ObjectiveThis paper aims to investigate the differential mechanisms underlying the staged therapeutic effects of Qijia Rougan formula on liver fibrosis using proteomic technology. MethodsThe staged rat model of liver fibrosis was established by subcutaneous injection of carbon tetrachloride （CCl₄） and olive oil. One hundred and four SD rats were randomized into thirteen groups：a normal group，a two-week model group，a four-week model group，a six-week model group，an eight-week model group，a two-week Qijia Rougan formula group，a four-week Qijia Rougan formula group，a six-week Qijia Rougan formula group，an eight-week Qijia Rougan formula group，a two-week compound Biejia Ruangan tablet group，a four-week Compound Biejia Ruangan Tablet group，a six-week Compound Biejia Ruangan Tablet group，and an eight-week compound Biejia Ruangan tablet group. After two weeks of drug intervention，liver tissue and abdominal aortic blood samples were collected from the rats for testing. Hematoxylin-eosin （HE） staining，Masson staining，and Picro Sirius red staining were used to observe pathological damage and collagen fiber deposition in liver tissues. Immunohistochemistry （IHC） was employed to detect the contents of fibrosis markers in liver tissues. The contents of liver function indicators in the serum were measured using a fully automated biochemical analyzer，and the levels of liver fibrosis indicators in the serum were assessed by enzyme-linked immunosorbent assay （ELISA）. Liver tissues from the normal group，each model group，and each Qijia Rougan formula group were subjected to label-free quantitative proteomic analysis to identify differential proteins among the groups，with key proteins validated by Western blot. Finally，bioinformatics analysis was performed on the differential proteins. Results（1） The staged rat model of liver fibrosis constructed with CCl₄ and olive oil showed pathological results at the 2^nd，4^th，6^th，and 8^th weeks of modeling that were consistent with the Metavir standards for the F1，F2，F3，and F4 stages. Compared with those in the normal control group，the protein expressions of α-smooth muscle actin （α-SMA） and Collagen Ⅰ were significantly increased in each stage （P<0.05）. The levels of liver function indicators in the serum，including alanine aminotransferase （ALT），aspartate aminotransferase （AST），alkaline phosphatase （ALP），direct bilirubin （DBIL），and total bilirubin （TBil） in each model group，were significantly elevated in each stage （P<0.01）. The levels of liver fibrosis indicators in the serum，including procollagen Ⅲ peptide （PⅢP），type Ⅳ collagen（Ⅳ-C），hyaluronic acid （HA），and laminin （LN） in each model group，were significantly increased in each stage （P<0.05，P<0.01）. This study successfully established a staged rat model of liver fibrosis. （2） Compared with the model groups at each stage，the administration groups showed a reduction in hepatocyte ballooning degeneration，a more orderly arrangement of hepatocytes，and a decrease of inflammatory cell infiltration. The blue-stained collagen fibers became significantly thinner and finer，with reduced and narrowed fibrous septa. The areas of collagen fibers and Picro Sirius red staining were reduced （P<0.05）. The positive areas of α-SMA and Collagen Ⅰ expression were significantly decreased （P<0.05）. The levels of ALT，AST，ALP，DBIL，and TBil in the rats of the model groups at each stage were significantly reduced （P<0.05，P<0.01）. The levels of PⅢP，Ⅳ-C，HA，and LN in the rats of the model groups at each stage were significantly decreased （P<0.05）. Among these，the improvements in all indicators were most significant in the F3 stage （P<0.01）.（3） The proteomic results show that a total of 165 differential proteins exhibit a callback trend when comparing the model groups at four stages with the normal group，and when comparing the Qijia Rougan formula group with the model group. Western blot analysis reveals that the levels of NAD（P）H：quinone oxidoreductase 1 （NQO1），mitogen-activated protein kinase 1 （MAPK1），arginase 1 （Arg1），and glutathione S-transferase α1 （GSTA1） were consistent with the proteomic results. Bioinformatics results reveal that 165 differentially expressed proteins are enriched in multiple signaling pathways. Notably，signaling pathways such as drug metabolism-cytochrome P450，arginine biosynthesis，and the peroxisome proliferator-activated receptor （PPAR） signaling pathway were found to be closely associated with liver fibrosis，suggesting that the Qijia Rougan formula may exert its staged regulatory effects on liver fibrosis by regulating these pathways. ConclusionThe Qijia Rougan formula may achieve staged regulation of liver fibrosis by regulating drug metabolism-cytochrome P450，arginine biosynthesis，and the PPAR signaling pathway.

Objective: To assess the nutritional supply status of school meals for primary and secondary school students in Henan Province, so as to provide a basis for scientific guidance of school meals. Methods: During 2021-2023, 115 lunch and dinner samples were collected from 25 primary and secondary schools in Zhoukou, Anyang and Luoyang of Henan Province by a direct selection method, and 13 nutrients were determined for each sample. The nutrient supply was evaluated based on Nutrition Guidelines of School Meals and Reference Intake of Dietary Nutrients for Chinese Residents (2023 Edition). Mann-Whitney U test was used to compare the differences of nutritional supply between urban and rural schools. Results: The median values for energy (709.77 kcal,1 kcal=4.18 kJ), fat energy supply ratio (0.27) and carbohydrate energy supply ratio (0.55) in the 66 lunches and dinners from primary school were within the recommended range. The supply of protein (28.39 g) and sodium (1 464.59 mg) was excessive. The median values of zinc (2.62 mg) and dietary fiber (5.19 g) were lower than the reference values. No statistically significant differences were observed in the supply of 13 nutrients between urban and rural primary schools( U = 427.00 -633.00, P > 0.05 ). Among 49 samples from secondary schools, the median value of energy supply (930.02 kcal), carbohydrate energy ratio ( 0.54 ) and fat energy supply ratio(0.25) were within the recommended range; and the median values of protein (38.82 g) and sodium (2 556.80 mg) were higher than the standard; and the median values of calcium (250.32 mg) and vitamin B1 (0.16 mg) were lower than the standard. Additionally, the differences in the level of vitamin B2 ( U =372.00) and zinc ( U =375.00) between the urban and rural secondary schools were statistically significant ( P <0.05). Conclusion Nutrient supply of primary and secondary school meals in three cities of Henan Province is inadequate and imbalanced, and the recipe need to be further optimized and improved.

Objective:To analyze the relationship between the optimal surgical margin value and clinical prognosis of transoral robotic surgery（TORS） in treating human papillomavirus（HPV） -positive oropharyngeal squamous cell carcinoma. Methods:A single-center, prospective, observational cohort study was conducted, enrolling patients with early and moderated stage（≤T3 stage） oropharyngeal carcinoma undergoing TORS between July 2020 and April 2024. The proposed optimal surgical margin cutoff value for TORS was set as 2 mm. The primary objectives were to evaluate the optimal clear margin for TORS and its association with overall survival（OS） and progression-free survival（PFS）. Logistic regression was used to analyze correlations between surgical margins and clinical variables, while Cox regression models assessed the impact of surgical margins on OS and PFS. Results:A total of 90 patients（60 males, 66.7%） were included, all had squamous cell carcinoma, with a mean age of 58.0±9.0 years（range: 39-84 years） old. The 1, 2 and 3-year OS rates were 92.3%, 89.9% and 85.0%, respectively, while the 1, 2 and 3-year PFS rates were all 90.1%. For surgical margins ≤2 mm, the 1, 2 and 3-year OS rates were 80.8%, 69.3% and 69.3%, respectively, and PFS rates were 77.9% across three time points. For surgical margins>2 mm, the 1, 2 and 3-year OS rates were 96.5%, 96.5% and 90.6%, respectively, with PFS rates of 94.6%. Logistic regression showed no correlation between surgical margins and tumor type, T/N stage, smoking, alcohol use, or gender（P>0.05）. Cox analysis identified surgical margins>2 mm as a significant factor improving PFS（HR=0.14, 95%CI 0.02-0.90, P=0.038）. Conclusion:This systematic analysis suggests setting a 2 mm and longer as clear surgical margin for TORS. Margins>2 mm are associated with superior postoperative PFS rate and prolonged PFS time in HPV-positive oropharyngeal carcinoma patients.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Carcinoma, Squamous Cell/virology* ; Human Papillomavirus Viruses/isolation & purification* ; Margins of Excision ; Oropharyngeal Neoplasms/virology* ; Papillomavirus Infections/virology* ; Prognosis ; Prospective Studies ; Robotic Surgical Procedures/methods*

Objective:To construct and evaluate a radiomics model for predicting perineural invasion in patients with intrahepatic cholangiocarcinoma (ICC).Methods:Clinical data of 144 patients with ICC undergoing surgery in the People’s Hospital of Zhengzhou University ( n=113) and the Affiliated Cancer Hospital of Zhengzhou University ( n=31) from January 2018 to June 2023 were retrospectively analyzed, including 80 males and 64 females, aged (58.8±10.1) years. The patients were randomly divided into a training set ( n=100) and a test set ( n=44) at a ratio of 7: 3. The former set was used to build the model for predicting perineural invasion, and the latter was used to evaluate the model. Enhanced CT images and clinical data of the patients were collected, and features related to perineural invasion were screened. A light gradient boosting machine was used to construct an imaging genomics model. The model was evaluated using the receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA). Results:Univariate and multivariate logistic regression analysis showed that none of the clinical features were associated with neural invasion in ICC patients (all P>0.05). Six, 25, 32, and 37 radiomics features were obtained by screening the intratumoral, 2 mm peritumoral, 5 mm peritumoral, and 8 mm peritumoral regions, respectively. The area under the ROC curve for predicting perineural invasion in ICC patients was 0.849 (95% CI: 0.774-0.923) in the training set and 0.745 (95% CI: 0.597-0.894) in the test set for the intratumoral model, 0.966 (95% CI: 0.938-0.995) and 0.750 (95% CI: 0.604-0.896) for the 5mm peritumoral model, 0.936 (95% CI: 0.892-0.980) and 0.792 (95% CI: 0.644-0.939) for the 2mm peritumoral model, and 0.961 (95% CI: 0.929-0.992) and 0.689 (95% CI: 0.526-0.853) for the 8mm peritumoral model. The area under the ROC curve, accuracy, sensitivity, and specificity of the combined intratumoral and 5mm peritumoral model for predicting perineural invasion were 0.927 (95% CI: 0.878-0.976), 88.0%, 84.5%, and 89.8% in the training set, and 0.849 (95% CI: 0.737-0.960), 77.3%, 85.2%, and 72.0% in the test set, respectively. The calibration curve showed a deviation between the calibration curve of the combined intratumoral and 5mm peritumoral model and the ideal line, but it could achieve basic consistency. DCA showed that when the threshold was between 0.18 and 0.70, the combined intratumoral and 5mm peritumoral model could bring clinical net benefit to patients when predicting neural invasion. Conclusion:The intratumoral and 5mm peritumoral imaging genomics model based on enhanced CT features can effectively predict neural invasion and offer clinical benefits in patients with ICC.

People with AIDS are highly prone to opportunistic infections,among which pulmonary infections are the most common,with high incidence and fatality rates.How to effectively prevent or reduce the occurrence of HIV-related lung infections(HRLI)is of great practical significance.From the perspective of traditional Chinese medicine(TCM),HRLI and AIDS share common pathological mechanisms,including qi deficiency,spleen defi-ciency,phlegm-heat or phlegm-dampness obstructing the lungs.Based on the TCM theory of"preventing disease before it occurs",the three-level prevention approach includes:the first step"preventing disease before it occurs"emphasizes strengthening the body's defense mechanisms by tonifying the spleen and benefiting qi,and nourishing the lungs and kidneys to strengthen the exterior and protect the body;the second step"preventing the progression of disease once it occurs"emphasizes the removal of pathogenic factors while strengthening the body's defenses.Based on the principles of syndrome differentiation and treatment in TCM,focuses are kept on the spleen tonifying and phlegm resolving,pathogenic factors dispelling and dampness eliminating,while protecting the spleen and nourishing other organs.The third step"preventing relapse after recovery"emphasizes replenishing deficiencies and enhancing immunity.Adopting the three-level prevention approach and early intervention can reduce the inci-dence of HRLI,which is a new idea for the prevention and treatment of HRLI with TCM.

Objective To investigate the effect of tritiated water on the immune system of zebrafish and its potential molecular mechanism. Methods Zebrafish embryos (2.5 to 3 hours post-fertilization [hpf]) were exposed to 3.7 × 10⁴ Bq/mL tritiated water (tritiated water group), and those exposed to E3 culture medium were used as the control group. The mortality rate, hatching rate, deformity rate, heart rate, body length, yolk sac area, neutrophil count in the tail, immune-related gene expression, and immune-related protein expression of zebrafish in the two groups were determined. Then transcriptome technology was used to further analyze the possible mechanism of tritiated water affecting the immune system of zebrafish. Results Compared with the control group, zebrafish at 72 hpf in the tritiated water group had no significant changes in the mortality rate, hatching rate, deformity rate, body length, and yolk sac area((t = 0.9045, 0.5000, 1.0000, 0.7238, 0.0337, P = 0.4169, 0.6433, 0.3739, 0.4785, 0.9735), but had significantly increased heart rate(t = 4.575，P = 0.002). At 4 days post-fertilization (dpf), the neutrophil count in the tail of zebrafish in the tritiated water group was significantly increased(t = 2.563，P = 0.0196), the mRNA expression of TNF-α was significantly decreased(t = 2.891, P = 0.045), the protein expression of nuclear factor-kappa B (NF-κB) was significantly increased(t = 3.848, P = 0.018), and the protein expression of NLRP3 was significantly decreased(t = 14.98, P = 0.001). At 7 dpf, the neutrophil count in the tail and the protein expression levels of NF-κB, NLRP3, and interleukin-1β were significantly decreased(t = 3.772, 7.048, 15.620, 4.423, P = 0.014, 0.002, 0.0001, 0.012). Transcriptome sequencing revealed that differentially expressed genes were mainly enriched in the “neutrophil activation” and “platelet activation pathways” at 4 dpf and in the “neutrophil apoptosis”, “ferroptosis”, and “necroptosis” pathways at 7 dpf. Conclusion Tritiated water exposure induces a temporally dynamic immune response in zebrafish, potentially affecting immune homeostasis by regulating neutrophil activation and apoptosis, as well as the expression of NF-κB and NLRP3.

OBJECTIVES: To evaluate the osteogenic efficacy of three-dimensional printing individualized titanium mesh (3D-PITM) as a scaffold material in guided bone regeneration (GBR). METHODS: 1) Patients undergoing GBR for alveolar bone defects were enrolled as study subjects, and postoperative healing complications were recorded. 2) Postoperative cone beam computed tomography (CBCT) scans acquired at least 6 months post-surgery were used to calculate the percentage of actual bone formation volume. 3) Alveolar bone specimens were collected during the first-stage implant surgery for histomorphometric analysis. This analysis quantitatively measured the proportions of newly formed bone and newly formed unmineralized bone within the specimens. Specimens were categorized into three groups based on healing complications (good healing group, wound dehiscence group, 3D-PITM exposure group) to compare differences in the proportions of newly formed bone and newly formed unmineralized bone. RESULTS: 1) Twelve patients were included. Guided bone regeneration failed in one patient, and 3D-PITM exposure occurred in three patients (exposure rate: 25%). 2) The mean percentage of actual bone formation volume in the 11 successful guided bone regeneration cases was 95.23%±28.85%. 3) Histomorphometric analysis revealed that newly formed bone constituted 40.35% of the alveolar bone specimens, with newly formed unmineralized bone accounting for 13.84% of the newly formed bone. Intergroup comparisons showed no statistically significant differences (P>0.05) in the proportions of newly formed bone or newly formed unmineralized bone between the good healing group and the wound dehiscence group or the 3D-PITM exposure group. CONCLUSIONS 3D-PITM enables effective bone augmentation. Radiographic assessment demonstrated favorable bone formation volume, while histological analysis confirmed substantial formation of newly formed mineralized bone within the surgical site.
Humans ; Printing, Three-Dimensional ; Titanium ; Cone-Beam Computed Tomography ; Bone Regeneration ; Osteogenesis ; Surgical Mesh ; Tissue Scaffolds ; Alveolar Process/surgery* ; Adult ; Male ; Middle Aged ; Female ; Wound Healing ; Guided Tissue Regeneration, Periodontal/methods* ; Alveolar Bone Loss/surgery*

Organoids are three-dimensional (3D) cellular structures formed through the differentiation and self-organization of pluripotent stem cells or tissue-derived cells, showing considerable potential in the research on disease mechanism, personalized medicine, and developmental biology. However, the development of organoids is limited by the complex composition, batch-to-batch variations, and immunogenicity of basement-membrane matrix in the current culture system, which hinders the clinical translation and in vivo applications of organoids. Hydrogels are highly hydrated 3D polymer network materials, with modifiable mechanical and biochemical properties by engineering, representing an ideal alternative to basement-membrane matrix. This article reviews the research progress in engineered hydrogels with defined composition currently used in organoid culture. We introduce the structural characteristics and engineering design considerations of hydrogels, emphasize the latest research progress and specific application cases, and discuss the future development of these engineered hydrogels, provide valuable insights for the further advancement and optimization of engineered hydrogels for organoid.
Hydrogels/chemistry* ; Organoids/cytology* ; Tissue Engineering/methods* ; Humans ; Animals ; Pluripotent Stem Cells/cytology* ; Cell Culture Techniques, Three Dimensional/methods* ; Tissue Scaffolds

Objective:To identify individuals with accelerated aging under the frailty index (FI) as a proxy indicator of biological age, and to investigate the associations of age at menarche, age at menopause, and reproductive lifespan with frailty status and multi-timepoint FI trajectories among Chinese adult women.Methods:The current study included 302 471 women from the China Kadoorie Biobank 2004-2008 baseline survey data. Their age at menarche and menopause were self-reported, and the duration of reproductive lifespan was calculated by subtracting the two ages. The baseline FI was constructed using 28 baseline variables, including diseases, symptoms, and anthropometric measurements. Frailty status was categorized into three groups: non-frail (FI≤0.10), pre-frail (0.10