Objective As one of the adult stem cells, adi pose-derived stem cells (ADSCs) have become an important seed cell source for tissue engineering recently. But whether the thawed cryopreserved ADSCs could be used to tissue engineered bone remains unknown. To investigate the effect of cryopreservation on the growth and osteogenesis of ADSCs invitro. Methods The ADSCs were isolated from the adipose aspirates by collagenase digestion method. For the experimental group, the 2nd generation cells were stored with a simple method of cryopreservation by slow cool ing with dimethyl sulphoxide as a cryoprotectant and rapid thawing. After cryopreserved in l iquid nitrogen for 4 weeks, ADSCs were recovered and cultured in osteogenic media, with non-cryopreserved ADSCs as the control group. The osteogenic differentiation was evaluated by alkal ine phosphatase (ALP) staining and Al izarin red O staining at 2 and 3 weeks respectively. The cell growth and osteogenesis of ADSCs were further determined using DNA assay and the ALP activity and calcium content were measured. Results The survival percentage of the cryopreserved cells was 90.44% ± 2.62%. The cell numbers and ALP activity increased with osteogenic induction time, and reach plateaus at 7 days and 11 days, respectively. The ALP staining and Al izarin red O staining results were both positive at 2 weeks and 3 weeks after osteogenic induction, respectively. And no significant difference in the cells number, ALP activity, and calcium content were found between experimental group and control group (P gt; 0.05). Conclusion Cryopreservation does not affect the growth and osteogenesis of ADSCs, and the cryopreserved ADSCs can be used as cell source for tissue engineered bone.
Objective Calcium phosphate bioceramics has a broad appl ication prospect because of good biocompatibil ity, but porous scaffolds with complex shape can not be prepared by the traditional methods. To fabricate porous calcium phosphate ceramics by rapid prototyping and to investigate the in vitro osteogenic activities. Methods The porous calcium phosphate ceramics was fabricated by rapid prototyping. The bone marrow mesenchymal stem cells (BMSCs)were isolated from bone marrow of Beagle canine, and the 3rd passage BMSCs were seeded onto the porous ceramics. The cell/ceramics composite cultured in osteogenic medium were taken as the experimental group (group A) and the cell/ceramics composite cultured in growth medium were taken as the control group (group B). Meanwhile, the cells seeded on the culture plate were cultured in osteogenic medium or growth medium respectively as positive control (group C) or negative control (group D). After 1, 3, and 7 days of culture, the cell prol iferation and osteogenic differentiation on the porous ceramics were evaluated by DNA quantitative analysis, histochemical staining and alkal ine phosphatase (ALP) activity. After DiO fluorescent dye, the cell adhesion, growth, and prol iferation on the porous ceramics were also observed by confocal laser scanning microscope (CLSM). Results DNA quantitative analysis results showed that the number of BMSCs in all groups increased continuously with time. Plateau phase was not obvious in groups A and B, but it was clearly observed in groups C and D. The CLSM observation indicated that the activity of BMSCs was good and the cells spread extensively, showing good adhesion and prol iferation on the porous calcium phosphate ceramics prepared by rapid prototyping. ALP quantitative analysis results showed that the stain of cells on the ceramics became deeper and deeper with time in groups A and B, the staining degree in group A were ber than that in group B. There was no significant difference in the change of the ALP activity among 4 groups at the first 3 days (P gt; 0.05); the ALP activity increased obviously in 4 groups at 7 days, group A was significantly higher than other groups (P lt; 0.05) and groups C, D were significantly higher than group D (P lt; 0.05). Conclusion The porous calcium phosphate ceramics has good cytocompatibil ity and the designed pores are favorable for cell ingrowth. The porous ceramicsfabricated by rapid prototyping has prominent osteogenic differentiation activity and can be used as a choice of scaffolds for bone tissue engineering.
ObjectiveTo develop a nomogram prediction model for predicting the objective response rate (ORR) in patients with unresectable hepatocellular carcinoma (uHCC) receiving transarterial chemoembolization (TACE) combined with targeted therapy and immunotherapy. MethodsClinicopathological data of uHCC patients treated with TACE combined with targeted therapy and immunotherapy at Leshan People’s Hospital from January 2018 to November 2023 were collected. Patients were randomly divided into a training set and a validation set at a 7∶3 ratio. Univariate and multivariate logistic regression analyses were used to identify independent risk factors for ORR. Variables with P<0.05 in the multivariate analysis, together with clinically important factors [such as Eastern Cooperative Oncology Group performance status (ECOG PS) score and portal vein tumor thrombus (PVTT)], were incorporated into the nomogram model. Model performance and clinical utility were assessed using the area under the receiver operating characteristic curve (AUC), concordance index (C-index), calibration curves, and decision curve analysis. ResultsA total of 105 patients with uHCC were enrolled, including 73 in the training set and 32 in the validation set. Multivariate logistic regression analysis identified alpha-fetoprotein (AFP), Barcelona Clinic Liver Cancer (BCLC) stage, and prothrombin time (PT) as independent predictors of ORR (P<0.05). The nomogram incorporating these factors along with ECOG PS score and PVTT achieved AUCs (95%CI) of 0.81 (0.71, 0.91) in the training set and 0.80 (0.64, 0.96) in the validation set. Bootstrap internal validation (1 000 resamples) yielded a mean C-index (95%CI) of 0.76 (0.63, 0.89). The Hosmer-Lemeshow test indicated good model fit (training set: χ2=5.64, P=0.58; validation set: χ2=3.89, P=0.69), and calibration curves showed close alignment with the ideal diagonal in both sets. Decision curve analysis demonstrated positive net clinical benefit within threshold probability ranges of 0.02–0.78 (training set) and 0.10–0.80 (validation set). ConclusionThe nomogram prediction model based on AFP, BCLC stage, PT, ECOG PS score, and PVTT effectively predicts ORR in uHCC patients receiving TACE combined with targeted therapy and immunotherapy, providing a reference for individualized treatment decisions.
ObjectiveTo evaluate the safety and efficacy of extra-sheath dissection via the Laennec membrane space combined with indocyanine green (ICG) fluorescence navigation in laparoscopic anatomic hepatectomy. MethodsA retrospective analysis was conducted on the clinical data of hepatocellular carcinoma patients who underwent laparoscopic anatomic hepatectomy at Leshan People’s Hospital between January 2022 and September 2025. The study group comprised patients who underwent the extra-sheath anatomical approach via the Laennec’s capsule space combined with ICG fluorescence navigation, while the control group consisted of patients who underwent the conventional intra-sheath Glissonean pedicle transection approach combined with intraoperative ultrasound. ResultsA total of 113 patients were included in this study, including 54 cases in the study group and 59 cases in the control group. Intraoperative blood loss [MD=–139.88 mL, 95%CI (–229.51, –50.25) mL], number of drainage tubes × drainage tube indwelling time [MD=–2.00 (tubes×d), 95%CI (–4.00, 0.00) (tubes×d)], postoperative ambulation time [MD=–0.58 d, 95%CI (–0.94, –0.22) d], alanine aminotransferase level on postoperative day 1 [MD=–129.83 U/L, 95%CI (–259.96, –5.71) U/L], aspartate aminotransferase level on postoperative day 1 [MD=–131.09 U/L, 95%CI (–259.12, –3.06) U/L], albumin level on postoperative day 1 [MD=1.64 g/L, 95%CI (0.21, 3.06) g/L], and hospitalization cost [MD=–5 523.10 yuan, 95%CI (–10 572.42, –471.77) yuan] of the study group were all better than those in the control group (P< 0.05). ConclusionExtra-sheath dissection via the Laennec membrane space combined with ICG fluorescence navigation can improve the safety and efficacy of laparoscopic anatomical hepatectomy, and is worthy to explore further its clinical application value.
ObjectiveTo investigate the prognostic value of the alkaline phosphatase + gamma-glutamyl transpeptidase / platelet (AGPR) in predicting overall survival in patients with hepatocellular carcinoma (HCC) after curative hepatectomy, and to develop an AGPR-based nomogram prediction model to enhance the accuracy of prognostic risk stratification and individualized treatment decision-making. MethodsThis multicenter retrospective study included patients who underwent radical resection for HCC at the Affiliated Hospital of Southwest Medical University (2013–2023) as the training and internal validation sets, and at Leshan People’s Hospital (2018–2024) as the external validation set. Baseline characteristics, preoperative laboratory tests, surgical details, and pathological findings were collected. Optimal cutoff values for AGPR and the pan-immune-inflammation value (PIV) were determined using receiver operating characteristic (ROC) curves. Independent prognostic factors for overall survival were identified using univariate and multivariate Cox regression, then a nomogram was developed. Model performance was assessed using the area under the ROC curve (AUC), concordance index (C-index), calibration, and decision curve analysis for clinical utility assessment. Patients were stratified into low- and high-risk groups based on the median nomogram score, and overall survival was compared. ResultsA total of 618 patients were included: 250 in the training set, 167 in the internal validation set, and 201 in the external validation set. Optimal cutoff values were 1.06 for AGPR and 316.79 for PIV. Multivariate Cox regression identified alpha-fetoprotein (AFP), AGPR, PIV, and tumor diameter as independent prognostic factors (P＜0.05). The nomogram predicting 1-, 3-, and 5-year overall survival rates achieved AUCs of 0.820, 0.734, and 0.727 in the training set; 0.784, 0.722, and 0.705 in the internal validation set; and 0.789, 0.703, and 0.702 in the external validation set. C-indices were 0.748, 0.701, and 0.692, respectively. The Hosmer-Lemeshow test indicated good calibration (P>0.05), and calibration curves aligned closely with the ideal diagonal. Decision curve analysis demonstrated clinical net benefit across all datasets. Low-risk patients had significantly better overall survival than high-risk patients (P＜0.05). ConclusionsAGPR is an independent prognostic factor for overall survival in HCC patients after radical resection. The nomogram incorporating AGPR, AFP, PIV, and tumor diameter exhibits favorable discrimination, calibration, and clinical utility, supporting its use for postoperative risk stratification and individualized treatment planning.
