Objective To investigate the causes and management strategies for lower limb ischemic necrosis following xenogeneic heterotopic heart transplantation from a multigene-edited pig to a rhesus monkey. Methods A xenogeneic heterotopic heart transplantation was performed on December 16, 2023, at the Institute of Experimental Animals of Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, using a quintuple-gene-edited pig as the donor and a rhesus monkey as the recipient. On postoperative day (POD) 9, the recipient monkey underwent left lower limb amputation due to ischemic necrosis. Blood samples were collected at various time points after transplantation for analysis of hematologic parameters, liver and renal function, myocardial enzymes, and coagulation profiles. Ultrasound and computed tomography (CT) were used to evaluate anastomotic patency and cardiac structure. Immunological assays, including complement-dependent cytotoxicity (CDC) and IgG/IgM antibody detection, combined with clinical observations, were employed to assess rejection type and therapeutic response. Results The recipient monkey survived for 46 days after transplantation. Echocardiography demonstrated preserved biventricular systolic function in the recipient’s native heart, with left ventricular ejection fraction (LVEF) consistently exceeding 50%. In the donor pig heart, left ventricular endocardial thickening was noted on POD 9, followed by right ventricular endocardial thickening on POD 24, while LVEF remained around 35%. No hyperacute or acute rejection was detected immunologically. CDC positivity ranged between 3.4% and 5.1%, with IgG/IgM antibody binding trends consistent with CDC results. Following amputation, the recipient exhibited elevated inflammatory markers, coagulopathy, and reactive thrombocytosis, which later normalized. Immunohistochemical staining of the necrotic limb revealed arterial and venous thrombosis; however, no T-cell or B-cell infiltration was observed in vascular structures, thrombi, nerves, muscles, fascia, or skin tissues, with CD3 and CD20 staining both negative. Conclusion Limb ischemia after xenogeneic heart transplantation may be associated with lower extremity vascular thrombosis triggered by local trauma in the context of transplantation-induced inflammatory activation and coagulation dysfunction. While no clear lymphocyte-mediated rejection was observed, further studies are needed to explore the potential role of non-lymphocyte-mediated immune mechanisms.

Objective To evaluate the efficacy of a combined immunosuppression regimen in modulating rejection in genetically modified pig-to-macaque xenogeneic heart transplantation.Methods Two xenogeneic heart transplantation models were constructed using genetically modified pigs and macaques.Dynamic monitoring of recipient peripheral blood immune parameters and observation of graft pathological changes were performed.Results Regimen 1,featuring B-cell depletion,T-cell inhibition,and C3 complement suppression,reduced lymphocyte levels but failed to control acute humoral rejection and macrophage infiltration.Regimen 2,adding C5 complement inhibition and interleukin-6 inhibition to Regimen 1,more effectively lowered lymphocyte levels,inhibited acute humoral rejection and complement activation,and decreased antibody deposition.However,a late-phase cytokine storm and residual T cells emerged.Conclusions Regimen 2 reduces the hyperacute and acute rejection risks through multi-target intervention.Yet,it requires balancing medication complexity and safety.This indicates the need to optimize cellular immune regulation and adjust the plan through dynamic multidimensional monitoring.

Objective To evaluate the efficacy of a combined immunosuppression regimen in modulating rejection in genetically modified pig-to-macaque xenogeneic heart transplantation.Methods Two xenogeneic heart transplantation models were constructed using genetically modified pigs and macaques.Dynamic monitoring of recipient peripheral blood immune parameters and observation of graft pathological changes were performed.Results Regimen 1,featuring B-cell depletion,T-cell inhibition,and C3 complement suppression,reduced lymphocyte levels but failed to control acute humoral rejection and macrophage infiltration.Regimen 2,adding C5 complement inhibition and interleukin-6 inhibition to Regimen 1,more effectively lowered lymphocyte levels,inhibited acute humoral rejection and complement activation,and decreased antibody deposition.However,a late-phase cytokine storm and residual T cells emerged.Conclusions Regimen 2 reduces the hyperacute and acute rejection risks through multi-target intervention.Yet,it requires balancing medication complexity and safety.This indicates the need to optimize cellular immune regulation and adjust the plan through dynamic multidimensional monitoring.

Objective:To construct a recombinant plasmid DNA carrying the decorin( DCN) gene and study its therapeutic effect on hypertrophic scars of rabbit ears. Methods:The human decorin gene fragment amplified by PCR was cloned into plasmid vector pUDK to construct the recombinant plasmid pDCN, which was identified by enzyme digestion and sequencing. pDCN was transfected into 293T cells, and the expression of DCN and TGF-β1 was detected. The therapeutic effect of pDCN on rabbit ear hypertrophic scar model was observed by hypertrophy index, pathological examination and immunohistochemical staining. Results:The decorin gene was successfully inserted into pUDK, which was examined by enzyme digestion and sequencing. The expression level of mRNA and protein of DCN was up-regulated in 293T cells post pDCN transfection, and the expression of TGF-β1 was suppressed. Then the rabbit ear hypertrophic scars were treated with different doses of pDCN, and the results showed that the hypertrophy index of the medium dose (200 μg/cm 2) pDCN group was significantly lower than that of the PBS group ( P<0.05), while there was no significant difference in the hypertrophy index of the low dose and high dose pDCN group compared with the PBS group. The expression of DCN in ears skin in the medium dose pDCN group was significantly higher than that in the PBS group ( P<0.05). The pathological examination showed that inflammatory cell infiltration and fibrous deposition in scar tissue were significantly reduced. These results indicated that the medium-dose pDCN could effectively inhibit the hyperplasia of hypertrophic scar in rabbit ears. Conclusions:pDCN, the plasmid carrying decorin gene, has therapeutic effects on hypertrophic scars of rabbit ears by inhibiting TGF-β1 expression.

Objective:To construct a recombinant plasmid DNA carrying the decorin( DCN) gene and study its therapeutic effect on hypertrophic scars of rabbit ears. Methods:The human decorin gene fragment amplified by PCR was cloned into plasmid vector pUDK to construct the recombinant plasmid pDCN, which was identified by enzyme digestion and sequencing. pDCN was transfected into 293T cells, and the expression of DCN and TGF-β1 was detected. The therapeutic effect of pDCN on rabbit ear hypertrophic scar model was observed by hypertrophy index, pathological examination and immunohistochemical staining. Results:The decorin gene was successfully inserted into pUDK, which was examined by enzyme digestion and sequencing. The expression level of mRNA and protein of DCN was up-regulated in 293T cells post pDCN transfection, and the expression of TGF-β1 was suppressed. Then the rabbit ear hypertrophic scars were treated with different doses of pDCN, and the results showed that the hypertrophy index of the medium dose (200 μg/cm 2) pDCN group was significantly lower than that of the PBS group ( P<0.05), while there was no significant difference in the hypertrophy index of the low dose and high dose pDCN group compared with the PBS group. The expression of DCN in ears skin in the medium dose pDCN group was significantly higher than that in the PBS group ( P<0.05). The pathological examination showed that inflammatory cell infiltration and fibrous deposition in scar tissue were significantly reduced. These results indicated that the medium-dose pDCN could effectively inhibit the hyperplasia of hypertrophic scar in rabbit ears. Conclusions:pDCN, the plasmid carrying decorin gene, has therapeutic effects on hypertrophic scars of rabbit ears by inhibiting TGF-β1 expression.

Previous studies have shown that the sciatic nerve has neurotrophic activity, and nerve regeneration, differentiation, and axon outgrowth can be modulated by different sciatic nerve preparations. However, numerous animals may have to be sacrificed to obtain enough sciatic nerves to make a sciatic nerve preparation. Some studies have demonstrated that the role of sciatic nerve preparations in neural differentiation depends on the neurotrophins that Schwann cells secrete, and these factors are highly conserved among different species. To reduce the use of experimental animals, in this study, we made a leachate by using the sciatic nerve of cattle and explored its effect on neuronal differentiation of rat PC12 cells (a useful model for studying neuronal differentiation). Results showed the neurite outgrowth of PC12 cells treated with the cattle sciatic nerve leachate for 3, 6, and 9 days was significantly improved, and the expressions of β3-tubulin and microtubule-associated protein 2 (two neuron-specific proteins) were increased. Moreover, the ERK1/2 signaling pathway was activated after PC12 cells were incubated with cattle sciatic nerve leachate for 9 days. Thus, a sciatic nerve leachate obtained from cattle can effectively induce neuronal differentiation of rat PC12 cells via ERK1/2 signaling pathway.
Animals ; Axons ; Cattle ; Microtubule-Associated Proteins ; Nerve Growth Factors ; Nerve Regeneration ; Neurites ; Neurons ; PC12 Cells ; Rats ; Schwann Cells ; Sciatic Nerve

To reveal the intrinsic characteristics and rules of change within the system for evaluation of human resources for military medical sciences .We intended to gain insights into the system for the evaluation of human resources for military medical sciences while analyzing the effect of the environment of military medicine , competitions for talents and regulations on the evaluation system .We also interpreted the system structure composed of the evaluation subject , evaluation object and evaluation activity as well as their interactions .We also made an attempt to study value evaluations , judgment functions and functions of selection , guidance , manipulation and supervision and the self-consistency mechanism between the above-mentioned functions .We also analyzed the features of this complicated system and rules of evolution .

Objective To compare the efficacy after conventional radiotherapy and late course accelerated fractionation radiotherapy for nasopharyngeal carcinoma (NPC).Methods A total of 200 NPC patients were enrolled and randomly assigned to conventional radiotherapy (CF) group with 99 cases or late course accelerated fractionation radiotherapy (LCAF) group with 101 cases,who received irradiation to 60Co γ or 6 MV X-rays.In the CF group,the total dose of nasopharynx was 70 Gy/35 fractions at 2 Gy daily.In the LCAF group,for the first two-thirds of the treatment,two daily fractions of 1.2 Gy were given to the primary lesion and the total dose was 48 Gy/40 fractions.For the last one third of the treatment,the dose per fraction was increased to 1.5 Gy and the total dose was 30 Gy/20 fractions.Results There were 25,16,25 in CF group and 16,13,18 patients in LCAF group who had recurrence of nasophaynx,cervical lymph nodes,and distant metastasis,respectively.The 5-year nasopharyngeal control and overall survival rates was 75.9％ and 87.6％ in CFgroup (x2 =4.066,P＜0.05),58.0％ and 74.1％(x2 =5.076,P ＜ 0.05) in LCAF group,respectively.Cervical lymph nodes local rates and distant metastasesfree rates at 5 years were 8 1.5％ and 90.0％ in CF group (P ＞ 0.05),74.1％ and 83.3％ (P ＞ 0.05) in LCAF group,respectively.Conclusions Compared with CF,LCAF can improve nasopharyngeal control and overall survival rates,but there are no significant difference in the recurrence rates of cervical lymph nodes and distant metastasis.

Objective To evaluate the volume of left ventricle (LV) based on PET-CT in left-sided breast cancer patients treated with adjuvant radiotherapy. To assess the impact of dose and volume in interrelations for heart and cardiac sub-structures using dose-volume histograms. Methods From October 2008 to February 2009, 14 patients with post-operatively confirmed left-sided breast cancer were enrolled in this study. Patients were scanned using contrast enhanced CT for simulation and FDG PET-CT have been applied to display the structure of left ventricle in each patient before radiotherapy (RT). The LV has been delineated based on PET-CT. Other critical sub-structures, such as left anterior descending coronary artery (LAD) have been contoured in each patient. The six-field simple intensity modulated radiotherapy (slMRT)technique has been created in all patients. Results The mean volumes of left ventricle based on PET-CT (LV-PET) were 112. 931 cm3. The volume of LV receiving ≥50% prescription dose highly correlated with the volume of heart receiving ≥50% prescription dose (R = 0. 869, P = 0. 000). There was less correlation between the volume of LAD and the volume of heart receiving ≥ 50% prescription dose ( R = 0. 220, P =0. 440). Conclusions The left ventricle could be delineated efficiently based on imaging of PET-CT. The volume of LV receiving higher dose in RT has high correlation with the volume of heart using sIMRT technique. It may pave the way for further exploring radiation induced cardiac injury in patients with breast cancer.

Objective To determine the change of tumor bed volume during whole breast irradiation by repeated computed tomography scanning and to analyze the dosimetric impact of boost-planning on different CT images. Methods From July 2008 to Jan 2009, sixteen patients with early-stage breast cancer underwent breast conservative surgery (BCS) were enrolled in the study. All patients received whole breast irradiation and tumor bed boost, no adjuvant chemotherapy was given. Two additional CT scans were acquired in addition to the planning CT ( CT1 ), one in the course of radiotherapy ( CT2 ) and the other before the boost (CT3). Tumor beds were contoured in all CT images. Three-dimensional conformal radiotherapy planning for tumor bed boost was done on CT1 and CT3 respectively. Results The mean tumor bed volume on CT1, CT2 and CT3 were 49.5 cm3, 25.6 cm3 and 22. 2 cm3 ( F = 5. 63, P = 0. 007 ),respectively. Further analysis found statistically significant difference between CT1 and CT2 ( q = 0. 03, P =0. 010), CT1 and CT3 ( q = 0. 01, P = 0. 004), but not between CT2 and CT3 ( q = 1.00, P = 0. 333 ). The average reduction of tumor bed volume from CT1 to CT3 was 43.4%. A reduction of 20% or above was found in 88% of the patients ( n = 14), 50% or above in 38% of the patients (n = 6). In the boost-planning, the volume of the ipsilateral breast receiving 100% prescribed dose (V100%) on CT1 and CT3 was 183.5 cm3 and 144. 5 cm3, respectively ( t = 3.06, P = 0. 008 ). Conclusions Volume of tumor bed is dynamically reduced in the course of whole breast irradiation after BCS, with more important reduction in the early weeks after the beginning of irradiation. A second CT scan before tumor bed boost is warranted.