Objective:To summarize the technical experience of using a contralateral free gracilis myocutaneous flap combined with an adductor magnus perforator flap for salvaging failure of breast reconstruction after transplantation with the free adductor magnus perforator flap.Methods:A retrospective analysis was conducted on patients with breast cancer who received contralateral posteromedial thigh flap (gracilis myocutaneous flap combined with adductor magnus perforator flap) transplantation for immediate unilateral breast reconstruction developed vascular crisis and caused flap necrosis at the Department of Oncology Plastic Surgery, Hunan Cancer Hospital, between December 2016 to December 2022. All patients with early-stage breast cancer received modified radical mastectomy. All patients were selected to undergo the immediate breast reconstruction surgery with unilateral free adductor magnus perforator flap transplantation. The proximal end of internal mammary vessels were used as the recipient vessels in all cases. After the emergency exploration confirmed the necrosis of the flap, a contralateral free gracilis myocutaneous flap combined with adductor magnus perforator flap were immediately harvested for salvage breast reconstruction. The vascular pedicle of the gracilis muscular branch and the adductor magnus perforator branch were anastomosed with the proximal and distal ends of the internal mammary vessels, respectively. Postoperative patient monitoring was conducted intensively, with follow-up assessments focusing on breast contour and donor site recovery.Results:A total of 5 patients were included, aged 26 to 42 years, with an average of (31.5±1.8) years. All salvaged breast reconstruction surgeries were successful, the salvaged flap measured 15.0 cm×6.0 cm×4.0 cm-17.0 cm×7.5 cm×5.5 cm. All the transplanted flaps survived after the operation, with satisfactory breast contour, good flap elasticity, and no contracture or deformation. The donor sites of both thigh flaps healed well, leaving only linear scars and no significant functional impairment to the lower limbs. Follow-up ranged from 9 to 24 months, with an average of 15.7 months. The breast shape was satisfactory, and there was no recurrence of breast cancer.Conclusion:The donor site morbidity of the posteromedial thigh flap is relatively small. When unilateral flap transplantation fails, immediate transfer of the contralateral flap can be chosen for salvage reconstruction. This approach ensures symmetrical donor site outcomes and is generally well-accepted by patients.

Objective:To explore the technical points and precautions of using bilateral free posteromedial thigh flap to reconstruct bilateral breasts.Methods:A retrospective analysis was conducted on the clinical data of patients who underwent bilateral mastectomy and bilateral free posteromedial thigh flap transplantation for bilateral breast reconstruction at Hunan Cancer Hospital from June 2020 to January 2023. The operation was carried out simultaneously by two groups of doctors. Breast surgeons performed modified radical mastectomy for breast cancer or subcutaneous mastectomy. The flap group prepared bilateral free posteromedial thigh flaps. The posteromedial thigh flaps were designed in three patterns: transverse, vertical and oblique. The intrathoracal vessels were used as recipient vessels in all cases. There were two types of vascular anastomosis method: ① For flaps with single vascular pedicle, the artery was anastomosed with the proximal end of the internal thoracic artery, and the only accompanying vein was anastomosed with the proximal end of the internal thoracic vein, or the two accompanying veins were anastomosed with the proximal and distal ends of the internal thoracic vein; ② For flaps with dual vascular pedicles, arteries were anastomosed with the proximal and distal ends of the internal thoracic artery, and the accompanying veins on both sides communicated with the proximal and distal ends of the internal thoracic vein, respectively. Normal distribution measurement data were expressed as Mean±SD.Results:A total of 9 female patients were enrolled, aged 29 to 43 years, with an average age of 38.3 years. Among them, bilateral breast cancer was considered in 4 cases, unilateral breast cancer with multiple nodules in the opposite breast was in 3 cases, and bilateral multiple nodules along with high-risk of developing breast cancer suggested by gene testing were found in 2 cases. Modified radical mastectomy for breast cancer and nipple-sparing subcutaneous mastectomy were performed in 7 and 11 breasts, respectively. There were three design method for the posteromedial thigh flap: horizontal design (6 thighs); vertical design (6 thighs); diagonal design (6 thighs). A total of 18 posteromedial thigh flaps were raised. Four different types of posteromedial thigh flaps were used: gracilis muscle flap (6 thighs); great adductor muscle perforator flap (5 thighs); great adductor muscle flap (5 thighs); femoral gracilis flap+ adductor magnus flap (2 thighs). The skin flaps harvested were (18.9±0.6) cm in length, (7.2±0.4) cm in width and (4.2±0.3) cm in thickness. The average weight of the skin flaps were 235 grams (185-355 grams). The aforementioned vascular anastomosis method ① and ② were carried out in 15 and 3 breasts, respectively. The patients were followed up for 6 to 32 months, with an average of 17.3 months. All skin flaps survived, and the reconstructed breasts presented with good appearance, good elasticity, and no skin flap contraction or deformation occurred; scarring on the donor area was linear and mild; there was no significant impact on lower limb movement. The breast shape was satisfactory, and there was no recurrence of breast cancer.Conclusion:The application of bilateral free posteromedial thigh flap transplantation for reconstruction of bilateral breasts has flexible flap preparation and vascular anastomosis forms, which can achieve satisfactory result of bilateral symmetry.

The free inferior epigastric artery perforator flap is currently a highly regarded autologous breast reconstruction technique. However, in clinical practice, the anatomical characteristics of the lower abdominal vasculature and surrounding tissue structures do not always permit ideal perforator flap harvesting. In many cases, it becomes necessary to preserve a larger rectus abdominis muscle cuff to ensure reliable blood supply to the flap through the perforating vessels. Compared with various forms of rectus abdominis myocutaneous flaps that all require incision of the anterior sheath and intramuscular vascular pedicle dissection, the superficial inferior epigastric artery (SIEA) perforator flap significantly reduces donor-site morbidity. Nevertheless, the anatomical variability of the superficial inferior epigastric vessels increases surgical uncertainty. Based on clinical observations, this study establishes a comprehensive clinical strategy algorithm. Primarily focusing on different configurations of free rectus abdominis flaps, it incorporates additional preparation of free SIEA perforator flaps when anatomical conditions permit. The objectives are to ensure reliable perfusion of the lower abdominal flap while minimizing donor-site damage and reducing surgical complexity.

Objective:To evaluate the efficacy and safety of remedial pedicled rectus abdominis myocutaneous flap transplantation for breast reconstruction.Methods:This retrospective study included 23 female breast cancer patients who underwent autologous breast reconstruction with a remedial pedicled rectus abdominis myocutaneous flap at Hunan Cancer Hospital from June 2022 to June 2023. The patients' ages ranged from 24 to 56 years, with a mean age of (43.2±1.3) years. All patients had previously undergone mastectomy and tumor resection before receiving the remedial pedicled rectus abdominis myocutaneous flap reconstruction. Follow-up was conducted through outpatient visits and telephone interviews, with a median follow-up period of 10 (6-15) months. Observation indicators included survival rate of flaps, patient satisfaction rate and aesthetic outcomes. Adverse reactions including but not limited to flap contracture and deformation, during and following the surgical procedure were also recorded.Results:The remedial pedicled rectus abdominis myocutaneous flap technique was utilized for autologous breast reconstruction in 23 patients. Specifically, this approach was chosen due to two primary reasons: the perforator distance from the vascular pedicle being excessively long in 18 cases, and suboptimal characteristics of the main pedicle in 5 cases. Two types of flap configurations were employed. For 15 patients, a unilateral remedial pedicled rectus abdominis myocutaneous flap was used, with an average flap size of (21.5±0.8) cm in length and (11.2±1.3) cm in width. All flaps demonstrated adequate blood supply without the need for additional vascular anastomosis. For the remaining 8 patients, a combination of a unilateral remedial pedicled rectus abdominis myocutaneous flap and a contralateral free lower abdominal flap was utilized, resulting in an average flap size of (29.3±0.7) cm in length and (12.0±1.8) cm in width. In these cases, the inferior epigastric vessels of the free lower abdominal flap were successfully anastomosed to the internal thoracic vessels of the recipient site, ensuring all flaps survived postoperatively. The survival rate of flaps in 23 patients was 100% (23/23), and the patient satisfaction rate was 100% (23/23). Postoperative evaluation revealed that the reconstructed breasts exhibited satisfactory shape and texture, with no instances of flap contracture or deformation. The reconstructed breasts were largely symmetrical with the contralateral side, leaving only linear scars at the donor and recipient sites. Importantly, there was no significant impact on abdominal wall function, and no adverse reactions such as flap contracture or impaired abdominal wall function were observed in any of the patients.Conclusion:When the characteristics of the perforating vessels in the lower abdomen are suboptimal, the use of a remedial pedicled rectus abdominis myocutaneous flap technique for breast reconstruction can ensure satisfactory outcomes and significantly enhance surgical safety.

Objective:To explore the clinical effects of lobulated supercharged pedicled rectus abdominis myocutaneous flap for repairing huge chest wall wounds.Methods:This study was a retrospective observational study. From January 2020 to June 2023, 9 patients with huge chest wall wounds remained after extended radical tumor resection, extended tumor resection, or lesion resection and conformed to the inclusion criteria were admitted to Hunan Cancer Hospital, including 3 males and 6 females, aged 31-59 years. The lobulated supercharged pedicled rectus abdominis myocutaneous flap was used to repair the huge chest wall wounds. The area of chest wall skin and soft tissue defects ranged from 19 cm×15 cm to 25 cm×21 cm, and the area of the harvested myocutaneous flap ranged from 25.0 cm×7.5 cm to 32.0 cm×13.0 cm. After repairing the rectus abdominis muscle and its anterior sheath with a polypropylene mesh, the incision in the donor site was directly sutured. The blood supply of the myocutaneous flap and the selection of blood vessels in recipient area for supercharging during the surgery, the survival of the myocutaneous flap and the healing of the donor area incision after the surgery were observed. The appearance and texture of the reconstructed chest wall, the scar formation in the abdominal donor area, and their impacts on function and appearance, and the tumor recurrence and metastasis were followed up.Results:The blood supply of the unilateral myocutaneous flap was poor in 7 patients, and that of the bilateral myocutaneous flaps was poor in 2 patients during the surgery. The recipient area vessels selected for supercharging of the myocutaneous flap were the internal thoracic vessels in 7 patients, the thoracodorsal vessels in 2 patients, and the thoracoacromial vessels in 2 patients. All the myocutaneous flaps survived after surgery. The donor area incisions of 7 patients healed smoothly after surgery; 2 patients had partial dehiscence in the incisions due to excessive incision tension, which healed after debridement and suturing. Follow-up for 8 to 12 months showed that the reconstructed chest wall had good appearance and soft texture; only a linear scar remained in the abdominal donor area, which had no obvious impact on abdominal breathing and the abdomen was aesthetically pleasing; no local tumor recurrence was observed; distant metastases occurred in 2 breast cancer patients.Conclusions:The lobulated supercharged pedicled rectus abdominis myocutaneous flap can effectively cover huge chest wall wounds, while maximally ensuring the blood supply of the myocutaneous flap to the greatest extent and safeguarding the success of chest wall reconstruction surgery.

Objective:To investigate the clinical application effects of the pedicled anterior intercostal artery perforator flap in breast reconstruction after breast-conserving surgery for breast cancer.Methods:This study was a retrospective observational study. From January to December 2023, 16 female breast cancer patients who met the inclusion criteria were hospitalized in the Department of Breast Surgery of the Affiliated Hospital of Southwest Medical University, with the age of (48±8) years. The pedicled anterior intercostal artery perforator flap was used for breast reconstruction of patients after breast-conserving surgery. After complete resection of tumor tissue, a "crescent-shaped" incision was designed at the inframammary fold. The pedicled anterior intercostal artery perforator flap was harvested based on the tumor location and the defect area after tumor removal. The flap was de-epithelialized, coapted, and rotated anterogradely or retrogradely to fill the defect. The donor site wound was closed with layered sutures. The following parameters were recorded: breast tissue loss volume during surgery, surgical duration, retention duration of the drainage tube, positive proportion of tumors in the breast incision margin tissue, breast loss ratio, flap survival, and incidence ratio of complications after operation. Patients were followed up for local recurrence or distant metastasis of tumor. At the last follow-up, the Ueda score was used to evaluate cosmetic outcomes of reconstructed breasts after breast-conserving surgery, and the Breast-Q scale version 2.0 was applied to assess patients' satisfaction and quality of life with breast reconstruction after breast-conserving surgery.Results:The breast tissue loss volume during surgery in this group of patients was 20-128 (59±34) cm3, the surgical duration was 105-200 (143±27) min, the retention duration of the drainage tube was 3-7 (4.6±1.0) d, and the positive proportion of tumors in the breast incision margin tissue was 1/16, with breast loss ratio of 0. After the surgery, the patient's transplanted flaps all survived. One patient had postoperative fat liquefaction in the surgical area, and the incidence ratio of postoperative complications was 1/16. The patients were followed up for 3-12 (11±4) months, and no local breast cancer recurrence or distant metastasis occurred. At the last follow-up, the cosmetic score of breast reconstruction after breast-conserving surgery were excellent in 6 cases, good in 8 cases, and fair in 2 cases, with an excellent and good ratio of 14/16. At the last follow-up, the highest score in the evaluation of patients' satisfaction with breast reconstruction and quality of life after breast-conserving surgery was the satisfaction with the surgeons, with a score of 59-100 (91±13), followed respectively by physiological health of the chest with a score of 60-100 (77±14), psychological health with a score of 35-100 (74±20), breast satisfaction with a score of 55-100 (73±13), satisfaction with information acquisition with a score of 53-100 (70±14), and sexual health with a score of 34-100 (70±23).Conclusions:The pedicled anterior intercostal artery perforator flap is safe and reliable for breast reconstruction after breast-conserving surgery for breast cancer, and can achieve high cosmetic effects and patient satisfaction. This flap is simple in design, easy to operate and highly reproducible, and is worthy of clinical promotion and application.

Objective To understand the membrane protein molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in the region,and provide some evidence for rational drug use or application of efflux pump inhibitors. Methods Collected Pseudomonas aeruginosa isolated from four hospitals in the region from October 2022 to August 2023,and used SYBR-PCR method to quantitatively detect the relative mRNA expression (RE) levels of 10 membrane protein coding genes,including mexA,B,C,D,E,F,X,Y,and oprD,M. Then categorized the strains into five groups based on ceftazidime,cefepime,imipenem,and meropenem resistance phenotype combination,including the compassionate group (Group Ⅰ),Group Ⅱ with full resistance,IPM,MEM resistant,CAZ and CFP sensitive groups (Group Ⅲ),IPM resistance,MEM non-resistance (sensitive or intermediate) group (Group Ⅳ),IPM,MEM resistance,CAZ and CFP non-resistance groups (Group V).The median RE of each membrane protein-coding gene was analyzed. Results A total of 108 strains of Pseudomonas aeruginosa were collected,with 24 strains in Group Ⅰ as controls and 84 strains in the carbapenem resistant group,including 32 strains in Group Ⅱ,22 strains in Group Ⅲ,13 strains in Group Ⅳ,and 17 strains in Group Ⅴ. The expression of mexD,mexE,mexF,mexX and mexY in the drug-resistant group was higher than that in the control group,and the differences were statistically significant (U=409.5～661.0,all P<0.05). There was no statistically significant difference in mexA,mexB,mexC,oprD and oprM with the control group (U=767.0～1004.5,all P>0.05). There was no significant difference in the expression of RE genes encoding various membrane proteins among strains from different hospitals (H=0.914～7.407,all P>0.05). Among the four different phenotypes,there was no statistically significant difference in the irregular distribution of mexA and oprM RE between each group and the control group (UmexA=95.0～264.0,UoprM=143.0～331.0). The mexC RE in each group was lower than that in the control group,but the differences were not statistically significant (U=134.0～344.5,all P>0.05). MeixE and meixY RE were both higher than the control group,and the differences were statistically significant (UmexE=48.0～230.0,UmexY=83.0～184.0). MeixB was lower than the control group in group Ⅳ (U=72.0),and the differences were statistically significant (all P<0.05). MeixD and meixF showed consistent expression,with higher expression in groups Ⅲ,Ⅳ and Ⅴ compared to the control group (UmeixD=34.0～102.0,UmeixF=65.0～113.0). MeixX was expressed higher in groups Ⅱ,Ⅳ and Ⅴ compared to the control group (U=164.0,58.0,111.0),while oprD was only expressed lower in group Ⅲ than in the control group (U=140.0),with statistically significant differences (all P<0.05). Although the expression of oprD in groups Ⅱ,Ⅳ and Ⅴ was lower than that in the control group,the differences were not statistically significant (U=381.0,102.0,144.0,all P>0.05). Conclusion ExCD,mexEF and mexXY are the main membrane protein combinations of CRPA efflux pumps in Kunming area. Upregulation of mexD,E,F,X,and Y membrane protein expression enhanced efflux. The correlation between mexAB oprM efflux pump and carbapenem resistance in CRPA in this area was low. The low expression of oprD played a role in the efflux mechanism in strains that do not produce β-lactase,but there was no significant difference in low expression in enzyme producing strains.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

Objective To understand the membrane protein molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in the region,and provide some evidence for rational drug use or application of efflux pump inhibitors. Methods Collected Pseudomonas aeruginosa isolated from four hospitals in the region from October 2022 to August 2023,and used SYBR-PCR method to quantitatively detect the relative mRNA expression (RE) levels of 10 membrane protein coding genes,including mexA,B,C,D,E,F,X,Y,and oprD,M. Then categorized the strains into five groups based on ceftazidime,cefepime,imipenem,and meropenem resistance phenotype combination,including the compassionate group (Group Ⅰ),Group Ⅱ with full resistance,IPM,MEM resistant,CAZ and CFP sensitive groups (Group Ⅲ),IPM resistance,MEM non-resistance (sensitive or intermediate) group (Group Ⅳ),IPM,MEM resistance,CAZ and CFP non-resistance groups (Group V).The median RE of each membrane protein-coding gene was analyzed. Results A total of 108 strains of Pseudomonas aeruginosa were collected,with 24 strains in Group Ⅰ as controls and 84 strains in the carbapenem resistant group,including 32 strains in Group Ⅱ,22 strains in Group Ⅲ,13 strains in Group Ⅳ,and 17 strains in Group Ⅴ. The expression of mexD,mexE,mexF,mexX and mexY in the drug-resistant group was higher than that in the control group,and the differences were statistically significant (U=409.5～661.0,all P<0.05). There was no statistically significant difference in mexA,mexB,mexC,oprD and oprM with the control group (U=767.0～1004.5,all P>0.05). There was no significant difference in the expression of RE genes encoding various membrane proteins among strains from different hospitals (H=0.914～7.407,all P>0.05). Among the four different phenotypes,there was no statistically significant difference in the irregular distribution of mexA and oprM RE between each group and the control group (UmexA=95.0～264.0,UoprM=143.0～331.0). The mexC RE in each group was lower than that in the control group,but the differences were not statistically significant (U=134.0～344.5,all P>0.05). MeixE and meixY RE were both higher than the control group,and the differences were statistically significant (UmexE=48.0～230.0,UmexY=83.0～184.0). MeixB was lower than the control group in group Ⅳ (U=72.0),and the differences were statistically significant (all P<0.05). MeixD and meixF showed consistent expression,with higher expression in groups Ⅲ,Ⅳ and Ⅴ compared to the control group (UmeixD=34.0～102.0,UmeixF=65.0～113.0). MeixX was expressed higher in groups Ⅱ,Ⅳ and Ⅴ compared to the control group (U=164.0,58.0,111.0),while oprD was only expressed lower in group Ⅲ than in the control group (U=140.0),with statistically significant differences (all P<0.05). Although the expression of oprD in groups Ⅱ,Ⅳ and Ⅴ was lower than that in the control group,the differences were not statistically significant (U=381.0,102.0,144.0,all P>0.05). Conclusion ExCD,mexEF and mexXY are the main membrane protein combinations of CRPA efflux pumps in Kunming area. Upregulation of mexD,E,F,X,and Y membrane protein expression enhanced efflux. The correlation between mexAB oprM efflux pump and carbapenem resistance in CRPA in this area was low. The low expression of oprD played a role in the efflux mechanism in strains that do not produce β-lactase,but there was no significant difference in low expression in enzyme producing strains.