Echinocandin B (ECB) is a key precursor of the antifungal drug anidulafungin. It is a secondary metabolite of Aspergillus nidulans, and its titer in fermentation is significantly affected by the ECB synthesis pathway and cell morphology. In this study, the key genes related to the transcription activation, hydroxylation, and cell morphology during ECB biosynthesis were investigated to increase the fermentation titer of ECB and to change the cell morphology of Aspergillus nidulans to reduce the viscosity of the fermentation broth. The results indicated that after overexpression of ecdB and ecdK, the ECB titer increased by 25.8% and 23.7%, respectively, compared with that of the wild-type strain, reaching (2 030.5±99.2) mg/L and (1 996.4±151.4) mg/L. However, the deletion of fksA associated with cell wall synthesis resulted in damage to the cell wall, affecting strain growth and product synthesis. The engineered strain overexpressing ecdB was fermented in a 50-L bioreactor, in which the ECB titer reached 2 234.5 mg/L. The findings laid a research foundation for the subsequent metabolic engineering of this strain.
Fermentation ; Aspergillus nidulans/genetics* ; Echinocandins/genetics* ; Bioreactors/microbiology* ; Fungal Proteins/biosynthesis* ; Metabolic Engineering

Objective To investigate the efficacy of Jianpiwenyang Gel(SSWYG)for treating chronic diarrhea and explore its therapeutic mechanism.Methods Eighty patients with chronic diarrhea of spleen and stomach weakness type were randomized into two groups for interventions with lifestyle adjustment and treatment with bifid triple viable capsules(control group,n=40)or naval application with SSWYG(treatment group,n=40)for one week,after which symptoms of chronic diarrhea were evaluated.The Chinese medicine system pharmacology analysis platform(TCMSP),GeneCards,NCBI,OMIM database and GEO database(GSE14841)were used to obtain the active ingredients and target proteins of SSWYG and chronic diarrhea-related targets.The key targets were obtained by topological analysis for Gene Ontology(GO)and KEGG analyses.The affinity and binding characteristics of SSWYG for specific targets were verified by molecular docking using AutoDock software.Results In both groups,gastrointestinal symptom rating scale(GSRS),Bristol Scale and TCM syndrome scores significantly improved after the treatments(P<0.05),and better effects were observed in the treatment group(P<0.05).Sixty-eight targets of SSWYG in treating chronic diarrhea were obtained,and 33 most probable ones were screened out by topological analysis.GO and KEGG analyses identified several chronic diarrhea-related pathways including the TNF and IL-17 pathways.Molecular docking study showed good affinity of the core components of SSWYG for the key targets CASP3,JNK,IL1B,IL6,and AKT1.JUN and CASP3 had the lowest binding energy and the highest stable binding energy with multiple major active ingredients of SSWYG.Conclusion SSWYG can significantly improve clinical symptoms of chronic diarrhea possibly by regulating the TNF and IL-17 as well as other pathways via CASP3 and JUN,suggesting a complex therapeutic mechanism of SSWYG involving multiple ingredients and targets and coordinated regulation of multiple pathways.

Objective:To explore the strategies and technical points for breast reconstruction using free lower abdominal skin flap transplantation through lateral chest incision.Methods:The data of patients who underwent unilateral breast reconstruction with free lower abdominal flap transplantation using lateral thoracic incision in Hunan Cancer Hospital from October 2022 to March 2023 were retrospectively analyzed. Lateral thoracic incision was used in the recipient area. Firstly suitable radical mastectomy was performed for patients with early onset of breast cancer or recurrence after breast-conserving surgery, and prosthetic dilator removal or breast capsule release was performed for patients with dilator removed or capsular contracture after breast cancer prosthesis reconstruction. After that, the free lower abdominal skin flap breast reconstruction was completed under the same incision. During the operation, blood vessel preparation in the recipient area, skin flap preparation in the donor area, blood vessel anastomosis and breast shaping were completed successively according to the actual situation of the patient. Postoperative observation of complications, follow-up evaluation of reconstructed breast appearance and donor site healing were carred out.Results:A total of 15 female patients were included, aged range from 24 to 57 years old, with an average of 42.3 years old. There were 9 cases of early breast cancer, 2 cases of breast cancer recurrence after breast conserving surgery, 2 cases of breast cancer with dilator removed after breast reconstruction, and 2 cases of capsule contracture after breast cancer prosthesis reconstruction. The length of the lateral chest incision was 7.5-11.2 cm, with an average of 8.7 cm. Six cases were performed with 3/4 of the lower abdominal skin flaps, using a unilateral inferior abdominal vascular pedicle. The volume of the flaps ranged from 19.0 cm × 10.0 cm × 4.5 cm to 23.0 cm × 13.0 cm × 6.0 cm. The weight ranged from 280 to 510 g, with an average of 370 g. Nine cases were performed by cutting the entire lower abdominal skin flap and using bilateral inferior abdominal vascular pedicle. The volume of the skin flaps ranged from 27.0 cm × 11.0 cm × 5.0 cm to 30.0 cm × 14.0 cm × 6.5 cm. The weight ranged from 420 to 730 g, with an average of 530 g. Nine cases were selected as the main trunk of the thoracic and dorsal blood vessels, 4 cases were selected as the anterior serratus branch of the thoracic and dorsal blood vessels, and 2 cases were selected as the main trunk of the thoracic and dorsal blood vessels combined with the anterior serratus branch. One flap experienced venous crisis and the surgery failed, while the remaining 14 flaps survived smoothly. Follow-up period was 6-17 months, with an average of 10.4 months. The reconstructed breast had a satisfactory appearance and texture, without skin flap contraction or deformation. The skin flap donor area and breast recipient area only had linear scars, and there was no significant impact on abdominal wall and shoulder joint function. There was no tumor recurrence or metastasis during follow-up.Conclusion:When the side chest incision is used to perform the related operations of breast cancer radical resection, the free lower abdominal skin flap can be transplanted into the same incision for breast reconstruction. The effect is satisfactory, and the damage of the affected area is further reduced.

Objective To explore the expression of serum fatty acid-binding protein 4(FABP4)and fibroblast growth factor 19(FGF19)in patients with β-thalassemia and their relationship with clinical prognosis.Methods A total of 112 cases ofβ-thalassemia patients diagnosed and treated in Qianjiang Hospital Affiliated to Chongqing University from January 2018 to August 2020 were selected as the case group,and 60 healthy individuals who underwent physical examinations during the same period were taken as the control group.Enzyme-linked immunosorbent assay was used to detect levels of serum FABP4 and FGF19 expression.Multivariate logistic regression analysis was used to analyze factors affecting the prognosis of patients with β-thalassemia.Receiver operating characteristic curve was used to analyze the prognostic value of FABP4 and FGF19 in patients with β-thalassemia.Results The serum FABP4 level(67.13±11.35 μg/L)in the case group was higher than that in the control group(22.01±4.16μg/L),while the serum FGF19 level(104.24±21.46 ng/L)was lower than that in the control group(218.01±36.79 ng/L),with significant differences(t=29.708,25.620,all P＜0.05).The serum FABP4 levels(54.20±12.63 μ g/L,66.83±10.5 μ g/L,79.72±11.05 μ g/L)in the mild group,intermediate group,and severe group were increased sequentially,while FGF19 levels(122.53±22.36 ng/L,103.16±20.37 ng/L,86.53±18.14 ng/L)were decreased sequentially,and the differences were significant(F=39.701,24.231,all P＜0.05).Compared to the survival group,serum FGF19 level(62.80±22.09 ng/L vs 110.16±20.69 ng/L),Hb and the proportion of heterozygous genotypes in the death group patients(β CD17/β N,β CD41-42/β N)was lower,while serum FABP4(116.69±12.30 ng/L vs 60.05±10.17 ng/L),ferritin and the proportion of cardiac enlargement were higher,with significant differences(t/x2=4.436～18.981,all P＜0.05).FGF19(OR=0.634,95％CI:0.451～0.891)was an independent protective factor for β-thalassemia patients(P＜0.001),and serum FABP4(OR=1.840,95％CI:1.193～2.838)was an independent risk factor for prognosis(P＜0.001).The area under the curve(95％CI)of serum FABP4 and FGF19 combination in prognosis evaluation for β-thalassemia patients was 0.897(0.853～0.951),which was greater than the single serum indicator detection of 0.842(0.801～0.879)and 0.814(0.762～0.858),with significant differences(Z=4.864,5.270,P=0.002,0.001).Conclusion The serum FABP4 expression is increased,but serum FGF19 expression is decreased in patients with β-thalassemia.The combination of serum FABP4 and FGF19 may have a high predictive value for the prognosis of patients with β-thalassemia.

Objective:To investigate the status of evidence-based nursing competence among clinical nurses in tertiary grade A hospitals of Anhui province, and analyze the influencing factors based on random forest model, so as to provide reference for improving the evidence-based nursing ability of clinical nurses and formulating intervention strategies.Methods:The convenience sampling method was used to select 543 clinical nurses from 4 tertiary grade A hospitals in Anhui Province from October to December 2022. The general data questionnaire, Evidence-based Nursing Competence Scale, Information Literacy Scale, and Nurse Innovation Ability Scale were used to investigate. The random forest model was used to evaluate the importance of the influencing factors. The Lasso regression analysis was used to complete the screening of the influencing factors. The influencing factors of the evidence-based nursing competence among clinical nurses were explored by multiple linear regression analysis.Results:A total of 543 valid questionnaires were retrieved. Among 543 clinical nurses, 55 males and 388 females, aged (32.34 ± 6.93) years old. Evidence-based nursing competence scored (45.49 ± 21.18) points, information literacy scored (73.50 ± 10.47) points, innovation ability scored (126.78 ± 21.99) points. The random forest model and Lasso regression analysis showed that the model achieved the best fit with 8 variables. In order of importance, the top 8 variables were information literacy (25.78%), innovation ability (22.37%), night shift per month (9.91%), educational background (9.19%), English proficiency (8.44%), professional title (6.71%), scientific research and innovation experience (5.17%), and professional attitude (4.50%). Multiple linear regression analysis showed that information literacy, innovation ability, and English proficiency were the influencing factors of evidence-based nursing competence among clinical nurses ( t=9.17, 7.31, 2.52, all P<0.05). Conclusions:The level of evidence-based nursing ability among clinical nurses in tertiary grade A hospitals of Anhui province needs to be improved. From the perspective of improving the information literacy among clinical nurses, nursing managers can increase the training of information retrieval ability and English ability, enhance their English literature reading skills, pay attention to the cultivation of nurses' innovation ability, stimulate the innovative consciousness and thinking among clinical nurses, formulate and implement targeted interventions, so as to gradually improve the level of evidence-based nursing ability among clinical nurses.

Objective To investigate the efficacy of Jianpiwenyang Gel(SSWYG)for treating chronic diarrhea and explore its therapeutic mechanism.Methods Eighty patients with chronic diarrhea of spleen and stomach weakness type were randomized into two groups for interventions with lifestyle adjustment and treatment with bifid triple viable capsules(control group,n=40)or naval application with SSWYG(treatment group,n=40)for one week,after which symptoms of chronic diarrhea were evaluated.The Chinese medicine system pharmacology analysis platform(TCMSP),GeneCards,NCBI,OMIM database and GEO database(GSE14841)were used to obtain the active ingredients and target proteins of SSWYG and chronic diarrhea-related targets.The key targets were obtained by topological analysis for Gene Ontology(GO)and KEGG analyses.The affinity and binding characteristics of SSWYG for specific targets were verified by molecular docking using AutoDock software.Results In both groups,gastrointestinal symptom rating scale(GSRS),Bristol Scale and TCM syndrome scores significantly improved after the treatments(P<0.05),and better effects were observed in the treatment group(P<0.05).Sixty-eight targets of SSWYG in treating chronic diarrhea were obtained,and 33 most probable ones were screened out by topological analysis.GO and KEGG analyses identified several chronic diarrhea-related pathways including the TNF and IL-17 pathways.Molecular docking study showed good affinity of the core components of SSWYG for the key targets CASP3,JNK,IL1B,IL6,and AKT1.JUN and CASP3 had the lowest binding energy and the highest stable binding energy with multiple major active ingredients of SSWYG.Conclusion SSWYG can significantly improve clinical symptoms of chronic diarrhea possibly by regulating the TNF and IL-17 as well as other pathways via CASP3 and JUN,suggesting a complex therapeutic mechanism of SSWYG involving multiple ingredients and targets and coordinated regulation of multiple pathways.

Objective:To explore the strategies and technical points for breast reconstruction using free lower abdominal skin flap transplantation through lateral chest incision.Methods:The data of patients who underwent unilateral breast reconstruction with free lower abdominal flap transplantation using lateral thoracic incision in Hunan Cancer Hospital from October 2022 to March 2023 were retrospectively analyzed. Lateral thoracic incision was used in the recipient area. Firstly suitable radical mastectomy was performed for patients with early onset of breast cancer or recurrence after breast-conserving surgery, and prosthetic dilator removal or breast capsule release was performed for patients with dilator removed or capsular contracture after breast cancer prosthesis reconstruction. After that, the free lower abdominal skin flap breast reconstruction was completed under the same incision. During the operation, blood vessel preparation in the recipient area, skin flap preparation in the donor area, blood vessel anastomosis and breast shaping were completed successively according to the actual situation of the patient. Postoperative observation of complications, follow-up evaluation of reconstructed breast appearance and donor site healing were carred out.Results:A total of 15 female patients were included, aged range from 24 to 57 years old, with an average of 42.3 years old. There were 9 cases of early breast cancer, 2 cases of breast cancer recurrence after breast conserving surgery, 2 cases of breast cancer with dilator removed after breast reconstruction, and 2 cases of capsule contracture after breast cancer prosthesis reconstruction. The length of the lateral chest incision was 7.5-11.2 cm, with an average of 8.7 cm. Six cases were performed with 3/4 of the lower abdominal skin flaps, using a unilateral inferior abdominal vascular pedicle. The volume of the flaps ranged from 19.0 cm × 10.0 cm × 4.5 cm to 23.0 cm × 13.0 cm × 6.0 cm. The weight ranged from 280 to 510 g, with an average of 370 g. Nine cases were performed by cutting the entire lower abdominal skin flap and using bilateral inferior abdominal vascular pedicle. The volume of the skin flaps ranged from 27.0 cm × 11.0 cm × 5.0 cm to 30.0 cm × 14.0 cm × 6.5 cm. The weight ranged from 420 to 730 g, with an average of 530 g. Nine cases were selected as the main trunk of the thoracic and dorsal blood vessels, 4 cases were selected as the anterior serratus branch of the thoracic and dorsal blood vessels, and 2 cases were selected as the main trunk of the thoracic and dorsal blood vessels combined with the anterior serratus branch. One flap experienced venous crisis and the surgery failed, while the remaining 14 flaps survived smoothly. Follow-up period was 6-17 months, with an average of 10.4 months. The reconstructed breast had a satisfactory appearance and texture, without skin flap contraction or deformation. The skin flap donor area and breast recipient area only had linear scars, and there was no significant impact on abdominal wall and shoulder joint function. There was no tumor recurrence or metastasis during follow-up.Conclusion:When the side chest incision is used to perform the related operations of breast cancer radical resection, the free lower abdominal skin flap can be transplanted into the same incision for breast reconstruction. The effect is satisfactory, and the damage of the affected area is further reduced.

Objective To establish a nomogram model for efficiently predicting overall survival(OS)and cancer-specific survival(CSS)in patients with CRCHBM.Method 2239 patients from 2010 to 2019 were retrospectively analyzed from the Surveillance,Epidemiology,and End Results Program(SEER)databases and Wuhan Union Hospital Cancer Center.SEER is randomly assigned to the training and internal validation cohorts,and the Wuhan database serves as the external validation.Cox regression analyses were used to determine the independent clinicopathological prognosis factors affecting OS and CSS,and a nomogram was constructed to predict OS and CSS.The clinical utility of columnar plots was assessed using calibration curves,area under the curve(AUC),and decision curve analysis(DCA).Result OS column line graphs were constructed based on nine independent predictors:age,tumor location,degree of differentiation,tumor size,TNM stage,chemotherapy,primary focus surgery,number of lymph nodes sampled,and serum carcinoembryonic antigen(CEA)level.The C-index of the nomogram to predict the 1-,3-,and 5-year OS were 0.764,0.790,and 0.805 in the training group,0.754,0.760,and 0.801 in the internal validation group,and 0.822,0.874,and 0.906 in the external validation group.CSS column line graphs were constructed based on 3 independent predictors of TNM staging,radiotherapy and chemotherapy.The 1-,3-,and 5-year CSS AUROC values of the training group were 0.791,0.757,and 0.782,respectively.0.682,0.709,0.625 in the internal validation group and 0.759,0.702,0.755 in the external validation group,respectively.The results of receiver operating characteristic curve(ROC),ROC and DCA showed that the use of our model was more effective in predicting OS and CSS than other single clinicopathological features.Conclusion In summary,the nomogram based on significant clinicopathological features can be conveniently used to predict OS and CSS individually in patients with CRCHBM.

Objective:To investigate the clinical effects of combined tissue flap transplantation in repairing giant chest wall defects.Methods:This study was a retrospective observational study. From August 2013 to December 2020, 31 patients with chest wall tumor or radiation ulcer after radical resection of chest wall tumor and conformed to the inclusion criteria were admitted to the Department of Breast Oncoplastic Surgery of Hunan Cancer Hospital, including 12 males and 19 females, aged 25-71 years. After resection of tumor or ulcer and wound debridement, the area of secondary chest wall defect was 300-600 cm 2 with length of 16-35 cm and width of 16-32 cm. According to the actual situation of the patients and the preoperative design, the chest wall defects were repaired with the flexible combination of perforator flaps and myocutaneous flaps from different donor sites, and the area of the combined tissue flap was 260-540 cm 2 with length of 20-30 cm and width of 13-20 cm. Free posteromedial thigh perforator flap+free anterolateral thigh myocutaneous flap were used in 2 patients, free deep inferior epigastric artery perforator flap+free anterolateral thigh myocutaneous flap were used in 5 patients, free deep inferior epigastric artery perforator flap+pedicled rectus abdominis myocutaneous flap+free anterolateral thigh myocutaneous flap were used in 7 patients, free deep inferior epigastric artery perforator flap+pedicled rectus abdominis myocutaneous flap+pedicled latissimus dorsi myocutaneous flap were used in 2 patients, and bilateral free anterolateral thigh myocutaneous flaps were used in 15 patients. For the remaining small area of superficial tissue defect after being repaired by combined tissue flaps, skin graft was used to repair or delayed local flap transfering was performed after the tissue flaps survived and edema subsided. The appropriate blood vessels in the donor and recipient sites were selected for anastomosis to reconstruct the blood supply of tissue flaps. The wounds in the donor sites of tissue flaps that can be directly sutured were sutured directly; for those that cannot be sutured directly, the skin grafting or delayed suture was performed. The anastomosis of blood vessels in the recipient sites, operation length, and postoperative hospital stay were recorded. The survivals of tissue flaps and skin grafts, the shape and texture of reconstructed chest wall, the wound healing, scar formation, and function of donor sites of tissue flaps, and the scar formation of the donor sites of skin grafts were observed after operation. Tumor recurrence and death of recurrent patients were followed up after operation. Results:The blood vessels in the recipient sites were anastomosed as follows: proximal internal thoracic vessels for 24 times, distal internal thoracic vessels for 12 times, trunk of thoracodorsal vessels for 4 times, anterior serratus branches of thoracodorsal vessels for 8 times, and thoracoacromial vessels for 12 times. The operation length was 6.0 to 8.5 hours, and the postoperative hospital stay was 9 to 21 days. Necrosis at the edge of partial tissue flaps occurred in 4 patients after operation, which healed after dressing change, and the tissue flaps and skin grafts of the other patients survived completely. The shape and texture of the reconstructed chest wall were good. Four patients had poor wound healing in the donor sites of abdominal tissue flaps, which healed after dressing change and local drainage. Only linear scar was left in the donor sites of all tissue flaps, and there was no obvious dysfunction in the donor sites of tissue flaps. Mild hypertrophic scar was left in the donor sites of skin grafts. During follow-up of 9 to 36 months after operation, 6 patients had tumor recurrence, and the recurrence time was 5 to 20 months after operation. After comprehensive treatment for patients with tumor recurrence, 3 patients died.Conclusions:Transplantation of combined tissue flaps in repairing the giant chest wall defects can shorten the time of total operation and hospital stay, and avoid multiple operations. After operation, patients had good chest wall appearance, with reduced tumor recurrence in patients with chest wall tumor.

Objective To investigate the expression of miR-142-5p in oral squamous cell carcinoma(OSCC)tis-sues and cell lines and its effects on oral squamous cell proliferation,migration,invasion and angiogenesis.Meth-ods Sixteen groups of oral tumour tissues and paraneoplastic tissues were collected,and qRT-PCR was applied to detect the expression of miR-142-5p in the tissues.The effects of miR-142-5p on cell proliferation,migration,and invasion were observed by cell counting kit-8(CCK-8),cloning,wound healing,Transwell,invasion assays,and the effect of miR-142-5p on angiogenesis was also detected by lumen formation assay.The expression of angiogene-sisrelated proteins vascular endothelial growth factor(VEGFA),vascular endothelial calreticulin(VE-cadherin),epithelial calreticulin(E-cadherin),matrix metalloproteinase 2(MMP2),and matrix metalloproteinase 9(MMP9)was detected by Western blot after overexpression of miR-142-5p.Results miR-142-5p was lowly ex-pressed in oral tumour tissues and cell lines.CCK-8 and clonogenic assays showed that miR-142-5p was inversely correlated with the proliferation of OSCC cells,wound healing and Transwell assays showed that miR-142-5p was inversely correlated with the migration of OSCC cells,and cell invasion assays showed that miR-142-5p was con-versely correlated with the invasion of OSCC cells.Analysis of lumen formation assay showed that overexpression of miR-142-5p reduced the tube length and nodes of HUVECs.Western blot assay showed that up-regulation of miR-142-5p inhibited the VEGFA,VE-cadherin,MMP2,MMP9 expression and promoted E-cadherin expression.Con-clusion Overexpression of miR-142-5p inhibites the proliferative,migratory and invasive effects of oral squamous carcinoma cells as well as angiogenesis,suggesting that miR-142-5p is a novel target for anti-tumour angiogenesis and against oral squamous carcinoma.