward typing as O type and weak antibodies on one side of reverse typing, 92 were identified as O type, 46 as ABO subtypes already named and recorded by ISBT (mainly including 18 cases of Ael02, 8 cases of AW37, and 9 cases of Bel03), 12 suspected as new alleles not yet recorded by ISBT (among which 5 were suspected to carry novel alleles that have not been recorded or reported in the literature), and 10 were suspected as weakened A/B antigens. Conclusion: In clinical transfusion, attention should be paid to cases with ABO forward-reverse discrepancy, where forward typing is O type and reverse typing shows weak antibodies on one side. These cases may involve ABO subtypes, and further testin is necessary to accurately determine the ABO blood group, achieve precise transfusion, and avoid blood wastage.

OBJECTIVE To explore the mechanism of Chaiqi yigan granules （CQYG） against liver cancer through the ferroptosis pathway. METHODS Network pharmacology combined with ferroptosis-related database was used to screen key targets and main effective components of CQYG against liver cancer via regulating ferroptosis； molecular docking technology was employed to analyze the binding ability of main active components to key targets. Human liver Huh-7 cells were divided into blank serum control （CON） group， CQYG drug-containing serum （CQYGKL） group， ferroptosis inducer （RSL3） group， mammalian target of rapamycin complex 1 （mTORC1） inhibitor （RMC-5552） group， mTORC1 agonist （CCT007093） group， and CCT007093+CQYGKL group. The levels of Fe 2+ ， malondialdehyde （MDA）， and glutathione （GSH） in the cells were detected in the former three groups； mRNA expressions of mammalian target of rapamycin （mTOR）， sterol regulatory element-binding protein 1 （SREBP1）， and stearoyl-CoA desaturase 1 （SCD1）， protein expressions of SREBP1 and SCD1 as well as phosphorylation levels of mTOR and ribosomal S6 kinase （S6K） proteins were detected in all groups. RESULTS Key targets of CQYG for anti-liver cancer through the ferroptosis pathway were mTOR， SREBP1， SCD1，etc. The main active components included quercetin， tanshinone Ⅱ A ， baicalein， etc. The binding energies of main active components to key targets were all less than －5 kJ/mol. Compared with CON group， the levels of Fe 2+ and MDA in the cells in CQYGKL group and RSL3 group were significantly increased， while the levels of GSH were significantly decreased （ P ＜0.05）. mRNA expressions of mTOR， SREBP1 and SCD1， protein expressions of SREBP1 and SCD1， as well as the phosphorylation levels of mTOR and S6K proteins were significantly decreased in the CQYGKL group， RSL3 group， and RMC-5552 group， whereas all the above indicators were significantly increased in the CCT007093 group （ P ＜0.05）. Compared with CCT007093 group， the changes in all the above indicators were significantly suppressed in the CCT007093+CQYGKL group （ P ＜0.05）. CONCLUSIONS CQYG may induce ferroptosis by inhibiting mTORC1/SREBP1/SCD1 axis， thereby exerting anti-liver cancer effects.

ObjectiveTo analyze the distribution characteristics of traditional Chinese medicine （TCM） constitution and factors associated with hyperuricemia （HUA） in the HUA population， providing a reference for persona-lized prevention and management of HUA. MethodsA cross-sectional study was conducted among individuals who had completed both serum uric acid （SUA） testing and TCM constitution identification during health examinations. According to SUA levels， participants were divided into a hyperuricemia group and a normal SUA group； according to the constitution identification results， participants were classified into balanced constitution， single biased constitution， and mixed constitution. The distribution characteristics of TCM constitution were compared between the two groups； participants in the hyperuricemia group were stratified by sex and age （≥18 years and ＜40 years， ≥40 years）， and the distribution characteristics of TCM constitution were compared across different sex and age groups. Logistic regression was used to analyze factors associated with HUA， with further stratified analyses by sex and age. ResultsA total of 10 920 participants were included， comprising 4170 cases （38.19%） in the hyperuricemia group and 6750 cases （61.81%） in the normal serum uric acid group. According to TCM constitution， there were 420 cases （3.85%） with balanced constitution， 6108 cases （55.93%） with single biased constitution， and 4392 cases （40.22%） with mixed constitution. In the hyperuricemia group， there were 80 cases （19.05%） with balanced constitution； 2275 cases （37.25%） with single biased constitution， among which damp-heat constitution （1205 cases， 52.97%）， phlegm-dampness constitution （500 cases， 21.98%）， and qi-stagnation constitution （181 cases， 7.96%） accounted for relatively high proportions； and 1815 cases （41.33%） with mixed constitution， among which mixed constitution involving phlegm-dampness （51.35%， 932 cases）， mixed constitution involving blood stasis （47.00%， 853 cases）， and mixed constitution involving damp-heat （41.21%， 748 cases） accounted for relatively high proportions. Logistic regression showed that qi-stagnation constitution （OR=1.68）， mixed constitution involving qi-stagnation （OR=1.53）， mixed constitution involving phlegm-dampness （OR=1.55）， and mixed constitution involving blood stasis （OR=1.50） were independently associated with HUA （P＜0.05）. Sex-stratified analysis showed that mixed constitution involving phlegm-dampness was positively associated with HUA risk in females； age-stratified analysis showed that， among individuals aged ≥18 and ＜40 years， qi-stagnation constitution， mixed constitution involving qi stagnation， and mixed constitution involving phlegm-dampness were positively associated with HUA risk （all P＜0.05）. ConclusionThe prevalence of HUA differs markedly by sex and age. Mixed constitution accounts for a higher proportion of HUA patients. Qi-stagnation constitution， mixed constitution involving qi stagnation， mixed constitution involving phlegm-dampness， and mixed constitution involving blood stasis are associated with HUA. Notably， the specific association of mixed constitution involving phlegm-dampness in females and qi-stagnation constitution in young adults is particularly prominent.

Objective:To investigate the effect of cytoskeleton-associated protein 4 (CKAP4) gene knockout on maxillary expansion osteogenesis and its regulatory mechanism on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSC).Methods:Nineteen wild type (WT) and nineteen CKAP4 gene knockout (Ckap4 -/-) mice aged 6-8 weeks were selected to establish a mouse model of rapid maxillary expansion. Samples were taken on the 7th and 14th day after the operation. Micro-CT and HE staining were used to evaluate bone regeneration. Tissue proteins in the modeled area were collected, and Western blotting analysis (WB) was used to detect the protein expression levels of alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN). BMSC were isolated from WT and Ckap4 -/- mice. The expression of surface markers CD29, Sca-1, CD44, CD45, CD34, and CD11b was detected by flow cytometry, and cell proliferation ability was detected by 5-ethynyl-2'-deoxyuridine (EdU). After 7 days of osteogenic induction, real-time fluorescence quantitative PCR (RT-qPCR) and WB were used to detect the expression levels of RUXN2, ALP, OCN, protein kinase B (AKT), and phosphorylated protein kinase B (p-AKT). After 21 days, alizarin red staining and cetyl pyridine chloride quantification were used to detect the differences in mineralized nodule formation in each group. In CKAP4 gene knockout BMSC, the small-molecule AKT agonist sc79 (4 μg/ml) was added as the intervention group (Ckap4 -/- +sc79), and dimethyl sulfoxide (DMSO) treatment was used as the control group (Ckap4 -/- +DMSO). After osteogenic induction, RT-qPCR, WB, and alizarin red staining were used to compare the osteogenic differentiation differences between the two groups of cells. Results:The micro-CT results showed that at 7 days and 14 days after surgery, the new bone volume in the Ckap4 -/- group [(0.070±0.010) and (0.146±0.019) mm 3] was significantly lower than that in the WT group [(0.094±0.006) and (0.196±0.013) mm 3] (both P<0.01). HE-stained histological sections showed that the area of new bone tissue in the Ckap4 -/- group at 7 days and 14 days after surgery [(0.101±0.008) and (0.158±0.010) mm 2] was also significantly lower than that in the WT group [(0.116±0.005) and (0.183±0.008) mm 2] (both P<0.05). WB was used to detect the tissue proteins in the maxillary modeling area of mice in the two groups 7 days after surgery. The results showed that the expression levels of ALP, RUNX2 and OCN in the Ckap4 -/- group were significantly lower than those in the WT group. BMSC from wild-type mice and CKAP4 knockout mice were both positively expressed for CD29, CD44, and Sca-1, and basically not expressed for CD45, CD34, and CD11b. EdU assay showed that there was no significant difference in the proliferation ability of cells in the two groups. After 21 days of osteogenic induction of BMSC, alizarin red staining results showed that the number of mineralized nodules in the Ckap4 -/- group was significantly less than that in the WT group. After adding sc79, the number of mineralized nodules increased significantly, which was consistent with the results of cetyl pyridine chloride quantification. After 7 days of osteogenic induction, It was found that the expression levels of ALP, RUNX2, and OCN in the CKAP4 -/-group (0.751±0.066, 0.484±0.040, 0.679±0.063) were significantly lower than those in the WT group (1.000±0.113, 1.000±0.081, 1.000±0.113) (all P<0.001). The results of WB were consistent with those of RT-qPCR. At the same time, the WB results showed that the level of p-AKT protein in the CKAP4 -/-group (0.518±0.114) was significantly lower than that in the WT group (1.000±0.234) ( P<0.05). After treatment with sc79 for 7 days of osteogenic induction, RT-qPCR was used to detect the gene expression levels of ALP, RUNX2, and OCN. The results showed that the expression levels in the CKAP4 -/-+sc79 group (2.755±0.353, 4.800±0.990, 2.524±0.137) were significantly higher than those in the CKAP4 -/-+DMSO group (1.000±0.078, 1.000±0.247, 1.000±0.175) (all P<0.001). Conclusions:CKAP4 knockout inhibits the osteogenic differentiation of BMSC by reducing the activity of the PI3K/AKT signaling pathway, thereby suppressing osteogenesis in maxillary expansion.

Objective To explore the MRI findings and clinical features of ovarian endometrioid carcinoma(OEC).Methods The preoperative data from 23 patients with surgical pathologic evidence of OEC were collected.The MRI signs[including shape,size,signal,enhancement mode,height of mural nodule,width,height-to-width ratio(HWR)],and apparent diffusion coefficient(ADC)value,as well as clinical features(including age,clinical manifestations,serological markers,pelvic effusion,lymph node metastasis,endometrial cancer,and endometriosis)were retrospectively analyzed.Results Among the 23 cases of OEC,21 exhibited cystic-solid masses with single cyst,and 20 cases displayed well-defined boundaries.The tumor sizes ranged from 4.3 to 17.5 cm.Lymph node metastasis was observed in 4 cases.Thirteen cases were accompanied by pelvic effusion.Additionally,6 cases(26%)with endometrial cancer,and 10 cases(43%)with endometriosis.Elevated CA125 levels were detected in 21 cases(91%),and elevated HE4 levels were found in 18 cases(78%).Regarding the cystic components of the tumors,both T2WI and fat suppression(FS)T2WI showed high signal intensity.In 9 cases,slightly high T1WI signal was detected with no enhancement,and mild enhancement was detected of the cyst wall.The mural nodule of the tumor were predominantly multifocal and wide-based,appearing isointense on T1WI and isoin-tense to slightly hyperintense on T2WI.They was measured with height of 1.8-10.3 cm,widths ranged from 2.7 to 10.6 cm,HWR of 0.68±0.15,high signal on diffusion weighted imaging(DWI),ADC values of(0.937±0.14)×10-3 mm2/s,and mainly showed progressive moderate to significant enhancement.Conclusion OEC typically exhibit a single cyst with multiple mural nodules.The mural nodules are predominantly multifocal and wide-based,with HWR of less than 0.69,limited diffusion,and moderate to signifi-cant enhancement.The enhancement of the parenchymal components is more pronounced than that of the cyst wall.Furthermore,OEC may be accompanied by pelvic effusion,endometrial cancer,endometriosis,and elevated serological markers.

Objective To explore the MRI and clinical features of squamous carcinoma transformation-mature teratoma(SCT-MT)of the ovary.Methods The pre-operative data from 7 patients with SCT-MT confirmed by surgery and pathology were collected.The MRI features(such as location,morphology,size,signal,boundaries,and the presence of a mural nodule,with or without fat or calcifi-cation,limited diffusion,transmural growth,and angle to the cyst wall)and clinical features(including age,clinical manifestations,serologi-cal markers,pelvic effusion,peripheral tissue infiltration,and lymph node metastasis)were retrospectively analyzed.Results Among the seven SCT-MT,all originated unilaterally,and were cystic-solid masses with a predominantly cystic component of round or round-like appearance.Six cases had well-defined boundaries,and six exhibited fat-fluid levels.The tumor sizes ranged from 9 cm to 17 cm.Seven cases showed mural nodules,without calcification and fat,with limited diffusion,and the mean apparent diffusion coefficient(ADC)value was(0.96±0.11)× 10-3 mm2/s.Six cases showed transmural growth,and the angle between the nodule and the cyst wall was obtuse in 5 cases,and the mural nodules were significantly enhanced.The seven SCT-MT patients ranged in age from 53 to 75 years old.Four patients had clinical manifestations of pain related to pelvic distension.Conclusion SCT-MT MRI typically presents as a unilateral large solid mass in the pelvic cavity,with a predominantly cystic component.The mural nodules within it lack calcification or fat,show limited diffusion,and may breech the wall and infiltrate adjacent structures,with significant enhancement.Furthermore,SCT-MT may be associated with older age and elevated serological markers.

Objective:To investigate the effect of cytoskeleton-associated protein 4 (CKAP4) gene knockout on maxillary expansion osteogenesis and its regulatory mechanism on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSC).Methods:Nineteen wild type (WT) and nineteen CKAP4 gene knockout (Ckap4 -/-) mice aged 6-8 weeks were selected to establish a mouse model of rapid maxillary expansion. Samples were taken on the 7th and 14th day after the operation. Micro-CT and HE staining were used to evaluate bone regeneration. Tissue proteins in the modeled area were collected, and Western blotting analysis (WB) was used to detect the protein expression levels of alkaline phosphatase (ALP), Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN). BMSC were isolated from WT and Ckap4 -/- mice. The expression of surface markers CD29, Sca-1, CD44, CD45, CD34, and CD11b was detected by flow cytometry, and cell proliferation ability was detected by 5-ethynyl-2'-deoxyuridine (EdU). After 7 days of osteogenic induction, real-time fluorescence quantitative PCR (RT-qPCR) and WB were used to detect the expression levels of RUXN2, ALP, OCN, protein kinase B (AKT), and phosphorylated protein kinase B (p-AKT). After 21 days, alizarin red staining and cetyl pyridine chloride quantification were used to detect the differences in mineralized nodule formation in each group. In CKAP4 gene knockout BMSC, the small-molecule AKT agonist sc79 (4 μg/ml) was added as the intervention group (Ckap4 -/- +sc79), and dimethyl sulfoxide (DMSO) treatment was used as the control group (Ckap4 -/- +DMSO). After osteogenic induction, RT-qPCR, WB, and alizarin red staining were used to compare the osteogenic differentiation differences between the two groups of cells. Results:The micro-CT results showed that at 7 days and 14 days after surgery, the new bone volume in the Ckap4 -/- group [(0.070±0.010) and (0.146±0.019) mm 3] was significantly lower than that in the WT group [(0.094±0.006) and (0.196±0.013) mm 3] (both P<0.01). HE-stained histological sections showed that the area of new bone tissue in the Ckap4 -/- group at 7 days and 14 days after surgery [(0.101±0.008) and (0.158±0.010) mm 2] was also significantly lower than that in the WT group [(0.116±0.005) and (0.183±0.008) mm 2] (both P<0.05). WB was used to detect the tissue proteins in the maxillary modeling area of mice in the two groups 7 days after surgery. The results showed that the expression levels of ALP, RUNX2 and OCN in the Ckap4 -/- group were significantly lower than those in the WT group. BMSC from wild-type mice and CKAP4 knockout mice were both positively expressed for CD29, CD44, and Sca-1, and basically not expressed for CD45, CD34, and CD11b. EdU assay showed that there was no significant difference in the proliferation ability of cells in the two groups. After 21 days of osteogenic induction of BMSC, alizarin red staining results showed that the number of mineralized nodules in the Ckap4 -/- group was significantly less than that in the WT group. After adding sc79, the number of mineralized nodules increased significantly, which was consistent with the results of cetyl pyridine chloride quantification. After 7 days of osteogenic induction, It was found that the expression levels of ALP, RUNX2, and OCN in the CKAP4 -/-group (0.751±0.066, 0.484±0.040, 0.679±0.063) were significantly lower than those in the WT group (1.000±0.113, 1.000±0.081, 1.000±0.113) (all P<0.001). The results of WB were consistent with those of RT-qPCR. At the same time, the WB results showed that the level of p-AKT protein in the CKAP4 -/-group (0.518±0.114) was significantly lower than that in the WT group (1.000±0.234) ( P<0.05). After treatment with sc79 for 7 days of osteogenic induction, RT-qPCR was used to detect the gene expression levels of ALP, RUNX2, and OCN. The results showed that the expression levels in the CKAP4 -/-+sc79 group (2.755±0.353, 4.800±0.990, 2.524±0.137) were significantly higher than those in the CKAP4 -/-+DMSO group (1.000±0.078, 1.000±0.247, 1.000±0.175) (all P<0.001). Conclusions:CKAP4 knockout inhibits the osteogenic differentiation of BMSC by reducing the activity of the PI3K/AKT signaling pathway, thereby suppressing osteogenesis in maxillary expansion.

BackgroundAttention deficit hyperactivity disorder （ADHD） is a neurodevelopmental disorder characterized by age-inappropriate inattention， excessive activities incongruous with setting， and emotional impulsivity. Subthreshold ADHD （sADHD） is clinically defined as the presence of ADHD symptoms that do not meet the full diagnostic criteria for ADHD. Children with sADHD exhibit deficits in executive function， demonstrate more conduct， learning， and anxiety-related problems compared to typically developing children， and show even poorer working memory performance than children diagnosed with ADHD. Currently， there is limited epidemiological research on sADHD in China， with few studies simultaneously investigating the prevalence of both ADHD and sADHD in children. ObjectiveTo investigate the prevalence of ADHD and sADHD among children aged 6–13 years in Chongqing， analyzing their distribution characteristics within this population， with the aim of providing references for developing preventive measures against both ADHD and sADHD. MethodsFrom October to November 2023， a total of 3 398 students in grades 1–6 from six primary schools in Jiangbei District， Chongqing were selected using a stratified cluster random sampling method. The occurrence of ADHD and sADHD was evaluated by using the short version （18-item version） of the Swanson， Nolan， and Pelham IV rating scales （SNAP-IV） and the Chinese vision of Schedule for Affective Disorder and Schizophrenia for School-aged Children-Present and Lifetime Version （K-SADS-PL）. ResultsThe ADHD detection rate among children in Chongqing was 1.90% （95% CI： 0.014–0.024）. Boys showed a significantly higher ADHD detection rate than girls （χ²=7.733， P=0.005）. No statistically significant differences were found in ADHD detection rates across different grades or age groups （χ²=7.347， 12.362， P>0.05）. The sADHD detection rate was 6.32% （95% CI： 0.054–0.072）. Similarly， boys exhibited significantly higher sADHD detection rates than girls （χ²=21.005， P<0.01）. Significant differences emerged across different grades （χ²=20.559， P=0.001）， while no statistically significant difference was observed in age groups （χ²=12.070， P=0.060）. ConclusionThe ADHD detection rates were comparable across all grade levels and age groups from 6–13 years old. Second-grade children demonstrated notably higher sADHD rates compared to other grades， while boys demonstrated higher prevalence rates than girls for both ADHD and sADHD. ［Funded by Science and Health Joint Medical Research Project in Jiangbei District， Chongqing City in the Second Half of 2023 （number， 2023JBKWLH022）］

Objective To explore the MRI and clinical features of squamous carcinoma transformation-mature teratoma(SCT-MT)of the ovary.Methods The pre-operative data from 7 patients with SCT-MT confirmed by surgery and pathology were collected.The MRI features(such as location,morphology,size,signal,boundaries,and the presence of a mural nodule,with or without fat or calcifi-cation,limited diffusion,transmural growth,and angle to the cyst wall)and clinical features(including age,clinical manifestations,serologi-cal markers,pelvic effusion,peripheral tissue infiltration,and lymph node metastasis)were retrospectively analyzed.Results Among the seven SCT-MT,all originated unilaterally,and were cystic-solid masses with a predominantly cystic component of round or round-like appearance.Six cases had well-defined boundaries,and six exhibited fat-fluid levels.The tumor sizes ranged from 9 cm to 17 cm.Seven cases showed mural nodules,without calcification and fat,with limited diffusion,and the mean apparent diffusion coefficient(ADC)value was(0.96±0.11)× 10-3 mm2/s.Six cases showed transmural growth,and the angle between the nodule and the cyst wall was obtuse in 5 cases,and the mural nodules were significantly enhanced.The seven SCT-MT patients ranged in age from 53 to 75 years old.Four patients had clinical manifestations of pain related to pelvic distension.Conclusion SCT-MT MRI typically presents as a unilateral large solid mass in the pelvic cavity,with a predominantly cystic component.The mural nodules within it lack calcification or fat,show limited diffusion,and may breech the wall and infiltrate adjacent structures,with significant enhancement.Furthermore,SCT-MT may be associated with older age and elevated serological markers.