【摘要】 目的 观察妊娠肝内胆汁淤积症（intrahepatic cholestasis of pregnancy，ICP）患者血清对体外培养的胎盘组织促肾上腺皮质激素释放激素（corticotropin-releasing hormone，CRH）的分泌水平的影响。 方法 收集2005年3月-7月在四川大学华西第二医院产科住院分娩的正常妊娠妇女胎盘组织及其血清（对照组）与ICP患者血清（ICP组）各10例。通过胎盘组织培养及放射免疫法测定其培养液中CRH水平。 结果 ICP组胎盘组织CRH分泌水平低于对照组，ICP组24、48、72、96 h分别为（84.95±34.98）、（74.57±29.93）、（71.16±27.26）、（81.07±37.18） pg/mL；对照组分别为（103.74±30.85）、（108.27±23.77） 、（109.20±23.81）、（118.15±26.84） pg/mL。两组比较，48h后差异有统计学意义（Plt;0.05）。 结论 ICP患者血清对体外培养的胎盘组织CRH分泌有抑制作用。【Abstract】 Objective To observe the effect of serum of pregnant patients with intrahepatic cholestasis (ICP) on the excretion level of corticotrophin-releasing hormone in the placental tissue in vitro. Methods Serum from 10 patients with ICP (ICP group) and from the healthy placental tissue of 10 normal people (control group) were collected from March to July, 2005. Cell culture and radioimmunoassay methods were used to investigate the corticotropin-releasing hormone (CRH) levels in placental tissue. Results The level of CRH in human placental tissue treated with sera of ICP was lower than that in the control group. 24, 48, 72, and 96 hours after treated with the serum, the levels of CRH in the ICP group were (84.95±34.98), (74.57±29.93), (71.16±27.26), and (81.07±37.18) pg/mL, respetively;while in the control group were (103.74±30.85), (108.27±23.77), (109.20±23.81), and (118.15±26.84) pg/mL, respectively. There was significant difference in the levels of CRH between ICP group and control group 48 hours after the culture (Plt;0.05). Conclusion The serum from the patients with ICP may inhibit the excretion of CRH in the placental tissue.
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.
目的 建立测定胎盘灌流液中格列苯脲浓度的高效液相色谱（HPLC）检测方法。方法 采用的色谱柱为Symmetry Shield RP C18（150 mm×4.6 mm，5 μm），柱温40℃，流动相为NaH2PO4缓冲盐（25 mmol/L，pH值5.2）︰乙腈=1︰1；内标为格列齐特，流速1.0 mL/min，检测波长228 nm，采用液-液萃取预处理方法测定胎盘灌流液中格列苯脲的浓度。 结果 格列苯脲浓度线性范围为2.0～25.0 μg/mL，线性方程为：y=0.226x+0.002，r=0.999 9 （n=6），日内相对标准偏差（RSD）＜3.1%，日间RSD＜9.5%，方法学回收率为95.32%～103.35%。 结论 HPLC检测方法灵敏、简便，可用于胎盘灌流液中格列苯脲浓度的检测。
【摘要】目的探讨瘢痕子宫不全破裂的早期诊断、处理及预防。方法2006年1月2009年1月发生瘢痕子宫不全破裂13例,术前临床症状加B超检查确诊,手术从原切口进入宫腔,取出胎儿,修剪原切口周围瘢痕组织,10可吸收线连续缝合浆肌层,再间断包埋缝合切口,术后常规预防感染,加强宫缩治疗;胎盘植入2例尽量取出胎盘,修整切口,活动性出血明显者用10可吸收线“8”字缝扎止血,术后加服米非司酮150 mg/d共3 d。结果母婴均痊愈出院。42 d后来院复查,B超探查8例子宫下段处有线状较强回声,肌层回声均匀,余未发现异常;胎盘植入2例，随防3个月血绒毛膜促性腺激素呈阴性。结论早期B超检查能提高瘢痕子宫不全破裂确诊率,确诊后急诊剖宫产,胎盘部分植入者加服米非司酮并预防感染。
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。
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.
目的 观察糖皮质激素对胎盘组织促肾上腺皮质激素释放激素（CRH）的分泌水平的影响。 方法 收集2006年1月－3月住院分娩的正常妊娠妇女的胎盘组织与妊娠肝内胆汁淤积症（ICP）患者胎盘及其血清各10例。分3组进行胎盘组织培养，即正常胎盘组、ICP胎盘组，正常胎盘组织加ICP患者血清组，分别用放射免疫法测定各组加与不加地塞米松胎盘组织培养液中CRH的水平。 结果 正常组与正常胎盘加地塞米松组培养24、48、72、96 h其CRH分泌水平分别为：（74.81 ± 27.92）、（63.71 ± 24.72）、（91.87 ± 41.64）、（98.90 ± 42.52） pg/mL；（66.94 ± 29.62）、（77.39 ± 31.84）、（61.89 ± 33.94）、（75.13 ± 36.98） pg/mL，两组比较差异有统计学意义（P＞0.05）。ICP组与ICP加地塞米松组培养上清液中CRH水平在24、48、72、96 h其CRH分泌水平分别为：（48.28 ± 16.56）、（60.20 ± 29.97）、（72.92 ± 31.65）、（69.22 ± 29.33）pg/mL；（41.81 ± 25.00）、（57.36 ± 39.75）、（57.72 ± 23.29）、（61.43 ± 20.77）pg/mL, 两组比较差异有统计学意义（P＞0.05）；正常胎盘加ICP血清培养组与正常胎盘加ICP血清加地塞米松培养组上清液中CRH水平在24、48、72、96 h其CRH分泌水平分别为：（84.9 ± 34.98）、（74.5 ± 29.93）、（71.1 ± 27.26）、（81.0 ± 37.18）pg/mL；（76.29 ± 33.11）、（63.70 ± 24.20）、（64.85 ± 28.39）、（67.65 ± 33.20）pg/mL，两组比较差异有统计学意义（P＞0.05）。3组加入地塞米松培养的胎盘组织，CRH分泌水平并无明显改变。 结论 地塞米松不影响体外培养胎盘组织CRH分泌。
ObjectiveTo investigate the effectiveness of human placental decidua basalis derived mesenchymal stem cells (PDB-MSCs) in repairing full-thickness skin defect of nude mice. MethodsHuman placenta samples were obtained from healthy donor mothers with written informed consent. PDB-MSCs were isolated through enzymic digestion and density gradient centrifugation; the 4th passage cells were identified by cellular morphology, cell adipogenic and osteogenic differentiation, and phenotype evaluation. Forty-two 4-5-week-old BALB/c female nude mice were randomly divided into experimental group (n=21) and control group (n=21). The 4th passage PDB-MSCs solution (200 μL, 5×106/mL) was injected into the mice of experimental group via caudal vein; the mice of control group were given equal volume of PBS. The full-thickness skin defect model of 1.5 cm×1.5 cm in size was made after 3 days. The wound healing was observed generally at 1, 2, 4, 7, 14, 18, 21, 25, and 30 days after operation, and the wound healing rate was calculated after wound decrustation. HE staining was used to observe the wound repair at 1, 7, 14, 21, and 31 days; immunofluorescent staining was used for cellular localization at 7, 14, and 31 days after operation. ResultsCells isolated from human placenta were MSCs which had multipotential differentiation ability and expressed MSCs phenotype. Animals survived to the end of the experiment. The general observation showed that the experimental group had a faster skin repairing speed than the control group; the time for decrustation was 12-14 days in experimental group and was 14-17 days after operation in the control group. The wound healing rate of experimental group was significantly higher than that of control group at 14, 18, and 21 days (t=4.001, P=0.016; t=3.380, P=0.028; t=3.888, P=0.018), but no significance was found at 25 and 30 days (t=1.565, P=0.193; t=1.000, P=0.423). HE staining showed lower inflammatory reaction, and better regeneration of the whole skin and glands with time in the experimental group. The immunofluorescent staining was positive in skin defect area of experimental group at different time points which displayed that human PDB-MSCs existed. ConclusionThrough enzymic digestion and density gradient centrifugation, PDB-MSCs can be obtained. Pre-stored PDB-MSCs can mobilize to the defect area and participate in repair of nude mice skin.
摘要:目的: 分析凶险型前置胎盘的临床特点, 预防产后出血和子宫切除的发生。 方法 :对11例凶险型前置胎盘与75例普通型前置胎盘的病例进行回顾性分析。 结果 :凶险型组与普通型组发生产前出血的量差异无统计学意义(Pgt;0.05);在发生胎盘植入、产后出血的量差异有统计学意义(Plt;0.05);子宫切除的发生率差异有统计学意义(Plt;0.05)。 结论 :凶险型前置胎盘对孕产妇有极大的威胁,应努力做好凶险型前置胎盘产后出血的抢救,减少子宫切除的发生。Abstract: Objective: To assess the clinical feature of dangerous placenta praevia in order to prevent postpartum hemorrhage and intrapartal hysterectomy. Methods : Retrospective analysis was done between the 11 cases of dangerous placenta praevia and ordinary placenta praevia . Results : There were no significant difference in blood volume antepartum (Pgt;0.05); There was significant difference in placenta increta and postpartum hemorrhage (Plt;0.05). Conclusion : Dangerous placenta praevia have great threat to gravid and puerperant, we should try our best to rescue postpartum hemorrhage about dangerous placenta praevia and reduce the incidence of intrapartal hysterectomy.