Construction of multifunctional coating with cationic amino acid-coupled peptides for osseointegration of implants.

Osseointegration is an important indicator of implant success. This process can be improved by coating modified bioactive molecules with multiple functions on the surface of implants. Herein, a simple multifunctional coating that could effectively improve osseointegration was prepared through layer-by-layer self-assembly of cationic amino acids and tannic acid (TA), a negatively charged molecule. Osteogenic growth peptide (OGP) and the arginine-glycine-aspartic acid (RGD) functional polypeptides were coupled with Lys6 (K6), the two polypeptides then self-assembled with TA layer by layer to form a composite film, (TA-OGP@RGD)n. The surface morphology and biomechanical properties of the coating were analyzed in gas and liquid phases, and the deposition process and kinetics of the two peptides onto TA were monitored using a quartz crystal microbalance. In addition, the feeding consistency and adsorption ratios of the two peptides were explored by using fluorescence visualization and quantification. The (TA-OGP@RGD)n composite membrane mediated the early migration and adhesion of cells and significantly promoted osteogenic differentiation and mineralization of the extracellular matrix in vitro. Additionally, the bifunctional peptide exhibited excellent osteogenesis and osseointegration owing to the synergistic effect of the OGP and RGD peptides in vivo. Simultaneously, the (TA-OGP@RGD)n membrane regulated the balance of reactive oxygen species in the cell growth environment, thereby influencing the complex biological process of osseointegration. Thus, the results of this study provide a novel perspective for constructing multifunctional coatings for implants and has considerable application potential in orthopedics and dentistry.

Expression and significance of LAT1 and p-s6 breast cancer / 临床与实验病理学杂志

Purpose To investigate the expression and significance of type L amino acid transporter 1 (LAT1 ) and phosphorylated s6 ribosomal protein (p-s6) in breast cancer tissues and their correlation. Methods LAT1 protein and p-s6 protein were detected by immunohistochemical EnVision two step method in 178 cases of breast cancer and 78 cases of benign breast lesion, and the relationship between the expression and clinicopathological parameters was analyzed. Results The positive rate of LAT1 in breast cancer was 36.5％, which was significantly higher than that of breast benign lesion tissues (23.1 ％, P＜ 0.05 ), the positive rate of p-s6 in breast cancer tissues was33.2％, which was significantly higher than that of breast benign lesion tissues (12.8％, P＜0.05). There was a positive correlation between the expression of LAT1 protein and p-s6 protein in breast cancer tissues (r = 0.345, P＜ 0.05). The expression of LAT1 protein in breast cancer was correlated with tumor diameter, axillary lymph node metastasis, TNM staging and HER-2 level (P＜ 0.05), but not associated with the patient's age, histological grade, ER, and PR levels (P＞ 0.05). The expression of p-s6 protein was related to axillary lymph node metastasis, TNM staging, age and ER level (P＜ 0.05), but not associated with tumor diameter, histological grade, PR and HER-2 levels (P＞ 0.05 ). Multivariate analysis showed that the expres sion of LAT1 protein was related to tumor diameter and expression level of HER-2. The expression of p-s6 protein was related to axillary lymph node metastasis. Conclusion The expression of LAT1 protein and p-s6 protein in breast cancer is up-regulated, and the expression of these two proteins is positively related, which implying that LAT1 and p-s6 might play a synergistic role in the development and progression of breast cancer.

Effect of silencing CD98hc expression on MDA-MB-231 breast cancer cells proliferation and invasion / 临床与实验病理学杂志

Purpose To investigate the effect of silencing of CD98hc on proliferation,migration and invasion of MDA-MB-231 breast cancer cells.Methods RNA interfere technology was used to silence CD98hc in MDA-MB-231 cells.MDA-MB-231 cells (CD98hc-shRNA) with stable low expression of CD98hc and Vector control was obtained.Western blot and qRT-PCR were used to verify the down-regulation of CD98hc.MTT assay was used to test the cell proliferation.Transwell migration and invasion experiment were used to assay cell migration and invasion.Results The expression levels of CD98hc was down-regulated by CD98hc-shRNA in the MDA-MB-231 breast cancer cells.Compared to that of blank cells (0.706 ± 0.013),vector control (0.724 ± 0.018),the cell proliferation potency of CD98hc-shRNA cells (0.580-0.035) was significantly inhibited (P ＜ 0.05),and the cell migration and invasion potency also were inhibited (P ＜ 0.05),there is on significant different between blank cells and vector control (P ＜ 0.05).Conclusion CD98hc might be involved in the regulation of breast cancer cell biological behaviors.

Prolactin Inhibits BCL6 Expression in Breast Cancer Cells through a MicroRNA-339-5p-Dependent Pathway / 한국유방암학회지

PURPOSE: Prolactin (PRL) plays a critical role in breast cancer progression by activating its cognate receptor and promotes the growth and differentiation of breast cancer cells. Studies have shown that B-cell lymphoma 6 (BCL6) is the target gene of microRNA-339-5p (miR-339-5p) and that BCL6 expression contributes to breast cancer progression. Herein, we identified PRL as a potent suppressor of BCL6 expression in human breast cancer cells. METHODS: Western blotting and quantitative reverse transcription-polymerase chain reaction were used to investigate molecular mechanisms underlying miR-339-5p expression and BCL6 manipulation in MCF-7, T47D, and SKBR3 breast cancer cells. Phenotypic changes in these breast cancer cell lines were assessed by performing cell viability (MTT), colony formation, migration, and invasion assays. RESULTS: PRL suppressed BCL6 protein and mRNA expression and upregulated miR-339-5p expression in MCF-7 and T47D breast cancer cells. Selective downregulation of miR-339-5p expression significantly reversed PRL-induced suppression of BCL6 mRNA and protein expression. Exogenous PRL stimulation significantly decreased the proliferation, colony formation, migration, and invasion of breast cancer cells, and suppression of miR-339-5p expression reversed these processes in vitro. CONCLUSION: These results indicated that PRL inhibited BCL6 expression and regulated breast cancer progression through a miR-339-5p-dependent pathway.

Expression of the LC3A protein in prostate cancer and its implications / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the expression of the LC3A protein in prostate cancer (PCa) and its clinicopathological significance. We detected the expression of the LC3A protein by immunohistochemistry in 54 cases of PCa and 14 cases of benign prostatic hyperplasia (BPH), and analyzed the correlation between the LC3A expression and the clinicopathological parameters in PCa. The positive signals of the LC3A protein were located in the cytoplasm and/or cell nuclei. The rate of its strongly positive expression was 90.7% in PCa, significantly higher than 14.3% in BPH (P < 0.01). The LC3A expression was also found in the cell nuclei of 22 cases of PCa, with no significant correlation to that in the cytoplasm (P > 0.05). The expression of LC3A was significantly correlated with Gleason scores (r = 0.297, P = 0.029 in cytoplasm; r = 0.288, P = 0.034 in cell nuclei), but not with the clinical stage, patient's age, androgen receptor (AR) level and preoperative levels of serum PSA and cPSA (all P > 0.05). LC3A was also expressed in the fibrocytes and smooth muscle cells in PCa and BPH. The positive rate of AR was 74.1% (40/54) in PCa and 64.3% (9/14) in BPH, with no significant difference between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>The expression of the LC3A protein might be involved in the development, differentiation, and prognosis of prostate cancer.</p>

Expression of CD147, matrix metalloproteinases and transforming growth factor beta1 in breast cancer / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of CD147, matrix metalloproteinase (MMP)-2, MMP-9, Transforming growth factor (TGFbeta1) and TGFbetaRI proteins and their relationships to breast cancer.</p><p><b>METHODS</b>The expression of CD147, MMP-2, MMP-9, TGFbeta1 and TGFbetaRI proteins was examined on tissue chips containing 160 cases of breast carcinomas by S-P immunohistochemical method.</p><p><b>RESULTS</b>The positive rates of CD147, MMP-2, MMP-9, TGFbeta1 and TGFbetaRI proteins were 87.5% (140/160), 96.9% (155/160), 95.0% (152/160), 73.7% (118/160) and 60.6% (97/160), respectively. The expression of CD147 was positively correlated with axillary lymph node metastasis, TNM staging and HER2 over expression (P < 0.01, P < 0.05 and P < 0.01, respectively), and inversely correlated with PR expression (P < 0.05). The patients' relapse-free survival was shorter in TGFbeta1-positive group than in TGFbeta1 negative group (P < 0.05). Both the expression of MMP-2 and MMP-9 were positively correlated with CD147 expression; and both the expression of TGFbeta1 and TGFbetaRI were positively correlated with CD147 expression (P < 0.01 and P < 0.05, respectively).</p><p><b>CONCLUSION</b>The expression of CD147 is considered closely correlated with tumor invasion, metastasis and prognosis in breast cancer, and has also a close correlation with MMP-2, MMP-9, TGFbeta1 and TGFbetaRI expression.</p>

Expression of TGF-beta1 and the mechanism of invasiveness and metastasis induced by TGF-beta1 in breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of MMP-2, TIMP-2, TGF-beta1 and TGF-betaRI and the relationship among them in breast cancer.</p><p><b>METHODS</b>The protein expression of MMP-2, TIMP-2, TGF-beta1 and TGF-betaRI was detected on tissue chips by S-P immunohistochemical staining in 160 cases of breast carcinoma.</p><p><b>RESULTS</b>The positive rates of TGF-beta1, TGF-beta1 mRNA, MMP-2, MMP-9, TIMP-1 and TIMP-2 expression were 73.7%, 56.2%, 96.9%, 95.0%, 87.5% and 89.4%, respectively. Axillary lymph node metastasis and TNM staging (P < 0.01 and P < 0.01, respectively) were positively correlated to the expression of TGF-beta1. Relase-free survival of TGF-beta1 positive group was lower than that of TGF-beta1 negative group (P = 0.023). The expression of MMP-2 or MMP-9 was positively correlated to that of TGF-beta1 (r = 0.170, P < 0.05; r = 0.221, P < 0.01) and was negatively correlated to that of TGF-beta1 mRNA (r = -0.126, P > 0.05;r = 0.019, P > 0.05).</p><p><b>CONCLUSION</b>The expression of TGF-beta1 may be closely correlated with the invasion and metastasis of breast cancer. TGF-beta1-induced invasiveness and metastasis of breast cancer cells are mediated by MMP-2 and MMP-9.</p>

Autologous fat grafting to the breast for cosmetic enhancement: experience in 66 patients with long-term follow up.

BACKGROUND: Autologous fat grafting to the breast for cosmetic enhancement remains controversial because the efficacy and fate of fat grafting to the breast are primarily unknown. In this report, we present our retrospective study in 66 patients who underwent autologous fat grafting to the breast for various cosmetic reasons and who were followed with sonography, mammography, or magnetic resonance imaging (MRI). METHODS: Sixty-six patients who desired cosmetic enhancement of the breast for various reasons underwent autologous fat transplantation between August 2000 and March 2005 in our institution. The cosmetic outcome was assessed by the plastic surgeons as well as the patients. The imaging features of fat necrosis, cyst formation, and calcification in these patients were carefully studied and biopsies of palpable lumps were evaluated histologically. RESULTS: All patients were followed from 13 to 61 months with an average of 37 months. Breast cosmetic contour was significantly improved in 28 patients (42.4%), improved in 24 patients (36.4%), and not improved in 14 patients (21.2%) as judged by the plastic surgeons. Twenty-seven patients (40.9%) were very satisfied, 26 patients (39.4%) were satisfied, and 13 patients (19.7%) were unsatisfied. Eleven patients (16.7%) developed liponecrotic cysts but only two patients elected to have the breast lump surgically removed. CONCLUSION: Autologous fat grafting to the breast can be a useful procedure for cosmetic enhancement in many patients who desire such a procedure. Patients with breast contour deformities after removal of silicon implants were found to be the best candidates for fat grafting. The primary long-term complication is the formation of liponecrotic cysts which have characteristically benign appearances in sonography, mammography or MRI.

Expression of a novel metastasis-inducing protein human anterior gradient-2 (AGR2) in breast cancer and its clinical and prognostic significance / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of a novel metastasis-inducing protein human anterior gradient-2 (AGR2) in breast cancer and its clinical and prognostic significance.</p><p><b>METHODS</b>AGR2 expression was assessed in 160 cases of breast cancer and 20 cases of benign breast diseases by immunohistochemistry using tissue chip technology. In addition the expression of ERa, PR and c-erbB-2 in breast cancer was also evaluated. Follow-up information of 5-year duration was available in 127 patients with breast cancer. Kaplan-Meier analysis and COX regression model were used to analyze the correlation between AGR2 expression and the follow-up clinical data.</p><p><b>RESULTS</b>The expression of AGR2 was significantly higher in breast cancers than that in benign diseases (68.3% vs. 25.0% , P < 0.01). There was a negative correlation between AGR2 expression and the histological grade of breast cancer (P <0.05) , whereas positive correlations was found between the expression of AGR2 and ERalpha (P <0.05), and between the expression of AGR2 and PR (P <0.01). In the subgroup of ERalpha-positive breast cancer, Logistic regression model demonstrated AGR2 and TNM stage were important factors affecting lymph node metastasis (both P < 0.01). Kaplan-Meier analysis demonstrated that a positive expression of AGR2 was associated with poor overall survival and relapse-free survival (both P <0.01). Moreover, COX regression model confirmed the expression of AGR2 as an independent prognostic factor among patients with ERa-positive breast cancer (P <0.01).</p><p><b>CONCLUSIONS</b>The abnormal expression of AGR2 may play a role in the pathogenesis and progression of breast cancer. The metastasis-inducing capability of AGR2 may be partly regulated through the ER pathway. Therefore, AGR2 may be a useful molecular marker for prognostication for patient with hormone-responsive breast cancer.</p>

Cadaveric comparison of two facial flap-harvesting techniques for alloplastic facial transplantation.

BACKGROUND: Functional and aesthetic reconstruction of severe facial deformities presents a major challenge, and the results are rarely satisfactory. Recent clinical success of composite tissue allograft transplantation and improvements in autoimmune regulation have initiated efforts to reconstruct severe facial deformities with alloplastic tissue. Few reports address the full facial flap dissection approach, where lengthy procedural times remain a limiting factor in achieving optimal graft survival. Extensive vascular anastomoses within facial tissues provide a unique opportunity to explore alternative graft harvesting strategies to optimise operative ischaemia. OBJECTIVE: The aim of the study was to shorten donor-graft harvesting time and reduce warm ischaemia. We evaluated alternative facial harvesting strategies through mock cadaveric facial transplantations. METHODS: Cadaveric dissections were performed to explore facial-scalp reconstruction alternatives. Six paired sub-superficial muscloaponeurotic system (SMAS) plane composite facial-scalp flaps were harvested using either a superficial temporal artery (STA) or a facial artery (FA) pedicle technique (Group I) or an external carotid artery (ECA) pedicle technique. Total harvesting times and lengths of vascular pedicles were measured. RESULTS: Harvesting time for a STA and FA pedicle total facial flap (mean=113min, range = 105-120 min, SD = 6 min) was shorter than that for an ECA pedicle flap (mean = 232 min, range = 225-240 min, SD = 6 min) (P<0.01). Mean pedicle lengths for the STA, the FA, the ECA, the external jugular vein, and the facial vein were 37 +/- 2.1, 35 +/- 1.8, 26 +/- 1.4, 52 +/- 3.0 and 42 +/- 2.6mm, respectively. Mean pedicle lengths for the supraorbital, supratrochlear, infraorbital, mental, and facial nerve were 15 +/- 1.5, 14 +/- 1.4, 24 +/- 1.2, 30 +/- 1.6 and 32 +/- 1.8mm, respectively. CONCLUSION: Compared with previously reported ECA pedicle total facial allograft harvesting techniques, an STA and FA pedicle flap provides a shorter harvesting time and potentially safer dissection method for facial transplantation by avoiding interference with the complicated anatomy of the carotid and submental triangle. Early graft ischaemic damage can be minimised by this harvesting technique, which significantly shortens harvesting time compared with previously described approaches, while maintaining adequate full facial perfusion.

[Anatomic study of supratrochlear artery and its application in nasal reconstruction].

OBJECTIVE: To study the anatomy of the cutaneous branch (CB) of supratrochlear artery and its relevance to the design of frontal flap in nasal reconstruction. METHODS: 10 fresh cadavers were dissected to study the position and course of the CB of supratrochlear artery (supraorbital rim and facial midline as landmark). The communication between the CB and supraorbital artery was also studied. 5 cases of ultra-thin frontal flaps and 11 cases of bi-flap( cutaneous flap and muscular flap) were designed on anatomic basis. The survival rate of flap, the stability and aesthetic appearance of the reconstructed nose were followed up. RESULTS: The supratrochlear artery gave off constant CB (1.18 +/- 0.36) cm from upper orbital rim and (1.35 +/- 0.34) cm from the midline of face. The CB passed in a subcutaneous plane and communicated with the bilateral muscular branch, CB of the opposite side and bilateral supraorbital artery. The supratrochlear artery only had CB with no muscular branch in 3 cases. All the flaps survived completely except one with blister on the nose tip which healed spontaneously. The postoperative aesthetic appearance was very satisfactory. CONCLUSIONS: The supratrochlear artery has constant CB. The frontal ultra-thin flap pedicled with the CB can improve the therapeutic effect of nasal reconstruction.

[Strategy study of harvesting total facial flap and donor choice for allograft transplantation in cadaver].

OBJECTIVE: To explore the operational strategy of harvesting total facial allograft by autopsy. METHODS: Twelve fresh human cadavers were dissected. They were divided into two groups randomly. The total facial-scalp flap of the group I was elevated by the bi-pedicle method, the group II was operated with single-pedicle method. Both were dissected at the deep plane of the SMAS. : the time of facial flap harvesting, length of the artery vein and nerve pedicles of the donor were measured and marked, after operation, in each group we transferred one facial allograft to another. Then the free graft of group I was poured through artery by methylthioninium chloride to study vascular territories. RESULTS: Mean harvesting time of the group I (46 +/- 11) minutes, group II (111 +/- 7) minutes, P < 0.01. Perfusion result shows that unilateral superficial temporal artery and the opposite side of the facial artery can supply blood for whole face. The pedicle was long enough for anastomosis. Post-operation appearance: the face looks like neither the donor nor the recipient primarily, It's mainly due to the characteristics of the skeleton and the soft tissues. CONCLUSIONS: The bi-pedicle method of the harvesting total facial allograft is concise, fast, safe can be widely applied in clinical.

[Research of the preservation of the composite facial allograft].

OBJECTIVE: To investigate the effective method of preserving composite facial allograft so as to attenuate ischemic injury. METHODS: The composite facial allografts were harvested from dog, perfused and preserved with 4 degrees C physiological sodium chloride and UW solution respectively. Immediately after the removal of the flap, after 12, 24, 48 h of preservation, MTT assay was used to determine the viability of several kinds of tissue, including skin, mucosa, muscle, bleed vessel, nerve and gland. The results of the two groups were compared in term of viability percentage. The pathology of several tissues were observed after 24 and 48 h of storage. RESULTS: The viability percentage of every tissue conserved in UW solution for 48 hours was more than 75%. There was significant difference between physiological sodium chloride group and UW group (P < 0.05). Some changes, including Porous arrangement of fibers in connective tissue of skin and mucosa, hyalinization of tissue around the hair follicle and edema of cell in hair follicle, enlargement of space between muscle bundles and unclearness of boundary of acinus could be seen in physiological sodium chloride group while no significant change in UW group. CONCLUSIONS: UW solution could be considered as preservation solution for composite facial allograft.

[Establishment of composite facial and scalp allograft transplantation model in canine].

OBJECTIVE: To develop an experimental model of composite facial and scalp allograft in canine in order to investigate technical and immunological aspects and functional recovery of facial muscles of this new approach to facial reconstruction. METHODS: (1) Anatomic study: Four mongrel dogs were used for anatomical dissection of the head and neck region and for harvesting flap experiment. (2) Autologous transplantation (group I): Three types composite facial and scalp autologous transplantation were performed in five mongrel dogs. Type I composite tissue flap (group I a n = 2) included bilateral external ear and orbicularis oculi muscle. Type II (group I b n = 1) included single-lateral external ear, orbicularis oculi muscle, external nose upper and lower lip. Type III (group I c n = 2) included single - lateral external ear and orbicularis oculi muscle. (3) Allograft transplantation (group II): In group II a (n = 2), two allograft transplantation were performed with type III composite facial and scalp . In group II b (n = 4), four allograft transplantation were performed with the modified type III composite facial and scalp which included single - lateral external ear, orbicularis oculi muscle and one third of inferior tarsal plate and palpebral conjunctiva. To prevent allograft rejection, Cyclosporin A (CsA) and Methylprednisolone (MP) or Prednisone (PS ) were combined used as immunosuppressive protocol . Dose of CsA was adjusted depending on its blood drug level. Electromyogram (EMG) of orbicularis oculi muscle was carried out at 4 weeks, 6 weeks, 12 weeks and 6 months postoperation. RESULTS: (1) The facial anatomic characteristic of dog is similar to that of human being, external carotid artery and external jugular vein afford good blood supply to composite facial and scalp. (2) The dogs in group I c were long-term surviving with leakage of salivary juice. (3) In group II a (n = 2), one dog presented rejection reaction at 28th day postoperation, the reversal of rejection was achieved by increasing the dose of CsA and prednisone and with topical clobetasol for 2 weeks, the dog survived indefinitely( > 309 days). In group II b (n = 4), there were three dogs survived indefinitely ( > 159 days, > 129 days, > 108 days) without complication, EMG showed the function of orbicularis oculi muscle was gradually improving. CONCLUSION: The modified type III composite facial and scalp allograft transplantation model is an ideal model for facial allograft transplantation study.