Objective:To update the classification of extremity primary lymphedema (PLE) based on age of onset, lymphatic anomalies and genetics.Methods:A prospective research method was adopted. Patients with lower and upper limb PLE who visited Department of Plastic & Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine from January 207 to December 2021 were selected. Sex, age of onset, location, family history and morbidity were documented. The lymphatic imaging findings of magnetic resonance lymphography (MRL), indocyanine green lymphography (ICGL) and lymphoscintigraphy (LSG), skin tissue histology and whole exome sequencing were evaluated. Descriptive statistical method was used for data statistics.Results:A total of 1 046 patients were included, among whom 1 013 had lower extremity involvement and 33 had upper extremity involvement. Divided by the age of onset, there were 237 cases of congenital (<1 year old) and 809 of late-onset (≥1 year old), with a ratio of about 1∶4. Among the late-onset patients, most patients began to suffer the disease during adolescence (11-20 years old), with a total of 276 cases. Among patients with congenital lower limb PLE, 12.7% (26/204) had a family history. Among those with late-onset of PLE, 6.6% (53/809) had a family history. A total of 225 patients underwent whole exome sequencing (208 cases in the lower limbs and 17 cases in the upper limbs), showing that 37 patients (17.8%, 37/208) with lower limb PLE were found to carry 38 pathogenic variants in FLT4, GJC2, CELSR1, PTPN14, FOXC2 and GATA2, only 1 patient (5.9%, 1/17) with upper limb PLE was found to carry a PIEZO1 compound heterozygote variant. Three major lymphatic anomalies were identified, in which segmental lymphatic dysfunction, characterized by delayed or partial demonstration of lymph vessels, was the most common type and associated with FLT4, GJC2, CELSR1, and PTPN14 mutations. The second most common type was lymphatic hyperplasia, which was associated with FOXC2 and GATA2 variants, followed by initial lymphatic aplasia or dysfunction and associated with FLT4 and PIEZO1 mutation.Conclusion:A classification of extremity PLE is proposed based on age of onset, lymphatic anomalies and genetics, which are segmental lymphatic dysfunction, lymphatic hyperplasia, and initial lymphatic aplasia or dysfunction.

Objective:To update the classification of extremity primary lymphedema (PLE) based on age of onset, lymphatic anomalies and genetics.Methods:A prospective research method was adopted. Patients with lower and upper limb PLE who visited Department of Plastic & Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine from January 207 to December 2021 were selected. Sex, age of onset, location, family history and morbidity were documented. The lymphatic imaging findings of magnetic resonance lymphography (MRL), indocyanine green lymphography (ICGL) and lymphoscintigraphy (LSG), skin tissue histology and whole exome sequencing were evaluated. Descriptive statistical method was used for data statistics.Results:A total of 1 046 patients were included, among whom 1 013 had lower extremity involvement and 33 had upper extremity involvement. Divided by the age of onset, there were 237 cases of congenital (<1 year old) and 809 of late-onset (≥1 year old), with a ratio of about 1∶4. Among the late-onset patients, most patients began to suffer the disease during adolescence (11-20 years old), with a total of 276 cases. Among patients with congenital lower limb PLE, 12.7% (26/204) had a family history. Among those with late-onset of PLE, 6.6% (53/809) had a family history. A total of 225 patients underwent whole exome sequencing (208 cases in the lower limbs and 17 cases in the upper limbs), showing that 37 patients (17.8%, 37/208) with lower limb PLE were found to carry 38 pathogenic variants in FLT4, GJC2, CELSR1, PTPN14, FOXC2 and GATA2, only 1 patient (5.9%, 1/17) with upper limb PLE was found to carry a PIEZO1 compound heterozygote variant. Three major lymphatic anomalies were identified, in which segmental lymphatic dysfunction, characterized by delayed or partial demonstration of lymph vessels, was the most common type and associated with FLT4, GJC2, CELSR1, and PTPN14 mutations. The second most common type was lymphatic hyperplasia, which was associated with FOXC2 and GATA2 variants, followed by initial lymphatic aplasia or dysfunction and associated with FLT4 and PIEZO1 mutation.Conclusion:A classification of extremity PLE is proposed based on age of onset, lymphatic anomalies and genetics, which are segmental lymphatic dysfunction, lymphatic hyperplasia, and initial lymphatic aplasia or dysfunction.

Objective:To investigate the effect of complex decongestive therapy(CDT) on elephantiasis and skin texture improvement.Methods:The clinical data of patients with elephantiasis of lower limbs who were admitted to the Department of Plastic Surgery, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2016 to December 2018 were retrospectively analyzed. The patients with elephantiasis of different causes were treated with CDT for 1 course (4 weeks). The skin texture parameters (skin water content, transepidermal water loss, skin fibrosis), limb circumference and segmental edema were measured before and after treatment and analyzed by paired t test. Pearson correlation analysis was used to detect the correlation between segmental edema and skin texture parameters before and after treatment. The subjective feelings of patients were recorded during follow-up. A self-designed online questionnaire was used to follow up the patient’s disease control status, including the limb circumferences, changes in quality of life (life, work, social and emotion), the adaptability to bandages and elastic socks. Data were classified and analysed as excellent, good, medium and poor.Results:A total of 20 patients were included, including 16 females and 4 males. The average age was 50.45 years (ranged 9-70 years). 5 cases were primary lymphedema and 15 cases were secondary lymphedema. The duration of the disease ranged from 2 years to 27 years, with an average of 8.4 years. After 1 course of treatment, the skin texture was significantly improved. The skin water content decreased from 55.6%±7.4% before treatment to 42.1%±7.4% after treatment ( P< 0.001). The transepidermal water loss decreased from (14.981±5.699) g·m -2·h -1 before treatment to (9.312±2.590) g·m -2·h -1 after treatment. The skin fibrosis decreased from (0.087±0.042) N before treatment to (0.065±0.033) N after treatment ( P<0.001). The circumference of the affected limb decreased from (11.09±3.14) cm before treatment to (4.82±2.83)cm after treatment ( P<0.001). The segmental edema decreased from (4.00±2.14)L before treatment to (1.21±0.78) L after treatment. Segmental edema was positively correlated with skin water content, transepidermal water loss, and skin fibrosis ( r=0.447, r=0.429, r=0.751, P<0.05). The patients were followed up for 6 months after treatment. The circumference control was excellent in 13 cases, good in 5 cases and moderate in 2 cases. The quality-of-life changes were excellent in 16 cases, good in 3 cases, medium in 1 case. The adaptability of elastic material was excellent in 9 cases, good in 8 cases, medium in 2 cases and poor in 1 case. Conclusions:CDT can reduce subcutaneous edema, effectively reduce limb volume, decrease skin fibrosis, improve skin barrier function, improve skin texture, restore the appearance of the limb and improve patients’ quality of life.

Objective:To investigate the effect of complex decongestive therapy(CDT) on elephantiasis and skin texture improvement.Methods:The clinical data of patients with elephantiasis of lower limbs who were admitted to the Department of Plastic Surgery, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2016 to December 2018 were retrospectively analyzed. The patients with elephantiasis of different causes were treated with CDT for 1 course (4 weeks). The skin texture parameters (skin water content, transepidermal water loss, skin fibrosis), limb circumference and segmental edema were measured before and after treatment and analyzed by paired t test. Pearson correlation analysis was used to detect the correlation between segmental edema and skin texture parameters before and after treatment. The subjective feelings of patients were recorded during follow-up. A self-designed online questionnaire was used to follow up the patient’s disease control status, including the limb circumferences, changes in quality of life (life, work, social and emotion), the adaptability to bandages and elastic socks. Data were classified and analysed as excellent, good, medium and poor.Results:A total of 20 patients were included, including 16 females and 4 males. The average age was 50.45 years (ranged 9-70 years). 5 cases were primary lymphedema and 15 cases were secondary lymphedema. The duration of the disease ranged from 2 years to 27 years, with an average of 8.4 years. After 1 course of treatment, the skin texture was significantly improved. The skin water content decreased from 55.6%±7.4% before treatment to 42.1%±7.4% after treatment ( P< 0.001). The transepidermal water loss decreased from (14.981±5.699) g·m -2·h -1 before treatment to (9.312±2.590) g·m -2·h -1 after treatment. The skin fibrosis decreased from (0.087±0.042) N before treatment to (0.065±0.033) N after treatment ( P<0.001). The circumference of the affected limb decreased from (11.09±3.14) cm before treatment to (4.82±2.83)cm after treatment ( P<0.001). The segmental edema decreased from (4.00±2.14)L before treatment to (1.21±0.78) L after treatment. Segmental edema was positively correlated with skin water content, transepidermal water loss, and skin fibrosis ( r=0.447, r=0.429, r=0.751, P<0.05). The patients were followed up for 6 months after treatment. The circumference control was excellent in 13 cases, good in 5 cases and moderate in 2 cases. The quality-of-life changes were excellent in 16 cases, good in 3 cases, medium in 1 case. The adaptability of elastic material was excellent in 9 cases, good in 8 cases, medium in 2 cases and poor in 1 case. Conclusions:CDT can reduce subcutaneous edema, effectively reduce limb volume, decrease skin fibrosis, improve skin barrier function, improve skin texture, restore the appearance of the limb and improve patients’ quality of life.

Objective:To investigate the effect of complex decongestive therapy(CDT) on elephantiasis and skin texture improvement.Methods:The clinical data of patients with elephantiasis of lower limbs who were admitted to the Department of Plastic Surgery, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2016 to December 2018 were retrospectively analyzed. The patients with elephantiasis of different causes were treated with CDT for 1 course (4 weeks). The skin texture parameters (skin water content, transepidermal water loss, skin fibrosis), limb circumference and segmental edema were measured before and after treatment and analyzed by paired t test. Pearson correlation analysis was used to detect the correlation between segmental edema and skin texture parameters before and after treatment. The subjective feelings of patients were recorded during follow-up. A self-designed online questionnaire was used to follow up the patient’s disease control status, including the limb circumferences, changes in quality of life (life, work, social and emotion), the adaptability to bandages and elastic socks. Data were classified and analysed as excellent, good, medium and poor.Results:A total of 20 patients were included, including 16 females and 4 males. The average age was 50.45 years (ranged 9-70 years). 5 cases were primary lymphedema and 15 cases were secondary lymphedema. The duration of the disease ranged from 2 years to 27 years, with an average of 8.4 years. After 1 course of treatment, the skin texture was significantly improved. The skin water content decreased from 55.6%±7.4% before treatment to 42.1%±7.4% after treatment ( P< 0.001). The transepidermal water loss decreased from (14.981±5.699) g·m -2·h -1 before treatment to (9.312±2.590) g·m -2·h -1 after treatment. The skin fibrosis decreased from (0.087±0.042) N before treatment to (0.065±0.033) N after treatment ( P<0.001). The circumference of the affected limb decreased from (11.09±3.14) cm before treatment to (4.82±2.83)cm after treatment ( P<0.001). The segmental edema decreased from (4.00±2.14)L before treatment to (1.21±0.78) L after treatment. Segmental edema was positively correlated with skin water content, transepidermal water loss, and skin fibrosis ( r=0.447, r=0.429, r=0.751, P<0.05). The patients were followed up for 6 months after treatment. The circumference control was excellent in 13 cases, good in 5 cases and moderate in 2 cases. The quality-of-life changes were excellent in 16 cases, good in 3 cases, medium in 1 case. The adaptability of elastic material was excellent in 9 cases, good in 8 cases, medium in 2 cases and poor in 1 case. Conclusions:CDT can reduce subcutaneous edema, effectively reduce limb volume, decrease skin fibrosis, improve skin barrier function, improve skin texture, restore the appearance of the limb and improve patients’ quality of life.

Objective:To investigate the effect of complex decongestive therapy(CDT) on elephantiasis and skin texture improvement.Methods:The clinical data of patients with elephantiasis of lower limbs who were admitted to the Department of Plastic Surgery, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2016 to December 2018 were retrospectively analyzed. The patients with elephantiasis of different causes were treated with CDT for 1 course (4 weeks). The skin texture parameters (skin water content, transepidermal water loss, skin fibrosis), limb circumference and segmental edema were measured before and after treatment and analyzed by paired t test. Pearson correlation analysis was used to detect the correlation between segmental edema and skin texture parameters before and after treatment. The subjective feelings of patients were recorded during follow-up. A self-designed online questionnaire was used to follow up the patient’s disease control status, including the limb circumferences, changes in quality of life (life, work, social and emotion), the adaptability to bandages and elastic socks. Data were classified and analysed as excellent, good, medium and poor.Results:A total of 20 patients were included, including 16 females and 4 males. The average age was 50.45 years (ranged 9-70 years). 5 cases were primary lymphedema and 15 cases were secondary lymphedema. The duration of the disease ranged from 2 years to 27 years, with an average of 8.4 years. After 1 course of treatment, the skin texture was significantly improved. The skin water content decreased from 55.6%±7.4% before treatment to 42.1%±7.4% after treatment ( P< 0.001). The transepidermal water loss decreased from (14.981±5.699) g·m -2·h -1 before treatment to (9.312±2.590) g·m -2·h -1 after treatment. The skin fibrosis decreased from (0.087±0.042) N before treatment to (0.065±0.033) N after treatment ( P<0.001). The circumference of the affected limb decreased from (11.09±3.14) cm before treatment to (4.82±2.83)cm after treatment ( P<0.001). The segmental edema decreased from (4.00±2.14)L before treatment to (1.21±0.78) L after treatment. Segmental edema was positively correlated with skin water content, transepidermal water loss, and skin fibrosis ( r=0.447, r=0.429, r=0.751, P<0.05). The patients were followed up for 6 months after treatment. The circumference control was excellent in 13 cases, good in 5 cases and moderate in 2 cases. The quality-of-life changes were excellent in 16 cases, good in 3 cases, medium in 1 case. The adaptability of elastic material was excellent in 9 cases, good in 8 cases, medium in 2 cases and poor in 1 case. Conclusions:CDT can reduce subcutaneous edema, effectively reduce limb volume, decrease skin fibrosis, improve skin barrier function, improve skin texture, restore the appearance of the limb and improve patients’ quality of life.

Objective: To elaborate the characteristics and advantages of Whole-Mount immune fluorescence staining by observing the lymphatic vessels of mice. Methods: The ear skin tissue, the hindlimb lymphatic vessels and the mesenteric lymphatic vessels were harvested from normal C57 mice. The tissue samples were subjected to whole-tissue immunofluorescence staining.These tissue samples were fixed by paraformaldehyde, blocked by bovine serum and incubated in primary and secondary antibodies. Then, the lymphatic vessels were observed and analyzed in these samples with a confocal laser-scanning microscope. Results: The capillary lymphatic vessels and lymphatic endothelial cells can be clearly showed in the ear skin. The valves and smooth muscles can be clearly showed in the hindlimb and mesenteric lymphatic vessels by Whole-Mount immunofluorescence staining. Conclusions The whole-tissue immunofluorescence staining technique can observe the external morphology of lymphatic vessels clearly and stereoscopically, and can deeply observe the internal structure of lymphatic vessels. This technique can provide more accurate study on physiology and pathology of lymphatic vessels.

Objective: To explore the relationship between protease-chymase secreted by mast cells, and activated transforming growth factor-β1(TGF-β1), in skin with secondary lymphedema(SLE)in lower extremity, so as toidentify the key factors in fibrosis of lymphedema. Methods: In this study, the affected limb skin of 7 SLE patients was includedas the experimental group, and normal skin tissue of the lower limb of 7 volunteers was used as controls. The skin samples were assayed by Masson staining, and the expressions of chymase and TGF-β1 were assayed by immunohistochemistry, immunofluorescent staining and enzyme-linked immunosorbent assay. Results: There was obvious fibrosis in the skin of lower extremity in patients with lymphedema. The number of MCs andthe expressions of chymase, latency-associated peptide TGF-β1 (LAP TGF-β1) and TGF-β1 were all significantly increased in fibrotic skin in lymphedema, compared with those in normal skin. At the same time, the chymase-containing mast cells accumulated in the lymphatic vessels, with higher expression of TGF-β1. Conclusions The expression of chymase and TGF-β1 was significantly increased in the fibrotic skin insecondary lower extremity lymphedema. The increased expression of chymase in the skin may activate more TGF-β1 expression, and the increased TGF-β1 may promote skin fibrosis in SLE.

The treatment of lymphedema has been difficult due to the lack of understanding of the etiology and pathophysiology of the disease, as well as the embryology and physiology of the lymphatic system. The previous surgical treatment mostly has the blindness. Complete decongestion therapy (CDT) is the most popular treatment currently. The ultimate goal of lymphedema is the targeted and individualized treatment. New technology of multimodality lymphatic imaging emerged in the recent years largely improves the diagnosis of lymphatic circulation disorders. The treatment of peripheral lymphedema is expected to have new achievement. The key points that should be noticed in peripheral lymphedema treatment are ① to distinguish between primary and secondary lymphedema as the function and structure of the lymphatic system are substantially different. The lymphatic vessel regeneration and self-repair has not be proved in primary lymphedema. ② lymphatic vessel (possibly lymph node too) undergo a series pathological and degenerative changes in the affected limb. ③ the pathological changes of lymphoedema tissue Including chronic inflammation, tissue fibrosis and fatty deposition, should be included in the treatment target. To decrease the expression and prevent the function of the key inflammatory factor may be a new way for the treat of lymphedema.

Objective: To evaluate the diagnostic value of indocyanine green(ICG)lymphography in primary lymphedema of extremities. Methods: 61 patients with limb lymphedema were enrolled in the study. The contralateral healthy limbs were used as controls. After intradermal injection of Indocyanine Green, dynamic observation of lymphatic vessels and lymph flow was performed using PDE near-infrared fluorescent instrument. The frequency of lymphatic contraction was calculated. Results: Lymphatic vessel images were clearly visualized in healthy limbs. Inguinal or axillary lymph nodes can be visualized when examined 30 minutes after injection. Lymphatic contraction frequency was 1(0.33-5.00)time per minute. 94.8% lymphedematous limb demonstrated delayed lymphatic vessel or lymph nodes. "Dermal backflow" and diffused shadow pattern can be observed in all affected limbs. Abnormal lymphatic vasculature and contraction can be visualized. The frequency of lymphatic contraction in the affected limb was higher compared to controls: 2.5(0.5-7.0) times per minute. Conclusions ICG lymphography possess high sensitivity for lymphedema diagnosis. ICG lymphography furthers our understanding of pathophysiological alterations of lymphatic disorders.