OBJECTIVE To explore the therapeutic effect of basic fibroblast growth factor (bFGF) coacervates on diabetic peripheral neuropathy (DPN) in diabetic rats.METHODS Poly (ethylene argininylaspartate diglyceride) (PEAD), heparin and bFGF were dissolved in saline at the mass ratio of 50:10:1 to obtain bFGF coacervates. The loading efficiency of bFGF in the coacervates was analyzed by Western blotting. The release profile of bFGF from the coacevates was detected by ELISA. Male SD rats were ip injected with streptozotocin 65 mg · kg- 1 to establish a diabetic model,and DPN occurred 8 weeks later. The DPN rats were randomly divided into free-coacervate group, bFGF group and bFGFcoacervate group. For bFGF group, bFGF 200 μg·kg-1 was im injected once daily for 3 d. In bFGF-coacervate group, bFGF coacervate solution (244 μL) equal to bFGF 200 μg · k - 1, was im given only once. DPN rats in free- coacervate group were im given the same volume of vehicle(PEAD + heparin) only once. Ten normal age peer rats were taken as normal control group.Footprint analysis was conducted each week to evaluate motor function. On the 30th day after treatment,the rats were sacrificed, and sciatic nerves of both sides were harvested for pathological observation through HE staining. Apoptosis in nerve tissue was detected by DAPI staining, and Ki67 and proliferating cell nuclear antigen (PCNA) protein levels were detected by Western blotting. RESULTS Western blotting and ELISA analysis indicated that bFGF-coacervates were well prepared at a mass ratio of 50:10:1,and controlled bFGF release for at least 35 d. The result of rat behavior evaluation and pathological index test indicated that, compared with normal control group, the sciatic function index (SFI) in free-coacervate group decreased significantly(P<0.01), the internal nerve fibers were accompanied by irregularity and serious demyelination, and there was a large number of apoptotic nuclei and low expressions of Ki67 and PCNA proteins (P<0.01).After injection with bFGF or bFGF-coacervates, the SFI increased progressively (P<0.05, P<0.01), and the proportion of fibers with myelin abnormalities and apoptotic cells was significantly reversed. Moreover, the levels of Ki67 and PCNA was evidently enhanced on the 30th day post- operation (P<0.05, P<0.01). Compared with bFGF group, the results of those detection indicators in bFGF-coacervate group were better (P<0.05, P<0.01). CONCLUSION PEAD and heparin complex can load bFGF with high efficiency, and control its release in a steady manner. For DPN rats,treatment with bFGF-coacervates is more effective than bFGF alone.

Objective: To study the clinical effect of nano-fat mixed granule fat transplantation in the treatment of cicatricial facial depression and atrophy, and to explore the related experimental mechanism. Methods: (1) From January 2012 to April 2018, 105 patients conforming to the inclusion criteria, with cicatricial facial depression and atrophy deformity who needed facial fat transplantation, were admitted to our unit. Their medical records were analyzed retrospectively. According to the patients′ wishes, 54 patients (12 males and 42 females) aged 10-59 years received traditional transplantation of pure autologous granule fat collected from abdomen/thigh and were included in simple transplantation group, while the other 51 patients (14 males and 37 females) aged 7-63 years received transplantation of autologous nano-fat mixed granule fat collected from abdomen/thigh and were included in mixed transplantation group. The treatment satisfaction of patients 3 and 6 months after operation was assessed by the facial fullness, symmetry, scar, and complications using self-made scales and photo data before and after operation. Six months after operation, the patients were assessed whether they needed to undergo a second operation, and the rate of second operation was calculated. During the second operation, the fat of patients transplanted in the first operation was collected, and the morphology of adipocytes and microangiogenesis was observed under a scanning electron microscope. (2) Adipose-derived stem cells (ADSCs) were isolated and cultured from abdominal fat of a 4-week-old male Sprague-Dawley (SD) rat. The 5th passage of cells were selected to observe cell morphology after cultured for 14 days, observe expression of vimentin and cytokeratin-18 by immunofluorescence method, identify osteogenic and adipogenic differentiation, and detect rates of CD29 and CD44 positive cells by flow cytometer (n=3). Eighteen 4-week-old male SD rats were divided into ADSCs transplantation group, simple scar group, and blank control group according to the random number table, with 6 rats in each group. Rats in ADSCs transplantation group and simple scar group were subcutaneously injected with 1 mL bleomycin which was dissolved in phosphate buffered saline (PBS) with a mass concentration of 1 mg/mL at the back to establish scar models. After 3 hours, rats in ADSCs transplantation group were injected with 1×10⁶ ADSCs suspended in 0.1 mL PBS at the same injection site, while rats in simple scar group were injected with 0.1 mL PBS. Rats in blank control group were injected with the same doses of PBS in the same place at the same two time points mentioned above. After continuous injection for 28 days in each group, the full-thickness skin tissue of the injected area of all rats was collected to observe the collagen fibers by Masson staining and expressions of α-smooth muscle actin (α-SMA) and transforming growth factor β₁ (TGF-β₁) by immunohistochemistry, and the positive cells were counted. Data were processed with Mann-Whitney U test, χ² test, one-way analysis of variance, and least significant difference test. Results: (1) Compared with the preoperative condition, the facial fullness and symmetry of patients in simple transplantation group were better in 3 months after operation, with scar color closer to the surrounding skin, and the filling volume of patients in this group decreased in 6 months after operation as compared with that in 3 months after operation. In mixed transplantation group, the facial fullness and symmetry of patients were better in 3 and 6 months after operation as compared with the preoperative condition, with scar color and texture closer to the surrounding skin, and the filling volume in 6 months after operation was not obviously reduced as compared with that in 3 months after operation. Fat liquefaction and subcutaneous nodule formation occurred respectively in 1 patient in simple transplantation group within 3 months after operation. The treatment satisfaction of patients in mixed transplantation group was significantly higher than that in simple transplantation group in 3 and 6 months after operation (Z=-2.566, -3.084, P<0.05 or P<0.01). Six months after operation, the second operation rate of patients in mixed transplantation group was 7.84% (4/51), which was significantly lower than 22.22% (12/54) in simple transplantation group (χ²=4.199, P<0.05). At the second operation, compared with those of simple transplantation group, the cells of fat transplanted in the first operation of patients in mixed transplantation group were more plump, without collapse or dryness, and the cells were closely arranged, with smaller gap; the tubular and the cord-like microvascular structure on the cell surface were more abundant, and the cell gap was full of network-like microvascular structure that grew into the adipose tissue. (2) The fifth passage of cells isolated and cultured from rat fat grew adherently to the wall, with long fusiform or spindle shape, showing shoal-of-fish-like growth. Vimentin and cytokeratin-18 were highly expressed in the cells. Cells showed osteogenic and adipogenic differentiation ability by induction. The positive expression rates of CD29 and CD44 were higher than 90.00%. The cells were identified as ADSCs. After 28 days of injection, the collagen fibers in the dermis of skin tissue at the injection area of rats in blank control group were finely arranged. In simple scar group, a large amount of collagen was deposited in the dermis of skin tissue at the injection area of rats, the fiber bundles were thick and loosely unevenly arranged, and a large number of inflammatory infiltration and scattered muscle fibers were observed. In ADSCs transplantation group, the collagen fibers in the dermis of skin tissue at the injection area of rats were thicker than those of blank control group, with still neat arrangement, and a small amount of scattered muscle fiber and inflammatory infiltration was observed. After 28 days of injection, the expression of α-SMA in ADSCs transplantation group was mainly in microvessels in the dermis of skin tissue at the injected area of rats, and the number of α-SMA and TGF-β₁ positive cells was (49±12) and (63±10) cells per 20-fold field of view, respectively, which was similar to (35±16) and (44±17) cells per 20-fold field of view of blank control group (P>0.05), all significantly less than (135±13) and (121±23) cells per 20-fold field of view of simple scar group (P<0.05). Conclusions Compared with those of autologous simple granule fat transplantation, autologous nano-fat mixed granule fat transplantation has better filling fullness in the treatment of patients with scar facial depression and atrophy. The filling effect lasts longer, and the improvement of scar texture is more obvious. As showed in the rat scar model experiment, the mechanism may be that ADSCs inhibit the expressions of α-SMA and TGF-β₁, thus inhibiting the formation of scar.

[摘 要] 目的：探讨内吞作用相关基因FCHO2在各亚型乳腺癌中的表达及其与乳腺癌患者的预后和免疫细胞浸润的相关性。方法：应用免疫组化法和bc-GenExMiner v5.0数据库数据分析FCHO2在各亚型乳腺癌组织中的表达，通过GEO和TIMER数据库数据分析FCHO2与各亚型乳腺癌患者预后和免疫细胞浸润的关系，利用STRING和GEPIA数据库数据分析与FCHO2的互作蛋白网络和其与互作蛋白的相关性，通过UALCAN和DAVID数据库数据对乳腺癌组织中FCHO2表达相关基因进行KEGG和GO分析。结果：免疫组化法结果显示，FCHO2在管腔型和HER2+乳腺癌组织中均呈高表达（均P<0.05），且与HER2和Ki67表达有关联（P=0.03和P=0.007）。FCHO2高表达的管腔型乳腺癌患者总生存期（OS）和无复发生存期（RFS）均明显缩短（均P<0.05）。FCHO2蛋白与EPS15等多种蛋白表达相关且构成蛋白-蛋白互作网络。KEGG和GO分析显示，乳腺癌组织中FCHO2相关表达基因主要与昼夜节律、自噬等生物学过程有关，涉及叉头框蛋白O（FoxO）和TGF-β等信号通路。FCHO2表达与各亚型乳腺癌组织中的免疫细胞浸润相关（均P<0.05）。结论：FCHO2在管腔型、HER2+乳腺癌组织中呈高表达，且与管腔型乳腺癌患者预后及免疫细胞浸润相关，其可能成为乳腺癌治疗的潜在靶点。

[摘 要] 目的：探讨内吞作用相关基因FCHO2在各亚型乳腺癌中的表达及其与乳腺癌患者的预后和免疫细胞浸润的相关性。方法：应用免疫组化法和bc-GenExMiner v5.0数据库数据分析FCHO2在各亚型乳腺癌组织中的表达，通过GEO和TIMER数据库数据分析FCHO2与各亚型乳腺癌患者预后和免疫细胞浸润的关系，利用STRING和GEPIA数据库数据分析与FCHO2的互作蛋白网络和其与互作蛋白的相关性，通过UALCAN和DAVID数据库数据对乳腺癌组织中FCHO2表达相关基因进行KEGG和GO分析。结果：免疫组化法结果显示，FCHO2在管腔型和HER2+乳腺癌组织中均呈高表达（均P<0.05），且与HER2和Ki67表达有关联（P=0.03和P=0.007）。FCHO2高表达的管腔型乳腺癌患者总生存期（OS）和无复发生存期（RFS）均明显缩短（均P<0.05）。FCHO2蛋白与EPS15等多种蛋白表达相关且构成蛋白-蛋白互作网络。KEGG和GO分析显示，乳腺癌组织中FCHO2相关表达基因主要与昼夜节律、自噬等生物学过程有关，涉及叉头框蛋白O（FoxO）和TGF-β等信号通路。FCHO2表达与各亚型乳腺癌组织中的免疫细胞浸润相关（均P<0.05）。结论：FCHO2在管腔型、HER2+乳腺癌组织中呈高表达，且与管腔型乳腺癌患者预后及免疫细胞浸润相关，其可能成为乳腺癌治疗的潜在靶点。