AIM:To observe the effect and mech-anism of cuttlebone extract regulating vascular en-dothelial growth factor(VEGF)/phosphatidylinosi-tol 3-kinase(PI3K)/protein kinase B(AKT)pathway on refractory wounds in rats.METHODS:Cuttle-bone extract(CE)was obtained by water extraction of cuttlebone.Fifty SD rats were randomly divided into negative Control group,Model group,Cuttle-bone extract low-dose(CE(L))group,Cuttlebone extract high-dose(CE(H))group,and cuttlebone ex-tract high-dose+inhibitor(CE(H)+LY294002)group.After the refractory wound model was successfully established,0.02％furacillin solution or cuttlebone extract solution were applied to the wound area of rats in each group,and the treatment was adminis-tered once a day.After 14 days of treatment for re-fractory wounds,the changes in wound healing,angiogenesis,inflammation and expression of relat-ed regulatory proteins were quantitatively ana-lyzed by measuring skin ulcer wound area,patho-logical sections,immunofluorescence staining,Eli-sa,Western blot,RT-qPCR and other methods.RE-SULTS:Compared with Model group,CE(L)and CE(H)groups can increase the number of epithelial cells and collagen,and promote the healing of re-fractory wound in rats.Serum VEGF,skin tissue mi-crovascular density,P-PI3K,P-AKT,VEGF protein ex-pression and mRNA expression levels of PI3K,Akt,VEGF and eNOS were increased(P＜0.05),while se-rum TNF-α and IL-6 levels were decreased(P＜0.05).LY294002 could partially reverse the repair-ing effect of high dose cuttlebone extract on refrac-tory wound(P＜0.05).CONCLUSION:Cuttlebone ex-tract can regulate the VEGF/PI3K/AKT signaling pathway,inhibit the inflammatory response of re-fractory wounds in rats,induce angiogenesis and promote wound healing.

Liver cancer is one of the most common malignant tumors worldwide. Currently, the available treatment methods cannot fully control its recurrence and mortality rate. Establishing appropriate animal models for liver cancer is crucial for developing new treatment technologies and strategies. The patient-derived xenograft (PDX) model preserves the tumor's microenvironment and heterogeneity, which makes it advantageous for biological research, drug evaluation, personalized medicine, and other purposes. This article reviews the development, preparation techniques, application fields, and challenges of PDX models in liver cancer, providing insights for the research and exploration of PDX models in diagnostic and therapeutic strategies of liver cancer.
Liver Neoplasms/drug therapy* ; Animals ; Humans ; Xenograft Model Antitumor Assays/methods* ; Mice ; Disease Models, Animal

Ulcerative colitis (UC) is a chronic inflammatory disorder with a complex etiology, characterized by intestinal inflammation and barrier dysfunction. Platycodon grandiflorus polysaccharides (PGP), the primary component of Platycodon grandiflorus, and hesperidin (Hesp), a prominent active component in Citrus aurantium L. (CAL), have both demonstrated anti-inflammatory properties. This study aims to elucidate the underlying mechanism of the synergistic effect of PGP combined with Hesp on UC, focusing on the coordinated interaction between the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathways. A mouse model of UC induced by dextran sulfate sodium (DSS) and a cell model using lipopolysaccharide (LPS)-induced RAW264.7/IEC6 cells were employed to investigate the in vitro and in vivo anti-inflammatory effects of PGP combined with Hesp on UC and its potential mechanism of action. The results indicated that compared to the effects of either drug alone, the combination of PGP and Hesp significantly modulated inflammatory factor levels, inhibited oxidative stress, regulated colonic mucosal immunity, suppressed apoptosis, and restored intestinal barrier function in vitro and in vivo. Further in vitro studies revealed that PGP significantly inhibited the PI3K/AKT signaling pathway, while Hesp significantly inhibited the JAK2/STAT3 signaling pathway. The use of inhibitors and activators targeting both pathways validated the synergistic effects of PGP combined with Hesp on the PI3K/AKT and JAK2/STAT3 signaling pathways. These findings suggest that PGP combined with Hesp exhibits a synergistic effect on DSS-induced colitis, potentially mediated through the phosphatase and tensin homolog (PTEN)/PI3K/AKT and interleukin-6 (IL-6)/JAK2/STAT3 signaling pathways.
Animals ; STAT3 Transcription Factor/genetics* ; Janus Kinase 2/genetics* ; Polysaccharides/administration & dosage* ; Colitis, Ulcerative/chemically induced* ; Mice ; Signal Transduction/drug effects* ; Proto-Oncogene Proteins c-akt/genetics* ; Drug Synergism ; Male ; Hesperidin/administration & dosage* ; Platycodon/chemistry* ; Phosphatidylinositol 3-Kinases/genetics* ; Disease Models, Animal ; RAW 264.7 Cells ; Mice, Inbred C57BL

L-citrulline is a nonprotein amino acid that plays an important role in human health and has great market demand. Although microbial cell factories have been widely used for biosynthesis, there are still challenges such as genetic instability and low efficiency in the biosynthesis of L-citrulline. In this study, an efficient, plasmid-free, non-inducible L-citrulline-producing strain of Escherichia coli BL21(DE3) was engineered by combined strategies. Firstly, a chassis strain capable of synthesizing L-citrulline was constructed by block of L-citrulline degradation and removal of feedback inhibition, with the L-citrulline titer of 0.43 g/L. Secondly, a push-pull-restrain strategy was employed to enhance the L-citrulline biosynthesis, which realized the L-citrulline titer of 6.0 g/L. Thirdly, the NADPH synthesis and L-citrulline transport were strengthened to promote the synthesis efficiency, which achieved the L-citrulline titer of 11.6 g/L. Finally, fed-batch fermentation was performed with the engineered strain in a 3 L fermenter, in which the L-citrulline titer reached 44.9 g/L. This study lays the foundation for the industrial production of L-citrulline and provides insights for the modification of other amino acid metabolic networks.
Citrulline/biosynthesis* ; Escherichia coli/genetics* ; Metabolic Engineering/methods* ; Fermentation ; NADP/biosynthesis*

Objective:To establish a tumor tissue xenograft (PDX) model derived from liver cancer patients and explore the factors affecting tumorigenicity of liver cancer in the PDX model.Methods:The hepatocellular carcinoma tissues were inoculated subcutaneously in the axilla of NPG mice using the tissue block method to establish a PDX model. The demographic characteristics and related clinical examination data of 60 hepatocellular carcinoma patients were collected using the electronic medical record system and comprehensive medical information system of Beijing You'an Hospital, affiliated to Capital Medical University. The hepatocellular carcinoma samples of 24 cases were sequenced using the Oak Wing TM-808 gene detection reagent and high-throughput sequencing technology. SPSS 17.0 statistical software was used for statistical analysis, and the count data were analyzed using the χ2 test. Results:The tumorigenicity rate of PDX samples from 60 patients with liver cancer was 35% (21/60). The average tumorigenic duration in the PDX-P0 generation was 110.71±50.45 days. There were statistically significant differences ( P<0.05) corresponding to Edmondson grade ( χ2=5.910, P=0.015) and Ki67 expression ( χ2=4.615, P=0.032) among PDX with tumorigenicity and without tumorigenicity between the liver cancer samples. There was no statistically significant difference in gene mutation (TOP25) among PDX with tumorigenicity and without tumorigenicity between liver cancer samples. Conclusion:The factors affecting the tumorigenicity of liver cancer in PDX models are complex. The high pathological grade and strong Ki67 expression may be the key factors for the completion of liver cancer in PDX models.

Objective:To investigate the clinical effect of reconstruction of the degloving injury of thumb and finger with free perforator flap of superficial peroneal artery.Methods:This is a retrospective study. From June 2020 to June 2023, 18 superficial peroneal artery perforator flaps from 14 calves were used to treat 15 degloved digital wounds of 14 patients in the Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital. There were 13 single digital degloving wounds and 1 two-digital degloving wounds. Of which, 3 digits were reconstructed with 2 free perforator flaps of ipsilateral superficial peroneal artery and 12 with a single free perforator flap of superficial peroneal artery. The size of wounds was 4.0 cm×2.0 cm-10.0 cm×4.0 cm, and the flaps were 5.0 cm×3.0 cm-12.0 cm×4.0 cm in size. The donor sites in calves were sutured layer by layer with absorptive sutures. Postoperative follow-ups were conducted through regular outpatient visits, phone calls or WeChat. Survival of flaps, postoperative complications, therapeutic effect of the flaps and patient satisfaction were observed.Results:There was no vascular compromise or wound infection of the flaps. All patients were included in the 4 to 36 months of postoperative follow-up, with an average of 9.86 months. All flaps had good appearance, without obvious swelling. Colour and texture of the flaps was close to the surrounding skin. The flaps had no obvious pigmentation or ulcer and scar hyperplasia at recipient site. Sensation of the flaps recovered to S 2 to S 3. There was no obvious scar hyperplasia, pain or dysfunction at donor sites. According to the comprehensive evaluation scale of flap, the scores were found at 81 to 91, with an average score of 85 in the 14 patients, and of whom 2 were excellent and 12 were good. Patient satisfaction was evaluated according to the Michigan Hand Outcomes Questionnaire (MHQ) and 10 patients were very satisfied and 4 were satisfied. Conclusion:Free perforator flap of superficial peroneal artery is one of the ideal flaps in reconstruction of degloving injury of thumb and fingers. It features constant and multiple perforators, reliable blood supply, high survival rate, flexible in design, thin and a small damage to the donor site.

Liver cancer imposes a substantial global disease burden.Hepatocellular carcinoma(HCC),as the primary pathological type of primary liver cancer,accounts for approximately 90％of all cases.As a minimally invasive treatment,thermal ablation causes minimal impact on liver function and trauma,serving as the first-line treatment option for early-stage HCC patients unsuit-able for surgical resection.However,clinical data indicate that 5-year recurrence rate of HCC pa-tients after thermal ablation treatment is as high as 50％～70％,and incomplete ablation remains a core factor leading to recurrence.Therefore,exploring and optimizing complete ablation strategies has become a critical direction for improving the efficacy of thermal ablation in HCC and enhanc-ing long-term survival rates of patients.This paper systematically reviews the current applications of thermal ablation in HCC treatment and focuses on discussing key technologies and strategies for achieving complete ablation,with the aim of providing theoretical foundations and technical refer-ences for improving the complete ablation rate of thermal ablation in clinical practice and formu-lating personalized diagnosis and treatment plans.

AIM:To observe the effect and mech-anism of cuttlebone extract regulating vascular en-dothelial growth factor(VEGF)/phosphatidylinosi-tol 3-kinase(PI3K)/protein kinase B(AKT)pathway on refractory wounds in rats.METHODS:Cuttle-bone extract(CE)was obtained by water extraction of cuttlebone.Fifty SD rats were randomly divided into negative Control group,Model group,Cuttle-bone extract low-dose(CE(L))group,Cuttlebone extract high-dose(CE(H))group,and cuttlebone ex-tract high-dose+inhibitor(CE(H)+LY294002)group.After the refractory wound model was successfully established,0.02％furacillin solution or cuttlebone extract solution were applied to the wound area of rats in each group,and the treatment was adminis-tered once a day.After 14 days of treatment for re-fractory wounds,the changes in wound healing,angiogenesis,inflammation and expression of relat-ed regulatory proteins were quantitatively ana-lyzed by measuring skin ulcer wound area,patho-logical sections,immunofluorescence staining,Eli-sa,Western blot,RT-qPCR and other methods.RE-SULTS:Compared with Model group,CE(L)and CE(H)groups can increase the number of epithelial cells and collagen,and promote the healing of re-fractory wound in rats.Serum VEGF,skin tissue mi-crovascular density,P-PI3K,P-AKT,VEGF protein ex-pression and mRNA expression levels of PI3K,Akt,VEGF and eNOS were increased(P＜0.05),while se-rum TNF-α and IL-6 levels were decreased(P＜0.05).LY294002 could partially reverse the repair-ing effect of high dose cuttlebone extract on refrac-tory wound(P＜0.05).CONCLUSION:Cuttlebone ex-tract can regulate the VEGF/PI3K/AKT signaling pathway,inhibit the inflammatory response of re-fractory wounds in rats,induce angiogenesis and promote wound healing.

Liver cancer imposes a substantial global disease burden.Hepatocellular carcinoma(HCC),as the primary pathological type of primary liver cancer,accounts for approximately 90％of all cases.As a minimally invasive treatment,thermal ablation causes minimal impact on liver function and trauma,serving as the first-line treatment option for early-stage HCC patients unsuit-able for surgical resection.However,clinical data indicate that 5-year recurrence rate of HCC pa-tients after thermal ablation treatment is as high as 50％～70％,and incomplete ablation remains a core factor leading to recurrence.Therefore,exploring and optimizing complete ablation strategies has become a critical direction for improving the efficacy of thermal ablation in HCC and enhanc-ing long-term survival rates of patients.This paper systematically reviews the current applications of thermal ablation in HCC treatment and focuses on discussing key technologies and strategies for achieving complete ablation,with the aim of providing theoretical foundations and technical refer-ences for improving the complete ablation rate of thermal ablation in clinical practice and formu-lating personalized diagnosis and treatment plans.

Objective:To investigate the clinical effect of reconstruction of the degloving injury of thumb and finger with free perforator flap of superficial peroneal artery.Methods:This is a retrospective study. From June 2020 to June 2023, 18 superficial peroneal artery perforator flaps from 14 calves were used to treat 15 degloved digital wounds of 14 patients in the Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital. There were 13 single digital degloving wounds and 1 two-digital degloving wounds. Of which, 3 digits were reconstructed with 2 free perforator flaps of ipsilateral superficial peroneal artery and 12 with a single free perforator flap of superficial peroneal artery. The size of wounds was 4.0 cm×2.0 cm-10.0 cm×4.0 cm, and the flaps were 5.0 cm×3.0 cm-12.0 cm×4.0 cm in size. The donor sites in calves were sutured layer by layer with absorptive sutures. Postoperative follow-ups were conducted through regular outpatient visits, phone calls or WeChat. Survival of flaps, postoperative complications, therapeutic effect of the flaps and patient satisfaction were observed.Results:There was no vascular compromise or wound infection of the flaps. All patients were included in the 4 to 36 months of postoperative follow-up, with an average of 9.86 months. All flaps had good appearance, without obvious swelling. Colour and texture of the flaps was close to the surrounding skin. The flaps had no obvious pigmentation or ulcer and scar hyperplasia at recipient site. Sensation of the flaps recovered to S 2 to S 3. There was no obvious scar hyperplasia, pain or dysfunction at donor sites. According to the comprehensive evaluation scale of flap, the scores were found at 81 to 91, with an average score of 85 in the 14 patients, and of whom 2 were excellent and 12 were good. Patient satisfaction was evaluated according to the Michigan Hand Outcomes Questionnaire (MHQ) and 10 patients were very satisfied and 4 were satisfied. Conclusion:Free perforator flap of superficial peroneal artery is one of the ideal flaps in reconstruction of degloving injury of thumb and fingers. It features constant and multiple perforators, reliable blood supply, high survival rate, flexible in design, thin and a small damage to the donor site.