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Objective@#Ischemia-reperfusion (IR) injury is a leading cause of flap compromise and organ dysfunction during free-tissue transfer, and remains a great challenge for plastic surgeons. Thioredoxin-1 (Trx-1) was proved to protect the IR flap by mitigating the oxidative stress, and inhibiting the activation of apoptosis signal-regulating kinase-1 (ASK-1) and mitogen-activated protein kinase (MAPK) pathway. The aim of this study is to investigate the distinction of Trx-1 expression, apoptosis indices in different layers of IR flaps, and the feasibility of tissue-layer-specific administration of Trx-1.@*Methods@#Ten patients′ specimens of IR flaps for DIEP breast reconstruction were collected and assessed for apoptosis and Trx-1 expression. Twenty mice were used to establish the IR flap model. The mice were sacrificed twenty-four hours after reperfusion. The flap tissues were harvested and tested by immunohistochemistry staining and TUNEL assay. The tissue-layer-specific dermoprotective effect of Trx-1 and the molecular mechanisms were assessed by an in vitro epithelial skin cell hypoxia-reoxygenation model. The statistics were conducted by t test and ANOVA using SPSS 20.0.@*Results@#Trx-1 expression and apoptotic cells were observed mainly located in the basal layer of epidermis and the papillary layer of dermis in human IR flaps and mice models. Trx-1 depletion was 24.19 %± 2.23% in the basal layer of epidermis and the papillary layer of dermis of patient IR flaps, decreasing significantly compared with 70.71% ± 6.38% in control group (t = 27.54, P< 0.001). Similar tissue-layer-specific down regulation of Trx-1 also displayed in mice IR flap models (19.83% ± 2.34% vs. 76.59% ± 4.88%; t = 34.71, P<0.001). The apoptotic index in human samples significantly increased from 1.32% ± 1.52% in control group to 43.71 %± 3.17% in IR group (t =38.23, P<0.001); while it was proved to be dramatically raised in mice models from 0.86% ± 1.15% in control group to 41.14 %± 4.21% in IR group (t= 36.96, P < 0.001). Western Blot analysis revealed Trx-1 down regulation and a significant increase in ASK-1, p-p38, and c-PARP abundance in the hypoxia-reoxygenation-treated HaCaT cells (P < 0.01). Supplementation of recombinant human Trx-1 significantly reduced the apoptosis-related protein expression.@*Conclusions@#The basal layer of epidermis and the papillary layer of dermis are the main damaged tissue layers in the early stage of skin flap ischemia-reperfusion injury. The IR flap can be protected by precisely replenishing the vulnerable layers with Trx-1.
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Objective To investigate the effect of cancer-associated fibroblasts (CAFs)-derived chemokine ligands 7 (CCL7) on the proliferation and invasion of triple-negative breast cancer (TNBC) cells. Methods The mRNA expression level and protein level of CCL7 in CAFs and paracancerous fibroblasts were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western Blot respectively. To confirm the paracrine level of CCL7 in CAFs and paracancerous fibroblasts, the protein levels of CCL7 in the corresponding conditional medium were detected through enzyme-linked immunosorbent assay (ELISA). The effect of CCL7 on the proliferation and invasion of MDA-MB-231 (TNBC cell line) was investigated by MTS assay and Transwell assay, respectively. Results In comparison with paracancerous fibroblasts, the mRNA expression level and protein level of CCL7 in CAFs were significantly increased (both P<0.01). There was an obviously increase of paracrine level of CCL7 in CAFs-conditional medium (P<0.01). The MTS assay and Transwell assay results indicated that CCL7 was more able to promote the proliferation and invasion of MDA-MB-231. Conclusion CAFs in the TNBC stroma can produce more chemokine CCL7, and CCL7 can promote the proliferation and invasion of TNBC cells
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Objective To evaluate satisfaction and complications of deep inferior epigastric perforator (DIEP) flaps breast reconstruction afer mastectomy in breast cancer patients.Methods The 26 consecutive breast cancer patients who underwent DIEP flap breast reconstruction were collected.General patient satisfaction and esthetic satisfaction with DIEP flap breast reconstruction were accessed using a study-specific questionnaire modified from the Michigan Breast Reconstruction Outcome Study.Results Flap success rate was 100％.The general satisfaction of DIEP breast reconstruction was 61.9％ (13/21),and the esthetic satisfaction was 66.7％ (14/21).Age (≤40 years old was 81.8％ [9/11],＞40 years old was 40.0％ [4/10],P=0.049) and previous abdominal wall surgery (no previous abdominal wall surgery was 75.0％ [12/16] and previous abdominal wall surgery was 20.0％ [1/5],P=0.027) were significant negative factors affecting general satisfaction;Body mass index (BMI ＜25 kg/m2 was 76.5％ [11/17],BMI 25-30 kg/m2 was 50.0％ [2/4],P=0.049) was a statistically significant factor associated with esthetic satisfaction.Four flaps presented venous congestion.One patient had fat necrosis and one had flap infection of the reconstructed breast.One patient had a decrease in abdominal rotational strength.Conclusions DIEP flap breast reconstruction will offer distinct advantages to patients,in terms of decreased donor-site morbidity and shorter recovery periods.
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Objective: To assess the expression of retinoic acid receptor beta 2 (RAR-β2) in breast tumor and to evaluate the relationship between RAR-β2 expression and tumorigenesis of breast cancer. Methods: Immu-nohistochemistry was used to detect the expression of RAR-β2 protein in specimens from 40 cases of breast cancer, 40 cases of atypical ductal hyperplasia, 40 cases of fibroadenoma and 20 cases of normal breast tissues. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression levels of RAR-β2 gene in 20 cases of breast cancer, 20 cases of atypical ductal hyperplasia, 20 cases of fibroadenoma, and 20 cases of normal breast tissues. Results: Immunohistochemical staining results revealed that the positive expression of RAR-β2 protein showed nuclear staining. The positive expression rate of RAR-β2 was 30% (12/40) in breast cancer, 17.5% (7/40) in atypical ductal hyperplasia, 87.5% (35/40)in fibroadenoma, and 95% (19/20) in normal breast tissues. The expression of RAR-β2 protein in breast cancer was significantly lower than that in normal breast tissues (X~2=26.30, P<0.001). The expression of RAR-β2 was not significantly different between atypical ductal hyperplasia and breast cancer (P>0.05). No correlation was found between the expression of RAR-β 2 protein and the tumor size, menopausal age, lymph node metastasis, clinical stage, histological grade, or protein expression of ER and PR in breast cancer tissues (P>0.05). Follow-up results showed that 3 out of 28 patients with negative RAR-β2 expression had visceral organ metastasis, but only one of the 12 RAR-β2 positive patients had osseous metastasis. RT-PCR analysis showed that the positive expression rate of RAR-β2 mRNA in breast cancer, atypical ductal hyperplasia, fibroadenoma and normal breast tissues was 25% (5/20), 35% (7/20), 85% (17/20) and 100% (20/20), respectively. The RAR-β2 mRNA expression rate in breast cancer was significantly lower than that in normal breast tissues (X~2=30.43, P<0.001). No significant difference in RAR-β2 mRNA expression was found between atypical ductal hyperplasia and breast cancer tissues (P>0.05). Conclusion: RAR-β2 gene may play a repressive role in the initiation of breast cancer, and the loss of the expression of RAR-β2 gene may be the initial step in breast carcinogenesis.
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Objective To study tumor immune escape in a gastric carcinoma cell line expressing human indoleamine 2,3-dioxygenase(IDO).Methods Human IDO gene was cloned by RT-PCR and the vector for pIRES_2-EGFP-IDO was constructed.BGC-823 cells were transfected with the plasmid using eleetroporation.The integrated INDO genes were detected by RT-PCR and Western blot.The enzyme activity of IDO were measured.T cells from gastric cancer patients were cecuhured with BGC-823 transfected with IDO or added with 1-MT circumstance,T cell-mediated cytotoxicity and proliferation were detected.Results Higher level expression of IDO mRNA and IDO protein Was detected in tumor cells transfected with IDO gene.The level of kynurenic acid was higher in transfected cells compared with no-transfeeted group (4.84±0.11)mg/L vs.(1.83±0.10)mg/L,P=0.000.The cytotoxicity ratio of the IDO transfected group and transfected group with 1-MT circumstance (1-MT group) was lower than control group (P<0.05).The inhibition rate of transfected group with 1-MT group Was higher than control group(P<0.05).Conclusion Gastric cancer cell lines encoded with IDO inhibits T cell-mediated cytotoxicity and proliferation.