Objective To investigate the relationship between the expressions of P-gp, GST-π and C-erbB-2 and clinicopathologic characteristics as well as prognosis in breast cancer. Methods The expressions of P-gp， GST-π and C-erbB-2 were detected by immunohistochemistry in 48 cases of breast cancer, and histopathologic characteristics as well as 5-year survival rate of these cases were analyzed. Results There was no significant difference in the expressions of P-gp and GST-π with age, histologic grade, number of lymph node metastasis and TNM stage of breast cancer ( P ＞ 0.05). There was significant difference in expression of C-erbB-2 with histologic grade, number of lymph node metastasis and TNM stage of breast cancer （ P ＜ 0.05）. Positive rate of P-gp expression in breast cancer with positive C-erbB-2 expression was remarkably higher than that in breast cancer with negative C-erbB-2 expression ( P ＜ 0.05) . Positive rate of GST-π and C-erbB-2 expression in survivals within 5 years was remarkably lower than that in deaths within 5 years ( P ＜ 0.01). Conclusion P-gp participates primary drug-resistance mechanism of breast cancer. The possibility of primary drug-resistance is higher in breast cancer with positive C-erbB-2 expression. The expression of C-erbB-2 helps to evaluate prognosis and the result of treatment in breast cancer.
Objective To investigate the protocols of combined culture of human placenta-derived mesenchymal stem cells (HPMSCs) and human umbilical vein endothelial cells (HUVECs) from the same and different individuals on collagen material, to provide the. Methods Under voluntary contributions, HPMSCs were isolated and purified from human full-term placenta using collagenase IV digestion and lymphocyte separation medium, and confirmed by morphology methods and flow cytometry, and then passage 2 cells were cultured under condition of osteogenic induction. HUVECs were isolated from fresh human umbilical vein by collagenase I digestion and subcultured to purification, and cells were confirmed by immunocytochemical staining of von Willebrand factor (vWF). There were 2 groups for experiment. Passage 3 osteoblastic induced HPMSCs were co-cultured with HUVECs (1 ∶ 1) from different individuals in group A and with HUVECs from the same individual in group B on collagen hydrogel. Confocal laser scanning microscope was used to observe the cellular behavior of the cell-collagen composites at 1, 3, 5, and 7 days after culturing. Results Flow cytometry showed that HPMSCs were bly positive for CD90 and CD29, but negative for CD31, CD45, and CD34. After induction, alizarin red, alkaline phosphatase, and collagenase I staining were positive. HUVECs displayed cobble-stone morphology and stained positively for endothelial cell marker vWF. The immunofluorescent staining of CD31 showed that HUVECs in the cell-collagen composite of group B had richer layers, adhered and extended faster and better in three-dimension space than that of group A. At 7 days, the class-like microvessel lengths and the network point numbers were (6.68 ± 0.35) mm/mm2 and (17.10 ± 1.10)/mm2 in group A, and were (8.11 ± 0.62) mm/mm2 and (21.30 ± 1.41)/mm2 in group B, showing significant differences between the 2 groups (t=0.894, P=0.000; t=0.732, P=0.000). Conclusion Composite implant HPMSCs and HUVECs from the same individual on collagen hydrogel is better than HPMSCs and HUVECs from different individuals in integrity and continuity of the network and angiogenesis.
Objective To investigate the effect of human placental-derived mesenchymal stem cells (PMSCs) on immunological rejection in mouse allogeneic skin transplantation. Methods The placenta fetal tissues from voluntary donors were used to isolate and culture the PMSCs, and the 3rd passage PMSCs were used in the experiment. Thirty Vr ∶ CD1 (ICR) mice at age of 1-2 days were used as skin donors for allogeneic skin transplantation. Thirty C57BL/6 mice at age of 6-8 weeks as recipients were made back skin defect of 12 mm in diameter and were randomly divided into 3 groups (n=10): group A, autograft; group B, allogeneic graft + PBS tail vein injection; and group C, allogeneic graft + human PMSCs (1 × 105 cells/mouse) tail vein injection. The flap survival was observed. At 7 days after skin transplantation, blood leukocyte counting, abdominal fluid macrophage activation, and the expression levels of interleukin 4 (IL-4), interleukin 17 (IL-17), and interferon γ (INF-γ) in blood and spleen were detected by ELISA and RT-PCR, respectively. Results The flap survival time was significantly longer in group A [(58.33 ± 4.04) days] than in groups B and C [(3.80 ± 0.92) days and (6.80 ± 0.82) days] (P lt; 0.05), and in group C than in group B (P lt; 0.05). At 7 days after transplantation, the blood leukocyte number was (6.32 ± 0.45) × 109/L in group A, (7.45 ± 0.52) × 109/L in group B, and (6.35 ± 0.39)× 109/ L in group C, and it was significantly more in group B than in groups A and C (P lt; 0.05). The macrophage activation rate of the abdominal fluid was 6.87% ± 2.40% in group A, 7.84% ± 0.44% in group B, and 15.98% ± 2.87% in group C; group C was significantly higher than groups A and B (P lt; 0.01). ELISA results showed that there was no significant difference in the concentrations of IL-4 among 3 groups (P gt; 0.05). Compared with group B, the concentrations of IL-17 and IFN-γ were significantly reduced in group C (P lt; 0.05), while the concentration of IFN-γ was significantly increased in group B when compared with group A (P lt; 0.05). RT-PCR results showed that there were significant differences in the expressions of IL-4, IL-17, and IFN-γ mRNA between groups B, C and group A (P lt; 0.05); the expressions of IL-4 and IFN-γ mRNA were significantly lower in group C than in group B (P lt; 0.05). Conclusion Human PMSCs transplantation can suppress the acute immunological rejection in allogeneic skin transplantation. The possible mechanism may be partially related to the inhibitory effect on the secretion of IL-17 and IFN-γ.
Objective To study the effect of hypoxia on the prol iferation of hBMSCs and human placental decidua basal is-MSCs (hPDB-MSCs), and to provide the theoretical basis for discovering the new seed cells source for tissue engineering. Methods Density gradient centrifugation method was adopted to isolate and culture hBMSCs and hPDB-MSCs,flow cytometry (FCM) was appl ied to detect cell surface marker. After establ ishing the experimental model of CoC12 chemical hypoxia, MTT method was appl ied to evaluate the prol iferation of hBMSCs and hPDB-MSCs at different time points (6, 12, 24, 48, 72, 96 hours) with various CoC12 concentration (0, 50, 75, 100, 125, 150, 175, 200 μmol/L). Results FCM analysis revealed that hPDB-MSCs and hBMSCs expressed CD9, CD29, CD44, CD105, CD106 and human leucocyte antigen ABC (HLA-ABC), but both were absent for CD34, CD40L and HLA-DR. Compared with hBMSCs, hPDB-MSCs expressed stage-specific embryonic antigen 1 (SSEA-1), SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81 better. The prol iferations of hPDB-MSCs and hBMSCs were inhibited within the first 12 hours under hypoxia condition, but promoted after 12 hours of hypoxia. Compared with the control group, the hBMSCs were remarkably prol iferated 24 hours after hypoxia with CoC12 concentration of 150 µmol/L (P lt; 0.05), while hPDB-MSCs were significantly prol iferated 12 hours after hypoxia with CoC12 concentration of 75 µmol/L (P lt; 0.05). Conclusion Compared with hBMSCs, hPDB-MSCs express more specific surface antigens of embryonic stem cells and are more sensitive to the prol iferation effects of chemical hypoxia, indicating it may be a new seed cells source for tissue engineering.
To investigate the value of plasma placental growth factor (PlGF) in percutaneous coronary angioplasty and stent implantation. Methods From May 2006 to March 2007, 61 patients (53 males and 8 females, mean age61 years) and 28 normal controls were included. All patients present with acute chest pain and underwent coronary angiography, the lesion severity of coronary arteries was assessed by Gensini coronary scoring system. Of them, 26 patients having serious coronary lesion underwent (percutaneous transluminal coronary angioplasty, PTCA) and stent implantation. Cardiovascular events were recorded after 30 days. Plasma PlGF was determined by ELISA. Results According to the angiography, the patients could be divided into CAD group (n=45) and Non- CAD group (n=16). Plasma PlGF level in CAD group was significantly higher than that in Non-CAD group and control group [(10.70 ± 0.49) ng/L vs (4.53 ± 0.64) ng/L vs (3.64 ± 0.36) ng/L, P lt; 0.001)], and there was no significant difference between the non-CAD group and control group (P gt; 0.05). A significant positive correlation was found between Gensini coronary score and plasma PlGF level (r=0.918, P lt; 0.01). Moreover, patients with cardiovascular events had a higher PlGF level than those without cardiovascular events after PTCA and stent implantation [(13.98 ± 3.39) ng/L vs (7.25 ± 2.96) ng/L, P lt; 0.01)]. Conclusion PlGF level has diagnostic value in patients with acute chest pain. The measurement of plasma PlGF might be helpful for early diagnosis of coronary artery disease. Patients with higher plasma PlGF level may have more severe coronary lesion. PlGF may be one of predictors for cardiovascular events after PCI.
Objective To explore a method to isolate, culture and multiplicate the placentaderived mesenchymal stem cells (PMSCs) and the bone marrow-derived mesenchymal stem cells (BMSCs) of rabbit，and to compare their biological characteristics. Methods PMSCs were isolated from placenta of 1fetation rabbitby Percoll density gradient centrifuge and cultured in vitro. BMSCs were isolated from hindlimb bone marrow blood of 1 new born rabbit by direct plates culturemethod. The 3rd passage PMSCs and BMSCs were observed by inverted phase contrast microscope. The stem cell marker (CD44, CD105, CD34 and CD40L) were examined by immunohistochemistry. The 2nd passage PMSCs and BMSCs were co-cultured with biomaterials，(1.0-1.5)×106 cells in one biomaterial， and then observed by aematoxylinstaining after 5 days,and by SEM after 3 days and 8 days. Results PMSCs and BMSCs were both uniformly spondle-shaped in appearance and showed active proliferative capacity. The proliferative ability of PMSCs were quite b and declined with passages. After cultured 10 passages in vitro, its growthslowed. Both PMSCs and BMSCs expressed CD44 and CD105,but did not express CD34 and CD40L immunoreactivity. PMSCs and BMSCs poliferated and adhered to the surface of biomaterials, and cell formed clumps and network; the cells proliferation and the matrix were seen in the pore after 5 days of culture. The observation ofSEM showed that many cells adhered to the biomaterials with spindle-shape and polygon after 3 days; and that PMSCs and BMSCs grew,arranged in layers andsecreted many matrices; the reticular collagen formed arround cells after 8 days. Conclusion PMSCs and BMSCs have similar biological characteristics and PMSCs can be served as excellent seedingcells for tissue engineering.
Abstract To look for a substitute material of microvessel, 30 rabbits were divided into 2 groups. A segment of 5mm of rabbit femoral artery was resected and a segment of 10mm of human placental artery was placed in the defect with both ends anastomosed. Twenty experimental rabbits received freezedried human placental vessel and 10 rabbits received fresh as control. After 24 hours, 2 weeks, 3 months, the arteries were explored. If the artery was found to be obstructed, the segment was removed for histologic examination. The results showed: in the experimental group, 2 weeks after operation, the rateof patency was 85%, and it decreased to 50% 3 months after the operation. Under the light and scanning electronic microscope, the transplanted vessels were decomposed and inside the unobstructed transplanted vessels in both groups a layer of fibrous tissue was formed as the new wall of vessel. It was concluded thatthe immunologic reaction could not be prevented but would only be put off for aperiod of time by cryochem. Further more study should be done in using placental atery as a substitute material in repairing vascular defect.
目的 探讨不典型胎盘早剥的临床特点。 方法 对2008年5月－2009年5月收治的55例胎盘早剥患者的临床资料进行回顾性分析。其中产前漏诊30例，疑诊15例，确诊10例。胎盘早剥的产前确诊率为18.2%，漏诊率为54.5%。所有患者均经产后证实。 结果 重度子痫前期（25.5%）、胎膜早破（12.7%）是胎盘早剥的主要发病诱因；阴道流血（52.7%）、腰腹痛（47.3%）及胎心异常（36.4%）是其常见的临床表现。胎盘早剥者，剖宫产率、胎儿窘迫及早产率均增加。 结论 不典型胎盘早剥病情隐匿。后壁胎盘、早剥面积小及B型超声检查阴性是漏诊的主要原因。对此患者应提高认识，动态监测，及时处理，以改善母婴结局。
目的 通过对1例原发于宫颈的胎盘部位滋养细胞肿瘤（PSTT）的资料及相关文献的复习，全面介绍胎盘部位滋养细胞肿瘤的临床特征、病理特点、治疗方式及预后。 方法 对收治1例罕见原发于宫颈部位的PSTT的临床病理资料进行分析，并以“胎盘部位滋养细胞肿瘤”为主题词查阅中国知网（CNKI）及PubMed等文献数据库进行文献复习。 结果 该例原发于宫颈部位的PSTT患者术后化学疗法后，目前情况良好，已随访3年，无复发及转移征象。2011年9月1日前，CNKI数据库共报道300余例PSTT病例，PubMed数据库共160余例，其中有2例原发于宫颈部位的PSTT。原发于宫颈部位的PSTT罕见，易误诊，往往需要宫颈活组织检查或宫颈搔刮才能确诊。 结论 原发于宫颈部位的PSTT的预后是否遵循PSTT尚需收集更多的病例证实。PSTT因其发病率低，临床表现多无特异性，通常需要通过诊断性刮宫、活组织检查甚至术后病理检查才能确诊。PSTT首选的治疗方式为手术，多数患者病灶清除后可治愈，对有高危因素的患者术后宜选择依托泊苷+甲氨蝶呤+放线菌素D+环磷酰胺+长春新碱（EMA-CO）方案或依托泊苷+甲氨蝶呤+放线菌素D+依托泊苷+顺铂（EMA-EP）方案进行化学治疗，以期改善预后。