In situ tumor vaccine has become an important strategy in cancer immunotherapy owing to its ability to induce immune responses locally and overcome tumor heterogeneity. However, the abnormal structure and mechanical properties of the tumor’s physical microenvironment significantly limit the efficiency of vaccine delivery and immune efficacy. In this review, the key factors in the tumor’s physical microenvironment, including solid pressure, interstitial fluid pressure, matrix stiffness, and tissue microstructure, are systematically discussed. Their obstructive roles in immune cell infiltration, antigen presentation, and immune activation are analyzed. The potential of approaches, such as radiotherapy, anti-angiogenic therapy, extracellular matrix degradation agents, nanomaterials, and hydrogel delivery platforms, in reshaping the tumor’s physical microenvironment is explored. This review aims to offer theoretical and practical guidance for optimizing in situ vaccine strategies through the regulation of the tumor’s physical microenvironment, ultimately advancing the precision and effectiveness of cancer immunotherapy.

BACKGROUND:Osteoporotic fracture is the most serious complication of osteoporosis.Previous studies have demonstrated that gut microbiota has a regulatory effect on skeletal tissue and that gut microbiota has an important relationship with osteoporotic fracture,but the causal relationship between the two is unclear. OBJECTIVE:To explore the causal relationship between gut microbiota and osteoporotic fractures using Mendelian randomization method. METHODS:The genome-wide association study(GWAS)datasets of gut microbiota and osteoporotic fracture were obtained from the IEU Open GWAS database and the Finnish database R9,respectively.Using gut microbiota as the exposure factor and osteoporotic fracture as the outcome variable,Mendelian randomization analyses with random-effects inverse variance weighted,MR-Egger regression,weighted median,simple model,and weighted model methods were performed to assess whether there is a causal relationship between gut microbiota and osteoporotic fracture.Sensitivity analyses were performed to test the reliability and robustness of the results.Reverse Mendelian randomization analyses were performed to further validate the causal relationship identified in the forward Mendelian randomization analyses. RESULTS AND CONCLUSION:The results of this Mendelian randomization analysis indicated a causal relationship between gut microbiota and osteoporotic fracture.Elevated abundance of Actinomycetales[odds ratio(OR)=1.562,95%confidence interval(CI):1.027-2.375,P=0.037),Actinomycetaceae(OR=1.561,95%CI:1.027-2.374,P=0.037),Actinomyces(OR=1.544,95%CI:1.130-2.110,P=0.006),Butyricicoccus(OR=1.781,95%CI:1.194-2.657,P=0.005),Coprococcus 2(OR=1.550,95%CI:1.068-2.251,P=0.021),Family ⅩⅢ UCG-001(OR=1.473,95%CI:1.001-2.168,P=0.049),Methanobrevibacter(OR=1.274,95%CI:1.001-1.621,P=0.049),and Roseburia(OR=1.429,95%CI:1.015-2.013,P=0.041)would increase the risk of osteoporotic fractures in patients.Elevated abundance of Bacteroidia(OR=0.660,95%CI:0.455-0.959,P=0.029),Bacteroidales(OR=0.660,95%CI:0.455-0.959,P=0.029),Christensenellacea(OR=0.725,95%CI:0.529-0.995,P=0.047),Ruminococcaceae(OR=0.643,95%CI:0.443-0.933,P=0.020),Enterorhabdus(OR=0.558,95%CI:0.395-0.788,P=0.001),Eubacterium rectale group(OR=0.631,95%CI:0.435-0.916,P=0.016),Lachnospiraceae UCG008(OR=0.738,95%CI:0.546-0.998,P=0.048),and Ruminiclostridium 9(OR=0.492,95%CI:0.324-0.746,P=0.001)would reduce the risk of osteoporotic fractures in patients.We identified 16 gut microbiota associated with osteoporotic fracture by the Mendelian randomization method.That is,using gut microbiota as the exposure factor and osteoporotic fracture as the outcome variable,eight gut microbiota showed positive causal associations with osteoporotic fracture and another eight gut microbiota showed negative causal associations with osteoporotic fracture.The results of this study not only identify new biomarkers for the early prediction of osteoporotic fracture and potential therapeutic targets in clinical practice,but also provide an experimental basis and theoretical basis for the study of improving the occurrence and prognosis of osteoporotic fracture through gut microbiota in bone tissue engineering.

Objective:s To investigate the risk factors for delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) in patients with acute carbon monoxide poisoning (ACOP) and to develop predictive models based on machine learning algorithms.Methods:Patients with ACOP hospitalized at the First Affiliated Hospital of Zhengzhou University from August 2019 to October 2024 were included, with the occurrence of DEACMP as the outcome measure. The dataset was randomly divided into training and validation sets at a ratio of 7:3. Lasso regression was used to select features influencing the outcome in training sets. Nine machine learning models—including Random Forest (RF), Extreme Gradient Boosting (XGBoost), and Support Vector Machine (SVM)—were constructed. Receiver operating characteristic (ROC) curves were plotted and the area under the curve (AUC) calculated for each model. Calibration curves were used to assess accuracy, and decision curve analysis (DCA) was applied to evaluate clinical utility. The SHapley Additive exPlanations (SHAP) method was employed to visualize and interpret the best-performing model.Results:A total of 264 ACOP patients were included, of whom 54 (20.5%) developed DEACMP. Lasso regression identified eight key feature variables. Based on these factors, predictive models were constructed, showing good AUC stability across the nine machine learning models in both training (0.92–0.99) and validation sets (0.85–0.91). The RF model performed best, with an AUC of 0.99 in the training set and 0.90 in the validation set; its calibration curve and DCA curve also demonstrated excellent performance. SHAP analysis of the RF model revealed the importance ranking of factors from highest to lowest as follows: Glasgow Coma Scale (GCS) score, duration of coma, age, history of coronary heart disease, CK-MB level, monocyte count, diastolic blood pressure (DBP), and drinking history.Conclusions:The RF model exhibited the highest predictive performance for DEACMP occurrence in ACOP patients. The influencing factors, ranked in order of importance from highest to lowest, are as follows: GCS score, duration of coma, age, history of coronary heart disease, CK-MB level, monocyte count, DBP, and drinking history.

Objective To investigate the mechanism of action of the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway in non-alcoholic fatty liver disease(NAFLD),and to explore the mechanism of Gegen QinLian Decoction for the treatment of NAFLD,using in vivo and in vitro experiments.Methods Rats were fed with high-fat chow for 24 weeks to induce NAFLD,and were then divided randomly into normal(C),model(M),high-,medium-,and low-dose Gegen QinLian Decoction(GGQLT-H,GGQLT-M,GGQLT-L),and metformin(Met)groups.From week 25 onwards,the rats were administered the corresponding drugs by gavage for 2 weeks according to the grouping,until sampling.Levels of the oxidative stress markers malondialdehyde(MDA)and glutathione(GSH)in the liver tissues were measured in each group using biochemical kits and ferrous iron(Fe2+)in rat liver tissues was detected using a Fe2+kit.Nrf2,heme oxygenase-1(HO-1),SLC7A11,glutathione synthetase(GSS),GPX4,and acyl coenzyme A synthetase 4(ACSL4)mRNA levels in rat liver tissues were measured by reverse transcription quantitative polymerase chain reaction.For cellular experiments lipid acc umulation was induced in HepG2 hepatocellular carcinoma cells using 1 mmol/L free fatty acid,to mimic the NAFLD in vitro model.Different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment.Lipid accumulation was detected in the cells in each group by Oil red O staining.The MDA and GSH contents of HepG2 cells in the different groups were determined using appropriate kits,and the ferrous contents were detected using a cell-specific ferrous kit.Expression levels of Nrf2,HO-1,SLC7A11,GSS,GPX4,and ACSL4 mRNA was detected in each group of cells using reverse transcription quantitative polymerase chain reaction.Results In the animal experiments,MDA and Fe2+liver levels were significantly higher in the M group than in the C group,while GSH levels were significantly lower(P＜0.01).GGQLT-H,GGQLT-M and Met groups showed significantly reduced MDA and Fe2+and elevated GSH levels compared with the M group(P＜0.01,P＜0.05).High-and medium-dose Gegen QinLian Decoction and metformin increased Nrf2,HO-1,GSS,and GPX4 mRNA and decreased ACSL4 mRNA expression levels(P＜0.01,P＜0.05).In cellular experiments,lipid droplets were significantly increased in the HepG2 cell M group compared with those in the C group,and lipid droplets were significantly reduced by Gegen QinLian Decoction and metformin.MDA and Fe2+levels were significantly increased and GSH levels were significantly decreased in the HepG2 M group compared with the levels in the C group(P＜0.01),while all doses of Gegen QinLian Decoction and metformin significantly decreased MDA and Fe2+levels(P＜0.01)and increased the GSH content(P＜0.01,P＜0.05).Nrf2,GSS,GPX4,and SLC7A11 mRNA expression levels in the GGQLT-H group,Nrf2,HO-1,and SLC7A11 in the GGQLT-L group,HO-1,SLC7A11,and GSS in the GGQLT-M group,and GSS,Nrf2,and HO-1 in the Met group were all significantly increased compared with the findings in the M group(P＜0.01,P＜0.05).ACSL4 mRNA expression levels were significantly decreased in the GGQLT-M and GGQLT-L groups and the Met group(P＜0.01,P＜0.05).Conclusions Gegen QinLian Decoction can improve NAFLD by inhibiting ferroptosis,and its mechanism may he related to regulation of the Nrf2/SLC7A 11/GPX4 signaling pathway.

Objective To investigate the mechanism of action of the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway in non-alcoholic fatty liver disease(NAFLD),and to explore the mechanism of Gegen QinLian Decoction for the treatment of NAFLD,using in vivo and in vitro experiments.Methods Rats were fed with high-fat chow for 24 weeks to induce NAFLD,and were then divided randomly into normal(C),model(M),high-,medium-,and low-dose Gegen QinLian Decoction(GGQLT-H,GGQLT-M,GGQLT-L),and metformin(Met)groups.From week 25 onwards,the rats were administered the corresponding drugs by gavage for 2 weeks according to the grouping,until sampling.Levels of the oxidative stress markers malondialdehyde(MDA)and glutathione(GSH)in the liver tissues were measured in each group using biochemical kits and ferrous iron(Fe2+)in rat liver tissues was detected using a Fe2+kit.Nrf2,heme oxygenase-1(HO-1),SLC7A11,glutathione synthetase(GSS),GPX4,and acyl coenzyme A synthetase 4(ACSL4)mRNA levels in rat liver tissues were measured by reverse transcription quantitative polymerase chain reaction.For cellular experiments lipid acc umulation was induced in HepG2 hepatocellular carcinoma cells using 1 mmol/L free fatty acid,to mimic the NAFLD in vitro model.Different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment.Lipid accumulation was detected in the cells in each group by Oil red O staining.The MDA and GSH contents of HepG2 cells in the different groups were determined using appropriate kits,and the ferrous contents were detected using a cell-specific ferrous kit.Expression levels of Nrf2,HO-1,SLC7A11,GSS,GPX4,and ACSL4 mRNA was detected in each group of cells using reverse transcription quantitative polymerase chain reaction.Results In the animal experiments,MDA and Fe2+liver levels were significantly higher in the M group than in the C group,while GSH levels were significantly lower(P＜0.01).GGQLT-H,GGQLT-M and Met groups showed significantly reduced MDA and Fe2+and elevated GSH levels compared with the M group(P＜0.01,P＜0.05).High-and medium-dose Gegen QinLian Decoction and metformin increased Nrf2,HO-1,GSS,and GPX4 mRNA and decreased ACSL4 mRNA expression levels(P＜0.01,P＜0.05).In cellular experiments,lipid droplets were significantly increased in the HepG2 cell M group compared with those in the C group,and lipid droplets were significantly reduced by Gegen QinLian Decoction and metformin.MDA and Fe2+levels were significantly increased and GSH levels were significantly decreased in the HepG2 M group compared with the levels in the C group(P＜0.01),while all doses of Gegen QinLian Decoction and metformin significantly decreased MDA and Fe2+levels(P＜0.01)and increased the GSH content(P＜0.01,P＜0.05).Nrf2,GSS,GPX4,and SLC7A11 mRNA expression levels in the GGQLT-H group,Nrf2,HO-1,and SLC7A11 in the GGQLT-L group,HO-1,SLC7A11,and GSS in the GGQLT-M group,and GSS,Nrf2,and HO-1 in the Met group were all significantly increased compared with the findings in the M group(P＜0.01,P＜0.05).ACSL4 mRNA expression levels were significantly decreased in the GGQLT-M and GGQLT-L groups and the Met group(P＜0.01,P＜0.05).Conclusions Gegen QinLian Decoction can improve NAFLD by inhibiting ferroptosis,and its mechanism may he related to regulation of the Nrf2/SLC7A 11/GPX4 signaling pathway.

Objective To establish a clinical-CT model,and to observe its value for evaluating lymphovascular invasion(LVI)and/or perineural invasion(PNI)in esophageal squamous cell carcinoma(ESCC).Methods Data of 156 ESCC patients were retrospectively analyzed.The patients were divided into positive group(n=58,LVI[+]and/or PNI[+])and negative group(n=98,LVI[-]and PNI[-])according to postoperative pathological results.Clinical and CT data were compared between groups.Logistic regression analysis was performed to establish a model,and its efficacy of evaluating ESCC LVI and/or PNI was analyzed.Results Significant differences of carcinoembryonic antigen(CEA),carbohydrate antigen 199(CA199),tumor thickness,tumor volume and CT venous phase value(CTV),the difference between CTV and CT plain phase value(CTP)(△CTV-P)and venous phase enhancement rate(V％)were found between groups(all P＜0.05),and the area under the curve(AUC)of the above parameters for evaluating ESCC LVI and/or PNI was 0.702,0.690,0.731,0.744,0.621,0.631 and 0.599,respectively.CEA,CA199,tumor thickness,tumor volume and CTV were all independent predictive factors for ESCC LVI and/or PNI.A combined model was established based on the above features,and its accuracy,sensitivity and specificity for evaluating ESCC LVI and/or PNI was 82.05％,65.52％and 91.84％,respectively,with AUC of 0.838,higher than that of each single parameter(all P＜0.05).Conclusion The established clinical-CT model could effectively evaluate ESCC LVI and/or PNI.

Objective:To identify diagnostic markers related to oxidative stress in chronic rhinosinusitis with nasal polyps (CRSwNP) by analyzing transcriptome sequencing data, and to investigate their roles in CRSwNP.Methods:Utilizing four CRSwNP sequencing datasets, differentially expressed genes (DEGs) analysis, weighted gene co-expression network analysis (WGCNA), and three machine learning methods for Hub gene selection were performed in this study. Subsequent validation was carried out using external datasets, as well as real-time quantitative polymerase chain reaction (Real-time qPCR), and immunofluorescence staining of clinical samples. Moreover, the diagnostic efficacy of the genes was assessed by receiver operating characteristic (ROC) curve, followed by functional and pathway enrichment analysis, immune-related analysis, and cell population localization. Additionally, a competing endogenous RNA (CeRNA) network was constructed to predict potential drug targets. Statistical analysis and plotting were conducted using SPSS 26.0 and Graphpad Prism9 software.Results:Through data analysis and clinical validation, CP, SERPINF1 and GSTO2 were identified among 4 138 DEGs as oxidative stress markers related to CRSwNP. Specifically, the expression of CP and SERPINF1 increased in CRSwNP, whereas that of GSTO2 decreased, with statistically significant differences ( P<0.05). Additionally, an area under the curve (AUC)>0.7 indicated their effectiveness as diagnostic indicators. Importantly, functional analysis indicated that these genes were mainly related to lipid metabolism, cell adhesion migration, and immunity. Single-cell data analysis revealed that SERPINF1 was mainly distributed in epithelial cells, stromal cells, and fibroblasts, while CP was primarily located in epithelial cells, and GSTO2 was minimally present in the epithelial cells and fibroblasts of nasal polyps. Consequently, a CeRNA regulatory network was constructed for the genes CP and GSTO2. This construction allowed for the prediction of potential drugs that could target CP. Conclusion:This study successfully identifies CP, SERPINF1 and GSTO2 as diagnostic and therapeutic markers related to oxidative stress in CRSwNP.

Objective To investigate the prevalence of Cryptosporidium infection and the distribution of parasite species and genotypes among HIV-positive individuals in Jiangxi Province. Methods HIV-positive individuals' sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023. Subjects' stool samples were collected, and genomic DNA was extracted from stool samples. Nested PCR assay was performed based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium, and Cryptosporidium gp60 gene was amplified in stool samples positive for the SSU rRNA gene. The second-round PCR amplification product was checked with 1.5% agarose gel electrophoresis, and the products of suspected positive amplifications were sequenced, followed by sequence alignment. The phylogenetic tree was created using the Neighbor-Joining method with the software MEGA 11.0, to characterize the species, genotypes and sub-genotypes of Cryptosporidium. Results A total of 382 HIV-positive individuals were enrolled, with two cases identified with Cryptosporidium infection (0.52% prevalence), and both cases had no abdominal pain or diarrhea. Following sequencing and sequence alignment, the gene sequences of these two Cryptosporidium isolates shared 99.76% and 99.88% similarity with the gene sequence of C. meleagridis isolates. Phylogenetic analysis based on the Cryptosporidium SSU rRNA gene sequence identified the species of these two Cryptosporidium-positive stool samples as C. meleagridis. Following nested PCR amplification of the Cryptosporidium gp60 gene, sequencing and sequence alignment, the two C. meleagridis isolates were characterized as III eA17G2R1 and III bA25G1R1a sub-genotypes, and the sub-genotype III bA25G1R1a was firstly described in humans. Conclusion The prevalence of Cryptosporidium is low among HIV-positive individuals in Jiangxi Province. The likelihood of Cryptosporidium infection cannot be neglected among HIV-positive individuals without diarrhea.

Lumbar peritoneal shunt (LPS) is the standard treatment for nonobstructive hydrocephalus. Shunt infection, overdrainage, bleeding, and cerebrospinal fluid leaks have been reported as LPS complications. We present a 70-year-old man who developed iatrogenic spondylodiscitis 2 weeks after LPS placement, experiencing severe back pain and neurological deficits. Despite the empiric antibiotics, his symptoms persisted. The patient underwent fully endoscopic debridement and drainage (FEDD) to address the infection without LPS removal. After the procedure, the patient experienced a significant reduction in pain. Even though pathogen cultures were negative, the empiric antibiotic treatment continued for 6 full weeks. The patient was able to ambulate with a thoracolumbar orthosis due to the lumbar kyphotic deformity. FEDD, in conjunction with effective antibiotics, offers rapid pain relief and functional improvement in iatrogenic spondylodiscitis, even with LPS placement. However, FEDD may not correct spinal deformities and is unsuitable for advanced spinal disease or instability. Early detection of spondylodiscitis is crucial for improved outcomes.

Extraforaminal lumbar disk herniation (E-LDH) constitutes a challenging subset of lumbar disk herniations. Minimally invasive methods like full-endoscopic lumbar discectomy provide benefits but carry risks of complications, such as postoperative dysesthesia and nerve root injury. A 62-year-old male with comorbidities presented with severe leg pain due to left L3/4 E-LDH. An L3 nerve block, guided by fluoroscopy, was performed with the patient in a right decubitus position. After alleviating the pain, the patient was placed in a prone position to undergo the extraforaminal full-endoscopic procedure under local anesthesia. The tip of the needle was targeted to the base of the superior articular process (SAP). Foraminoplasty was precisely performed with a high-speed diamond burr at the base of the SAP, facilitating the removal of the extraforaminal portion of the herniated disc. The procedure yielded immediate relief (visual analogue scale score reduced from 10 to 0). There was no leg pain or associated back pain at the 6-month follow-up. This approach demonstrates the feasibility of an extraforaminal full-endoscopic technique for E-LDH, overcoming challenges associated with the uncooperative patient due to the severe leg pain. However, the technique demands technical proficiency and warrants further research to establish its broader applicability.