OBJECTIVE: To explore the construction of a canine model of vascularized allogeneic spinal cord transplantation (vASCT) and preliminarily evaluate its therapeutic efficacy for spinal cord injury (SCI). METHODS: Sixteen female Beagle dogs aged 8-12 months were randomly selected, with 8 dogs serving as donors for the harvesting of spinal cord tissue with a vascular pedicle [dorsal intercostal artery (DIA) at the T₁₀ level and accompanying vein]. The remaining 8 dogs underwent a 1.5-cm-length spinal cord defect at the T₁₀ level, followed by transplantation of the donor spinal cord tissue for repair. Polyethylene glycol (PEG) was applied to both ends to spinal cord graft; then, using a random number table method, the dogs were divided into an experimental group (n=4) and a control group (n=4). The experimental group received immunosuppressive intervention with oral tacrolimus [0.1 mg/(kg∙d)] postoperatively, while the control group received no treatment. The operation time and ischemia-reperfusion time of two groups were recorded. The recovery of hind limb function was estimated by Olby score within 2 months after operation; the motor evoked potentials (MEP) was measured through neuroelectrophysiological examination, and the spinal cord integrity was observed through MRI. RESULTS: There was no significant difference in the operation time and ischemia-reperfusion time between the two groups (P>0.05). All dogs survived until the completion of the experiment. Within 2 months after operation, all dogs in the control group failed to regain the movement function of hind limbs, and Olby scores were all 0. In the experimental group, the movement and weight-bearing, as well as walking abilities of the hind limbs gradually recovered, and the Olby scores also showed a gradually increasing trend. There was a significant difference between the two groups from 3 to 8 weeks after operation (P<0.05). Neuroelectrophysiological examination indicated that the electrical signals of the experimental group passed through the transplanted area, and the latency was shortened compared to that at 1 month after operation (P<0.05), showing continuous improvement, but the amplitude did not show significant improvement (P>0.05). The control group was unable to detect any MEP changes after operation. MRI examination showed that the transplanted spinal cord in the experimental group survived and had good continuity with normal spinal cord tissue, while no relevant change was observed in the control group. CONCLUSION The vASCT model of dogs was successfully constructed. This surgical procedure can restore the continuity of the spinal cord. The combination of tacrolimus anti-immunity is a key factor for the success of transplantation.
Animals ; Dogs ; Female ; Spinal Cord/blood supply* ; Spinal Cord Injuries/surgery* ; Transplantation, Homologous ; Disease Models, Animal ; Recovery of Function ; Plastic Surgery Procedures/methods* ; Tacrolimus ; Immunosuppressive Agents

BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.

Objective To analyze the efficacy of vascularized lymph node transplantation(VLNT)combined with lymphatic venous anastomosis(LVA)in the treatment of secondary lower extremity lymphedema assisted by super microsurgical techniques.Methods A retrospective analysis was performed for 15 patients with secondary lower limb lymphedema who underwent VLNT+LVA surgery in the Department of Reconstructive and Reconstructive Microsurgery of Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine from July 2021 to July 2023,and compared the circumference and volume of each segment of the lower limb between the preoperative and postoperative 90 days.LVA according to the results of ICG examination,3-5 parts of the lower limb were selected for"Z"shaped surgery,and the lymphatic vessels and venules were anastomosed under the microscope in the subcutaneous fat layer.VLNT confirmed and labeled the saphenous branch of the descending knee artery and its accompanying veins in the popliteal fossa,and the latissimus dorsi lymph node flap was incised,and the flap vessels were anastomosed with the saphenous branch of the descending knee artery and its accompanying veins.Postoperative observation of flap vascularization.The circumference and volume of the affected limb were measured before surgery and 90 days after surgery.Results A total of 15 patients with lower extremity lymphedema were included without serious complications.Statistical analysis showed that the circumference of all levels of the affected limb and the volume of the affected limb were improved 90 days after operation compared with those before surgery.Among them,the limb circumference and volume at each level from the highest point of the dorsum of the foot to 52 cm above the ankle improved 90 days after the operation compared with those before the operation.Conclusion LNT+LVA treatment for secondary lower extremity lymphedema can effectively control edema and improve the function of the affected limb.

Objective To analyze the efficacy of vascularized lymph node transplantation(VLNT)combined with lymphatic venous anastomosis(LVA)in the treatment of secondary lower extremity lymphedema assisted by super microsurgical techniques.Methods A retrospective analysis was performed for 15 patients with secondary lower limb lymphedema who underwent VLNT+LVA surgery in the Department of Reconstructive and Reconstructive Microsurgery of Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine from July 2021 to July 2023,and compared the circumference and volume of each segment of the lower limb between the preoperative and postoperative 90 days.LVA according to the results of ICG examination,3-5 parts of the lower limb were selected for"Z"shaped surgery,and the lymphatic vessels and venules were anastomosed under the microscope in the subcutaneous fat layer.VLNT confirmed and labeled the saphenous branch of the descending knee artery and its accompanying veins in the popliteal fossa,and the latissimus dorsi lymph node flap was incised,and the flap vessels were anastomosed with the saphenous branch of the descending knee artery and its accompanying veins.Postoperative observation of flap vascularization.The circumference and volume of the affected limb were measured before surgery and 90 days after surgery.Results A total of 15 patients with lower extremity lymphedema were included without serious complications.Statistical analysis showed that the circumference of all levels of the affected limb and the volume of the affected limb were improved 90 days after operation compared with those before surgery.Among them,the limb circumference and volume at each level from the highest point of the dorsum of the foot to 52 cm above the ankle improved 90 days after the operation compared with those before the operation.Conclusion LNT+LVA treatment for secondary lower extremity lymphedema can effectively control edema and improve the function of the affected limb.

BACKGROUND:Previously,a SENP1 gene-silenced human alveolar epithelial cell line was successfully constructed in vitro,and the role of SENP1 in hyperoxic lung injury was investigated at the cellular level.OBJECTIVE:To construct a mouse model of alveolar type II epithelial cell-specific knockout of SENP1 gene based on the Cre-loxP recombinase system.METHODS:SENP1flox/-mice were self-crossed to obtain SENP1flox/flox and SENP1flox/-mice;Sftpc-Cre+/+mice were crossed with wild-type mice to obtain more Sftpc-Cre+/-mice.Sftpc-Cre+/+or offspring Sftpc-Cre+/-mice were crossed with SENP1flox/-or offspring SENP1flox/flox mice to obtain SENP1flox/-Sftpc-Cre+/-double heterozygous mice.SENP1flox/-Sftpc-Cre+/-mice were then crossed with SENP1flox/flox mice to obtain SENP1flox/floxSftpc-Cre+/-mice.The genomic DNA was extracted by tail clipping and amplified by PCR.The amplified product was subjected to agarose gel electrophoresis to determine the mouse genotypes.Lung tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were subjected to immunofluorescence double-labelling and western blot assay to verify the knockdown effect of SENP1 gene.Heart,liver,lung and kidney tissues of SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice were stained with hematoxylin-eosin to observe the histomorphology of each organ in the two groups of mice.RESULTS AND CONCLUSION:SENP1flox/floxSftpc-Cre+/-mice were correctly screened by agarose gel electrophoresis.Immunofluorescence double-labeling experiments showed that the mean fluorescence intensity of SENP1 was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with that of SENP1flox/flox mice(P<0.01)and no significant co-localization of SENP1 and Sftpc was observed(P<0.01).Western blot results showed that SENP1 protein expression was reduced in lung tissues of SENP1flox/floxSftpc-Cre+/-mice compared with SENP1flox/flox mice(P<0.001).Hematoxylin-eosin staining showed no significant alterations in the histomorphology of heart,liver,lung and kidney tissues in SENP1flox/flox and SENP1flox/floxSftpc-Cre+/-mice.This study successfully constructed alveolar type II epithelial cell-specific knockout SENP1 gene mice using the Cre-loxP recombinase system,which provides a good tool for the subsequent study of the role of SENP1 gene in lung diseases such as bronchopulmonary dysplasia and idiopathic pulmonary fibrosis,in which alveolar type II epithelial cells are the main damage cells.

The Wnt signaling pathway plays an important role in maintaining liver homeostasis and liver regeneration.In healthy livers,the Wnt signaling pathway is mostly inactive,but it is continuously overactivated during cell renewal or regeneration processes,as well as in certain pathological conditions,diseases,precancerous states,and cancers.Persistent liver cell injury often leads to chronic liver diseases such as liver fibrosis,liver cirrhosis,and liver cancer.This article summarizes the basic structural features of the Wnt signaling pathway and analyzes its important role in the pathological progression of various liver diseases,so as to provide new ideas for the prevention and treatment of liver diseases in clinical practice.

Objective Analyze the characteristics of delayed apheresis platelet donors due to health consultation or ex-amination,and the influencing factors of recall in Guangzhou,and to provide evidence for recruitment strategy.Methods 5 960 delayed apheresis platelet donors from January fisrt,2022 to June 30th,2022 were collected in Guangzhou Blood Center and fol-lowed up to August31th,2023.Logistic regression analysis was used to analyze the influence of sex,age,times of blood donation and length of blood donation on recall.Results Among the 5 960 delayed platelet donors,164(2.75%)were permanently un-suitable for blood donation.The most common reason was allergy.Among the 5 796(97.25%)delayed blood donors,the most common reason was abnormal blood routine.Analysis of factors associated with delayed donor recall found that male donors were more likely than female donors[OR =1.16,95%CI(1.01,1.34)],blood donors aged 26-35[OR =1.65,95%CI(1.41,1.92)],36-45[OR=1.90,95%CI(1.55,2.31)]or 46-60[OR =2.63,95%CI(1.96,3.53)]compared with those aged 18-25,blood donors who were blocked forless than 14 days[OR =4.22,95%CI(2.73,6.52)]orfor 15-89 days[OR =5.24,95%CI(3.42,8.03)]compared with those who were blocked for more than or equal to 90 days,the recall rate of repeat donors was higher than that of first-time donors[OR=6.78,95%CI(5.62,8.19)].Conclusion In order to improve the recall rate of apheresis platelet donors,we should pay more attention to the education of young donors and first-time donors,also consider a voluntary recall by text message or phone after a donor has been prevented from donating blood.

BACKGROUND: Hepatitis B surface antigen (HBsAg) clearance is vital for a functional cure of hepatitis B virus (HBV) infection. However, the incidence and predictors of HBsAg seroclearance in patients co-infected with HBV and human immunodeficiency virus (HIV) remain largely unknown in Guangdong, China. METHODS: Between 2009 and 2019, patients co-infected with HBV/HIV undergoing antiretroviral therapy (ART) in Guangzhou Eighth People's Hospital affiliated to Guangzhou Medical University were retrospectively reviewed with the endpoint on December 31, 2020. The incidence and risk factors for HBsAg seroclearance were evaluated using Kaplan-Meier and multivariate Cox regression analyses. RESULTS: A total of 1550 HBV/HIV co-infected patients were included in the study, with the median age of 42 years and 86.0% (1333/1550) males. Further, 98.3% (1524/1550) received ART containing tenofovir disoproxil fumarate (TDF) plus lamivudine (3TC). HBV DNA was examined in 1283 cases at the last follow-up. Over the median 4.7 years of follow-up, 8.1% (126/1550) patients achieved HBsAg seroclearance, among whom 50.8% (64/126) obtained hepatitis B surface antibody, 28.1% (137/488) acquired hepatitis B e antigen seroconversion, and 95.9% (1231/1283) undetectable HBV DNA. Compared with patients who maintained HBsAg positive, cases achieving HBsAg seroclearance showed no differences in age, gender, CD4 + T cell count, alanine aminotransferase (ALT) level, or fibrosis status; however, they presented lower HBV DNA levels, lower HBsAg levels, and higher rates of HBV genotype B at the baseline. Multivariate analysis showed that baseline HBsAg <1500 cutoff index (COI) (adjusted hazard ratio [aHR], 2.74, 95% confidence interval [95% CI]: 1.48-5.09), ALT elevation >2 × upper limit of normal during the first six months after receiving ART (aHR, 2.96, 95% CI: 1.53-5.77), and HBV genotype B (aHR, 3.73, 95% CI: 1.46-9.59) were independent predictors for HBsAg seroclearance (all P <0.01). CONCLUSIONS Long-term TDF-containing ART has high anti-HBV efficacy including relatively high overall HBsAg seroclearance in HBV/HIV co-infected patients. Lower baseline HBsAg levels, HBV genotype B, and elevated ALT levels during the first six months of ART are potential predictors of HBsAg seroclearance.
Male ; Humans ; Adult ; Hepatitis B Surface Antigens ; Hepatitis B virus/genetics* ; HIV Infections/drug therapy* ; HIV ; DNA, Viral ; Incidence ; Coinfection/drug therapy* ; Retrospective Studies ; Tenofovir/therapeutic use* ; Lamivudine/therapeutic use* ; Hepatitis B, Chronic/drug therapy*

Objective:To investigate the effect of vascularized lymph node transfer (VLNT) combined with lymphatico-venous anastomosis (LVA) in the treatment of lymphedema after breast cancer surgery.Methods:The data of patients with upper limb lymphedema after breast cancer surgery who were treated in the Department of Reconstructive and Reconstructive Microsurgery, Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine from July 2021 to July 2022 were retrospectively analyzed. According to different treatment methods, the patients were divided into LVA group and VLNT combined LVA group. Indocyanine green (ICG) near-infrared lymphography was performed on all affected limbs before surgery. In the LVA group, according to the results of ICG lymphography, 4 to 5 levels of the affected limb were selected, Z shaped incisions were made and dissescted until the subcutaneous fat layer. End-to-end or end-to-side anastomosis was performed between lymphatic vessels and subcutaneous venules under the microscope. In the VLNT combined LVA group, the branches of brachial artery and vein in the axillary region were marked. The inguinal flap with the superficial iliac circumflex vessel pedicle and 4-5 lymph nodes was dissected. End-to-end anastomoses of the superficial iliac circumflex vessel pedicle with the branches of brachial artery and vein were performed in the axillary region of the affected limb. LVA was performed according to ICG lymphography, the same as in the LVA group. The skin and soft tissue condition of the affected limb and the blood supply of the flap in the VLNT combined LVA group were observed after operation. The circumference of the upper arm (from the wrist to 32 cm above the wrist, every 4 cm, a total of 9 levels of circumference) and upper limb volume were measured before and after operation. SPSS 24.0 was used for data processing and analysis. Measurement data were expressed Mean±SD. The data before and after operation in the same group were compared by paired samples t test, and the comparison between the two groups was conducted by independent samples t test. P<0.05 was considered statistically significant. Results:A total of 14 female patients were enrolled, with 7 patients in each group. All cases were unilateral lymphedema. There were no significant differences in age, stage of disease, limb circumference and limb volume between the two groups before operation ( P>0.05). After operation, the skin and soft tissue condition of the affected limbs were good, and no complications such as erysipelas, cellulitis, or lymphangitis occurred. All flaps in the VLNT combined LVA group survived successfully, and the operation wounds healed well. There were no complications such as infection and necrosis of the flaps. One year after operation, the circumference and volume of the affected limb in the two groups were improved to varying degrees, and the circumference of the affected limb (wrist, upper wrist 4, 8, 12, 16, 20, 24, 28, 32 cm levels) in the VLNT combined LVA group was significantly smaller than that before operation ( P<0.01). In the LVA group, the circumference of the affected limb (wrist, upper wrist 4, 8, 12, 16, 20 cm levels) after operation was significantly smaller than that before operation ( P<0.05). The limb volumes of both groups were significantly reduced ( P<0.05). The comparison between the two groups showed that the reduction degree of postoperative affected limb cricumference (at the levels of 24, 28 and 32 cm above the wrist) and volume in the VLNT combined LVA group were more significant than those in the LVA group ( P<0.05). Conclusion:Compared with LVA alone, VLNT combined with LVA is more effective in the treatment of patients with upper limb lymphedema after breast cancer surgery.

Objective:To investigate the effect of vascularized lymph node transfer (VLNT) combined with lymphatico-venous anastomosis (LVA) in the treatment of lymphedema after breast cancer surgery.Methods:The data of patients with upper limb lymphedema after breast cancer surgery who were treated in the Department of Reconstructive and Reconstructive Microsurgery, Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine from July 2021 to July 2022 were retrospectively analyzed. According to different treatment methods, the patients were divided into LVA group and VLNT combined LVA group. Indocyanine green (ICG) near-infrared lymphography was performed on all affected limbs before surgery. In the LVA group, according to the results of ICG lymphography, 4 to 5 levels of the affected limb were selected, Z shaped incisions were made and dissescted until the subcutaneous fat layer. End-to-end or end-to-side anastomosis was performed between lymphatic vessels and subcutaneous venules under the microscope. In the VLNT combined LVA group, the branches of brachial artery and vein in the axillary region were marked. The inguinal flap with the superficial iliac circumflex vessel pedicle and 4-5 lymph nodes was dissected. End-to-end anastomoses of the superficial iliac circumflex vessel pedicle with the branches of brachial artery and vein were performed in the axillary region of the affected limb. LVA was performed according to ICG lymphography, the same as in the LVA group. The skin and soft tissue condition of the affected limb and the blood supply of the flap in the VLNT combined LVA group were observed after operation. The circumference of the upper arm (from the wrist to 32 cm above the wrist, every 4 cm, a total of 9 levels of circumference) and upper limb volume were measured before and after operation. SPSS 24.0 was used for data processing and analysis. Measurement data were expressed Mean±SD. The data before and after operation in the same group were compared by paired samples t test, and the comparison between the two groups was conducted by independent samples t test. P<0.05 was considered statistically significant. Results:A total of 14 female patients were enrolled, with 7 patients in each group. All cases were unilateral lymphedema. There were no significant differences in age, stage of disease, limb circumference and limb volume between the two groups before operation ( P>0.05). After operation, the skin and soft tissue condition of the affected limbs were good, and no complications such as erysipelas, cellulitis, or lymphangitis occurred. All flaps in the VLNT combined LVA group survived successfully, and the operation wounds healed well. There were no complications such as infection and necrosis of the flaps. One year after operation, the circumference and volume of the affected limb in the two groups were improved to varying degrees, and the circumference of the affected limb (wrist, upper wrist 4, 8, 12, 16, 20, 24, 28, 32 cm levels) in the VLNT combined LVA group was significantly smaller than that before operation ( P<0.01). In the LVA group, the circumference of the affected limb (wrist, upper wrist 4, 8, 12, 16, 20 cm levels) after operation was significantly smaller than that before operation ( P<0.05). The limb volumes of both groups were significantly reduced ( P<0.05). The comparison between the two groups showed that the reduction degree of postoperative affected limb cricumference (at the levels of 24, 28 and 32 cm above the wrist) and volume in the VLNT combined LVA group were more significant than those in the LVA group ( P<0.05). Conclusion:Compared with LVA alone, VLNT combined with LVA is more effective in the treatment of patients with upper limb lymphedema after breast cancer surgery.