BACKGROUND:The use of autologous or allogeneic transplantation materials for tissue repair and reconstruction is limited and carries limitations in clinical applications.Transgenic stem cells and tissue engineering materials offer new therapeutic possibilities.OBJECTIVE:To investigate the in vitro biological characteristics of enhanced green fluorescent protein recombinant lentivirus transfected into rabbit bone marrow mesenchymal stem cells and the biommineralization characteristics of transgenic tissue engineering materials constructed using decalcified bone matrix.METHODS:Rabbit bone marrow mesenchymal stem cells were obtained through adherent culture and density gradient centrifugation.The fifth-generation bone marrow mesenchymal stem cells were transfected with enhanced green fluorescent protein recombinant lentivirus at the optimal multiplicity of infection(MOI=100).The differences in cell proliferation capacity,cell phenotype,cell cycle,alkaline phosphatase expression,osteogenic transcription factor(Runx2)expression,and osteocalcin gene expression after osteogenesis were observed between the transfected cells and non-transfected cells in vitro.The micromorphology and elemental energy spectrum analysis of the transgenic tissue engineering materials constructed from cells and decalcified bone matrix scaffold were also observed.RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells transfected with enhanced green fluorescent protein recombinant lentivirus showed that cell proliferation at 24 and 48 hours after transfection was slower than that of untransfected cells(P<0.05).After 72 hours,the cell phenotype remained unchanged,and the expressions of alkaline phosphatase,Runx2,and osteocalcin after osteogenic induction were not significantly different from those of untransfected bone marrow mesenchymal stem cells(P>0.05).The transgenic tissue engineering materials constructed in vitro using enhanced green fluorescent protein recombinant lentivirus transfected bone marrow mesenchymal stem cells and decalcified bone matrix showed good biocompatibility on the decalcified bone matrix scaffold.Based on fluorescence expression intensity,it was estimated that the target gene would exert its maximum biological function in about 2 weeks,and calcium phosphate deposits would continue to appear,demonstrating superior biomineralization characteristics.

Objective To investigate the relationship between clinical efficacy and lymphocyte subset variation in the treatment of middle and late stage hepatocellular carcinoma(HCC)undergoing initial transarterial chemoembolization(TACE).Methods A retrospective analysis was conducted on the clinical data of 33 patients with middle and late stage HCC who received initial TACE treatment.Lymphocyte subsets were measured before treatment and one month after treatment.Short-term efficacy was evaluated based on the modified response evaluation criteria in solid tumors(mRECIST),and then the patients were divided into remission group[18 cases,including complete remission(CR)and partial remission(PR)]and non-remission group[15 cases,including stable disease(SD)and progressive disease(PD)].The characteristics of lymphocyte subset variation characteristics before and after treatment were compared between the two groups.Results In the remission group,the absolute count and percentage of CD8+cells significantly increased,while the CD4/CD8 ratio and the percentage of CD19+cells significantly decreased after treatment(P＜0.05).Conversely,in the non-remission group,the percentage of CD8+cells significantly decreased,while the CD4/CD8 ratio and the percentage of CD19+cells significantly increased after treatment(P＜0.05).Significant differences were observed between the two groups in the changes of CD8+cells percentage,CD4/CD8 ratio,and CD19+cells percentage before and after treatment(P＜0.05).Conclusion The CD8+cells percentage,CD4/CD8 ratio,and CD19+cells percentage may serve as potential molecular markers for predicting treatment outcomes in HCC patients undergoing TACE.A decrease in the CD8+cells percentage accompanied by an increase in the CD4/CD8 ratio and CD19+cells percentage after the initial TACE treatment may indicate poor therapeutic outcomes.Timely adjustment of individualized treatment plans is recommended to improve patient prognosis.

Objective To explore the technical characteristics and clinical application value of Endosize intelligent surgical planning software in guiding the treatment of Stanford type B aortic dissection(TBAD)with single-branch Castor stent.Methods A retrospective analysis was conducted on the clinical data of 37 patients with TBAD who were manually measured to have insufficient proximal anchoring zones.Preoperative computed tomography angiography(CTA)images of the patients were analyzed using Endosize software for precise endovascular measurements and three-dimensional visual reconstruction of the aortic anatomy in the interventional treatment area.The C-arm angle was preset,and the stent type,size,and surgical pathway were precisely planned.Intraoperatively,digital subtraction angiography(DSA)was used to verify the consistency between the preoperative plan and the actual surgical plan,and the postoperative outcomes and perioperative complications were analyzed.Comprehensive evaluation was performed in combination with early follow-up results.Results The measurement results of the Endosize software were highly consistent with DSA results.Among the 37 patients,3 had sufficient proximal anchoring zones,and 34 had insufficient proximal anchoring zones.The consistency rate between manual measurements and software measurements was 91.89%.All 34 patients with insufficient proximal anchoring zones successfully underwent treatment with single-branch Castor stent.The stent type and size planned preoperatively were completely consistent with those actu-ally used,and the technical success rate was 100%.No adverse events occurred during the perioperative period.The mean follow-up duration was(6.09±4.42)months,with no deaths during this period.All branch stents remained patent,no endoleaks occurred,thrombosis occurred in all of the dissection false lumen,and aortic morphology was remodeled.Conclusion Endosize intelligent surgical planning software can provide detailed preoperative information for endovascular treatment,and it demonstrates significant clinical application value in guiding the treatment of TBAD with insuffi-cient proximal anchoring zones using single-branch Castor stent.

Objective To investigate the relationship between clinical efficacy and lymphocyte subset variation in the treatment of middle and late stage hepatocellular carcinoma(HCC)undergoing initial transarterial chemoembolization(TACE).Methods A retrospective analysis was conducted on the clinical data of 33 patients with middle and late stage HCC who received initial TACE treatment.Lymphocyte subsets were measured before treatment and one month after treatment.Short-term efficacy was evaluated based on the modified response evaluation criteria in solid tumors(mRECIST),and then the patients were divided into remission group[18 cases,including complete remission(CR)and partial remission(PR)]and non-remission group[15 cases,including stable disease(SD)and progressive disease(PD)].The characteristics of lymphocyte subset variation characteristics before and after treatment were compared between the two groups.Results In the remission group,the absolute count and percentage of CD8+cells significantly increased,while the CD4/CD8 ratio and the percentage of CD19+cells significantly decreased after treatment(P＜0.05).Conversely,in the non-remission group,the percentage of CD8+cells significantly decreased,while the CD4/CD8 ratio and the percentage of CD19+cells significantly increased after treatment(P＜0.05).Significant differences were observed between the two groups in the changes of CD8+cells percentage,CD4/CD8 ratio,and CD19+cells percentage before and after treatment(P＜0.05).Conclusion The CD8+cells percentage,CD4/CD8 ratio,and CD19+cells percentage may serve as potential molecular markers for predicting treatment outcomes in HCC patients undergoing TACE.A decrease in the CD8+cells percentage accompanied by an increase in the CD4/CD8 ratio and CD19+cells percentage after the initial TACE treatment may indicate poor therapeutic outcomes.Timely adjustment of individualized treatment plans is recommended to improve patient prognosis.

BACKGROUND:The use of autologous or allogeneic transplantation materials for tissue repair and reconstruction is limited and carries limitations in clinical applications.Transgenic stem cells and tissue engineering materials offer new therapeutic possibilities.OBJECTIVE:To investigate the in vitro biological characteristics of enhanced green fluorescent protein recombinant lentivirus transfected into rabbit bone marrow mesenchymal stem cells and the biommineralization characteristics of transgenic tissue engineering materials constructed using decalcified bone matrix.METHODS:Rabbit bone marrow mesenchymal stem cells were obtained through adherent culture and density gradient centrifugation.The fifth-generation bone marrow mesenchymal stem cells were transfected with enhanced green fluorescent protein recombinant lentivirus at the optimal multiplicity of infection(MOI=100).The differences in cell proliferation capacity,cell phenotype,cell cycle,alkaline phosphatase expression,osteogenic transcription factor(Runx2)expression,and osteocalcin gene expression after osteogenesis were observed between the transfected cells and non-transfected cells in vitro.The micromorphology and elemental energy spectrum analysis of the transgenic tissue engineering materials constructed from cells and decalcified bone matrix scaffold were also observed.RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells transfected with enhanced green fluorescent protein recombinant lentivirus showed that cell proliferation at 24 and 48 hours after transfection was slower than that of untransfected cells(P<0.05).After 72 hours,the cell phenotype remained unchanged,and the expressions of alkaline phosphatase,Runx2,and osteocalcin after osteogenic induction were not significantly different from those of untransfected bone marrow mesenchymal stem cells(P>0.05).The transgenic tissue engineering materials constructed in vitro using enhanced green fluorescent protein recombinant lentivirus transfected bone marrow mesenchymal stem cells and decalcified bone matrix showed good biocompatibility on the decalcified bone matrix scaffold.Based on fluorescence expression intensity,it was estimated that the target gene would exert its maximum biological function in about 2 weeks,and calcium phosphate deposits would continue to appear,demonstrating superior biomineralization characteristics.

Objective To explore the technical characteristics and clinical application value of Endosize intelligent surgical planning software in guiding the treatment of Stanford type B aortic dissection(TBAD)with single-branch Castor stent.Methods A retrospective analysis was conducted on the clinical data of 37 patients with TBAD who were manually measured to have insufficient proximal anchoring zones.Preoperative computed tomography angiography(CTA)images of the patients were analyzed using Endosize software for precise endovascular measurements and three-dimensional visual reconstruction of the aortic anatomy in the interventional treatment area.The C-arm angle was preset,and the stent type,size,and surgical pathway were precisely planned.Intraoperatively,digital subtraction angiography(DSA)was used to verify the consistency between the preoperative plan and the actual surgical plan,and the postoperative outcomes and perioperative complications were analyzed.Comprehensive evaluation was performed in combination with early follow-up results.Results The measurement results of the Endosize software were highly consistent with DSA results.Among the 37 patients,3 had sufficient proximal anchoring zones,and 34 had insufficient proximal anchoring zones.The consistency rate between manual measurements and software measurements was 91.89%.All 34 patients with insufficient proximal anchoring zones successfully underwent treatment with single-branch Castor stent.The stent type and size planned preoperatively were completely consistent with those actu-ally used,and the technical success rate was 100%.No adverse events occurred during the perioperative period.The mean follow-up duration was(6.09±4.42)months,with no deaths during this period.All branch stents remained patent,no endoleaks occurred,thrombosis occurred in all of the dissection false lumen,and aortic morphology was remodeled.Conclusion Endosize intelligent surgical planning software can provide detailed preoperative information for endovascular treatment,and it demonstrates significant clinical application value in guiding the treatment of TBAD with insuffi-cient proximal anchoring zones using single-branch Castor stent.

Objective To evaluate the effect of human bone morphogenetic protein 2 (hBMP-2)/Bone Mesenchymal Stem Cells (BMSCs)/demineralized bone matrix(DBM) on repairing rabbits’femoral head after necrosis and to explore the new treatments for femoral head necrosis. Methods Femoral head necrosis models was established by clinical core decom?pression combined with liquid nitrogen frozen. Then, animals were randomly devided into 4 groups (n=12 per group):Group A were not implanted anything as control group, Group B were implanted with DBM. Group C were implanted with hBMP-2/DBM. Group D were implanted with hBMP-2/BMSCs/DBM. Four rabbits from each group were sacrificed at 4,8 and 12 weeks after surgery to evaluate the the repairing effect of Osteonecrosis of the femoral head (ONFH) through X-ray examina?tion, observation of the specimen and HE staining. Results X-ray revealed defect of femoral head in Group A without clear bone formation. There is a little fibrous hyperplasia and no obvious osteogenic response. By contrast, the femoral head defect areas became fuzzy in group B, group C and group D with new bone trabeculars. And the regenerate phenomenons of group D were significantly better than that of group B and group C of the same time point. As to the Lane-Sandhu X Ray scores, it is lower in group A than that in group B;It is lower in group C than that in group D(P<0.05). There is no statistical difference between Group B and Group C. General observation of the specimen revealed that the femoral head of group A collapsed with drilling holes. The femoral heads of group B and group C showed no collapse but the drilling holes existed. Femoral head in group D was not collapsed and the drilling holes disappeared. HE staining showed that bone trabeculars became ne?crotic and fragmented in Group A with a lot of air trapped cells. There were newborn immature bone trabeculars and osteo? blasts in group B and group C. Group D were of large number of bone cells, fat cells, and newborn mature bone trabeculars. The ratio of empty lacuna is higher in Group A than that in Group B;it is higher in Group C than that in Group D(P<0.05). Conclusion hBMP-2/BMSCs/DBM can induce BMSCs differentiation into osteoblasts after being implanted. It has good re?pairing effect on ONFH with good application prospect.

Objective To observe the biocompatibility of rabbit bone marrow mesenchymal stem cells (BMSCs)combined with allogeneic decalcified bone matrix(DBM)after transfecting adenoviral recombinant human bone morphogenetic protein-2(Ad-rhBMP-2).Methods The rabbit allogeneic DBM material was prepared according to the Ursit method.After transfecting Ad-BMP-2 on rabbit bone marrow mesenchymal stem cells,the immunohistochemical was used to detect the expression of BMP-2 in the transfected cells;after 48 h of transfection,the cells were planted on the allograft DBM,then the scanning electron microscopy was used to observe the cell growth and adhesion condition on material,and the proliferation condition of BMSCs was detected by MTT. Results After 48 h of adenoviral transfection,BMSCs could express BMP-2 successfully.The scanning electron microscopy showed that the cells after transfection adhered well and massively proliferated on DBM material.The MTT assay showed that the prolifer-ation condition of the cells after transfection planted on DBM was normal,which showed no statistically significant difference when compared with the control group (P >0.05).Conclusion The Ad-BMP-2 transfection on BMSCs is well biocompatible to allogene-ic DBM.

Objective To study the effects of short-term cryopreservation on the proliferation ,bio-characteristics and osteogenic capability of rabbit BMSCs and lay the foundation for the further study .Methods The 5th generation of rabbit BMSCs were cryo-preservated by liquid nitrogen for 30 days ,and then thawing the cells to culture it to the 10th generation in vitro ,put the non-cryo-preservated and the same generation BMSCs as the control group .The MTT ,cell cycle ,cell surface marker ,the content of ALP and the immunohistochemical staining for collagen typeⅠ and alizarin red staining for calcium after differentiation induced by osteogene-sis were used to evaluate the proliferation ,bio-characteristic and osteogenic differentiation capability of rabbit BMSCs .Results The growth incubation period of BMSCs after cryopreservated was extended ,but it gradually recover through serial passage .The prolif-eration index and the proportion of BMSCs in G1 phase were 42 .9% ± 3 .4% and 57 .0% ± 3 .4% respectively .The positive rate of cell surface marker of CD44 was 93 .62% ± 1 .05% without expression of CD45 .The contents of ALP(U/gprot) were 6 .73 ± 1 .92 and 15 .99 ± 4 .36 in BMSCs after 7th day and 14th day with osteogenic induction ,respectively .The collagen typeⅠ and alizarin red staining for calcium indicated positive at BMSCs after 14th day and 21th day with osteogenic induction ,respectively .These results showed no significant difference compared with the control group (P>0 .05) .Conclusion Short-term cryopreservation has no obvi-ous impacts on the proliferation ,bio-characteristic and osteogenic capability of BMSCs and it could be used for further study .