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Objective: To observe the effect of peritumoral electroacupuncture on the induction of vascular normalization in a mouse breast cancer model., Methods: A subcutaneous graft model of breast cancer was established with 4T1 breast cancer cell line in female BALB/c mice aged 6-8 weeks. The mice were randomly assigned to three groups, a tumor-bearing group (TG), peritumoral electroacupuncture tumor-bearing group (EATG), and bevacizumab tumor-bearing group (BTG), with 18 mice in each group. The TG mice did not receive any intervention, the EATG mice received peritumoral electroacupuncture for 30 minutes, and the BTG mice were intraperitoneally injected with bevacizumab at 10mg/kg. Immunofluorescence was performed to assess the expression of CD31/alpha smooth muscle actin (α-SMA) and hypoxia-inducible factor 1-alpha (HIF-1α) in the tumor tissue at various points of time, including before intervention and 3 days and 5 days after intervention. Then, 3 days after intervention, observation of morphological changes of the microvessels in the tumor tissue was performed through Hematoxylin and Eosin (HE) staining and scanning electron microscope., Results: There was no significant difference in the expression of CD31, α-SMA, and HIF-1α in the tumor tissues of all groups before experimental intervention ( P >0.05). On day 3 of the experimental interventions, the CD31 and HIF-1α expression levels in the tumor tissues of the EATG and BTG mice were significantly reduced ( P <0.01), while α-SMA expression levels were significantly increased ( P <0.01) in both groups. On day 5 of the experimental interventions, the CD31 and HIF-1α expression levels in the tumor tissues of the EATG and BTG mice were still significantly lower than those in the TG mice ( P <0.01), while the α-SMA expression level was significantly higher than that in the TG group ( P <0.05). On day 3 of the experimental interventions, H&E staining showed visible microvessels in the tumor tissues of all 3 groups. In addition, scanning electron microscopic observation showed that the tumor microvessel walls of the TG mice were rough and defective, and that obvious deformities appeared in the lumen. In contrast, the walls of the microvessels of the EATG and BTG mice were generally intact and there was no obvious deformities in the lumen., Conclusion: Peritumoral electroacupuncture may induce microvasculature normalization by decreasing microvascular density and increasing pericyte coverage of the neovasculature, thereby improving hypoxic microenvironment of breast cancer in mice., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Sciences).)

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide. Microvascular invasion (MVI) is considered the major risk factor for postoperative recurrence and metastasis in HCC. The diagnosis of MVI relies on the postoperative pathological assessment of the tumor tissues. Seeking non-invasive methods and biomarkers for evaluation of MVI before surgery has important clinical implications for guiding surgical treatment and improving patients' survival. Recent studies have reported the applications of radiomics technique in prediction of MVI in HCC and showed promising results. Herein we summarized the research progress in CT- or MRI-based radiomics models for prediction of MVI in HCC to provide helpful thinking for further research in this field.

Hepatocellular carcinoma (HCC) is a kind of highly aggressive tumor of the digestive system. Several studies have confirmed that microvascular invasion (MVI) is an independent risk factor for early recurrence and poor prognosis of HCC after surgery. Currently, pathological examination is the gold standard for diagnosing MVI. This paper summarizes concept, prognosis, preoperative prediction and treatment plan based on literature review of MVI in HCC.

This research is to explore the perfusion time-intensity curve parameters of a lung adenocarcinoma xenograft into nude mouse model with contrast enhanced ultrasonography(CEUS);and to investigate the angiogenesis features of tumor at different growth time.Twenty one lung adenocarcinoma xenografted nude mice were divided into three groups and inculcated with human lung adenocarcinoa.Time window for examining CEUS were respectively in7-day,14-day and 28-day.The perfusion parameters including rise time(RT),peak intensity(PI),area under the curve(AUC)of lung tumor were obtained on CEUS images by using off-line software Q lab.Immunohistochemically staining for CD34 was used to observe the microvessel density(MVD).The 7-day group had the highest AUC and PI;AUC and PI of 14-day and 28-day group decreased gradually(P<0.05).RT was increased as tumor growth.In tumor with necrosis,AUC and PI of non-necrosis part were also larger than necrosis part(P<0.05).Immunohistochemically staining for CD34 of all tumors reflected that the density of microvessels in necrosis tumor was significantly higher than those without necrosis(7.50±3.44 vs.12.44±5.74,P=0.034).Pearson correlation indicated that PI was positively related with MVD(r=0.668,P=0.008).Lung adenocarcinoma perfusion characteristic can be accessed from time-intensity curve parameters by using noninvasively and non-radiative contrast enhanced ultrasonography.Time-intensity curve parameters including AUC,PI and RT may reflect tumor angiogenesis.

Through textual research of the literature on tiny collateral and perforator vessel, the two concep- tions were compared and the similarity was analyzed in terms of definition, anatomical level, quantity, the flow of qi and blood and clinical application, etc. It is considered that the tiny collateral in Neijing (Inner Canon of Yellow Emperor) and the perforator vessel of modern medicine are different names of one physical structure. It is proposed that the combination of the tiny collateral theory of CM and the research findings of perforator vessel of western medicine would deepen the understanding of the tiny structure of human skin and promote the development of both TCM and modern medicine.

Objective: To explore the correlation between metabolic tumour volume (MTV) and microvessel density (MVD) and blood-borne metastasis in colorectal carcinoma., Methods: Thirty-six patients with CRC conformed by pathology underwent PET-CT examination before operation. SUVmax and MTV were obtained by PET VCRA software. The blood vessels were identified with CD34 immunohistochemical staining, and the MVD was recorded. The correlation between SUVmax and MTV with histological differentiation, T stage, MVD and blood-borne metastasis was analyzed., Results: The SUVmax, MTV and MVD in patients with blood-borne metastasis were 5.15 ± 5.41, (22.99 ± 18.63) cm³ and 14.17 ± 3.63, and were 10.65 ± 3.79, (16.95 ± 11.82) cm³ and 11.27 ± 3.69, respectively, in patients with non-blood-borne metastasis. The differences of SUVmax, MTV and MVD between blood-borne metastasis and non-blood-borne metastasis patients were statistically significant (all P > 0.05). Pearson correlation analysis found that there was no linear correlation between SUVmax and MVD, and the SUVmax was not statistically significant between high and low MVD groups (t = 0.919, P = 0.364). But there was a linear correlation between MTV and MVD (r = 0.621, P = 0.000), and the MTV was statistically significant between high and low MVD groups (t = 3.567, P = 0.001). The receiver-operating characteristic curves showed that MTV could be used to predict blood-borne metastasis of CRC, and the best cutoff value for MTV was 14.975 cm³, and the sensitivity, specificity, negative predictive value and positive predictive value were 85.7%, 54.5%, 72.3% and 64.2%, respectively. There were no significant relationships between SUVmax, MTV, MVD, blood-borne metastasis and histological differentiation (P > 0.05). With the increased T stage, the MTV, MVD and the probability of blood-borne metastasis were also increased (all P < 0.05)., Conclusions: There are correlations between MTV and MVD and blood-borne metastasis in CRC. The risk of blood-borne metastasis in patients with MTV > 14.975 cm³ is higher, and needs to take more effective intervention.

Objective: To investigate the clinicopathologic characteristics of meningioangiomatosis (MA)., Methods: Five cases of MA were evaluated morphologically by HE and immunohistochemistry on formalin-fixed paraffin-embedded tissue. Clinical information was also obtained. The literature was reviewed. The clinical pathology and biological behavior of MA were discussed., Results: Five cases of MA were reported, arising in three males and two females, with an age range of 16 to 26 years at diagnosis. All five subjects had intractable seizure disorders, and the duration of illness ranged from 8 months to 18 years. The lesions were resected from the frontal lobe in four patients, and from the temporal lobe in one. All the lesions were confined to the cortex, firm in consistency, without capsules and had poor blood supply. There was focal involvement of the overlying leptomeninges. Microscopically, they showed characteristic features of MA, such as proliferating microvessels with perivascular cuffs of spindle-cell within the cortex. Some had numerous calcifications, others showed acidophilic granular bodies. The cells were positive for EMA and vimentin by immunohistochemistry, and for reticulin by histochemical staining., Conclusions: MA is a rare, benign hamartomatous lesion of the central nervous system. It usually presents as plaque-like or nodular mass in the cerebral cortex and the overlying leptomeninges, consisting of meningovascular proliferation and leptomeningeal calcification. In some cases the lesion may show perivascular proliferation of elongated spindle-shaped cells. MA usually affects children and young adults, and is located in the frontal or temporal lobes with variable involvement of the overlying leptomeninges. Clinically, most of sporadic cases have a long history of intractable seizures despite multiantiepileptic drugs. MA has also been reported to coexist with arteriovenous malformations,meningiomas and other tumorous lesions.

Objective: To observe the inhibition of NK4 protein in the proliferation of human Raji lymphoma xenografts in nude mice, and to explore its molecular mechanism., Methods: Models of human Raji lymphoma xenograft transfected with HGF gene were established by subcutaneous inoculation in nude mice. After establishment of the models, the mice received continuous NK4 protein via tail vein for 4 weeks, and the weight and tumor growth were monitored every week. After 8 weeks, the expression of HGF mRNA and c-Met mRNA of tumor tissues was measured by real-time fluorescent quantitation PCR. The apoptotic index (AI) and microvessel density (MVD) were evaluated by terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) and immunohistochemistry, respectively., Results: The models of human Raji lymphoma xenograft were successfully established. Although the animal weights of all groups declined, especially in the groups with NK4 protein injection, there was no statistical significance (P > 0.05). The tumor volume in HGF gene transfected group was larger than those of the control groups (P < 0.01), and there was no statistical significance among the control groups (P > 0.05). However, the tumor volume of the NK4 protein injection group decreased significantly (P < 0.01). Expression of HGF mRNA and c-Met mRNA in HGF gene transfected group increased significantly after injection of NK4 protein (P < 0.01). AI in HGF gene transfected group (33.5% ± 12.3%) was significantly lower than that of control groups (89.1% ± 22.3% vs. 81.9% ± 27.0%, P < 0.05), but became significantly higher (119.1% ± 18.9%) after NK4 protein injection (P < 0.01). MVD in HGF gene transfected group (28.5 ± 2.0) was higher than that of control groups (12.2 ± 1.4, 13.8 ± 1.3, P < 0.01), although declined (15.5 ± 2.5) after NK4 protein injection (P < 0.01)., Conclusions: NK4 protein suppresses significantly the growth of human Raji lymphoma xenografts transfected with HGF gene. The pathogenesis may be involved in promoting tumor cell apoptosis and restraining tumor angiogenesis through competitive interrupting HGF/Met signal pathway.

Objective: To investigate the expression of platelet derived growth factor-BB (PDGF-BB) and microvessel density (MVD) marked with CD34 in clear cell renal cell carcinoma (CCRCC) and explore their relevance to clinicopathologic features and prognoses of patients., Methods: Expressions of PDGF-BB and CD34 in the tissue samples of 100 clear cell renal cell carcinomas were detected by immunohistochemical (IHC) SP staining. The microvessel density (MVD) was counted using Weidner's method. For PDGF-BB assessment, the staining intensity and the proportion of positive tumor cells were analyzed. Staining was considered immunoreactive when brown granules were identified in the cytoplasm or nuclei of tumor cells. Staining intensity and the proportion of positively stained tumor cells in lesions was scored for further analysis. Statistical analysis was performed using the software SPSS 18.0., Results: The MVD value marked by CD34 in the 100 cancer tissues was (105.49 ± 37.95) profiles/HPF. The median value of MVD in the entire cohort was used as the cut-off point for low MVD group (42 cases) and high MVD group (58 cases). The MVD of the low and high MVD groups was (75.12 ± 22.41) profiles/ HPF and (135.86 ± 22.91) profiles/HPF, respectively, with a statistically significant difference (P < 0.001). MVD was significantly correlated with the tumor T staging, histopathological grading and postoperative metastasis in CCRCC (P < 0.05, respectively). Among the 100 CCRCC cases, there were 38 cases with low PDGF-BB expression and 62 cases with high PDGF-BB expression, and the expression of PDGF-BB was significantly correlated with tumor diameter, T staging, histopathological grading and postoperative metastasis in the CCRCC (P < 0.05, respectively). Kaplan-Meier survival analysis showed that the cancer specific survival (CSS) in CCRCC patients with high expression of MVD and PDGF-BB was significantly better than that in the group with low MVD and low PDGF-BB expression (P < 0.001, respectively). Expression of PDGF-BB protein was positively associated with the MVD assessed by Spearman's correlation and factor analysis (r = 0.461, P < 0.001)., Conclusion: Significantly increased MVD and PDGF-BB expression detected in CCRCC patients indicate a better tumor grading and staging, and a longer survival time.

Objective: To investigate the therapeutic effect of thyroid hormone in nude mice bearing human pancreatic cancer xenograft., Methods: A BALB/c nude mouse model bearing pancreatic cancer was established with human pancreatic cancer cell line Bx-PC3. The mouse models were divided randomly into 5 groups, namely the control group treated with distilled water, high and low concentrations of thyroid hormone (T3) groups, and high and low concentration of propylthiouracil (PTU) groups. After intervention for 21 days, the changes in body weight and xenograft tumor volume and weight were measured, and the serum T3 concentration was detected by ELISA assay. The expression of proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) were detected using immunohistochemistry., Results: The body weight of nude mice in T3 groups was significantly reduced after intervention, while that in PTU groups showed no obvious changes. Compared with PTU groups and control group, T3 groups showed significantly reduced tumor volume and weight (P<0.05) with also reduced PCNA expression and MVD, but these effect did not exhibit a dose dependence (P>0.05)., Conclusion: Thyroid hormone can inhibit the growth of human pancreatic cancer in nude mice by suppressing the proliferation and angiogenesis of the tumor cells, suggesting the potential value of thyroid hormone in pancreatic cancer therapy.

Objective: To explore the feasibility of volume perfusion CT imaging to dynamically monitor and evaluate the response of rabbit VX2 soft-tissue tumor to antiangiogenic treatment., Methods: To establish an experimental animal model of VX2 soft tissue tumor on 20 New Zealand white rabbits. Twenty rabbits were randomly divided into 2 groups. The therapy group was treated with recombinant human endostatin (3 mg·kg⁻¹·d⁻¹) for 7 days, and the control group received saline in the same dose only. Four times of CT volume perfusion scan were performed before treatment and on the second, forth, seventh days of treatment, respectively. The value of blood flow (BF), blood volume (BV), mean transit time (MTT), and permeability (PMB) in the VX2 tumors were measured after scanning. The microvessel density (MVD) and expression of vascular endothelial growth factor (VEGF) in the tumors were determined using immunohistochemical staining., Results: The tumor volume of the therapy group was (1.36 ± 0.73) cm³ on the forth day of treatment and (1.69 ± 0.68) cm³ on the seventh day of the treatment. The tumor volume of the control group was (2.35 ± 0.62) cm³ on the fourth day of treatment and (3.87 ± 0.93) cm³ on the seventh day of the treatment (P < 0.05). On the seventh day of treatment, tumor necrosis ratio of the therapy group and the control group was (25.58 ± 5.51)% and (42.93 ± 4.34)%, respectively (P < 0.05). Comparing the perfusion parameters between the two groups on the same day, and the second, forth, seventh days of treatment, the value of PMB of the therapy group was (70.36 ± 23.46) ml·100 ml⁻¹·min⁻¹, (79.64 ± 13.68) ml·100 ml⁻¹·min⁻¹ and (84.76 ± 3.55) ml·100 ml⁻¹·min⁻¹, respectively, and that in the control group was (26.61 ± 6.47) ml·100 ml⁻¹·min⁻¹, (33.74 ± 16.47) ml·100 ml⁻¹·min⁻¹ and (30.47 ± 10.64) ml·100 ml⁻¹·min⁻¹, respectively (P < 0.05). The value of BF in the therapy group and control group was (71.19 ± 12.21) ml·100 ml⁻¹·min⁻¹ and (43.56 ± 12.21) ml·100 ml⁻¹·min⁻¹, respectively, on the seventh day of treatment (P < 0.05). The parameters on different days in the same group were compared. In the control group, the value of BF on the seventh day of treatment was significantly lower than that before and on the second and forth days of treatment (P < 0.05). However, in the therapy group, the value of PMB on the second, forth, and seventh days of treatment was significantly higher than that before treatment (P < 0.05). MVD of tumor in the control group was increased gradually, whereas increased on the first day and then decreased more in the therapy group. The VEGF expressions did not differ significantly between the experimental and control groups., Conclusions: Volume perfusion CT is helpful to quantify the tumor perfusion and evaluate the functional changes of tumor vasculature, and then evaluate the early therapeutic effect of antiangiogenic treatment.

This study was aimed to analyze the correlation between CT perfusion parameters, e. g. perfusion (PF), blood volume (BV), peak enhancement image (PEI), time to peak (TTP), and the microvessel density (MVD) of cervical cancer. CT perfusion scans were carried out in 31 patients with cervical cancer. After their surgical resections, we obtained 31 cases of cervical cancer specimens, and 15 cases of adjacent normal cervical specimens as control group, then these 46 specimens were used in MVD detection through immunohistochemical CD34 staining. The MVDs of the tumor group were significantly higher than those in the control group, and the former seems to increase with the clinical stages with a positive correlation. The PF and PEI values have positive correlations with corresponding MVD values. The PF value of squamous carcinoma group has correlation with corresponding MVD value. The PF and PEI values of poorly differentiated group have correlations with corresponding MVD values. The PF, PEI and BV values of II-III stage group are positively correlated with corresponding MVD values. The PF value of lymph metastasis or non-lymph metastasis group both have correlations with corresponding MVD values. We found that CT perfusion parameters were positively correlated with corresponding MVD values in cervical cancer. Our results suggest that CT perfusion is a new method which could reflect tumor angiogenesis, histological malignant grade, invasion, and clinical stage. It will help us determine the tumor staging and prognosis.

Objective: To explore the relationship between neuropilin 2 (NRP-2) and lymphangiogenesis and lymphatic metastasis of human colorectal carcinoma (CRC), as well as the expression of NRP-2 in CRC tissues., Methods: A total of 55 cases of CRC, adjacent and normal tissues of surgical resection were randomly selected at our hospital from March 2010 to January 2012. All pathological findings were confirmed by histopathology. The expression of NRP-2 was detected with reverse transcription (RT)-PCR and immunohistochemistry in parenchymatous and surrounding malignant tissues. Then lymphangiogenesis was marked with D2-40 monoclonal antibody and microlymphatic density (MLD) counted., Results: Significant differences of MLD existed between those of tumor region (39 ± 19) and tumor margin (53 ± 26, P < 0.01). Both the number and shape of lymphangiogenesis were different between the parenchymatous and surrounding tissues. The expression of NRP-2 had a positive correlation with MLD both at the protein level (r = 0.325, P < 0.05) and at the gene level (r = 0.545, P = 0.000). And it was also correlated with the differentiation degree, infiltrative degree, lymphovascular invasion, lymph node metastasis and Dukes tumor staging (all P < 0.05)., Conclusion: The expression of NRP-2 may regulate lymphangiogenesis and it may play an important role in the incidence and development of CRC.

Objective: To investigate the inhibitory effect of compound cantharides capsules on the proliferation of xenografts of human hepatocellular carcinoma HepG(2215) in mice and their mechanism of action., Methods: One hundred healthy Balb/c mice (5-week old, male:female 1:1) were used in this study. Mouse models of human HepG(2215) hepatocarcinoma were established. The tumor-bearing mice were divided into five groups randomly. The control group A received daily intragastric administration of physiologic saline. The intervention groups B1, B2 and B3 were treated with compound cantharides capsule in a dose of 12.5 mg×kg(-1)×d(-1), 25 mg×kg(-1)×d(-1) and 37.5 mg×kg(-1)×d(-1), respectively, for 10 consecutive days. The group C had intraperitoneal injection of cyclophosphamide (25 mg×kg(-1)×d(-1)) for 10 consecutive days. The mice were sacrificed after the completion of administration. The tumors were taken out, the tumor volume was measured, the inhibitory rate of body weight was calculated, and the serum AFP concentration and the level of HBV DNA were determined. The survival of each group mice was analyzed. The levels of mRNA expression of apoptosis-related genes were assayed by quantitative RT-PCR. Apoptosis in the tumor cells was assayed with TUNEL staining. Flow cytometry was used to detect the levels of CD3(+), CD19(+), CD4(+) and CD8(+), and microvessel density (MVD) of the tumors was assessed by immunohistochemistry., Results: After completion of the treatment, the inhibition rate of tumor growth of the groups B1, B2 and B3 was 29.8%, 38.7% and 48.1%, respectively, and that of the group C was 52.4%, with a significant difference among the groups (P < 0.05). The median survival time of the groups A, B1, B2, B3 and C was (30.0 ± 3.2) days, (49.0 ± 5.1) days, (50.0 ± 5.2) days, (57.5 ± 6.5) days and (49.0 ± 4.7) days, respectively. The median survival time of the group B3 was significantly longer than that of other groups (P < 0.05). The serum AFP level in the groups A, B1, B2, B3 and C was (492.7 ± 48.5) ng/ml, (281.2 ± 25.6) ng/ml, (194.3 ± 18.7) ng/ml, (170.1 ± 15.8) ng/ml and (138.7 ± 12.5) ng/ml, respectively, indicating that it was significantly inhibited in the group C. The inhibition rate of HBV DNA replication of the groups B1, B2, B3 and C was (46.0 ± 5.1)%, (65.5 ± 6.9)%, (81.3 ± 7.8)% and (19.5 ± 2.1)%, respectively, showing that compound cantharides capsules inhibited HBV DNA replication in a dose-dependent manner. The apoptosis rate of the groups A, B1, B2, B3 and C was (0.27 ± 0.03)%, (7.18 ± 2.12)%, (9.17 ± 2.42)%, (11.27 ± 3.03)% and (5.44 ± 2.45)%, respectively, and that of the group B3 was significantly higher than that of the groups A, B1, B2 and C (P < 0.05). The expression level of bax mRNA was significantly higher than that of the group C (P < 0.05). The drug could significantly decrease the bcl-2 mRNA expression level, more remarkably along with the increasing dose of cantharides, and it was significantly lower than that in the group C (P < 0.05). The levels of CD4(+), CD8(+), CD3(+) and CD19(+) were significantly higher than that in the groups A and C (P < 0.05). The value of MVD of the group B3 was significantly lower that that of groups A and C (P < 0.05)., Conclusion: Compound cantharides capsules may inhibit the replication of HBV DNA in HepG(2215) cells, inducing apoptosis in the tumor cells, enhancing the immune function to inhibit the growth of liver cancer cells in mice, and significantly prolong the median survival time of tumor-bearing mice.

Objective: To observe the effect of rosiglitazone (RGZ) and all-trans-retinoic acid (ATRA) on the growth of myeloma xenograft in nude mice and to explore the influence of RGZ and ATRA on VEGF expression and angiogenesis in the tumor., Methods: VEGF gene expression in myeloma cell line U266 cells was analyzed by semi-quantitative RT-PCR after incubation with RGZ, ATRA, or RGZ + ATRA for 24 h. Myeloma xenograft was established by subcutaneous injection of 10(7) U266 cells in the scapula area of 4-week old nude mice. 7 days later, the nude mice were administered with RGZ, ATRA or RGZ + ATRA, respectively, by intraperitoneal injection once every day for 21 days. The control mice were given equal volume of normal saline instead of the drug. On the 21(st) day of treatment, the mice were sacrificed and the tumors were taken off, and the tumor volume and weight were measured. The tumors were examined by histopathology with HE staining, and microvessel density (MVD), CD34 and VEGF expression in the tumors were analyzed by immunohistochemical staining., Results: VEGF mRNA was highly expressed in U266 cells and was decreased in a dose-dependent manner after incubation with RGZ. The VEGF mRNA level was further more decreased after RGZ + ATRA treatment. Xenografts of U266 cells were developed in all nude mice. The volume and weight of xenografts in the RGZ group were (785 ± 262) mm(3) and (1748 ± 365) mg, respectively, significantly lower than those of the control group (both P < 0.01). More significant inhibition was in the RGZ + ATRA group, (154 ± 89) mm(3) and (626 ± 102) mg, respectively, both were P < 0.05 vs. the RGZ group. RGZ inhibited the angiogenesis in U266 xenografts and immunohistochemical staining showed that the tumor MVD and VEGF expression were significantly decreased by RGZ treatment, and further more inhibited in the RGZ + ATRA group. VEGF protein was expressed in all xenografts in the nude mice. Its immunohistochemical staining intensity was 2.20 ± 0.40 in the control group, significantly higher than that of 1.48 ± 0.37 in the RGZ group (P < 0.01), and that of RGZ + ATRA group was 0.58 ± 0.26, further significantly lower than that of the RGZ group (P < 0.01). CD34 was expressed in all xenografts, most highly in the control group and lowest in the RGZ + ATRA group. The microvessel density (MVD) was highest in the control group (56.4 ± 15.2), significantly lower in the RGZ group (44.6 ± 11.2) (P < 0.05), and lowest in the RGZ + ATRA group (21.5 ± 8.6, P < 0.01)., Conclusions: The growth of myeloma cells can also be inhibited by RGZ and ATRA in nude mice in vivo. In addition to differentiation and apoptosis induction, RGZ can inhibit the formation of myeloma xenograft probably also through the downregulation of VEGF expression and subsequent angiogenesis.

Objective: To explore the expression of CD4(+), IL-17 and Foxp3 in non-small cell lung cancer (NSCLC) and their relationship with microvessel density (MVD)., Methods: The expressions of CD4(+), IL-17, Foxp3, CD31 and CD34 in the cancer tissues of 102 NSCLC cases were detected by immunohistochemisty. The relationship among the expressions of CD4(+), IL-17, Foxp3 and MVD was analyzed. The count data were analyzed using χ(2) test. The measurement data were analyzed using single-factor analysis of variance, and significance level α = 0.05., Results: Among the factors affecting CD31 expression, there was a statistically significant difference between the strong positive Foxp3 expression (++) and negative (-) and positive expressions (+) in the NSCLC cancer tissues (P < 0.05), and between the expressions in stage I and III cancer tissues (P < 0.05). Among the factors affecting CD34 expression, there was a significant difference between positive IL-7 expression (+) and strong positive IL-7 expression (++) (P < 0.05), between negative Foxp3 expression (-) and strong positive Foxp3 expression (++) (P < 0.05), and between the CD34 expressions in stage I and III and between those in stage II and III NSCLC cancer tissues (P < 0.05)., Conclusions: CD4, IL-17and Foxp3 may be involved in the tumor suppression caused by host immune response, and are related with the NSCLC invasion and metastasis.

Objective: Erythropoietin and erythropoietin receptor (EPO-R) are expressed in many kinds of tumors. The EPO/EPO-R signaling is involved in tumor cell proliferation, invasion and angiogenesis. The aim of this study was to detect the expression of EPO-R in non-small cell lung cancer (NSCLC), and explore its correlation with angiogenesis., Methods: The expression patterns of EPO and EPO-R in 31 cases of NSCLC tissues were detected by immunohistochemistry, and that in benign lung lesions of 21 patients as control. To analyze the correlation of EPO/EPO-R expression patterns and clinicopathological factors. CD34 was used to label the vascular endothelial cells and calculate the microvessel density (MVD)., Results: The positive rates of EPO and EPO-R expression in NSCLC were 67.7% and 96.8%, respectively, significantly higher than those in the control ones. The positive rates of EPO and EPO-R expression in adjacent tissues were 19.4% and 35.5%, and in benign lesions were 9.5% and 19.0%, respectively (P < 0.001). The expression patterns of EPO/EPO-R were not related with pTNM stage, histological type, histological grade and lymph node metastasis (P > 0.05). Increased MVD was correlated with poor differentiation, lymph node metastasis, and advanced stage., Conclusions: High expression of EPO/EPO-R in NSCLC patients suggest that they may be involved in tumorigenesis. EPO/EPO-R expression and MVD are closely related, and they might be an endogenous stimulant of angiogenesis during the progression of non-small cell lung cancer. It may provide evidence for clinical diagnosis.

Objectives: To dynamically observe how glioma stem cells promote the tumor formation and angiogenesis, and to study the correlation between the distribution of glioma stem cells and microvessels within different growth stages of subcutaneous tumor., Methods: Stem cell medium culture and magnetic activated cell sorting were carried out to obtain CD133+ cells from C6 rat glioma cell line. Sprague Dawley (SD) rat ears model were established to observe glioma stem cells promoting blood vessel formation. Subcutaneous glioma model of C6 and immunohistochemical staining of hypoxia inducible factor-1α (HIF-1α) and CD133 were used to investigate the relationship between distribution of glioma stem cells and microvessels. Expressions of CD133 protein in each stage of the subcutaneous tumor were detected by Western blot., Results: Isolation and identification of glioma stem cells deprived from C6 glioma cell line successfully, the establishment of ears model showed real-time dynamic observation of CD133+ cells involved in angiogenesis and tumor formation. SD rat model of subcutaneous glioma showed the initial of tumor formation, CD133+ cells scattered. With tumor growth, CD133+ cells began to tend to capillaries, in late distributed clusters in perivascular. Meanwhile as tumor growth, CD133 protein expression was gradually increased: the values of Western blot analysis of CD133 expression on 6, 9, 12, 15, 20 d were 0.208±0.004, 0.282±0.003, 0.360±0.004, 0.564±0.135, 0.756±0.007, the differences were significant between different groups (F=2601.681, P<0.01). At a high magnification, the CD133 scores with immunohistochemical staining on 6, 9, 12, 15 d were 0.8±0.4, 2.4±0.5, 4.0 ± 0.7, 6.0±0.7; HIF-1α scores were 0.8±0.4, 2.8±0.8, 5.0±0.7, 6.8±0.4. By Spearman rank correlation analysis found that the relationship between CD133 and HIF-1α expression was positively correlated (r=0.921, P<0.01)., Conclusions: Glioma stem cells promote angiogenesis more than non-stem cells; HIF-1α and its downstream gene product might mediate the distribution of glioma stem cells around the perivascular.

Objective: To compare the effect of rh-endostatin on micrangium in tumor and myocardial tissue in nude mice., Methods: Nude mice were randomized into 4 groups (10 mice in each group), blank control group (without tumor burden, received NS 100 µl×d(-1) injection), drug control group (without tumor burden, received rh-endostatin 400 µg×d(-1) injection), model group (with tumor burden, received NS 100 µl×d(-1) injection) and treatment group (with tumor burden, received rh-endostatin 400 µg×d(-1) injection) for 28 days. The tumor volume and body weight of the mice were measured before and after administration. The expression of CD34, MMP-2, MMP-9, HIF-1α and VEGF in the myocardium and tumor were detected by immunohistochemistry. The vascular structure was observed by immunoenzymatic CD34 and Masson double staining., Results: The increase of tumor volume of the treatment group [(48.18 ± 37.31) mm(3)] was significantly lower than that in the model group [(113.80 ± 73.27) mm(3)). The changes of body weight was not significant different among the four groups. After treated with rh-endostatin, the expressions of MMP-9 and VEGF in tumors were significantly down-regulated, but the expressions of MMP-2 and HIF-1α in the tumor were not. The microvessel density (MVD) in the tumors of treatment group was significantly decreased compared with that of model group. The proportion of tumor vessels covered by collagen in the treatment group was increased compared with that of the model group. However, MVD and micrangium in myocardium were not changed significantly., Conclusion: Rh-endostatin can decrease the expression of MMP-9, VEGF and MVD, inhibit the tumor growth and normalize tumor micrangium in tumor but not weaken the MMPs and MVD of mature micrangium in myocadium.

Objective: To study the cell apoptosis and the expression of connective tissue growth factor (CTGF) in the retina of diabetic rats and to explore their contributions to the changes of microcirculation., Methods: It was a experiment study. Fifty-five adult male Wistar rats were divided into two groups, normal control group (CON, 10 rats) and diabetes mellitus group (DM, 45 rats). The 30 surviving rats in the DM group were further divided into 3 groups based on the time of observation, 2 month (DM2), 4 month (DM4) and 6 month (DM6) groups, with 10 rats in each group. Cell apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL). Expression of CTGF was determined by immunohistochemical study. Retinal vessels were observed by retinal digest stretched periodic acid Schiff (PAS) staining., Results: Immunohistochemical staining and TUNEL staining revealed negative results in normal control group. Retinal cell apoptosis index increased gradually from DM2 to DM6, the differences between any two groups were statistically significant (t(2-4, 2-6, 4-6) = 21.432, 50.843, 29.410; P < 0.05). Expression of CTGF in the retina increased from DM2-DM6, the differences between any two groups were statistically significant (t(2-4, 2-6, 4-6) = 15.345, 26.316, 10.971; P < 0.05). PAS staining of retinal blood vessels obtained negative results in the CON and DM2 groups. Part of retinal capillaries were slightly stiff and narrow in DM4 group. Retinal capillaries in DM6 group were trunk stiff and were narrowed obviously. The number of pericytes was reduced in DM4, and progressed following the course of diabetes. The number of pericytes in the DM2 group did not different from that in the CON group (t = 0.875, P = 0.387). The number of pericytes in the DM4 and DM6 group were significantly decreased as compared to the CON group (t = 3.367, 6.667; P < 0.05). Retinal cellular apoptosis index had a significant positive correlation to the expression of CTGF (r = 0.958, P < 0.05). Number of pericytes was significantly correlated (negative correlation) with retinal cellular apoptosis index and the expression of CTGF (r = -0.540, -0.595; P < 0.05)., Conclusions: The appearances of cellular apoptosis and fibrosing factor CTGF in the retina of diabetic rats occurred earlier than the changes of microcirculation and the number of capillary pericytes.

Objective: To study the effect of Eupolyphaga fibrinolyric protein (EFP) on microvessel density (MVD) and the expression of vascular endthelial growth factor in transplantation S180 and H22 mice., Methods: The MVD in tumor was measured with immunohistochemical SP method and the VEGF level in serum was measured with ELISA method., Results: Compared with the control group, EFP could significantly reduce the microvessel density and decrease the expression of vascular endothelial growth factor., Conclusion: EFP has the effect of anti-angiogenesis.

Objective: To investigate the regulatory effect of jianpi jiedu Recipe (JJR) on the microvessel density (MVD) and cyclooxygenase-2 (COX-2) in long-term infection of Helicobacter pylori induced gastric cancer of C57BL/6 mice, thus providing experimental bases for its treatment of the H. pylori correlated gastropathy., Methods: C57BL/6 mouse gastric cancer model induced by H. pylori infection was established by gastrogavage of H. pylori standard strain SS1. Mice were divided into the control group, the model group, low dose JJR group, and the high dose JJR group, 40 in each group. Mice were sacrificed after 72-week medication. Changes of the gastric mucosa MVD of mice in each group were detected by immunohistochemical method. Expressions of COX-2 mRNA and protein were detected by Real-time fluorescent quantitative polymerase chain reaction and immunohistochemical method., Results: The occurrence rate of gastric cancer in the control group, the model group, the low dose JJR group, and the high dose JJR group was 0, 22.2%, 11.1%, and 10.0%, respectively. The gastric mucosa MVD (number/cm2) of mice in each group was 2.50 +/- 1.54, 18.56 +/- 2.62, 14.61 +/- 3.60, and 7.39 +/- 1.75, respectively. The gastric mucosa MVD in the model group increased more obviously than that in the control group (P < 0.01). The gastric mucosa MVD significantly decreased in the low dose JJR group and the high dose JJR group (P < 0.01). Expressions of COX-2 mRNA and protein in the model group increased more obviously than those in the control group (P < 0.01). Low dose JJR and high dose JJR could decrease their expressions in a dose dependent manner., Conclusions: H. pylori infection could increase the gastric mucosa MVD of C57BL/6 mice and promote COX-2 expressions, which might play a promoting effect in the incidence of H. pylori induced gastric cancer. JJR could decrease the gastric mucosa MVD and inhibit COX-2 expressions, which might be one of its important mechanisms of preventing and treating gastric cancer.

Objective: To investigate the expression and clinical significance of MMP9 and MVD in the carcinogenesis of squamous cell carcinoma of external auditory canal and middle ear., Method: Immunohistochemical SP method was used to detect the expression of MMP9 and MVD proteins in 26 squamous cell carcinoma tissues of external auditory canal and middle ear and 20 normal external ear canal skin tissues., Result: The positive rate of MMP9 in squamous cell carcinoma tissues of external auditory canal and middle ear was 73.1% (19/26) lower than that in the normal external ear canal skin tissues 25.0% (5/20). The positive rates of CD34 were 33.58 +/- 3.04 and 22.50 +/- 5.22, respectively. The positive rates of MMP9 and CD34 were correlated with the histological grade and tumor grade, but had no relationship with age and sex. The positive rates between MMP9 and CD34 were related (r=0.42, P<0.05)., Conclusion: MMP9 may be involved in the carcinogenesis of squamous cell carcinoma of external auditory canal and middle ear, and may play an important role in the invasion and metastasis of squamous cell carcinoma of external auditory canal and middle ear. MMP9 and CD34 play a cooperative role in the process of squamous cell carcinoma of external auditory canal and middle ear.

Objective: To explore possible correlations between renal Th1/Th2 ratio and renal microvascular injury in children with Henoch-Sch-nlein purpura nephritis (HSPN)., Methods: Thirty-two children with HSPN were enrolled. They were classified into four groups by renal pathology: HSPN class II (n=8), HSPN class IIIa (n=7), HSPN class IIIb (n=10) and HSPN class IV/V (n=7). Five patients undergoing nephrectomy due to trauma were used as the controls. INFγ, IL-4 and CD34 in the renal tissues were measured by immunohistochemical analysis. INFγ was used as a marker of Th1, IL-4 was used as a marker of Th2 and CD34 was used as a marker of microvessel. The renal microvessel density was evaluated according to the Weidner standard. The relationships among the local Th1/Th2 ratio, renal pathological grade, microvessel score and microvessel density were studied., Results: Immunohistochemical analysis showed a lower expression of INFγ and a higher expression of IL-4 in the HSPN groups than in the control group. The local Th1/Th2 ratio in the HSPN groups decreased and correlated significantly with the renal pathological grade. There were significant differences among four HSPN subgroups (P<0.05). Compared with the control group, the renal microvessel density in the HSPN class II and class IIIa groups increased significantly (P<0.05), but it decreased in the HSPN class IV/V group (P<0.05). The renal microvessel scores in the HSPN class IIIa, class IIIb and class IV/V groups increased significantly compared with those in the control and the HSPN classⅡ. The increased renal microvessel scores were associated with more severe renal pathological changes. A negative correlation was found between the local Th1/Th2 ratio and the microvessel density in kidneys (r=-0.921, P<0.01)., Conclusions: The decrease of Th1/Th2 ratio in kidneys might be responsible for renal microvascular injury in children with HSPN.

Objective: To investigate the expression of CD133 and CD34 in different parts of glioblastoma and its margin and explore the invasive path of glioma stem cells within the tumor and surrounding tissue., Methods: The surgical specimens were collected from the core of mass, junctional zones between tumor and peritumoral edematous areas and edematous areas in 52 patients with glioblastoma. Immunohistochemical cell staining and Western blot were employed to evaluate the expression of CD133 in different specimens while immunohistochemistry was used to detect the CD34-microvessel postforming. A correlation analysis was performed between them., Results: The expression of CD133 was not detected in the control groups while the scores were 7.3 ± 1.4, 5.2 ± 1.1, 2.7 ± 1.0 in junctional zones, tumor cores and edematous areas with immunohistochemistry and 0.79 ± 0.03, 0.38 ± 0.01, 0.20 ± 0.04 with Western blot respectively. There were significant differences between different groups (P < 0.05). Under light microscope, the CD133-positive cells frequently forming perivascular pseudorosettes were dense in junctional zones and mostly clustered near the microvessels in tumor cores and scattered in edematous areas. At a high magnification (200×), the CD34-MVD (/HP) values were 31.3 ± 4.0, 21.8 ± 2.6, 15.3 ± 2.4, 4.7 ± 1.5 respectively in junctional zones, tumor cores, edematous areas and control tissues. Significant differences were also found in these groups (P < 0.05). The expression level of CD133-positive cell was positively correlated with the distribution of CD34-microvessels (r = 0.948, P < 0.05)., Conclusion: Glioma stem cells tend to assemble in the junctional zones where the microvessels are enriched. There is probably an intimate nourishing relationship with the microvessels. The distribution of glioma stem cells may be related with the invasiveness within glioblastoma.

Objective: To investigate the expression of chemokine receptor 4 (CXCR4) in laryngeal squamous carcinoma and its correlation with clinicopathology character and the building of microvessel density (MVD). To evaluate its role in the carcinogenesis and progression in laryngeal squamous carcinoma., Method: The expression of CXCR4 in fresh laryngeal squamous carcinoma tissues and adjacent normal tissues from 42 patients were examined by RT-PCR, immunohistochemical staining and Image-pro-plus software. The numbers of regeneration blood vessels in the laryngeal squamous carcinoma was counted by antibody against factor V associated antigen and immunohistochemical analysis., Result: The positive expression rate of CXCR4 in tumor samples was significantly higher than that in normal ones (P < 0.01). In tumor samples, the expression of CXCR4 were not associated with age, sex, tumor site and T stage (P > 0.05), while it were higher in tumors of grade III, IV than in grade I, II of pathology classification (P < 0. 01). The expression of CXCR4 were significantly higher in tumors with cervical lymph node metastasis than that in tumor without cervical lymph node metastasis (P < 0.01). The expression of CXCR4 protein and CXCR4 mRNA were at the same level. The expression level of CXCR4 in the laryngeal squamous carcinoma tissue was positively correlated with vascularization., Conclusion: The higher expression of CXCR4 may play a key role in the invasion and metastasis of laryngeal squamous carcinoma, and were correlated with micro-vascularization.

Objective: To study expression and activation of p38 mitogen activated protein kinase (MAPK) in vascular endothelial cells dysfunction in preeclampsia., Methods: From Sept. 2009 to Mar. 2010, 54 pregnant women underwent deliveries in the First Affiliated Hospital of Chongqing Medical University were enrolled in this study, including 20 patients in mild preeclampsia group, 16 patients in severe preeclampsia group and 18 women with term cesarean section without perinatal complications as control group. Placental endothelial cells were labeled by CD₃₄ to assay microvessel density (MVD) of each group. Immunohistochemical SP and western blot were used to detect localization and expression of p-p38 MAPK protein, respectively. The levels of sera soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (sEng) were measured by ELISA., Results: (1) The MVD of placenta were 103 ± 3 in control group, 81 ± 5 in mild preeclampsia group and 63 ± 4 in severe group, respectively, which showed statistical difference among each group (P < 0.05). (2) The expression of p38 MAPK protein were 0.84 ± 0.05 in control group, 0.90 ± 0.14 in mild group and 0.86 ± 0.18 in severe group, which did not reach remarkable difference among each group (P > 0.05). The expression of p-p38 MAPK protein were 0.13 ± 0.05 in control group, 0.59 ± 0.12 in mild group and 1.16 ± 0.18 in severe group, which show statistical difference among each group (P < 0.05). (3) The localization of p-p38 was in trophoblast, endothelial cells and a few stromal cells in placenta. (4) The level of sFlt-1 and sEng: (1) The concentration of sFlt-1 and sEng were (5.2 ± 0.3) and (10.9 ± 0.4) µg/L in control group, (12.5 ± 1.2) and (20.4 ± 5.3) µg/L in mild group and (19.3 ± 3.0) and (29.5 ± 3.7) µg/L in severe group. When drawing paired comparison in those groups, the differences showed statistical difference (P < 0.05). (2) There were positive correlations between p-p38 MAPK protein levels and the concentrations of serum sFlt-1, sEng in preeclampsia groups (r = 0.68, P < 0.05; r = 0.87, P < 0.05)., Conclusions: The remarkable activation of the p38 MAPK in the placenta of patients with preeclampsia induced the increased levels of sFlt-l and sEng in maternal serum, which confer the injury of vascular endothelial cells that caused the significant decline of MVD in placentas. p38 MAPK signaling might be one of the key pathways in vascular endothelial cell dysfunction in preeclampsia.

Objective: To study the in vivo interference effects of vascular endothelial growth factor (VEGF) short hairpin RNA (shRNA) on xenografts of drug-resistant tongue cancer cells., Methods: Drug-resistant tongue caner cells Tca/Cisplatin (DDP) were injected subcutaneously into nude mice to establish xenograft models, which were randomly divided into non-transfected group, mock control group, control group transfected with scrambled sequence plasmid, interference group transfected with VEGF-shRNA expression plasmid. Liposome-mediated plasmid transfection was done in the latter three groups every three days. Xenografts were observed and tumor growth curve was measured. After the 10th transfection, tumors were anatomized and weigh. Microvessel density was detected by immunohistochemical staining. In situ hybridization assay was used to test VEGF mRNA, and immunohistochemistry to test VEGF, P-glycoprotein (P-gp), B cell lymphoma/leukemia-2 (bcl-2) and extracellular signal-regultaed kinase 2 (ERK-2) protein., Results: Tumor growth in VEGF-shRNA interference group was significantly slow. Tumor weight was (0.4781 ± 0.0860) g, microvessel density (7.35 ± 1.31)/view, VEGF mRNA (0.0767 ± 0.0234), VEGF protein (0.1301 ± 0.0433), P-gp (0.1517 ± 0.0184), bcl-2 (0.1218 ± 0.0251) and ERK-2 protein (0.1178 ± 0.0291) in VEGF-shRNA interference group; all of them were less than those in the other three groups (P < 0.05)., Conclusions: Inhibition targeting VEGF may become a potential therapy for drug-resistant tongue cancer.

Objective: To investigate the expression and correlation of Aquaporin-1 (AQP1), VEGF and MVD in nasopharyngeal tumor and non-tumor tissues., Method: Thirty nasopharyngeal biopsy samples were collected and examined with both immunohistochemistry and real-time fluorescence quantitative PCR (RT-PCR) to detect the expression level of AQP1 and VEGF in tissues. MVD were then calculated., Result: Both immunohistochemistry and RT-PCR showed higher expression of AQP1 and VEGF in tumor tissues than in non-tumor tissues. The two factors were positively correlated., Conclusion: In nasopharyngeal carcinoma, AQP1 may participate in angiogenesis by VEGF. Thus it may facilitate the tumor's growing and metastasis.

Objective: To study the expression of vascular endothelial growth factor(VEGF) and microvessel density (MVD), in nasal polyps and its significance., Method: The expression of VEGF, in nasal polyps from 50 patients and inferior turbinates from 10 patients were studied with immunohistochemical methods, measuring their grey score, the relations between expression microvessel density were analyzed., Result: (1) The differences between the VEGF expression in nasal polyp gland and vascular were significant (P < 0.05). (2) The differences between the microvessel quantities in nasal polyps and inferior turbinates were significant (P < 0.05). (3) The association between VEGF expression in vascular of nasal polyps and microvessel density was significant (r = -0.664 (< 0. 05)., Conclusion: In nasal polyps, VEGF, expression increased. Those results show that the abnormal expression of VEGF and MVD may play an impotent role in the development of nasal polyps. There was an important value for proper treatment and prediction of nasal polyps.

Objective: To investigate the effects of silencing heparanase (HPA) on growth, angiogenesis and metastasis of human gastric carcinoma transplanted in nude mice., Methods: Human gastric carcinoma SGC-7901 cells and those cells with silenced HPA (gastric carcinoma SGC-7901-HPA(-)) were separately transplanted subcutaneously in 6 nude mice. The time, size and speed of tumor growth were recorded. RT-PCR and Western-blot were used to detect the expression of HPA mRNA and protein in the subcutaneous tumors of the two groups. Immunohistochemical staining was used to detect microvessel density (MVD) in the subcutaneous tumors of the two groups. Cells of the subcutaneous transplanted tumors of the two groups were separately injected into the peritoneal cavity of nude mice, 6 mice each. The growth of metastatic tumors in nude mice was observed., Results: Human gastric carcinoma SGC-7901 cells and SGC-7901-HPA(-) cells were subcutaneously inoculated in nude mice, and tumors appeared at 4 days and 7 days after inoculation, respectively. The MVD was (20.69 ± 1.20)/HP and (11.35 ± 1.94)/HP, respectively (P < 0.05). The expressions of HPA mRNA and protein of the subcutaneously transplanted SGC-7901-HPA(-) tumor were decreased. Four voluminous metastatic tumors caused by SGC-7901 cells occurred in 3 mice in the liver, right kidney, omentum and intestine. Two smaller abdominal metastatic tumors of SGC-7901-HPA(-) cells were found in the liver and right kidney., Conclusion: Silencing HPA can inhibit the tumor growth, angiogenesis and metastasis of human gastric cancer in nude mice. It suggests that HPA might become a new target for prevention and treatment of gastric cancer.

Objective: To investigate the possibility of microvessel density (MVD) and blood vessel invade (BVI) as the indexes in predicting prognosis of rectal carcinoma at stages I to II., Methods: Tumor tissues from 380 patients who underwent resection of stage I or II rectal cancer were analyzed for MVD and BVI by immunohistochemical S-P method with anti-CD105 and anti-CD 34 antibody. Binary and multivariable Cox regression was applied to indicate independent factors associated with overall survival., Results: CD105 was present in the neovascularity of the cancer tissue but not in the normal tissue, while CD34 was present in the tumor tissue and the normal tissue. BVI on CD34 staining was significantly higher than that on HE staining. Multivariable analysis revealed that TNM stage, CD34-BVI, histologic type, and CD105-MDV were independent risk factors to predict the possibility of poor prognosis of stage I or II rectal cancer. CD34-BVI or CD105-MVD positivity had a hazard ratio of 4.483 (95% confidence interval 2.861-7.026) for mortality., Conclusion: The expressions of CD34-BVI and CD105-MVD are independent factors to predict the possibility of poor survival of stage I or II rectal carcinoma. Detection of CD105-MVD combined with CD34-BVI may help predict clinical outcome and design further individualized adjuvant treatment.

Objective: The effect of human bone marrow mesenchymal stem cells (hMSCs) on tumor neovascularization were studied., Methods: hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. hMSCs-EGFP were obtained by pLEGFP-N1 retroviral vector. Flow cytometry was used to detect the cell surface antigen and the differentiation potential of hMSCs-EGFP was investigated under conditioned media. The effect of hMSCs on tumor neovascularization were observed by establishing solid tumor models in BALB/C nude mice. In addition, effect of the conditioned medium used for tumor cells and endothelial cells (EC) cultivation was collected, to detect its effect on the growth and migration rates of hMSC. hMSCs were induced to differentiate into EC in vitro and the migratory effect on HUVEC was also evaluated., Results: hMSCs-EGFP, like hMSC, exhibited a fibroblast-like morphological feature, and both had the similar cell surface antigens. They could be induced into osteocytes or adipocytes under the conditioned media. The results not only suggested that hMSCs contributed to tumor neovascularization, but also indicated that most of vessels were host-derived angiogenesis mediated by hMSCs. The mean vascular density (MVD) in suspension group (13.67 ± 1.53) was strikely higher than that in MCF-7 group (5.33 ± 1.42), which showed statistical significance (P < 0.05). Only very few vessels were attributed to hMSCs transdifferentiation into ECs. Tumor cells and ECs can promote hMSCs proliferation and migration through paracrine action. Furthermore, hMSCs were positive for CD31 after 2 weeks induction and HUVEC migration can be facilitated by hMSCs., Conclusion: MSCs have the effect of promoting tumor neovascularization.

Objective: To study the changes in bulbar conjunctiva microcirculation (BCM) and the therapeutic effect of Pentoxifylline on BCM disturbance after high-voltage electrical burn (HEB) in rabbits., Methods: Forty-five rabbits were divided into control group (C), electrical burn group (EB), and Pentoxifylline treatment group (PT) according to random number table, with 15 rabbits in each group. Model of HEB was reproduced in rabbits from EB and PT groups with voltage regulator and experimental transformer. Rabbits in C group were sham injured with the same devices without electrification. Changes in BCM were observed with microcirculation microscope at 15 minutes before HEB and 5 minutes, 1, 2, 4, 8 hour(s) post HEB (PHM or PHH), including: (1) morphology of microvessels, such as the discernible, diameters of arterioles, venules, and capillaries, the unevenness in caliber, and ischemic area; (2) dynamic changes in microvascular blood flow, such as blood flow speed in arterioles, venules, and capillaries, erythrocyte aggregation, and microthrombi formation; (3) condition of tissues surrounding microvessel, such as bleeding and exudation. Measurement data were processed with t test; enumeration data were processed with Fisher's exact test., Results: (1) Morphology of microvessel: discernible of microvessels in EB and PT groups was decreased, but that of PT group was better than that of EB group. At PHM 5, diameter of arterioles, venules and capillaries was respectively (7.3+/-2.5), (12.3+/-2.4), (3.5+/-0.7) microm in EB group, all narrower than those of the control group [(14.6+/-3.1), (27.2+/-3.5), (9.0+/-1.4) microm, with t value respectively 5.23, 13.66, 14.04, P values all below 0.05]. Diameters of the microvessels in PT group [(10.2+/-3.8), (21.5+/-3.1), (7.1+/-1.2) microm] were larger than those in EB group (with t value respectively 2.21, 8.99, 10.18, P values all below 0.05). Diameters of arterioles, venules and capillaries in EB and PT groups recovered to the before HEB size at PHH 1. From PHH 2 to 8, arterioles and capillaries decreased gradually in caliber, venules dilated gradually in EB and PT groups, but the changes in PT group were not obvious. Thickness of microvessel was observed uneven in EB group at PHM 5, which lasted until PHH 8. Ischemia of the tissue was observed in EB group at PHM 5, which improved at PHH 2. Situation in PT group was better. (2) Dynamic changes in microvascular blood flow: at PHM 5, blood flow speed in arterioles, venules and capillaries was respectively (202+/-53), (198+/-44), (46+/-12) microm/s in EB group, all slower than those of the control group [(544+/-37), (359+/-32), (220+/-19) microm/s, with t value respectively 20.47, 11.51, 30.02, P values all below 0.05], and those of PT group [(335+/-42), (260+/-35), (119+/-23) microm/s] were faster than those of EB group (with t value respectively 7.55, 4.26, 14.85, P values all below 0.05). Blood flow speed in EB and PT groups recovered to the before HEB level at PHH 1. From PHH 2 to 8, blood flow speed decreased gradually in EB and PT groups, but that of PT group was faster than that of EB group. Erythrocyte aggregation in venules and capillaries was observed in EB group at PHM 5, which eased up at PHH 1, but aggregated at PHH 2, lasting until PHH 8. Obvious microthrombi were observed in EB group at PHH 2, which increased gradually. These changes were less obvious in PT group. (3) Condition of surrounding tissues of microvessel: in EB group, exudation was observed around microvessels at PHH 1, bleeding at PHH 2, with a worsening tendency. Changes in those in PT group were less obvious., Conclusions: HEB causes disturbance in BCM, but it can be ameliorated by Pentoxifylline.

Background and Objective: Radiation sensitivity is closely related to tissue oxygen, and rh-endostatin can induce the high level of oxygen content in tumor by "normalizing" tumor angiogenesis which is associated with radiotherapy sensitivity. The aim of this study is to observe the effect of combination of radiotherapy with rh-endostatin in the rats with lung cancer., Methods: Immediate lewis cancerous ascetic injection method was used to make rats tumors bearing model, then the rats was divided into four groups randomly: group A was treated with saline; group B was treated with rh-endostatin; group C was treated with irradiation and group D was treated with rh-endostatin and irradiation. After all rats were treated, inhibition rates and the tumor growth curve were calculated. Immunohistochemisty was adopted to check the expressions of vascular endothelial growth factor (VEGF) and microvessel density (MVD)., Results: Compared with group A, the growth rates of the tumors in the other group were obviously slower, and the tumor weights were significantly different form group A (P < 0.05). Compared with the other groups, the tumor weights of group D were obviously reduced (P < 0.05). Compared with group A, VEGF and MVD of other three groups were reduced (P < 0.05), and group D were significantly cut down., Conclusion: Combination with radiotherapy and rh-endostatin could inhibit the lung cancer significantly in rats. The possible mechanisms are to decrease the expression ofVEGF and inhibit the production of angiogenesis.

Objective: To analyze the histologic characteristics of macrochilia secondary to port-wine stain and to elucidate the possible mechanism., Methods: Twenty-one cases of macrochilia secondary to venular malformation were included and the histology of the lesions was observed by light microscope., Results: Histological examination revealed vascular abnormalities and a number of widely distributed hamartomatous changes in macrochilia secondary to venular malformation. The average vessel diameter is (39.8 +/- 15.7) microm. The degree of hamartomatous change: mild (1 case), moderate (7 cases) and severe (13 cases)., Conclusions: The complex hamartomatous changes suggest a genetically determined, multilineage developmental field defect in the pathogenesis of venular malformation.

Objective: To investigate the distribution patterns of microvascular density(MVD) and lymph vessel density (LVD) in nasopharyngeal carcinoma, and to discuss their relationships with tumor recurrence-metastasis and prognosis., Method: One hundred and six patients with pathologically proved nasopharyngeal carcinoma were included in this study. Immunostainings for the vascular endothelial marker CD34 and the lymph vessel marker D2-40 were performed. The microvascular and lymph vessels within the tumor and the peritumoral area were respectively quantified with computer assisted morphometric analysis., Result: Both microvascular and lymph vessels were positive in the specimen of 106 nasopharyngeal carcinoma patients, with MVD and LVD higher in the peritumoral area than in the tumor. Univariate analysis showed that intra-tumor MVD,intra-tumor and peritumor LVD were correlated with nasopharyngeal carcinoma recurrence-metastasis (P < 0.05), while no significant correlation was found between tumor recurrence-metastasis and age, gender, T stage, lymph node stage, clinical stage and peritumor MVD (P > 0.05). Multivariate analysis demonstrated that intra-tumor MVD and peri-tumor LVD were two independent risk factors of nasopharyngeal carcinoma recurrence-metastasis. Higher intra-tumor MVD was associated with adverse prognosis of nasopharyngeal carcinoma (P < 0.05)., Conclusion: High MVD and LVD may be of significance in predicting poor prognosis in patients with nasopharyngeal carcinoma. Intra-tumor and peri-tumor microvessels may play different roles in the tumor recurrence and metastasis. Intra-tumor MVD and peri-tumor LVD should be the therapeutic targets in the nasopharyngeal carcinoma.

Objective: The aim of this study was to evaluate the expressions of CD34, CD31 and microvessel density (MVD) in different subtypes of renal cell carcinoma (RCC) as well as the relationship between MVD and clinicopathological factors., Methods: Expressions of CD31 and CD34 were detected in 149 patients with RCC using SP immunohistochemical staining. The MVD was studied by Weidner's method., Results: The expressions of CD31 and CD34 in the clear cell renal cell carcinoma (CCRCC) (98.35 +/- 55.05, 128.04 +/- 46.44) were significantly higher than those in chromophobe renal cell carcinoma (ChRCC) (30.70 +/- 17.72, 48.55 +/- 14.09) and papillary renal cell carcinoma (PRCC) (21.60 +/- 9.38, 38.12 +/- 10.98) (P < 0.01). The MVD value marked by CD31 (30.70 +/- 17.72, 21.60 +/- 9.38) was much lower than that marked by CD34 (48.55 +/- 14.09, 38.12 +/- 10.98) between ChRCC and PRCC (P < 0.01). Smaller and immatured microvessels and even single endothelial cells could be clearly seen. The MVD values marked by CD31 and CD34 were negatively correlated with the pathological grades (r(CD34) = -0.618, P < 0.01; r(CD31) = -0.442, P < 0.01) and clinical stages (r(CD34) = -0.283, P < 0.05; r(CD31) = -0.256, P < 0.05) in CCRCC. But no association was found in non-CCRCC (P > 0.05)., Conclusion: MVD is significantly correlated with different types of endothelial labeling. The microvascular endothelial cells could be shown clearly by its related antigen labeling such as CD34 and CD31. CD34 is more sensitive than CD31. The MVD of CCRCC is significantly higher than that in non-CCRCC. The expressions of CD31 and CD34 are not correlated with tumor grade and stage in ChRCC and PRCC, while there is a negative correlation in CCRCC.

Background and Objective: Studies have shown that nitric oxide (NO) derived from endothelial nitric oxide synthase (eNOS) is expressed widely in tumor tissues and regulates tumor angiogenesis. However, the results are controversial. This study was to investigate the effect of NO on tumor angiogenesis and its mechanism., Methods: C57BL/6 mice inoculated with Lewis lung cancer cells were randomly divided into three groups. Mice in the NO group were inoculated with lung cancer cells transfected with eNOS gene, mice in the L-NAME group with L-NAME, an eNOS antagonist, and mice in the control group with normal saline. Plasma concentration of NO and the number of endothelial progenitor cells (EPCs) in peripheral blood were detected . Tumor vessel density, CD133+ cells, and the expression of VEGF-VEGFR in tumor tissues were also measured., Results: Four weeks after inoculation of Lewis cells, tumor volume was significantly larger in control group [ (3022 +/- 401) mm(3)] than in the L-NAME group [ (1204 +/-97) ) mm(3)] and in the eNOS group [(1824 +/- 239) mm(3)] (P<0.01). eNOS protein and NO production increased significantly in Lewis lung cancer cells transfected with eNOS gene. But the number of CD133-positive cells and vessel density in tumors were significantly lower in the eNOS group than in the control group [(48+/-19) / HPF vs. ( 103 +/- 27)/ HPF, (19+/- 7) HPF vs. (31 +/- 9) HPF, P<0.05]. The number of EPCs in peripheral blood was not statistically different between each group. The levels of NO in blood and tumor tissue significantly decreased after the treatment of L-NAME, while the tumor vessel density reduced to 12+/- 5/ HPF (P<0.01, vs. the control group; P<0.05, vs the eNOS transfected group). The number of EPCs in blood and that of CD133-positive cells in tumor tissue were significantly smaller in the L-NAME group than in the control group (P<0.05)., Conclusion: No derived from eNOS inhibits angiogenesis and tumor growth, which may be due to its suppression on either the mobilization or homing of EPCs via VEGF binding to VEGFR.

Objective: To study the function of MT1-MMP in tumor angiogenesis and elucidate the possible way of action that MT1-MMP contributes to angiogenesis., Methods: MT1-MMP was transfected into human breast carcinoma cell line MCF-7 cells. Semi-quantitative RT-PCR and immunofluorescence staining were employed to detect the expression of VEGF in the transfected and non-transfected MCF-7 cells. Tumor growth, microvessel density and expression of VEGF in nude mice were detected through in vivo tumorigenicity assay., Results: In MT1-MMP stable transfected MCF-7 cells, mRNA expression of VEGF(189), VEGF(165), and VEGF(121) and immunofluorescence intensity were significantly elevated (P < 0.001). In vivo tumorigenicity assay in the nude mice showed that MT1-MMP promoted tumor growth. The MVD in the MT1-MMP-transfected cells-transplanted tumor tissue was significantly elevated (P < 0.05). Immunohistochemical assay showed that there was a strong immunostaining of VEGF in those tumor tissues., Conclusion: MT1-MMP can induce tumor angiogenesis through up-regulation of VEGF expression. This function of MT1-MMP may open a new approach for clinical anti-tumor research and anti-tumor drug development.

Objective: To investigate the expression of Ets-1 in gastric carcinoma, para-cancerous tissue and metastatic lymph nodes, and to determine the relationship between Ets-1 expression and clinicopathological features, angiogenesis and survival of patients with gastric carcinoma., Methods: Gastric carcinoma tissue microarray was used to determine Ets-1 protein expression by SP immunohistochemical staining in 189 advanced gastric cancer, 54 papacancerous tissues, 41 metastatic lymph nodes and 32 control tissues., Results: The positive rates for Ets-1 expression of the carcinoma, paracancerous and control tissues were 71.4%, 29.6% and 18.8%, respectively, with a significant difference among the three groups (P < 0.01). In the cancer tissues, the positive rate of Ets-1 protein expression was significantly associated with depth of invasion and lymph node metastasis (P < 0.01), but not associated with degree of differentiation, Lauren's histological type, sex, age, and size of tumor (P > 0.05). The positive rates for Ets-1 expression of the 41 gastric cancer and 41 metastatic lymph nodes were significantly different (P < 0.05). In metastatic lymph nodes, the positive rate for Ets-1 expression was higher. The MVD in Ets-1 positive tumors was higher than that in the Ets-1 negative tumors, with a significant difference (P < 0.05). Kaplan-Meier survival analysis showed that the survival time of Ets-1-negative patients was longer than that of Ets-1-positive patients (P < 0.05). Cox regression analysis showed that Ets-1 expression was not an independent prognostic factor of gastric carcinoma., Conclusion: A higher expression of Ets-1 is involved in carcinogenesis, development, invasion, and metastasis of gastric cancer. Ets-1 plays an important role in angiogenesis in gastric cancer. Ets-1 is a useful marker for predicting the outcome for patients with gastric carcinoma, though it is not an independent prognostic indicator.

Objective: To investigate the expression of COX-2 and VEGF in clear cell renal cell carcinoma (CCRCC) and their correlation with tumor angiogenesis., Methods: Envision immunohistochemistry was used to determine the expression of COX-2 and VEGF, and microvessel density (MVD) was marked by CD34 in 80 CCRCC tissues and 20 normal kidney tissues. The relationship between the above mentioned markers were analyzed., Results: Expressions of COX-2 and VEGF were noted in both CCRCC and normal kidney tissues. The positive rates of COX-2 and VEGF were significantly higher in CCRCC than in normal kidney (P < 0.05); The expression of COX-2 was correlated with TNM stage (P < 0.05), histological grade (P < 0.05) and lymph node metastasis (P < 0.05) in CCRCC, but not with age (P = 0.663) and diameter of tumor (P = 0.528). Both COX-2 expression (r = 0.851, P < 0.01) and VEGF expression (r = 0.736, P < 0.01) were significantly associated with MVD in CCRCC. There was a positive correlation between expression of cox-2 and that of VEGF in CCRCC., Conclusion: COX-2 expression is correlated with tumor angiogenesis in CCRCC. It is likely that VEGF is one of the most important mediators in the COX-2 angiogenic pathway.

Objective: To investigate the relationship between vascular endothelial growth factor (VEGF) expression and microvessel injury of renal interstitium in children with Henoch-Schönlein purpura nephritis (HSPN)., Methods: Thirty-two children with HSPN and who had not received glucocorticoid or immunodepressants treatment before hospitalization were enrolled. Five children undergoing nephrectomy due to renal trauma were used as the control group. Renal samples were stained by hematoxylin and eosin and renal pathological changes were evaluated semi-quantitatively. CD34 and VEGF expression was detected by immunohistochemistry. CD34 was used as the marker for endothelial cells of renal microvessels. The microvessel density (MVD) was calculated by CD34 immunostaining., Results: Compared with the control and the renal pathological grade II HSPN groups, MVD in the grade III and above HSPN groups decreased significantly, with an obvious reduction in MVD with the increased renal pathological grade (p<0.05). The renal microvessel score in the grades IIIa, IIIb, IV, and V HSPN groups decreased obviously compared with that in the control group. The renal microvessel score decreased with the increased renal pathological grade (p<0.05). VEGF expression in the grade II HSPN group was higher (p<0.05), while that in the grades IV and V HSPN group was lower than that in the control group (p<0.05). VEGF expression in the HSPN group showed a significant reduction with the increased renal pathological grade (p<0.05). There was a positive correlation between VEGF expression and MVD in renal tissue in the HSPN group (r=0.935, p<0.01)., Conclusions: The decreased expression of VEGF may be responsible for the renal pathological damage and microvessel injury in HSPN.

Objective: The aim of this cohort study was to investigate the clinical outcome and prognostic factors in patients after resection for ductal adenocarcinoma of the pancreatic head., Methods: patients with pancreatic head cancer undergoing curative resection (R0) between 1997 and 2002 were included in this study. Univariate and multivariate analyses were performed to examine factors affecting clinical outcome and recurrence of the cancer., Results: Surgical procedures consisted of 58 (43.3%) extended pancreaticoduodenectomies (EPD), 47 (35.1%) pancreaticoduodenectomies (PD) and 29 (21.6%) pylorus-preserving pancreaticoduodenectomies (PPPD). The results showed that 81.3% (109/134) of patients had a recurrence during the study period, mainly retroperitoneal combined with distant metastasis (53.7%). The median postoperative survival time was 24.7 months. The 1-, 3- and 5-year overall survival rates for the study population were 67.1%, 38.5% and 17.6%, respectively. Univariate analysis showed that preoperative abdominal and/or back pain, tumor size > 2 cm, lymph node involvement and vascular invasion, and CA19-9 level were all significant predictors for poor survival. Multivariate analysis also showed that preoperative abdominal and/or back pain, tumor size > 2 cm, lymph node involvement and vascular invasion were all significant predictors for poor survival., Conclusion: Our results suggest that preoperative abdominal and/or back pain, tumor size > 2 cm, lymph node involvement and vascular invasion are significant predictors for poor survival in patients with pancreatic head cancer.

Background and Objective: Lymphatic microvessels may play an important role in tumor metastasis. This study detected lymphatic microvessel density (LVD) in supraglottic laryngeal carcinoma to find its correlation to clinicopathologic characteristics, and to assess its prognostic significance in supraglottic laryngeal carcinoma., Methods: Clinical data of 51 supraglottic laryngeal carcinoma patients who underwent primary lesion resection were reviewed. Immunohistochemical staining was performed with podoplanin monoclonal antibody to recognize lymphatic microvessels. The correlations of LVD to clinicopathologic characteristics and prognosis were analyzed., Results: The mean LVD was 3.25+/-2.37, with a median of 3, in the 51 specimens of supraglottic laryngeal carcinoma. LVD was significantly higher in the patients with cervical lymph node metastasis than in those without (P=0.009). The overall 5-year survival rate was 54.9%. The 5-year survival rate was significantly lower in the patients with poorly differentiated tumor, stages T3-4 disease, lymph node metastasis, stages III-IV disease and high LVD than in their counterparts. Cox regression analysis revealed that LVD, T classification, and differentiation were independent factors for survival., Conclusion: LVD is related to lymph nodes status, and is an independent factor for prognosis of supraglottic laryngeal carcinoma.

Objective: To determine the significance of MSCT perfusion scan on differentiation of NSCLC and to investigate its possible mechanisms., Methods: Forty four NSCLC patients underwent CT perfusion scan by MSCT. Among them, 22 cases were selected to detected the two-dimensional tumor microvascular architecture phenotype (2D-TMAP), the relationships between CT perfusion parameters (BF, BV, PEI, TIP), and the differentiation of NSCLC were analysed by using the correlation analysis and trend test. Spearman correlation analysis was used to study the relationships between CT perfusion parameters, differentiation, and 2D-TMAP., Results: The total BF, BV and PEI decreased with decreasing differentiation of NSCLC (P<0.05). The total PEI showed a positive correlation with the total MVD (P<0.05). There were negative correlations between the surrounding area BF, the total BF, BV, and PEI, the uncomplete lumen of the surrounding area MVD, and expression of PCNA, respectively (P<0.05). There were positive correlations between degree of differentiation and the uncomplete lumen of the surrounding area MVD (P<0.05). It was the same as degree of differentiation and expression of PCNA, VEGF, respectively. There were positive correlations between the uncomplete lumen of the surrounding area MVD and expression of VEGF, ephrinB2, EphB4, and PCNA, respectively (P<0.05)., Conclusion: Perfusion parameters reflect the difference of density of vassels with mature functional lumen. Careful evaluation of the differences of blood flow pattern in pulmonary space-occupying lesions by MSCT perfusion scan can be used to identify the degree of NSCLC differentiation.

Objective: To investigate the expression of EphA2 and EphrinA1 and its relationship with angiogenesis in renal cell carcinoma and its relevance to clinicopathologic features., Methods: The expression of the EphA2 and EphrinA1 was detected by immunohistochemistry (IHC) in the tissues samples from 68 renal cell carcinomas and 24 normal kidneys, and quantitatively analyzed. The microvessel density (MVD) was determined by CD34 immunostaining of microvascular endothelial cells. Statistical analysis was performed using the software SPSS (version 13.0)., Results: The expression of EphA2, EphrinA1 and MND in the cancerous tissues were significantly higher (P<0.01) than that in the normal ones. Significantly increased expression of EphA2, EphrinA1 and MVD (P<0.01) was detected in cancer tissues with higher grade differentiation, more advanced stage and more lymph node metastasis, respectively (P<0.05 for each group). Expression of the EphA2 and EphrinA1 protein was shown to be positively associated with the MVD assessed by Spearman's correlation and factor analysis (r=0.555, r=0.485, P<0.01). The MVD was also significantly correlated with the diameter of the tumor (P<0.01)., Conclusion: EphA2 and EphrinA1 are highly expressed in renal cell carcinoma, and positively correlated with histological differentiation, clinical stage and angiogenesis in the cancer.