ObjectiveTo investigate the mechanism of Forsythiae Fructus-Lonicerae Japonicae Flos（FF） in the treatment of acute lung injury（ALI） by investigating the effects of FF on serum metabolomics of rats with ALI. MethodsThirty male SD rats were acclimated for 1 week， and 6 rats were randomly selected as the blank group. The other 24 rats were injected with lipopolysaccharide（LPS） solution by tracheal drip to establish an ALI model. After successful model establishment， the rats were randomly divided into the model group， the FF low-dose group（3.0 g·kg^-1）， the FF high-dose group（6.0 g·kg^-1）， and the dexamethasone group（5 mg·kg^-1）， with six rats in each group. The FF low- and high-dose groups and the dexamethasone group were received daily oral administration of the corresponding drug solution， and the blank group and the model group were gavaged with an equal amount of saline， treatment was administered continuously for 3 d. The pathological conditions of rat lung tissues were evaluated by hematoxylin-eosin（HE） staining， wet/dry mass ratio（W/D） of the lung tissues， and protein concentration in rat bronchoalveolar lavage fluid（BALF）. Metabolomic analysis of rat serum was performed by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， combined with multivariate statistical analysis， the potential biomarkers of FF in treating ALI were screened by variable importance in the projection（VIP） value>1， P<0.05 from t-test， and log₂fold change（FC）>1 or log₂FC<-1. Kyoto Encyclopedia of Genes and Genomes（KEGG） database combined with MetaboAnalyst were used for pathway analysis of the screened differential metabolites. The protein expression levels of sphingosine-1-phosphate（S1P）， phosphatidylinositol 3-kinase（PI3K）， protein kinase B1（Akt1）， and phosphorylated Akt1（p-Akt1） were examined by Western bolt. The expression levels of interleukin（IL）-6， IL-1β， and tumor necrosis factor（TNF）-α in BALF were detected by enzyme-linked immunosorbent assay（ELISA）. ResultsCompared with the blank group， rats in the model group showed ALI pathological features such as alveolar lumen dilatation， interstitial hemorrhage and massive inflammatory cell infiltration， and the protein concentration in BALF and W/D of the lung tissues were significantly elevated（P<0.01）. Compared with the model group， the low- and high-dose groups of FF as well as the dexamethasone group exhibited reduced pulmonary bronchial hemorrhage in rats， and the protein concentration in BALF and W/D were significantly decreased（P<0.05）， and the lung injury was significantly alleviated. Analysis of rat serum metabolomics revealed that FF downregulated 38 biomarkers. Pathway enrichment analysis showed that FF primarily exerted therapeutic effects through 7 key metabolic pathways， including arginine biosynthesis， sphingomyelin metabolism， alanine， aspartate and glutamate metabolism， taurine and hypotaurine metabolism， α-linolenic acid metabolism， niacin and nicotinamide metabolism， and retinol metabolism. The results of Western bolt and ELISA showed that， compared with the blank group， the model group exhibited significantly elevated expression levels of S1P， PI3K， Akt1 and p-Akt1 proteins in the lung tissues， as well as increased expression levels of IL-6， IL-1β and TNF-α in BALF（P<0.01）. Compared with the model group， the expression levels of the aforementioned indicators were significantly downregulated in the low- and high-dose FF groups as well as the dexamethasone group（P<0.05， P<0.01）. ConclusionFF may play a role in ALI by regulating amino acid metabolism and lipid metabolism， and its mechanism may be related to the inhibition of S1P/PI3K/Akt1 signaling pathway to attenuate the inflammatory response caused by ALI.

Objective:To investigate the effect and mechanism of type ⅩⅦ collagen (COL17) on hair growth in mice with androgenetic alopecia (AGA).Methods:Forty-eight C57BL/6J mice were used to establish AGA model (the back hair of the mice was removed and dihydrotestosterone solution was applied) and divided into 6 groups of 8 mice each by random number table. Negative control group, injection of saline in the depilated area (single point injection of 0.05 ml, 5 points in total); positive control group, topical application of 5% minoxidil tincture in the depilated area, 1 ml/d; COL17 low, medium and high concentration groups, injection of 0.5, 1.0 and 2.0 mg/ml COL17 in the depilated area respectively (single point injection of 0.05 ml, 5 points in total); type Ⅲ and ⅩⅦ collagen (COL3+ COL17) combined high concentration group, injection of 2.0 mg/ml COL3 and COL17 in the depilated area (single point injection of 0.05 ml, 5 points in total). The total treatment time was 21 days, during which the hair growth of mice in each group was observed and recorded. After 21 days, the skin and subcutaneous tissue in the depilated area of the mice were taken to make pathological sections for HE staining, and the number and morphological changes of hair follicles were observed; fresh skin tissue in the depilated area of the mice was taken for total RNA sequencing analysis, and the differentially co-expressed genes were annotated by gene ontology (GO) functional annotation, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and gene set enrichment analysis (GSEA).Results:After 21 days of treatment, compared with the negative control group, the depilation area on the back of the mice in the positive control group, COL17 high concentration group, and COL3+ COL17 combined high concentration group was significantly reduced, and HE staining showed that the number of hair follicles was also significantly increased. Pearson correlation analysis, principal component analysis and cluster heat map between groups showed that COL17 high concentration group had high gene correlation with the positive control group ( R2=0.95, P=0.024), and the gene expression was relatively close, with 3 882 differentially expressed genes (1 705 up-regulated and 2 177 down-regulated) in the two groups, while COL3+ COL17 combined high concentration group had the highest gene correlation with the positive control group ( R2=0.96, P=0.001), and the gene expression was the closest, with 1 289 differentially expressed genes (385 up-regulated and 904 down-regulated). KEGG analysis showed that compared with the negative control group, the positive control group, COL17 high concentration group and COL3+ COL17 combined high concentration group of mice all upregulated Wnt signaling pathway, cell adhesion molecules and hedgehog signaling pathway related to hair growth. GO enrichment analysis suggested that COL17 high concentration group and COL3+ COL17 combined high concentration group had upregulated genes related to skin development and hair cycle. GSEA enrichment analysis found that COL17 high concentration group had upregulated genes related to fibroblast proliferation and interleukin-1 secretion, while COL3+ COL17 combined high concentration group had upregulated genes related to fibroblast migration, clearance of apoptotic cells and accelerated metabolism of reactive oxygen species. Conclusion:Local injection of 2.0 mg/ml COL17 has a certain promoting effect on hair growth in AGA model mice, and the effect is more significant after combined injection of 2.0 mg/ml COL3. Activation of Wnt signaling pathway is one of the main mechanisms of COL17 promoting hair growth.

Objective:To investigate the effect and mechanism of type ⅩⅦ collagen (COL17) on hair growth in mice with androgenetic alopecia (AGA).Methods:Forty-eight C57BL/6J mice were used to establish AGA model (the back hair of the mice was removed and dihydrotestosterone solution was applied) and divided into 6 groups of 8 mice each by random number table. Negative control group, injection of saline in the depilated area (single point injection of 0.05 ml, 5 points in total); positive control group, topical application of 5% minoxidil tincture in the depilated area, 1 ml/d; COL17 low, medium and high concentration groups, injection of 0.5, 1.0 and 2.0 mg/ml COL17 in the depilated area respectively (single point injection of 0.05 ml, 5 points in total); type Ⅲ and ⅩⅦ collagen (COL3+ COL17) combined high concentration group, injection of 2.0 mg/ml COL3 and COL17 in the depilated area (single point injection of 0.05 ml, 5 points in total). The total treatment time was 21 days, during which the hair growth of mice in each group was observed and recorded. After 21 days, the skin and subcutaneous tissue in the depilated area of the mice were taken to make pathological sections for HE staining, and the number and morphological changes of hair follicles were observed; fresh skin tissue in the depilated area of the mice was taken for total RNA sequencing analysis, and the differentially co-expressed genes were annotated by gene ontology (GO) functional annotation, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and gene set enrichment analysis (GSEA).Results:After 21 days of treatment, compared with the negative control group, the depilation area on the back of the mice in the positive control group, COL17 high concentration group, and COL3+ COL17 combined high concentration group was significantly reduced, and HE staining showed that the number of hair follicles was also significantly increased. Pearson correlation analysis, principal component analysis and cluster heat map between groups showed that COL17 high concentration group had high gene correlation with the positive control group ( R2=0.95, P=0.024), and the gene expression was relatively close, with 3 882 differentially expressed genes (1 705 up-regulated and 2 177 down-regulated) in the two groups, while COL3+ COL17 combined high concentration group had the highest gene correlation with the positive control group ( R2=0.96, P=0.001), and the gene expression was the closest, with 1 289 differentially expressed genes (385 up-regulated and 904 down-regulated). KEGG analysis showed that compared with the negative control group, the positive control group, COL17 high concentration group and COL3+ COL17 combined high concentration group of mice all upregulated Wnt signaling pathway, cell adhesion molecules and hedgehog signaling pathway related to hair growth. GO enrichment analysis suggested that COL17 high concentration group and COL3+ COL17 combined high concentration group had upregulated genes related to skin development and hair cycle. GSEA enrichment analysis found that COL17 high concentration group had upregulated genes related to fibroblast proliferation and interleukin-1 secretion, while COL3+ COL17 combined high concentration group had upregulated genes related to fibroblast migration, clearance of apoptotic cells and accelerated metabolism of reactive oxygen species. Conclusion:Local injection of 2.0 mg/ml COL17 has a certain promoting effect on hair growth in AGA model mice, and the effect is more significant after combined injection of 2.0 mg/ml COL3. Activation of Wnt signaling pathway is one of the main mechanisms of COL17 promoting hair growth.

Pleomorphic adenoma (PA) is the most common benign tumour in the salivary gland and has high morphological complexity. However, the origin and intratumoral heterogeneity of PA are largely unknown. Here, we constructed a comprehensive atlas of PA at single-cell resolution and showed that PA exhibited five tumour subpopulations, three recapitulating the epithelial states of the normal parotid gland, and two PA-specific epithelial cell (PASE) populations unique to tumours. Then, six subgroups of PASE cells were identified, which varied in epithelium, bone, immune, metabolism, stemness and cell cycle signatures. Moreover, we revealed that CD36⁺ myoepithelial cells were the tumour-initiating cells (TICs) in PA, and were dominated by the PI3K-AKT pathway. Targeting the PI3K-AKT pathway significantly inhibited CD36⁺ myoepithelial cell-derived tumour spheres and the growth of PA organoids. Our results provide new insights into the diversity and origin of PA, offering an important clinical implication for targeting the PI3K-AKT signalling pathway in PA treatment.
Humans ; Adenoma, Pleomorphic/genetics* ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; Transcriptome ; Myoepithelioma

Objective:To review clinical outcomes of extended V-Y latissimus dorsi myocutaneous flap in reconstruction of chest wall defects.Methods:Patients receiving chest wall defect reconstruction with extended V-Y latissimus dorsi myocutaneous flap at Fudan University Zhongshan Hospital from January 2016 to December 2018 were enrolled. Patients were followed up for more than 6 months to detect complications such as blood supply disorder of the flap, wound dehiscence, seroma or tumor recurrence.Results:A total of 16 patients, aged 28-78 years, with an average age of 54.8 years, were included. The chest wall defect size ranged from 10 cm×15 cm to 19 cm×26 cm. Among these cases, 1 case had venous congestion in the distal triangle area of the flap, 1 case had partial wound dehiscence of the flap, 2 cases developed seroma on the back and 2 cases under the flap. All cases healed after changing dressing and drain placement. All the other cases had complete flap survival and primary wound healing. All cases were followed up from 7 to 29 months, with good flap appearance and function and patient satisfaction with results.Conclusions:Chest wall defect reconstruction with V-Y latissimus dorsi myocutaneous flap is safe and reliable. It can repair a large chest wall wound and allow primary closure of the donor site simultaneously, which is worthy of being popularized in chest wall reconstruction.

Objective:To review clinical outcomes of extended V-Y latissimus dorsi myocutaneous flap in reconstruction of chest wall defects.Methods:Patients receiving chest wall defect reconstruction with extended V-Y latissimus dorsi myocutaneous flap at Fudan University Zhongshan Hospital from January 2016 to December 2018 were enrolled. Patients were followed up for more than 6 months to detect complications such as blood supply disorder of the flap, wound dehiscence, seroma or tumor recurrence.Results:A total of 16 patients, aged 28-78 years, with an average age of 54.8 years, were included. The chest wall defect size ranged from 10 cm×15 cm to 19 cm×26 cm. Among these cases, 1 case had venous congestion in the distal triangle area of the flap, 1 case had partial wound dehiscence of the flap, 2 cases developed seroma on the back and 2 cases under the flap. All cases healed after changing dressing and drain placement. All the other cases had complete flap survival and primary wound healing. All cases were followed up from 7 to 29 months, with good flap appearance and function and patient satisfaction with results.Conclusions:Chest wall defect reconstruction with V-Y latissimus dorsi myocutaneous flap is safe and reliable. It can repair a large chest wall wound and allow primary closure of the donor site simultaneously, which is worthy of being popularized in chest wall reconstruction.

Objective:To investigate the causes and treatment of implant malposition after breast augmentation surgery.Methods:A retrospective study was conducted to investigate the patients with implant malposition after breast augmentation treated or consulted in our department from January 2000 to December 2020. A total of 95 breasts in 74 patients were treated during the period. The age of the patients was from 19-50 years, with the mean of 31.2 years. Causes of these complications were analyzed and the patients were managed in different ways according to the times after primary breast augmentation.Results:Twenty-one malposition breasts occurred within one week after primary surgery, and 17 breasts were successfully treated with manual reduction and fixation. 4 breasts were treated with operation after failed with manual reduction. The patients had good and bilateral symmetric breast morphology during the 6 months follow-up after the second operation. 25 malposition breasts were treated from one week to one month after primary surgery. Reoperation was performed through the original incision, separating the adhesive cavity to reposition the implant. Those patients were successfully treated. No re-occurrence was detected at 6 months follow-up after treatment. 49 malposition breasts were treated more than 3 months after primary surgery. Excessive capsule wrapping the shifted implant was removed to recreate a fresh wound while the adhesive cavity was separated to reset the implant. Bandage fixation was necessary. 7 implants were exchanged. 2 breasts were over corrected and re-operated successfully.Conclusions:The causes of implant malposition at different time after breast augmentation are various. Different methods can be used for effective treatment.

Objective To explore the method and clinical effects of treatment for correcting severe contracture nasal deformity after rhinoplasty.Methods Totally 19 patients with severe contracture nasal deformity after rhinoplasty were treated with a new procedure in our hospital since March 2014.The soft tissue was treated with manual traction preoperatively to improve nasal skin and mucosa extensibility and flexibility.The costicartilage was taken as structure transplantation to build the nasal support structure.Results This group of patients were followed up for 5-27 months,contracture of nasal deformity was corrected and the shape of the nose was improved with no obvious contracture deformation.Conclusions Nasal shape is improved by nasal scaffold and coated tissue.It is a simple and feasible method to modify the condition of soft tissue by manual traction and autogenous costicartilage support reconstruction to ensure the long-term efficacy of backing;two combination therapy for correction of iatrogenic contracture nasal deformity can achieve better long-term nasal prolonged postoperative effect,which is suitable for clinical promotion.

Objective To investigate the influence factors of tumor diameter and related prognostic factors on the prognosis of hilar cholangiocarcinoma.Methods The retrospective case-control study was conducted.The clinicopathological data of 240 patients who underwent resection of hilar cholangiocarcinoma in the West China Hospital of Sichuan University between January 1995 and January 2013 were collected,including 104 patients with tumor diameter ≤ 2 cm (8 with tumor diameter ≤ 1 cm and 96 with 1 cm ＜ tumor diameter ≤ 2 cm),85 with 2 cm ＜ tumor diameter ≤ 3 cm and 51 with tumor diameter ＞ 3 cm (40 with 3 cm ＜ tumor diameter ≤ 4 cm and 11 with tumor diameter ＞ 4 cm).Observation indicators:(1) surgical situations;(2) follow-up situations;(3) risk factors analysis affecting the prognosis of patients;(4) correlation analysis between related prognostic indicators and tumor diameter.The follow-up using outpatient examination and telephone interview was performed to detect the survival up to August 2016.The survival curve and survival rate were respectively drawn and calculated by the Kaplan-Meier method,and the Log-rank test was used for survival analysis.The prognostic factors and correlation between related prognostic indicators and tumor diameter were respectively analyzed using the COX proportional hazard model and logistic regression model.Results (1) Surgical situations:240 patients underwent successful resection of hilar cholangiocarcinoma and lymph node dissection.Of 73 patients with postoperative complications,1 died of intraperitoneal infection induced to systemic infection and multiple organ failure,1 diel of renal failure,and other patients were cured by symptomatic treatment.(2) Follow-up situations:240 patients were followed up for 12.0-98.0 months,with a median time of 47.4 months.The overall median survival time,1-,3-and 5-year overall survival rates were respectively 30.6 months,81％,47％ and 29％.The median survival time and 5-year survival rate were 46.5 months,34％ in patients with tumor diameter ≤ 2 cm and 30.5 months,30％ in patients with 2 cm ＜ tumor diameter ≤ 3 cm and 13.8 months,20％ in patients with tumor diameter ＞ 3 cm,respectively,with a statistically significant difference (x2 =17.83,P＜0.05).Results of further analysis showed the median survival time and 5-year survival rate were 31.3 months,38％ in patients with tumor diameter ≤ 1 cm and 46.5 months,34％ in patients with 1 cm ＜ tumor diameter ≤ 2 cm,respectively,with no statistically significant difference (x2=1.16,P＞O.05).The median survival time and 1-year survival rate were 14.7 months,62％ in patients with 3 cm ＜ tumor diameter ≤ 4 cm and 13.0 months,55％ in patients with tumor diameter ＞ 4 cm,respectively,with no statistically significant difference (x2 =2.34,P＞O.05).(3) Risk factors analysis affecting the prognosis of patients:univariate analysis showed that tumor diameter,surgical margin,lymph node metastasis,vascular invasion and histological differentiation were the related factors affecting patients' prognosis [hazard ratio (HR)=1.456,8.714,1.737,2.246,1.665;95％ confidence interval (C I):1.212-1.748,5.558-13.663,1.311-2.301,1.494-3.378,1.375-2.016,P ＜ 0.05].The multivariate analysis showed that 2 cm ＜ tumor diameter ≤ 3 cm,tumor diameter ＞ 3 cm,R1 resection,lymph node metastasis and low-differentiated tumor were the independent risk factors affecting poor prognosis of patients (HR =1.559,1.868,7.410,1.521,2.274,95％ CI:1.125-2.160,1.265-2.759,4.497-12.212,1.136-2.037,1.525-3.390,P＜0.05).(4) Correlation analysis between related prognostic indicators and tumor diameter:the results of univariate analysis showed that there was a correlation between lymph node metastasis,vascular invasion,histological differentiation and T staging of American Joint Committee on Cancer (AJCC) and tumor diameter of 2 cm as a cut-off point (x2 =6.063,4.950,8.770,9.069,P＜0.05).There was a correlation between surgical margin,lymph node metastasis,vascular invasion and histological differentiation and tumor diameter of 3 cm as a cut-off point (x2=10.251,9.919,5.485,15.632,P＜0.05).The results of multivariate analysis showed that lymph node metastasis and T staging of AJCC were independent related factors affecting tumor diameter of 2 cm as a cut-off point[odds ratio (OR) =1.882,2.104,95 ％CI:1.075-3.293,1.220-3.631,P＜0.05];surgical margin and lymph node metastasis were independent related factors affecting tumor diameter of 3 cm as a cut-off point (OR=3.187,2.211,95 ％CI:1.377-7.379,1.133-4.314,P＜0.05).Conclusions The 2 cm ＜ tumor diameter ≤ 3 cm,tumor diameter ＞ 3 cm,R1 resection,lymph node metastasis and low-differentiated tumor are the independent risk factors affecting the prognosis of patients with hilar cholangiocarcinoma.Three cm (T staging in De Oliveira staging system) as the second cut-off point is feasible,meanwhile,2 cm cut-off point may be become another potential tumor dividing point described in De Oliveira staging system.

Objective:To investigate the targeted motion and controllable release of tumor drugs based on micromotor. Methods:The directional movement of Janus micro-capsules was achieved through an external magnetic field,and the controllable release of tumor drugs was induced by near-infrared laser.Results:During the same period, the movement speed of the Janus capsules micromotor was the fastest(36.8 μm·s-1,approximately equalled to 3 body length·s-1) in 15% H2O2solution. Under the control of the external magnetic field, the Janus capsules micromotor could move along the scheduled trajectory close to the area of HeLa cells. Through the irradiation of near-infrared laser, the Janus capsules micromotor was broken and released the loaded drugs quickly. Conclusion:The Janus capsule micromotor studied in the paper can be used for targeted drug delivery safely and effectively,therefore,it shows good application prospect in the field of tumor diagnosis and treatment.