BACKGROUND:How to effectively promote bone regeneration and bone reconstruction after bone injury has always been a key issue in clinical bone repair research.The use of biological and degradable materials loaded with bioactive factors to treat bone defects has excellent application prospects in bone repair. OBJECTIVE:To investigate the effect of polylactic acid-glycolic acid copolymer(PLGA)composite scaffold modified by lysine-grafted graphene oxide nanoparticles(LGA-g-GO)on osteogenic differentiation and new bone formation. METHODS:PLGA was dissolved in dichloromethane and PLGA scaffold was prepared by solvent evaporation method.PLGA/GO composite scaffolds were prepared by dispersing graphene oxide uniformly in PLGA solution.LGA-g-GO nanoparticles were prepared by chemical grafting method,and the PLGA/LGA-g-GO composite scaffolds were constructed by blending LGA-g-GO nanoparticles at different mass ratios(1%,2%,and 3%)with PLGA.The micromorphology,hydrophilicity,and protein adsorption capacity of scaffolds of five groups were characterized.MC3T3 cells were inoculated on the surface of scaffolds of five groups to detect cell proliferation and osteogenic differentiation. RESULTS AND CONCLUSION:(1)The surface of PLGA scaffolds was smooth and flat under scanning electron microscope,while the surface of the other four scaffolds was rough.The surface roughness of the composite scaffolds increased with the increase of the addition of LGA-g-GO nanoparticles.The water contact angle of PLGA/LGA-g-GO(3%)composite scaffolds was lower than that of the other four groups(P<0.05).The protein adsorption capacity of PLGA/LGA-g-GO(1%,2%,and 3%)composite scaffolds was stronger than PLGA and PLGA/GO scaffolds(P<0.05).(2)CCK-8 assay showed that PLGA/LGA-g-GO(2%,3%)composite scaffold could promote the proliferation of MC3T3 cells.Alkaline phosphatase staining and alizarin red staining showed that the cell alkaline phosphatase activity in PLGA/LGA-g-GO(2%,3%)group was higher than that in the other three groups(P<0.05).The calcium deposition in the PLGA/GO and PLGA/LGA-g-GO(1%,2%,and 3%)groups was higher than that in the PLGA group(P<0.05).(3)In summary,PLGA/LGA-g-GO composite scaffold can promote the proliferation and osteogenic differentiation of osteoblasts,and is conducive to bone regeneration and bone reconstruction after bone injury.

Objective:To explore the diagnostic value of shear wave elastography(SWE)for clinically significant prostate cancer(csPCa).Methods:We retrospectively analyzed the clinical data of 359 cases with suspected prostate cancer(PCa)in Baoji Central Hospital from June 2017 to July 2023.All the patients underwent the following examinations in the order of serum prostate-spe-cific antigen(PSA)testing,transrectal ultrasonography(TRUS),measurement of the stiffness of the entire prostate gland by SWE,and TRUS-guided prostate puncture biopsy.The stiffness of the entire prostate gland was defined as the average of Young's modulus at both sides of the base,middle,and apex of the prostate,including the maximum Young's modulus(Emax),mean Young's modulus(Emean),and minimum Young's modulus(Emin).We analyzed the correlation of the parameters of the stiffness of the entire prostate gland with the pathological results,focusing on their diagnostic performance for csPCa.Results:Of the 359 cases,189 were diag-nosed by pathological puncture biopsy as BPH,26 as non-csPCa,and 144 as csPCa.The PSA level,Emax,Emean and Emin were significantly higher in the csPCa than those in the BPH and non-csPCa groups(all P＜0.01),but showed no statistically significant difference between the BPH and non-csPCa groups(all P＞0.05).The area under the receiver operating characteristic curve(AUC),optimal cut-off value,sensitivity,specificity,positive predictive value(PPV),negative predictive value(NPV)and accura-cy of Emax in the diagnosis of csPCa were 0.852,143.92 kPa,72.22％,84.65％,75.91％,81.98％and 79.67％;those of Emean were 0.868,82.42 kPa,67.36％,91.16％,83.62％,80.66％and 81.62％;and those of Emin were 0.682,32.73 kPa,47.22％,89.30％,73.91％,71.54％and 72.14％,respectively.In the non-csPCa group,Emax,Emean and Emin were found be-low the optimal cut-off value in 73.08％(19/26),92.31％(24/26)and 88.46％(23/26),respectively.Conclusion:The stiff-ness of the entire prostate gland measured by SWE contributes to the diagnosis of csPCa,reduces unnecessary detection of non-csPCa,and provides some reference for its active surveillance.

To screen new antigens for novel tuberculosis(TB)vaccine research,we used bioinformatics to predict the B and T cell epitopes of Rv2318,and evaluated the immunogenicity of Rv2318 and its T/B epitope peptides(Rv2318p).The recombinant plas-mids pET32a-Rv2318 and pET32a-Rv2318p were constructed through gene synthesis methods.The recombinant proteins were ex-pressed in a prokaryotic system and purified with nickel affinity chromatography.Proteins were identified with SDS-PAGE and western blotting.BALB/c mice were immunized subcutaneously with the recombinant proteins to evaluate immunogenicity.Sera were collected,and antigen specific antibody titers were evaluated with ELISA.Splenocytes were isolated,and cytokines and T cell proliferation were analyzed with ELISA and flow cytometry,respectively.Rv2318 included two epitope fragments,aa10-130 and 350-410.SDS-PAGE and western blotting indicated that the target proteins were expressed and purified correctly,and their relative molecular weights were-approximately 68 kD and 42 kD,respectively.Rv2318 and Rv2318p induced stronger humoral immune responses than observed in the control groups(P<0.000 1,n=6).Compared with Rv2318,Rv2318p showed significantly greater enhancement of specific IgG and IgG subclass antibodies(P<0.000 1,n=6).In addition,Rv2318p increased the ratio of IgG2a/IgG1,thus indicating that it primarily induced a cellular immune response biased toward the Th1 type.Cytokine experiments revealed that IFN-γ,IL-2,IL-6,and IL-4 significantly increased after immunization with Rv2318p(P all<0.01,n=6),particularly Th1 type cytokines(IFN-γ and IL-2).Furthermore,Rv2318 increased the expression of only IL-2 and IL-6,particularly IL-6(P all<0.01,n=6).Although Rv2318 in-duced more IFN-γ,we observed no significant difference between Rv2318 and PBS immunized mice.Importantly,Rv2318p stimu-lated mice to express IFN-γ at 842 pg/mL,approximately 3 times the level elicited by Rv2318.Whereas both proteins increased the proportions of CD4+and CD8+T cells,Rv2318p promoted greater proliferation of T lymphocytes.These data indicated that both Rv2318 and its epitope peptides enhanced humoral and cellular immune responses,whereas the epitope peptides notably triggered a stronger Th1 type cellular response.In conclusion,the recombinant protein Rv2318 and its epitope peptides showed favorable immunogenicity,and the immunogenicity of Rv2318p was superior to that of Rv2318.This study provides a theoretical basis for TB vaccine development.

Objective:To analyze the predictive factors associated with late-onset sepsis(LOS) in very low birth weight infants,and to develop a risk prediction score scale applicable to these infants three days postnatal.This will provide valuable insights for early diagnosis and timely intervention.Methods:Very low birth weight infants admitted to the Children's Hospital of Fudan University from January 1,2022,to June 30,2024,were selected as research subjects.These infants were categorized into two groups:the LOS group and the non-LOS group,based on whether they developed LOS.LASSO regression analysis,alongside univariate and multivariate regression analyses,was employed to identify predictive factors for LOS in this population.A Logistic model was constructed using the optimal combination of predictive variables,and a risk assessment scale was subsequently developed.The prediction performance of the model was evaluated using the Hosmer-Lemeshow chi-square test and the receiver operating characteristic curve.Results:A total of 444 cases of very low birth weight infants were included,of which 185 had LOS and 259 did not.After screening the variables,seven independent factors were included into the model:birth weight,gestational age,tracheal intubation,abnormal skin color,abdominal distension,elevated C-reactive protein levels,and right hand perfusion index.A predictive scoring scale was developed based on the regression coefficients of each variable,with corresponding risk scores assigned as follows:1,4,3,2,1,1,and 2; a score of ≥3.5 indicated high-risk groups.The Hosmer-Lemeshow test results demonstrated that χ2 = 7.602( P = 0.473).The area under the receiver operating characteristic curve was 0.792 ( P<0.001),with a sensitivity of 73.5% and specificity of 71.0%. Conclusion:The risk score scale developed in this study exhibits significant predictive capability,providing valuable insights for clinical medical personnel to assess the risk of LOS in very low birth weight infants during the early postnatal period.

Gastric cancer remains one of the most prevalent and lethal malignancies of the digestive tract worldwide,underscoring the urgent need for more effective targeted therapeutic strategies.Poly(ADP-ri-bose)polymerase(PARP)inhibitors have demonstrated remarkable efficacy in tumors with homologous recombination repair(HRR)deficiency;however,their clinical application in gastric cancer remains limited.Clinical evidence suggests that patients harboring Helicobacter pylori infection in combination with HRR gene mutations exhibit a significantly elevated risk of developing gastric cancer,thereby supporting the potential benefit of PARP inhibition in this setting.In this study,a kinase inhibitor library was screened in combination with the PARP inhibitor olaparib in gastric cancer cells.And we identify the cy-clin-dependent kinase 8/19(CDK8/19)inhibitor Senexin A as a compound that synergistically enhances the cytotoxic effect of PARP inhibition(P<0.05).Phenotypic validation using CCK-8 and colony for-mation assays demonstrated that the combination treatment significantly suppressed cellular proliferation and clonogenic potential compared to either monotherapy(P<0.0001).Mechanistically,alkaline comet assays revealed a significant increase in DNA damage in the combination treatment group relative to either single-agent group(P<0.0001),suggesting that the synergistic effect results from the exacerbation of DNA damage via impaired DNA repair mechanisms.In addition,treatment with CDK8/19 inhibitors a-lone markedly increased the formation of γH2AX and 53BP1 foci in irradiated gastric cancer cells(P<0.0001),indicating inhibition of DNA damage repair pathways.Transcriptome sequencing further re-vealed that CDK8/19 inhibition impacts critical cellular pathways,including DNA repair,cell cycle reg-ulation,and RNA splicing.Co-immunoprecipitation assays confirmed that inhibition of CDK8/19 kinase activity significantly reduces the phosphorylation level of PARP1,suggesting a potential regulatory inter-action.Immunohistochemical analysis of tumor and adjacent non-tumor tissues from gastric cancer pa-tients demonstrated that CDK8 is significantly overexpressed in tumor tissues,supporting its potential as both a prognostic biomarker and a therapeutic target.Collectively,this study elucidates a mechanistic ba-sis by which CDK8/19 inhibition enhances the sensitivity of gastric cancer cells to PARP inhibitors.These findings provide a strong rationale for the combined use of CDK8/19 and PARP inhibitors as a tar-geted therapeutic strategy and offer promising translational implications for advancing personalized medi-cine in gastric cancer treatment.

Objective:To summarize the technical difficulties, key points and cosmetic effects of Ti-loop patch application in breast shaping of immediate reconstruction with omental flap transplantation after mastectomy.Methods:The clinical data of 90 cases with breast cancer who underwent immediate breast reconstruction by using laparoscopically harvested omental flap alone or combined with prosthesis after nipple and areola complex sparing mastectomy from May. 2020 to Jan. 2023 in Hunan Cancer Hospital was retrospectively analyzed. All patients were female between 16 and 55 years old with the average age of 42. Their courses of the disease range from 5 to 26 months with the mean of 15.5 months. These patients received mastectomy with conservation of nipple and areola, and then the greater omentum with right pedicle were harvested laparoscopically. Ti-loop patch was used to hang and fix the greater omentum as a support. 27 patients accepted immediate breast reconstruction with pedicled greater omentum alone, while 63 subjects with greater omentum and gel prosthesis. Among them, contralateral breast contraction and lift was operated on 9 cases.Results:All the pedicled greater omentum successfully survived, and wound surfaces of donor and recipient sites healed at stage I. Axillary lymph node dissection and postoperative radiotherapy were performed on 14 patients with positive pathological examination of sentinel lymph nodes. Breasts of two patients were found to be smaller after postoperative radiotherapy. All 90 subjects were followed up for 5 to 32 months, and the average follow-up period was 16.4 months. The reconstructed breasts were good in shape and tactile sensation. Only punctate scars were observed in donor sites of abdomen. All patients recovered with no upper limb lymphedema, no short and long term complications in abdomen, and only one patient had papillary recurrence without distant metastasis.Conclusions:The omental flap is hard to be sutured and shaped in the reconstructed breast, while easily to be avulsed and form hematoma. The application of Ti-loop patch, which could semi-surround and wrap the greater omentum, and suspend and fix it on the chest wall in upper pole of the breast as a support, can significantly improve the reconstruction effect.

Objective To investigate the differential diagnostic value of nomogram based on CT imaging features for bone island(BI)and osteoblastic metastases(OBM).Methods A retrospective analysis was conducted on the clinical and imaging data of 374 patients with spinal BI and OBM who underwent chest or abdominal CT scans.There were 290 patients with 445 lesions in BI group,and 84 patients with 449 lesions in OBM group.Multivariate logistic regression analysis was used to screen relevant variables,including age,gender,maximum CT value,average CT value,length and width,density uniformity,margin,and location of each lesion,to establish a nomo-gram prediction model.The receiver operating characteristic(ROC)curve and area under the curve(AUC)were used to evaluate the differential diagnostic performance of the nomogram model for BI and OBM.Results Univariate analysis revealed that statistically significant differences between the two groups in terms of age,maximum CT value,average CT value,spinous process sign,proximity to the bone cortex,vertebral location,length,and length/width ratio(P<0.05).Multivariate logistic regression analysis showed that average CT value,spinous process sign,proximity to the bone cortex,vertebral location,length,and length/width ratio were independent factors differentiating for BI and OBM(P<0.05).The AUC of ROC curves for the training and validation sets of the nomogram model,constructed based on average CT value,spinous process sign,proximity to the bone cortex,vertebral location,length,and length/width ratio,were 0.976 and 0.935,respectively.The sensitivity and specificity of the model's training set were 0.942 and 0.902,respectively,while those of the validation set were 0.932 and 0.799,respectively.Conclusion The nomogram model based on average CT value,spinous process sign,proximity to the bone cortex,vertebral location,length,and length/width ratio can effectively differentiate BI from OBM,providing valuable assistance for clinical decision-making.

Objective To explore the advantage of advanced diffusion weighted imaging(DWI)over single index DWI in distin-guishing low-grade transitional zone prostate cancer(TZ-PCa)from benign prostatic hyperplasia(BPH)with inflammation.Methods Thirty-two patients with low-grade TZ-PCa(low-grade TZ-PCa group)and 38 patients with BPH with inflammation(BPH with inflammation group)confirmed by pathology were selected.All patients underwent routine MR sequence,single index DWI,and advanced DWI[including introvoxel incoherent motion diffusion weighted imaging(IVIM-DWI),stretched-exponential model(SEM),diffu-sion kurtosis imaging(DKI)]sequences scans.The related parameters of the lesion area[including apparent diffusion coefficient(ADC),diffusion coefficient(D),distributed diffusion coefficient(DDC),and mean kurtosis(MK)values]were measured.Independ-ent sample t-tests were used to compare differences in parameters values between the two groups.The receiver operating characteris-tic(ROC)curve was used to calculate the area under the curve(AUC)of varous parameters values in the differential diagnosis of low-grade TZ-PCa and BPH with inflammation.The difference in AUC between advanced DWI and single index DWI was compared using Z-test.Results The ADC,D,and DDC values of the low-grade TZ-PCa group were lower than those of the BPH with inflammation group(P<0.05).The MK value of the low-grade TZ-PCa group was higher than that of the BPH with inflammation group(P<0.05).The D,DDC,and MK values,and a combination of these parameters(D+DDC+MK)exhibited high diagnostic effi-cacy(all AUC>0.9).The diagnostic efficacy of these parameters was significantly higher than that of the ADC value(AUC 0.902,0.904,0.966,0.994 vs 0.725,all P<0.05).The combined param-eter demonstrated the highest AUC.Although the diagnostic efficacy of the combined parameter was not statistically different from the individual D,DDC,and MK values(P>0.05),the diagnostic specificity was significantly improved compared to the individual parame-ters.Conclusion Advanced DWI(IVIM-DWI,SEM,DKI)has a higher diagnostic efficacy in distinguishing low-grade TZ-PCa from BPH with inflammation than single index DWI.Combining the parameters(D+DDC+MK)further improves diagnostic specificity.

Objective:To explore the impacts of 131I-NaI radiotherapy on the promotion of glycolysis and 18F-fluorodeoxyglucose ( 18F-FDG) uptake in pancreatic cancer cells via the induction of proviral integration moloney murineleukemia virus 2 (PIM2) and 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3 (PFKFB3). Methods:In the cell experiments, human pancreatic carcinoma cells-1 (PANC-1) were randomly divided into four groups: a control group, an HJ-PI01 group, a 131I-NaI group, and an HJ-PI01 + 131I-NaI group. Their aerobic glycolysis capacity was assessed by measuring glucose uptake, lactate production, and extracellular acidification rate (ECAR). In vivo animal experiments, 12 nu/nu female nude mice were given 100 μl (1 × 10 7 cells) of cell suspension through subcutaneous injection into the left lower limbs. When the tumor volume reached approximately 60 mm 3, these mice were divided into four groups (a control group, a HJ-PI01 group, a 131I-NaI group, and an HJ-PI01+ 131I-NaI group) using a random number table, with three mice in each group. After 14 days of treatment, 18F-FDG PET/CT imaging was performed to calculate the maximum standardized uptake value (SUV max) of the xenografts. Following PET/CT imaging, the tumor tissues were harvested and analyzed for PIM2, PFKFB3, and Ki-67 expressions using immunohistochemistry. Results:In cell experiments, compared to the control group, the HJ-PI01 group exhibited significant reduction in glucose uptake, lactate production, PFKFB3 protein expression, and ECAR in PANC-1 cells ( t = 4.59-13.98, P < 0.05). In contrast, the 131I-NaI group showed significant increases in these parameters ( t = 3.36-13.97, P < 0.05). Compared to the 131I-NaI group, the HJ-PI01+ 131I-NaI group showed significant reduction in glucose uptake, lactate production, PFKFB3 protein expression, and ECAR ( t = 5.14-20.87, P < 0.05). In the animal experiments, compared to the control group, the three groups displayed significant decrease in SUV max of 18F-FDG uptake in tumors ( t = 16.48, 22.49, 32.64, P < 0.001). Moreover, the HJ-PI01 + 131I-NaI group exhibite significantly lower SUV max than the 131I-NaI group ( t = 10.16, P < 0.001). Immunohistochemical analysis revealed that the HJ-PI01+ 131I-NaI group, compared to the 131I-NaI group, showed significantly lower Ki-67 expression and the PIM2/PFKFB3 signaling pathway in tumor tissues ( t = 3.27, 10.73, 14.85, P < 0.05). Conclusions:Glycolysis enhancement of PANC-1 cells, mediated by the PIM2/PFKFB3 signaling pathway inhibition, can significantly improve the anticancer capacity of 131I-NaI, providing a novel strategy for radiotherapy in pancreatic cancer.

AIM To evaluate the quality of Rubi Fructus.METHODS UPLC-Q-TOF-MS/MS was adopted in the component identification,after which the HPLC fingerprints were established,cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were used for chemical pattern recognition.and the contents of chlorogenic acid,ferulic acid,ellagic acid,isoquercitrin,kaempferol-3-O-rutinoside,astragalin,tiliroside quercetin,kaempferol were determined.RESULTS Total 34 constituents were identified.There were 19 common peaks in the fingerprints for 31 batches of medicinal materials with the similarities of more than 0.8.Wild varieties and cultivated varieties,and medicinal materials from different producing areas could be distinguished;4 principal components demonstrated the accumulative variance contribution rate of 84.142％;8 differential components were screened,2 of which were ellagic acid and astragalin.Ellagic acid and astragalin displayed higher contents in the wild varieties than those in the cultivated varieties(P＜0.05,P＜0.01).CONCLUSION UPLC-Q-TOF-MS/MS,HPLC fingerprints combined with content determination can be used for the quality control of Rubi Fructus.