Study on the expression levels of CXCR4, CXCL12, CD44, and CD147 and their potential correlation with invasive behaviors of pituitary adenomas / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To evaluate the factors of CXCR4, CXCL12, CD44, and CD147 as early potential diagnostic biomarkers by determining their expression levels in invasive and non-invasive pituitary adenomas.</p><p><b>METHODS</b>Fresh pituitary adenoma specimens were collected from 35 pituitary adenoma (21 invasive and 14 non-invasive) patients who underwent surgical treatment in our Neurosurgery Department between January and April of 2009. The expression levels of CXCR4, CXCL12, CD44, and CD147 were evaluated firstly by flow cytometry, fluorescence microscopy in single cell suspensions, and then by immunohistochemical staining of paraffin tissue sections.</p><p><b>RESULTS</b>Flow cytometric analyses showed that the percentage of CXCR4- and CXCL12-positive cells from invasive pituitary adenomas (IPA) was significantly higher in the single cell suspensions than that from non-invasive pituitary adenomas (nIPA) (P<0.05). Immunohistochemical staining revealed that CXCR4 and CXCL12 staining index scores of the invasive pituitary adenomas were significantly higher than those of the non-invasive pituitary adenomas (P<0.05). In contrast, neither flow cytometry nor immunohistochemical staining demonstrated significant difference between CD44 and CD147 expression levels, respectively.</p><p><b>CONCLUSION</b>Expression levels of CXCR4 and CXCL12 are correlated with the invasiveness of pituitary adenomas. Therefore, rather than CD44 and CD147, CXCR4 and CXCL12 may potentially serve as biomarkers for early detection of pituitary adenomas.</p>

Preparation of brain targeted immunoliposomes / 药学学报

To prepare a kind of effective non-viral transduction vector, which can deliver exogenous gene into the brain, this vector can be injected through vein system and has the ability to penetrate blood brain barrier. Several groups of materials proportion, type of oil phase, water-oil ratio, phosphatides-cholesterol ratio, temperature of steaming, ultrasonic temperature and time were compared for optimization. Well-constructed immunoliposomes encapsuling LacZ gene were infused into rats through tail vein. 48 h after injection, expression product beta-galactosidase of LacZ gene was detected by histochemistry staining to convince the validity of immunoliposomes as non-viral vectors. The best proportion of synthesis immunoliposomes is as following: phosphatides-cholesterol ratio is 1:1, lipids/drug is 100:1, the type of oil phrase is dichloromethane, oil-water ratio is 4:1, temperature of steaming is 30 degrees C, ultrasonic temperature and time is 10 degrees C and 5 min. At last, 10% trehalose was added as a stabilizer. The entrapment rate is 87.24% and antibody coupling rate is 69%. When immunoliposomes were infused into rats, the expression of LacZ gene could be observed in the brain and periphery organs. Through the best proportion of materials, gene delivering immunoliposomes had been synthesized successfully. This non-viral vector can deliver exogenous gene penetrating blood brain barrier and express in the brain, and will be well-used in the field of gene therapy of cerebral diseases.

Ferumoxide labeled Flk1+ CD31- CD34- human bone marrow mesenchymal stem cells and its in vivo tracing in the brains of Macaca Fascicularis / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the method for labeling Flk1+ CD31- CD34- human bone marrow mesenchymal stem cells (hBMSCs) with ferumoxide-PLL and evaluate the feasibility of its tracing after transplantation into the brains of Macaca Fascicularis.</p><p><b>METHODS</b>The hBMSCs were incubated with ferumoxide-PLL. Trypan blue staining, Prussian blue staining, and transmission electron microscope were performed to show intracellular iron, marking efficiency, and the vigor of the labeled cells. After the hBMSCs were transplanted into the brains of cynomolgus monkeys by stereotaxis, magnetic resonance imaging (MRI) was performed to trace the cells in vivo. Cell survival and differentiation were studied with immunohistochemistry, Prussian blue staining, and HE staining.</p><p><b>RESULTS</b>The marking efficiency of the ferumoxide-PLL was 96%. Iron particles were found intracytoplasmic of the hBMSCs by Prussian blue staining and transmission electron microscopy. The relaxation rates of labeled cells in MRI were 4.4 and 4.2 times higher than those of the unlabeled cells. Hypointensity area was found by MRI three weeks after transplantation. Many hBMSCs and new vessels were found in the transplantation zone by pathological and immunofluorescence methods.</p><p><b>CONCLUSIONS</b>Ferumoxide-PLL can effectively label hBMSCs and thus increase its contrast in MRI results. The cells can survive in the brains of cynomolgus monkeys. The labeled hBMSCs can be traced in vivo by MRI.</p>

In vivo tracking of bone marrow mesenchymal stem cells labeled with superparamagnetic iron oxide after cerebral ischemia in rats by magnetic resonance imaging / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the feasibility of in vivo tracking of bone marrow mesenchymal stem cells (BMSCs) labeled with superparamagnetic iron oxide (SPIO) by magnetic resonance imaging (MRI) in rats after cerebral ischemia, and to analyze the influence of stem cell therapy on the volume of cerebral infarction.</p><p><b>METHODS</b>The samples of rat bone marrow were collected. BMSCs separated by density gradient centrifugation were cultivated and harvested until the third passage. BMSCs were labeled with SPIO, which was mixed with poly-L-lysine. The labeling efficiency was evaluated by Prussian blue staining. Transient middle cerebral arterial occlusion (MCAO) was performed successfully in 18 adult Sprague-Dawley rats that scored from 6 to 12 by the modified neurological severity test. The 18 rats were then randomly divided into group A, B, and C, with 6 rats in each group and Group C was regarded as control group. BMSCs were injected into the contralateral cortex of ischemia in group A, ipsilateral corpora striata in group B, while D-Hank's solution was injected into ipsilateral corpora striata (group C) 24 hours after MCAO. MRI was performed 1 day after MCAO, 1 day and 14 days after transplantation. The volume of infarcted brain tissue was measured and analyzed. Prussian blue staining of brain tissues was performed to identify the migration of BMSCs.</p><p><b>RESULTS</b>The labeling efficiency of BMSCs with SPIO was 96%. The transplanted BMSCs migrated to the ischemic hemisphere along the corpus callosum and to the border of the infarction, which was confirmed by MRI and Prussian blue staining. The changes of infarction volume were not significantly different among these three groups.</p><p><b>CONCLUSIONS</b>MRI is feasible for in vivo tracking of BMSCs labeled with SPIO in rats. The stem cell therapy may not be able to affect the volume of cerebral infarction.</p>

Distribution and expression of peroxisome proliferator activated receptor gamma in human pituitary adenomas / 中国医学科学院学报

<p><b>OBJECTIVE</b>To evaluate the distribution and expression of peroxisome proliferator activated receptor gamma (PPAR-gamma) in human pituitary adenomas.</p><p><b>METHODS</b>Thirty eight consecutive surgically resected pituitary adenomas and 5 normal pituitary tissues were enrolled in the study. Immunohistochemistry was used to confirm the distribution of PPAR-gamma. Expression of PPAR-gamma was evaluated by Western blot.</p><p><b>RESULTS</b>PPAR-gamma immunoreactivity was located in the nucleoli of pituitary adenoma cells. PPAR-gamma was expressed in all human pituitary adenomas and normal pituitary tissues. Its expression in pituitary adenomas was significantly higher than in normal pituitary tissues (P < 0.01), and its expression in ACTH-secreting adenomas was significantly higher than in any other type of pituitary adenomas (P < 0.05).</p><p><b>CONCLUSIONS</b>PPAR-gamma may play an important role in the generation, growth, and invasion of human pituitary adenomas. It may become a novel therapeutic target for these tumors.</p>

Factors influencing the recurrence of Cushing's disease treated by transsphenoidal surgery / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore factors influencing the recurrence of patients with Cushing's disease after transsphenoidal surgery.</p><p><b>METHODS</b>We retrospectively analyzed the clinical data of 182 patients treated by transsphenoidal surgery with Cushing's disease in our department in PUMC Hospital from 1992 to 2002.</p><p><b>RESULTS</b>The recurrence rates were significantly different when patients had different radiological findings (P = 0.001), operative methods (P = 0.001), histological findings (P = 0.04), and postoperative cortisol levels (P = 0.02); however, such difference was not found in term of tumor size (P = 0.43).</p><p><b>CONCLUSION</b>Radiological findings, operative methods, histological findings, and postoperative cortisol estimates may be the factors influencing the recurrence of patients treated by transsphenoidal surgery.</p>

Expression of galectin-3 in invasive prolactinomas / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the expression of galectin-3 (Gal-3) in prolactinomas.</p><p><b>METHODS</b>Expressions of Gal-3 were evaluated by immunohistochemistry using polyclonal antibody in 16 invasive prolactinomas and 16 prolactinomas.</p><p><b>RESULTS</b>Gal-3 was expressed both in invasive prolactinomas and noninvasive prolactinomas while significantly higher expression seen in the invasive prolactinomas (P < 0.05).</p><p><b>CONCLUSION</b>Gal-3 expression may be used as a useful indicator to determine the invasiveness and prognosis of prolactinomas.</p>

Elevated soluble epidermal growth factor receptor level in pituitary adenoma and carcinoma / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate effect of the soluble epidermal growth factor receptor (sEGFR/sErbB1) level in the peripheral blood in development, invasiveness, apoplexy of each type of pituitary tumor.</p><p><b>METHODS</b>The sEGFR level was determined in peripheral serum from 190 patients with pituitary diseases by enzyme linked immunosorbent assay. The sEGFR levels were measured in 10 pituitary Rathke's pouch, 18 pituitary hyperplasia, 161 pituitary adenomas including 30 microadenomas, 83 large adenomas, 48 giant adenomas, 1 pituitary carcinoma, and 28 healthy controls.</p><p><b>RESULTS</b>In the patients with pituitary hyperplasia, microadenoma, large adenoma, giant adenoma, and pituitary carcinoma, the sEGFR level was 188.92 +/- 32.62, 209.83 +/- 19.01, 333.20 +/- 69.33, 405.85 +/- 37.38, and 617.45 fmol/mL independently. They were all significantly higher than patients with pituitary Rathke's pouch (156.78 +/- 18.24 fmol/mL, P < 0.001) and healthy control group (159.11 +/- 40.50 fmol/mL, P < 0.05). The sEGFR level in pituitary carcinoma was higher than pituitary adenoma. In patients with pituitary adenoma, the sEGFR level was positive correlated to the size of pituitary adenomas (r=0.998), the significant difference was observed for the sEGFR level in each group of the patients with pituitary adenomas (P < 0.001). Furthermore, in patients with pituitary ACTH-secreting microadenomas, the serum sEGFR levels in invasiveness (295.00 +/- 77.80 fmol/mL) was higher than that in non-invasiveness (210.60 +/- 16.4 fmol/mL, P < 0.05). In patients with pituitary ACTH-secreting, PRL-secreting, GH-secreting, and non-functioning large adenomas, the serum sEGFR levels in invasiveness (407.86 +/- 28.50, 399.25 +/- 30.10, 386.00 +/- 13.08, and 369.25 +/- 36.70 fmol/mL) was higher than that in non-invasiveness (335.25 +/- 63.49, 300.64 +/- 47.57, 297.00 +/- 61.93, and 269.30 +/- 25.68 fmol/mL) respectively (P < 0.05). In patients with invasive pituitary PRL-secreting, GH-secreting, and non-functioning giant adenomas, the serum sEGFR levels not significantly different in between invasiveness (417.50 +/- 35.94, 409.50 +/- 69.14, and 417.50 +/- 44.13 fmol/mL) and non-invasiveness (386.00 +/- 49.64, 417.50 +/- 44.03, and 409.51 +/- 35.17 fmol/mL) (P > 0.05). In patients with pituitary large adenomas, the sEGFR levels in pituitary apoplexy (377.48 +/- 39.18 fmol/mL) was higher than that in non-pituitary apoplexy (343.18 +/- 68.17 fmol/mL, P > 0.05).</p><p><b>CONCLUSIONS</b>The increased level of peripheral serum sEGFR is concomitant with development, proliferous size of the adenomas in patients with pituitary adenomas. In addition, the elevated levels of serum sEGFR occur in pituitary apoplexy as clinical active tumors, and the non-invasive ACTH secreting adenomas. The sEGFR levels could be differentiated helpfully between pituitary adenomas and non-pituitary adenomas. These data suggest that serum sEGFR could be as a referable marker of the size and activation of proliferation in pituitary adenoma.</p>

Expressive level of vascular endothelial growth factor in peripheral blood in patients with pituitary adenomas / 中国医学科学院学报

<p><b>OBJECTIVE</b>To discuss whether vascular endothelial growth factor (VEGF) in the peripheral blood can reflect the biological activities of pituitary adenomas.</p><p><b>METHODS</b>The concentrations of VEGF in peripheral blood were measured with ELISA in 203 patients with pituitary adenomas, 22 patients with pituitary hyperplasia, 7 patients with pituitary Rathke' pouch and 3 patients with pituitary abscess.</p><p><b>RESULTS</b>The serum VEGF levels were (366.8 +/- 211.1) pg/ml and (286.8 +/- 107.6) pg/ml in patients with pituitary adenomas and pituitary hyperhasia, respectively, which were higher than those in patients with pituitary Rathke' pouch [(180.5 +/- 61.7) pg/ml], patients with pituitary abscess [(147.5 +/- 46.3) pg/ml] and the health control [(180.8 +/- 56.2) pg/ml] (P < 0.05). In patients with pituitary adenomas, the VEGF levels were (380.0 +/- 234.5) pg/ml in macroadenomas and (380.1 +/- 2870.3) pg/ml in giant adenomas, higher than those in microadenomas [(294.6 +/- 111.6) pg/ml] and in pituitary hyperhasia respectively (P < 0.05). The serum VEGF levels were not significantly different in pituitary adenoma in terms of invasive growth, apoplexy, cyst and hormone secretory functions (P > 0.05).</p><p><b>CONCLUSIONS</b>The upregulation of serum VEGF expression may reflect the biological activities of pituitary adenoma. However, it may not be associated with pituitary Rathke' pouch, pituitary abscess, adenoma with invasiveness, apoplexy, cyst and hormone secretory function. The serum VEGF levels could be helpful in differentiating pituitary adenoma from pituitary Rathke' pouch and pituitary abscess.</p>

Measurement of soluble CD44v6 in peripheral blood as assistant diagnosis of invasive pituitary adenomas / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the value of measuring the concentration of soluble CD44 splice variant 6 (sCD44v6) in peripheral blood in patients with invasive and non-invasive pituitary adenomas.</p><p><b>METHODS</b>The concentrations of sCD44v6 in peripheral blood were measured with ELISA in 68 patients with invasive pituitary adenomas and 100 patients with non-invasive pituitary adenomas.</p><p><b>RESULTS</b>The serum concentration of sCD44v6 in patients with invasive pituitary adenomas was lower than that in patients with non-invasive pituitary adenomas, while the latter was lower than that in healthy controls. The serum concentrations of sCD44v6 were (44.63 +/- 7.21), (34.53 +/- 6.41), and (26.34 +/- 4.95) ng/ml in patients with invasive microadenoma, macroadenoma, and giant adenoma, and (60.78 +/- 9.61), (57.78 +/- 10.00), and (37.22 +/- 5.17) ng/ml in patients with non-invasive microadenoma, macroadenoma, and giant adenoma, lower than that in the healthy control group (68.73 +/- 6.00) ng/ml. Significant differences were observed among groups (P < 0.005). The concentration of sCD44v6 in peripheral blood decreased as the tumor size increased (P < 0.01), which was particularly significant in invasive pituitary adenomas. The positive rate in the patients with invasive pituitary adenomas reached 89.71%.</p><p><b>CONCLUSION</b>Serum concentration of sCD44v6 in the peripheral blood is inversely correlated with tumor size and its invasive growth, which may provide certain value in the early diagnosis, treatment and prognosis of invasive pituitary macroadenoma and giant adenoma.</p>

Measurement of epidermal growth factor receptor concentration in the pre- and postoperative serum in patients with meningiomas / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate effect of the serum concentration of epidermal growth factor receptor (EGFR) in the pre- and postoperative peripheral blood in patients with meningiomas.</p><p><b>METHODS</b>Using ELISA, the EGFR concentration were measured in the pre- and postoperative serum in 53 patients with meningiomas. In this study, the patients were divided into preoperative group, postoperative group (including 42 total resection, 11 subtotal resection), and 28 healthy control.</p><p><b>RESULTS</b>In 53 patients with meningiomas, concentration of preoperative serum EGFR was (352.93 +/- 66.18) fmol/ml, to show higher than control group (159.11 +/- 40.50) fmol/ml (P < 0.0001); Concentration of postoperative serum EGFR was (220.74 +/- 70.63) fmol/ml, to show lower than preoperative group (P < 0.001). Including in 42 patients with meningomas by total resection, serum EGFRs of the 38 patients were decreased (191.20 +/- 32.13) fmol/ml, the 4 patients with peritumoral edema were decreased (248.75 +/- 10.31) fmol/ml, to show lower than preoperative group (P < 0.001); the 11 patients with subtotal resection were decreased (322.14 +/- 89.53) fmol/ml, not to show different than preoperative group (P < 0.05).</p><p><b>CONCLUSIONS</b>In 92.16% patients with meningiomas, the more poison of the tumor is resected, the less the serum EGFR concentration is detected postoperatively, while the total resection of mangionmas resulted in a lowest level of EGFR. These results support the concept that human meningiomas may be autocrine EGFRs. The measurement of serum EGFR can be useful to patients with meningiomas for follow-up after surgery.</p>

Biological effect of hepatocyte growth factor gene transfection to rat cerebral ischemic model:the experimental study / 中华放射学杂志

Objective To investigate hepatocyte growth factor(HGF)gene expression and biological effect after gene transfection into penumbra tissue in rat cerebral ischemic model.Methods Human HGF cDNA was ligated to pIRES2-EGFP vector.The recombinant plasmid was transfected into the penumbra tissue with liposome.Brains of treated and control animals were analyzed 7 days later.Expression of HGF protein was determined by fluorescence microscopy and immunohistochemistry.Vessel numbers were quantified.Changes of cerebral blood flow(CBF)was detected by CT perfusion.Results Enzymatic digestion and electrophoresis confirmed that HGF fragment had been correctly cloned into BamH I and Sal I sites of pIRES2-EGFP.After HGF gene transfection,expression of HGF in transfected neurocytes was observed with fluorescence microscopy and immunohistochemistry.The number of vessels was significantly increased in penumbra tissue transfected with HGF vector as compared with control vector(46.71?7.11, 20.43?3.21,18.00?3.27,respective,F = 74.447;P