1.Exploration of the indications for pedicled transverse rectus abdominis myocutaneous flap combined with deep inferior epigastric artery perforator flap for breast reconstruction
Yutong YUAN ; Boyang XU ; Su FU ; Shangshan LI ; Dali MU ; Minqiang XIN ; Weiwei CHEN ; Wenyue LIU ; Zhaohan CHEN ; Xingyi DU ; Xiaomu MA ; Ao FU ; Yiye OUYANG ; Chunjun LIU ; Jie LUAN
Chinese Journal of Plastic Surgery 2023;39(9):939-946
Objective:This study aimed to explore the clinical application value of the indication selection criteria for bi-pedicled deep inferior epigastric arterial perforator flap(DIEP) versus pedicled transverse rectus abdominis myocutaneous flap(TRAM) combined with DIEP for breast reconstruction.Methods:The clinical data of patients who underwent delayed bi-pedicled abdominal flap breast reconstruction after breast cancer surgery in the Department of Oncoplastic and Reconstructive Breast Surgery and Department of Mammoplasty, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2008 to April 2022 were retrospectively analyzed. According to the indications for each procedure, corresponding blood supply source was selected. Subsequently, patients were divided into two groups: bi-pedicled DIEP group and pedicled TRAM+ DIEP combined flap group according to the type of blood supply source. The indication selection criteria for pedicled TRAM+ DIEP combined flap was as follows: lack of two groups of recipient vessels; no dominant perforator in the abdominal donor site; previous abdominal liposuction or abdominal surgery with large dissection range and severe scar; patients were unable to tolerate prolonged surgery or had high risk factors for microsurgery. A comparison of breast reconstruction surgery characteristics was conducted between the two groups. Independent-samples t-test, rank sum test, Chi-square test and Fisher’s exact probability method were used to analyze various data including age, body mass index (BMI), time interval between breast cancer surgery and breast reconstruction surgery, history of radiotherapy, history of chemotherapy, history of smoking, history of lower abdominal surgery, history of hypertension, original surgical incision type, length and width of flap, hospitalization length, preoperative and postoperative conditions, as well as postoperative complications. Statistical significance was defined at P<0.05. Results:A total of 54 patients who underwent unilateral breast reconstruction were included in this study with a mean age of 42.2 years (range, 30-59 years). The pedicled TRAM+ DIEP combined flap group consisted of 21 patients with a mean age of 42.7 years (range, 33-56 years) while the bi-pedicled DIEP group comprised 33 patients with a mean age of 41.8 years (30-59 years). Out of the 54 patients, a total of 38 were eventually followed up, comprising 14 in the pedicled TRAM+ DIEP combined flap group and 24 in the bi-pedicled DIEP group. The average follow-up duration for the pedicled TRAM+ DIEP combined flap group was 42 months (range, 6-69 months). Abdominal bulge occurred in one patient, while another patient experienced abdominal hernia. The average follow-up duration for the bi-pedicled DIEP group was 47 months (6-179 months), with no reported cases of abdominal bulge or abdominal hernia. There were no statistically significant differences observed between the two groups regarding age, BMI, time interval between breast cancer surgery and breast reconstruction surgery, history of radiotherapy or chemotherapy, history of smoking, history of lower abdominal surgery, history of hypertension, original surgical incision type, length and width of flap, hospitalization length or time to ambulation (all P>0.05). However, there was a statistically significant difference noted in duration of surgery between the pedicled TRAM+ DIEP combined flap group and bi-pedicled DIEP group [6.0(5.5-6.5) hours vs. 8.5(8.0-8.8) hours] ( P<0.01). Comparison analysis revealed no statistically significant differences in terms of partial flap necrosis (0/21 vs. 1/33), abdominal incision dehiscence (2/21 vs. 2/33), abdominal bulge (1/14 vs. 0 /24) or abdominal hernia (1/1 vs. 0/24) between the two groups ( P>0.05). Conclusion:For patients who needed bilateral vascular pedicled lower abdominal wall for breast reconstruction, utilizing a pedicled TRAM+ DIEP combined flap did not increase surgical complication risks. The proposed indication selection criteria for using a pedicled TRAM + DIEP combined flap outlined in this study could serve as guidance when choosing methods for bi-pedicled abdominal flap breast reconstruction.
2.Study on improving the effect of large-volume fat grafting breast augmentation using filtration-adsorption technique
Ningdan MA ; Jie LUAN ; Su FU ; Chunjun LIU ; Dali MU ; Minqiang XIN ; Boyang XU ; Lin CHEN
Chinese Journal of Plastic Surgery 2023;39(11):1213-1221
Objective:To preliminarily evaluate the outcome of fat grafting breast augmentation using filtration-adsorption technique.Methods:Patients undergoing autologous fat grafting (AFG) breast augmentation between July of 2020 and March of 2021 were retrospectively reviewed at Breast Plastic and Reconstructive Surgery Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College. Patients with complete data who followed up for at least 3 months were included. The lipoaspirates were processed with filtration-adsorption technique and AFG breast augmentation were performed (filtration-adsorption group), and then sedimentation method which was commonly used clinically was set as the control group (sedimentation group). During the follow-up, breast palpation and ultrasound inspection were performed, and the breast volumetric change between preoperative and postoperative was quantitatively compared by three-dimensional scanning technique. Demographic and surgical details were statistically analyzed. Postoperative breast volume retention rates and complication rates were calculated and compared. Normally distributed continuous variables were described in the form Mean±SD, and t-test was used for analysis. Non-normal distributed continuous variables were described in the form of M ( Q1, Q3) and analyzed by Mann-Whitney U test. The classified data were analyzed by Fisher exact probability test. Results:A total of 20 female patients (40 breasts) were included, including 10 patients (20 breasts) aged (31.4±2.5) years in sedimentation group and 10 patients (20 breasts) aged (28.5±4.4) years in the filtration-adsorption group. The fat injection volume in the filtration-adsorption group was 151.1 (125.0, 175.0) ml, and 165.0 (151.3, 196.3) ml in sedimentation group, respectively, with no statistical significance ( P>0.05). The breast volume retention rate in filtration-adsorption group was (62.93±14.06)%, which was significantly higher than that in sedimentation group (24.97±11.02)% ( P<0.01). During postoperative breast ultrasound examination, 2 breasts in sedimentation group had palpable nodules (< 6 mm), 1 breast had ultrasonically detectable but clinically inaccessible nodules (< 4 mm), while only 1 breast in the filtration-adsorption group had an oil cyst (4 mm×2 mm). The complication rate in the filtration-adsorption group was lower than that in sedimentation group[5.0%(1/20) vs. 15.0%(3/20), P>0.05]. Conclusion:The filtration-adsorption technique is an ideal method for fat grafting, which would multiply the volume retention rate in large-volume AFG breast augmentation and reduce the rate of complications.
3.Exploration of the indications for pedicled transverse rectus abdominis myocutaneous flap combined with deep inferior epigastric artery perforator flap for breast reconstruction
Yutong YUAN ; Boyang XU ; Su FU ; Shangshan LI ; Dali MU ; Minqiang XIN ; Weiwei CHEN ; Wenyue LIU ; Zhaohan CHEN ; Xingyi DU ; Xiaomu MA ; Ao FU ; Yiye OUYANG ; Chunjun LIU ; Jie LUAN
Chinese Journal of Plastic Surgery 2023;39(9):939-946
Objective:This study aimed to explore the clinical application value of the indication selection criteria for bi-pedicled deep inferior epigastric arterial perforator flap(DIEP) versus pedicled transverse rectus abdominis myocutaneous flap(TRAM) combined with DIEP for breast reconstruction.Methods:The clinical data of patients who underwent delayed bi-pedicled abdominal flap breast reconstruction after breast cancer surgery in the Department of Oncoplastic and Reconstructive Breast Surgery and Department of Mammoplasty, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2008 to April 2022 were retrospectively analyzed. According to the indications for each procedure, corresponding blood supply source was selected. Subsequently, patients were divided into two groups: bi-pedicled DIEP group and pedicled TRAM+ DIEP combined flap group according to the type of blood supply source. The indication selection criteria for pedicled TRAM+ DIEP combined flap was as follows: lack of two groups of recipient vessels; no dominant perforator in the abdominal donor site; previous abdominal liposuction or abdominal surgery with large dissection range and severe scar; patients were unable to tolerate prolonged surgery or had high risk factors for microsurgery. A comparison of breast reconstruction surgery characteristics was conducted between the two groups. Independent-samples t-test, rank sum test, Chi-square test and Fisher’s exact probability method were used to analyze various data including age, body mass index (BMI), time interval between breast cancer surgery and breast reconstruction surgery, history of radiotherapy, history of chemotherapy, history of smoking, history of lower abdominal surgery, history of hypertension, original surgical incision type, length and width of flap, hospitalization length, preoperative and postoperative conditions, as well as postoperative complications. Statistical significance was defined at P<0.05. Results:A total of 54 patients who underwent unilateral breast reconstruction were included in this study with a mean age of 42.2 years (range, 30-59 years). The pedicled TRAM+ DIEP combined flap group consisted of 21 patients with a mean age of 42.7 years (range, 33-56 years) while the bi-pedicled DIEP group comprised 33 patients with a mean age of 41.8 years (30-59 years). Out of the 54 patients, a total of 38 were eventually followed up, comprising 14 in the pedicled TRAM+ DIEP combined flap group and 24 in the bi-pedicled DIEP group. The average follow-up duration for the pedicled TRAM+ DIEP combined flap group was 42 months (range, 6-69 months). Abdominal bulge occurred in one patient, while another patient experienced abdominal hernia. The average follow-up duration for the bi-pedicled DIEP group was 47 months (6-179 months), with no reported cases of abdominal bulge or abdominal hernia. There were no statistically significant differences observed between the two groups regarding age, BMI, time interval between breast cancer surgery and breast reconstruction surgery, history of radiotherapy or chemotherapy, history of smoking, history of lower abdominal surgery, history of hypertension, original surgical incision type, length and width of flap, hospitalization length or time to ambulation (all P>0.05). However, there was a statistically significant difference noted in duration of surgery between the pedicled TRAM+ DIEP combined flap group and bi-pedicled DIEP group [6.0(5.5-6.5) hours vs. 8.5(8.0-8.8) hours] ( P<0.01). Comparison analysis revealed no statistically significant differences in terms of partial flap necrosis (0/21 vs. 1/33), abdominal incision dehiscence (2/21 vs. 2/33), abdominal bulge (1/14 vs. 0 /24) or abdominal hernia (1/1 vs. 0/24) between the two groups ( P>0.05). Conclusion:For patients who needed bilateral vascular pedicled lower abdominal wall for breast reconstruction, utilizing a pedicled TRAM+ DIEP combined flap did not increase surgical complication risks. The proposed indication selection criteria for using a pedicled TRAM + DIEP combined flap outlined in this study could serve as guidance when choosing methods for bi-pedicled abdominal flap breast reconstruction.
4.Study on improving the effect of large-volume fat grafting breast augmentation using filtration-adsorption technique
Ningdan MA ; Jie LUAN ; Su FU ; Chunjun LIU ; Dali MU ; Minqiang XIN ; Boyang XU ; Lin CHEN
Chinese Journal of Plastic Surgery 2023;39(11):1213-1221
Objective:To preliminarily evaluate the outcome of fat grafting breast augmentation using filtration-adsorption technique.Methods:Patients undergoing autologous fat grafting (AFG) breast augmentation between July of 2020 and March of 2021 were retrospectively reviewed at Breast Plastic and Reconstructive Surgery Center, Plastic Surgery Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College. Patients with complete data who followed up for at least 3 months were included. The lipoaspirates were processed with filtration-adsorption technique and AFG breast augmentation were performed (filtration-adsorption group), and then sedimentation method which was commonly used clinically was set as the control group (sedimentation group). During the follow-up, breast palpation and ultrasound inspection were performed, and the breast volumetric change between preoperative and postoperative was quantitatively compared by three-dimensional scanning technique. Demographic and surgical details were statistically analyzed. Postoperative breast volume retention rates and complication rates were calculated and compared. Normally distributed continuous variables were described in the form Mean±SD, and t-test was used for analysis. Non-normal distributed continuous variables were described in the form of M ( Q1, Q3) and analyzed by Mann-Whitney U test. The classified data were analyzed by Fisher exact probability test. Results:A total of 20 female patients (40 breasts) were included, including 10 patients (20 breasts) aged (31.4±2.5) years in sedimentation group and 10 patients (20 breasts) aged (28.5±4.4) years in the filtration-adsorption group. The fat injection volume in the filtration-adsorption group was 151.1 (125.0, 175.0) ml, and 165.0 (151.3, 196.3) ml in sedimentation group, respectively, with no statistical significance ( P>0.05). The breast volume retention rate in filtration-adsorption group was (62.93±14.06)%, which was significantly higher than that in sedimentation group (24.97±11.02)% ( P<0.01). During postoperative breast ultrasound examination, 2 breasts in sedimentation group had palpable nodules (< 6 mm), 1 breast had ultrasonically detectable but clinically inaccessible nodules (< 4 mm), while only 1 breast in the filtration-adsorption group had an oil cyst (4 mm×2 mm). The complication rate in the filtration-adsorption group was lower than that in sedimentation group[5.0%(1/20) vs. 15.0%(3/20), P>0.05]. Conclusion:The filtration-adsorption technique is an ideal method for fat grafting, which would multiply the volume retention rate in large-volume AFG breast augmentation and reduce the rate of complications.
5.DNA super-barcoding of several medicinal species in Gentiana from Yunnan province.
China Journal of Chinese Materia Medica 2021;46(20):5260-5269
Gentiana is an important but complicated group in Gentianaceae. The genus covers numerous medicinal plants which are difficult to be identified. In the present study, several medicinal species in Gentiana from Yunnan province, including G. rigescens, G.rhodantha, and G. delavayi, were sequenced using the Illumina HiSeq 2500 system. Three complete chloroplast genome sequences were obtained after assembly and annotation. According to several published genome sequences of G. crassicaulis, the DNA super-barcoding of species in Gentiana was preliminarily carried out. The results revealed that chloroplast genomes of the three species were conservative with short lengths(146 944, 148 992, and 148 796 bp, respectively). The genomes encoded 114 genes, including 78 protein-coding genes, 30 tRNA genes, 4 rRNA genes, and 2 pseudogenes. Furthermore, these medicinal species in Yunnan province were identified using DNA super-barcoding based on chloroplast genomes. The results showed that the Gentiana species could be gathered into monophyletic branches with a high support value(100%). It indicated that DNA super-barcoding possessed obvious advantages in discriminating species in complicated genera. This study is expected to provide a scientific basis for the identification, utilization, and conservation of Gentiana species.
China
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DNA
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Genome, Chloroplast/genetics*
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Gentiana/genetics*
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Phylogeny
6.Clinical application and planning strategy of bipedicled deep inferior epigastric perforator flaps in delayed breast reconstruction
Boyang XU ; Su FU ; Minqiang XIN ; Dali MU ; Chunjun LIU ; Jie LUAN
Chinese Journal of Plastic Surgery 2021;37(7):712-718
Objective:Introducing clinical application, planning strategy, and surgical tips of bipedicled deep inferior epigastric perforator (DIEP) flaps in delayed breast reconstruction.Methods:A retrospective study of patients who underwent delayed breast reconstruction using bipedicled DIEP flaps from April 2015 to April 2019 at the facility of the authors was carried out. Preoperative CT angiograph was performed for evaluation of deep inferior epigastric perforators. Intraoperative indocyanine green angiography was performed for real-time flap perfusion assessment. Baseline data, information of mastectomy and adjuvant therapy, surgical details, and follow-up records were reviewed and collected for statistical analysis.Results:A total of 31 patients were enrolled, with years of 43.0 on average (range, 30 to 55). Among them 16 patients received radiotherapy (51.6%), 9 patients had lower abdominal scars due to previous surgery (29.0%), 2 patients had body mass index (BMI) lower than 20 kg/m 2(6.5%). Measurements of flaps ranged from 20 cm×8 cm to 20 cm×38 cm. The operative time was 593±94 minutes. Of the 31 flaps enrolled, 24 were DIEP/DIEP flaps, 6 were DIEP/SIEA flaps, one was SIEA/SIEA flap. Intraflap anastomoses were performed in 6 flaps. The other 25 flaps were anastomosed to separate recipients. There was no flap loss encountered. Marginal necrosis of the flap happened in one patient, fat liquefaction of the donor site occurred in four patients, which was treated with conservative debridement. Patients were followed up for 13 months on average (range, 10 to 60 months). No long-term complication was observed during the follow-up. All patients reported satisfying aesthetic outcomes. Conclusions:Bipedicled DIEP flaps provide sufficient tissue quantity and reliable blood supply. Patients with severe breast deficiency, lacking lower abdominal fat, or having abdominal midline scars were appropriate to perform this type of flaps for delayed breast reconstruction.
7.Clinical application and planning strategy of bipedicled deep inferior epigastric perforator flaps in delayed breast reconstruction
Boyang XU ; Su FU ; Minqiang XIN ; Dali MU ; Chunjun LIU ; Jie LUAN
Chinese Journal of Plastic Surgery 2021;37(7):712-718
Objective:Introducing clinical application, planning strategy, and surgical tips of bipedicled deep inferior epigastric perforator (DIEP) flaps in delayed breast reconstruction.Methods:A retrospective study of patients who underwent delayed breast reconstruction using bipedicled DIEP flaps from April 2015 to April 2019 at the facility of the authors was carried out. Preoperative CT angiograph was performed for evaluation of deep inferior epigastric perforators. Intraoperative indocyanine green angiography was performed for real-time flap perfusion assessment. Baseline data, information of mastectomy and adjuvant therapy, surgical details, and follow-up records were reviewed and collected for statistical analysis.Results:A total of 31 patients were enrolled, with years of 43.0 on average (range, 30 to 55). Among them 16 patients received radiotherapy (51.6%), 9 patients had lower abdominal scars due to previous surgery (29.0%), 2 patients had body mass index (BMI) lower than 20 kg/m 2(6.5%). Measurements of flaps ranged from 20 cm×8 cm to 20 cm×38 cm. The operative time was 593±94 minutes. Of the 31 flaps enrolled, 24 were DIEP/DIEP flaps, 6 were DIEP/SIEA flaps, one was SIEA/SIEA flap. Intraflap anastomoses were performed in 6 flaps. The other 25 flaps were anastomosed to separate recipients. There was no flap loss encountered. Marginal necrosis of the flap happened in one patient, fat liquefaction of the donor site occurred in four patients, which was treated with conservative debridement. Patients were followed up for 13 months on average (range, 10 to 60 months). No long-term complication was observed during the follow-up. All patients reported satisfying aesthetic outcomes. Conclusions:Bipedicled DIEP flaps provide sufficient tissue quantity and reliable blood supply. Patients with severe breast deficiency, lacking lower abdominal fat, or having abdominal midline scars were appropriate to perform this type of flaps for delayed breast reconstruction.
8.Effects of age on the fat grafts with adipose-derived stem cells for fat survival: an experimental study
Yaping QU ; Jie LUAN ; Dali MU ; Su FU ; Qian WANG ; Zifei LI ; Tong LIU
Chinese Journal of Plastic Surgery 2020;36(8):905-915
Objective:The aim of this study is to compare the effects of age on the biological properties of adipose-derived stem cells(ASCs) and fat survival of ASC-assisted lipotransfer. To identify the effect of age factors on the biological characteristics of human ASCs and compare the effects of ASCs-assisted subcutaneous fat transplantation/lipotransfer on nude mice at different ages.Methods:Human lipoaspirates were obtained from 30 healthy female patients (aged from 18 to 65 years) acquiring the abdominal liposuction. Samples were divided into three groups according to donor age: group A, 18-29 years; group B, 30-49 years; group C, 50-65 years. Stromal vascular fraction cells were isolated from the harvested adipose tissue using collagenase. The yield and cell viability of SVF were tested using the Muse cell count and viability assay. ASCs were cultured and harvested at the second passage. MSC surface markers of ASCs were examined by the flow cytometry. The cell proliferation of ASCs from different age was determined by the CCK-8 assay. The scratch test was used for assessing the ASCs migration ability. The adipogenic differentiation potential of ASCs was analyzed by induction of lipid formation in vitro. The expression levels of PPAR-γ and CEBP-α genes were detected by RT-PCR assay. The survival of adipocytes in the grafts was analyzed by perilipin-A immunofluorescence staining. The fat survival of ASCs-enriched grafts from different age was measured in animal models. The weight and residual volume of fat grafts were compared in different groups after three months. The histologic analysis was evaluated by cell integrity and necrosis tissue in fat grafts. The vessel density was measured using the CD31 immunohistochemical staining. Data were analyzed by SPSS software version 21.0 with one-way ANOVA to compare the difference of multiple groups. A value of P< 0.05 was considered statistically significant. Results:The yield and cell viability of SVF isolated from lipoaspirates were: group A, (7.06±1.28)×10 5/ml and 82.46%±2.81%; group B, (6.90±0.32)×10 5/ml and 82.01%±3.85%; group C, (6.40±0.62)×10 5/ml and 77.82%±3.45%, respectively. No significant difference was found in different age groups. SVF viability was decreased with increasing age. The expression of positive surface markers CD90, CD44, CD105 and CD73 of ASCs in each group was above 95%, and the expression of negative surface markers was below 2%, all of which met the criteria for the expression level of mesenchymal stem cell surface markers. Moreover, there was a decline in cell proliferation and migration of ASCs with increasing age. No significant difference was found in the adipogenic differentiation of ASCs in three groups. The fat grafts were harvested three months after cell-assisted lipotransfer. The graft weight was(0.18±0.02) g in group A, (0.17±0.02) g in group B, (0.15±0.01) g in group C, (0.13±0.03) g in control group, respectively; F=9.274, P<0.001. The residual volume of grafts was(262.88±17.69)/mm 3 in group A, (263.83±25.96)/mm 3 in group B, (240.06±25.08)/mm 3 in group C, (201.81±31.48)/mm 3 in the control group; F=12.95, P<0.001. There were significant differences in the weight and residual volume of fat grafts in different age groups( F=5.231, P=0.012; F=3.364, P=0.049). HE staining result showed that compared with the blank control group, ASC-assisted groups had uniform distribution of adipocytes, less fibrous connective tissue and necrotic tissue. There was a statistically significant difference in the proportion of fat integrity and necrotic tissue between the groups ( F=3.434, P=0.027; F=9.314, P<0.001). Results of the histologic analysis showed no significant difference in the proportion of fat cell integrity and necrotic tissue in each group( F=0.282, P=0.756; F=0.421, P=0.661). Immunofluorescence staining result showed that, compared with the control group, a higher number of perilipin-positive adipocytes were observed in ASCs-assisted fat grafting from different age groups, with uniform distribution. The vessel density of fat grafts was (15.70±4.16)/mm 2 in group A, (17.03±8.30)/mm 2 in group B; (16.68±6.71)/mm 2 in group C, (11.50±4.04)/mm 2 in control group; F=3.523, P=0.019. Conclusions:The proliferation and migration of human ASCs decreased with age, but age did not affect the adipogenic differentiation potential of ASCs. ASCs from different ages effectively improved the fat survival of grafts. ASCs-assisted fat grafting was more effective in young people than in elder.
9.Effects of age on the fat grafts with adipose-derived stem cells for fat survival: an experimental study
Yaping QU ; Jie LUAN ; Dali MU ; Su FU ; Qian WANG ; Zifei LI ; Tong LIU
Chinese Journal of Plastic Surgery 2020;36(8):905-915
Objective:The aim of this study is to compare the effects of age on the biological properties of adipose-derived stem cells(ASCs) and fat survival of ASC-assisted lipotransfer. To identify the effect of age factors on the biological characteristics of human ASCs and compare the effects of ASCs-assisted subcutaneous fat transplantation/lipotransfer on nude mice at different ages.Methods:Human lipoaspirates were obtained from 30 healthy female patients (aged from 18 to 65 years) acquiring the abdominal liposuction. Samples were divided into three groups according to donor age: group A, 18-29 years; group B, 30-49 years; group C, 50-65 years. Stromal vascular fraction cells were isolated from the harvested adipose tissue using collagenase. The yield and cell viability of SVF were tested using the Muse cell count and viability assay. ASCs were cultured and harvested at the second passage. MSC surface markers of ASCs were examined by the flow cytometry. The cell proliferation of ASCs from different age was determined by the CCK-8 assay. The scratch test was used for assessing the ASCs migration ability. The adipogenic differentiation potential of ASCs was analyzed by induction of lipid formation in vitro. The expression levels of PPAR-γ and CEBP-α genes were detected by RT-PCR assay. The survival of adipocytes in the grafts was analyzed by perilipin-A immunofluorescence staining. The fat survival of ASCs-enriched grafts from different age was measured in animal models. The weight and residual volume of fat grafts were compared in different groups after three months. The histologic analysis was evaluated by cell integrity and necrosis tissue in fat grafts. The vessel density was measured using the CD31 immunohistochemical staining. Data were analyzed by SPSS software version 21.0 with one-way ANOVA to compare the difference of multiple groups. A value of P< 0.05 was considered statistically significant. Results:The yield and cell viability of SVF isolated from lipoaspirates were: group A, (7.06±1.28)×10 5/ml and 82.46%±2.81%; group B, (6.90±0.32)×10 5/ml and 82.01%±3.85%; group C, (6.40±0.62)×10 5/ml and 77.82%±3.45%, respectively. No significant difference was found in different age groups. SVF viability was decreased with increasing age. The expression of positive surface markers CD90, CD44, CD105 and CD73 of ASCs in each group was above 95%, and the expression of negative surface markers was below 2%, all of which met the criteria for the expression level of mesenchymal stem cell surface markers. Moreover, there was a decline in cell proliferation and migration of ASCs with increasing age. No significant difference was found in the adipogenic differentiation of ASCs in three groups. The fat grafts were harvested three months after cell-assisted lipotransfer. The graft weight was(0.18±0.02) g in group A, (0.17±0.02) g in group B, (0.15±0.01) g in group C, (0.13±0.03) g in control group, respectively; F=9.274, P<0.001. The residual volume of grafts was(262.88±17.69)/mm 3 in group A, (263.83±25.96)/mm 3 in group B, (240.06±25.08)/mm 3 in group C, (201.81±31.48)/mm 3 in the control group; F=12.95, P<0.001. There were significant differences in the weight and residual volume of fat grafts in different age groups( F=5.231, P=0.012; F=3.364, P=0.049). HE staining result showed that compared with the blank control group, ASC-assisted groups had uniform distribution of adipocytes, less fibrous connective tissue and necrotic tissue. There was a statistically significant difference in the proportion of fat integrity and necrotic tissue between the groups ( F=3.434, P=0.027; F=9.314, P<0.001). Results of the histologic analysis showed no significant difference in the proportion of fat cell integrity and necrotic tissue in each group( F=0.282, P=0.756; F=0.421, P=0.661). Immunofluorescence staining result showed that, compared with the control group, a higher number of perilipin-positive adipocytes were observed in ASCs-assisted fat grafting from different age groups, with uniform distribution. The vessel density of fat grafts was (15.70±4.16)/mm 2 in group A, (17.03±8.30)/mm 2 in group B; (16.68±6.71)/mm 2 in group C, (11.50±4.04)/mm 2 in control group; F=3.523, P=0.019. Conclusions:The proliferation and migration of human ASCs decreased with age, but age did not affect the adipogenic differentiation potential of ASCs. ASCs from different ages effectively improved the fat survival of grafts. ASCs-assisted fat grafting was more effective in young people than in elder.
10. Isolation and epidemiological significance of Yersinia pestis phages in indicator animals from the house mouse plague foci of Yunnan Province
Youhong ZHONG ; Chao SU ; Biao DUAN ; Yibo DING ; Qingchun DU ; Cunjuan DUAN ; Danni ZHAO ; Wei LI ; Peng WANG ; Hesong WU
Chinese Journal of Endemiology 2019;38(11):861-867
Objective:
To investigate whether plague phages were present in the indicator animals of plague foci in Yunnan Province, and to explore their epidemiological significance.
Methods:
Anus swabs were collected from indicator animals (dogs or cats) of the 41 plague affected villages in 26 towns of 10 cities (counties, districts) of Yunnan plague foci from November of 2015 to March of 2018. The

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