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。
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.
ObjectiveTo systematically evaluate the changes in placental protein expressions in gestational diabetes mellitus (GDM) and their correlations with maternal insulin resistance (IR). Methods PubMed, Cochrane Library, Scopus, Web of Science, Embase, China National Knowledge Infrastructure, VIP database, Wanfang Database and CBMdisc were searched for case-control studies published from January 2009 to November 2021, which reported the placental protein expressions in GDM and their correlations with IR. Two researchers independently reviewed the literature, extracted data and evaluated the literature quality. RevMan 5.4 software was used for meta-analysis, and descriptive analysis was performed on data that cannot be combined. ResultsA total of 19 studies were included, comprising 2 012 patients. The results of meta-analysis showed that: the expression level of retinol binding protein 4 (RBP4) [standard mean difference=2.11, 95% confidence interval (CI) (1.64, 2.58), P<0.000 01] and the positive rate of protein tyrosine phosphatase-1B (PTP1B) [relative risk (RR)=1.56, 95%CI (1.29, 1.88), P<0.000 01] were up-regulated, and the positive rate of insulin receptor substrate 1 (IRS-1) [RR=0.69, 95%CI (0.60, 0.78), P<0.000 01] was down-regulated. The protein expression levels of RBP4 (P<0.000 01) and PTP1B (P<0.000 01) were positively correlated with homeostasis model assessment of insulin resistance (HOMA-IR), while the protein expression levels of IRS-1 (P<0.000 01) and APN (P=0.002) were negatively correlated with HOMA-IR, and glucose transporter 4 (GLUT 4) was not correlated with HOMA-IR (P=0.79). Descriptive analysis found that the expression levels or positive rates of adipocytokines (leptin, resistin), oxidative stress markers (xanthione oxidase, malondialdehyde, 8-isoprostaglandin),inflammatory factors (tumor necrosis factor α, Toll-like receptor 4, Galectin-3, Galectin-2, migration inhibitory factor),fetuin-A, forkhead box transcription factor 1, forkhead box transcription factor 3a and estrogen receptor α in GDM placenta were up-regulated and all were positively correlated with HOMA-IR. The expression levels or positive rates of insulin signaling pathway proteins [phosphoinositide 3-kinase (PI3K), protein kinases B (AKT), phospho-protein kinases B (p-AKT), GLUT 4] were down-regulated, PI3K and AKT were negatively correlatedwith HOMA-IR, while p-Akt had no correlation with HOMA-IR. ConclusionsThe dysregulation of placental protein expressions may mediate maternal IR exacerbation, thus promote the occurrence and development of GDM and other pregnancy complications. The causal relationship and regulatory mechanism are still unclear, which need to be further studied.
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.
【摘要】目的探讨骨桥蛋白(OPN)及其受体整合素ανβ3在子痫前期(preeclampsia,PE)患者胎盘组织中的表达及其意义。方法2008年11月2009年9月,采用免疫组织化学方法检测20例PE患者(轻度及重度PE各10例)和14例正常足月孕妇(对照组)胎盘组织中OPN及ανβ3蛋白表达水平。采用RTPCR检测各组孕妇胎盘组织中的OPN、αν和β3的mRNA的表达水平。结果PE组孕妇胎盘组织中OPN及ανβ3蛋白表达低下,与对照组相比,差异有统计学意义(Plt;0.05);重度PE组OPN及ανβ3蛋白表达水平更低,与轻度PE组比较,差异有统计学意义(Plt;0.05)。PE组孕妇胎盘组织中OPN mRNA水平明显低于对照组,两组差异有统计学意义(Plt;0.05);重度PE组OPN mRNA水平显著降低,与轻度PE组比较,两组差异有统计学意义(Plt;0.05);但αν和β3 mRNA的表达水平三组间比较差异无统计学意义。结论OPN及其受体整合素ανβ3在PE胎盘组织中的低表达可能在子痫前期的发病过程中起重要作用。
ObjectiveTo discuss the value of diffusion weighted imaging (DWI) in the diagnosis of placenta increta. MethodsThe clinical data of 42 patients with placenta increta admitted to Sichuan Provincial Hospital for Women and Children between May 2012 and January 2014 were retrospectively analyzed. All the patients were examined by prenatal magnetic resonance scans and DWI scans for subsequent comparison between ADC of the local convex placental region and ADC of the normal placental region and between the results of the two imaging methods. ResultsADC of the implantation area was significantly different from that of the normal placenta, so it could be used as a quantitative index. DWI had a higher sensitivity of diagnosis than conventional MRI. ConclusionCompared with conventional magnetic resonance imaging, DWI is more valuable in the clinical diagnosis of placenta increta, which provides a reliable basis for clinical treatment.
【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.
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.
ObjectiveTo comprehensively analyze and compare the biological difference between bone marrow mesenchymal stem cells (BMSCs) and placenta-derived MSCs (PMSCs) in hypoxia and to extend the knowledge for seed cells selection. MethodsThe domestic and foreign related literature about the effects of hypoxia microenvironment on proliferation, apoptosis, differentiation, paracrine secretion, migration, and homing ability of BMSCs and PMSCs were summarized and analysed. ResultsPMSCs proliferated much faster and more sensitive to the hypoxia than BMSCs; in addition, PMSCs showed stronger survivability. Similar to BMSCs, PMSCs can home to hypoxic-ischemic tissues efficiently, secrete a lot of growth factors and differentiate into tissue-specific cells to stimulate tissue regeneration. ConclusionPMSCs as the seed cells will have broad application prospects in the regenerative medicine.
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。