Diagnostic performance of a new two-dimensional shear wave elastography expression using siemens ultrasound system combined with ACR TI-RADS for classification of benign and malignant thyroid nodules: A prospective multi-center study.

Objective: The present study aimed to evaluate the efficacy of a new two-dimensional shear wave elastography (2D-SWE) method using a Siemens ultrasound system and its combination with the American College of Radiology Thyroid Imaging Reporting and Data System (ACR TI-RADS) for the differential diagnosis of benign and malignant thyroid nodules. Methods: Conventional ultrasound images and 2D-SWE (E-whole-mean and E-stiffest-mean) were prospectively analyzed in 593 thyroid nodules from 543 patients. Nodules were divided into diameter (D) ≤10 mm and D > 10 mm groups and graded using ACR TI-RADS. The receiver operating characteristic curve was plotted using pathological findings as the gold standard. Diagnostic performance was compared among 2D-SWE, ACR TI-RADS, and their combination. Results: The area under the curve (AUC) for E-whole-mean was higher than that for E-stiffest-mean (0.858 vs. 0.790, P < 0.001), which indicated that it was the better 2D-SWE parameter for differentiating malignant nodules from benign nodules with an optimal cut-off point of 11.36 kPa. In the all-sizes group, the AUC for E-whole-mean was higher than that for ACR TI-RADS (0.858 vs. 0.808, P < 0.001). The combination of E-whole-mean and ACR TI-RADS resulted in a higher AUC (0.929 vs. 0.858 vs. 0.808, P < 0.001), sensitivity (87.0% vs. 80.3% vs. 85.2%), specificity (85.1% vs. 74.0% vs. 73.6%), accuracy (86.3% vs. 78.1% vs. 81.1%), positive predictive value (91.5% vs. 85.1% vs. 85.6%), and negative predictive value (78.0% vs. 67.0% vs. 72.9%) compared to E-whole-mean or ACR TI-RADS alone. The AUC for the combination of 2D-SWE and ACR TI-RADS was superior to that for E-whole-mean or ACR TI-RADS alone in both D ≤ 10 mm and D > 10 mm groups (P < 0.001). Conclusion: As the better 2D-SWE parameter, E-whole-mean had a higher diagnostic power than ACR TI-RADS and enhanced the diagnostic performance of ACR TI-RADS when identifying benign and malignant thyroid nodules. The combination of E-whole-mean and ACR TI-RADS improved the diagnostic performance compared to using ACR TI-RADS alone, providing a new and reliable method for the clinical diagnosis of thyroid nodules.

Convolutional neural network based on automatic segmentation of peritumoral shear-wave elastography images for predicting breast cancer.

Objective: Our aim was to develop dual-modal CNN models based on combining conventional ultrasound (US) images and shear-wave elastography (SWE) of peritumoral region to improve prediction of breast cancer. Method: We retrospectively collected US images and SWE data of 1271 ACR- BIRADS 4 breast lesions from 1116 female patients (mean age ± standard deviation, 45.40 ± 9.65 years). The lesions were divided into three subgroups based on the maximum diameter (MD): ≤15 mm; >15 mm and ≤25 mm; >25 mm. We recorded lesion stiffness (SWV1) and 5-point average stiffness of the peritumoral tissue (SWV5). The CNN models were built based on the segmentation of different widths of peritumoral tissue (0.5 mm, 1.0 mm, 1.5 mm, 2.0 mm) and internal SWE image of the lesions. All single-parameter CNN models, dual-modal CNN models, and quantitative SWE parameters in the training cohort (971 lesions) and the validation cohort (300 lesions) were assessed by receiver operating characteristic (ROC) curve. Results: The US + 1.0 mm SWE model achieved the highest area under the ROC curve (AUC) in the subgroup of lesions with MD ≤15 mm in both the training (0.94) and the validation cohorts (0.91). In the subgroups with MD between15 and 25 mm and above 25 mm, the US + 2.0 mm SWE model achieved the highest AUCs in both the training cohort (0.96 and 0.95, respectively) and the validation cohort (0.93 and 0.91, respectively). Conclusion: The dual-modal CNN models based on the combination of US and peritumoral region SWE images allow accurate prediction of breast cancer.

Nonylphenol Promoted Epithelial-Mesenchymal Transition in Colorectal Cancer Cells by Upregulating the Expression of Regulator of Cell Cycle.

Nonylphenol (NP) is a widely used chemical, which has been considered a kind of endocrine-disrupting chemical and is involved in the occurrence and development of many types of cancers. Our recent studies demonstrated that NP exposure is related to colorectal cancer (CRC) progression. In this study, we also found epithelial-mesenchymal transition (EMT) promoted by NP treatment in CRC cells. However, the mechanism of NP on tumor metastasis is still unclear. In this study, we focused on the effect of the regulator of cell cycle (RGCC) induced by NP treatment. The cancer genome atlas (TCGA) analysis suggested that the expression of RGCC increased in CRC tissues, and our clinical samples showed that the expression of RGCC in tumor tissues is positively correlated with the serum level of NP in CRC patients. Further studies revealed that overexpression of RGCC could enhance the NP-induced EMT process in CRC cells and activate ERK signaling pathways. Inhibiting ERK signaling by ERK inhibitors or the knockdown of RGCC could attenuate the NP-induced EMT process. In addition, both RGCC overexpression and NP treatment could activate ERK pathways and attenuate the effect of ERK inhibitors on the EMT process in CRC cells. Altogether, this study demonstrated that NP could induce cell invasion and migration by increasing the expression of RGCC to enhance the EMT process, which might be through the activation of ERK signaling pathways. This finding supported a potential target for studying NP exposure-related colorectal cancers.

Evaluation of Tissue Stiffness Around Lesions by Sound Touch Shear Wave Elastography in Breast Malignancy Diagnosis.

The aim of the study described here was to assess the evaluation of tissue stiffness around lesions by sound touch shear wave elastography (STE) in breast malignancy diagnosis. This was an institutional ethics committee-approved, single-center study. A total of 90 women with breast masses examined with conventional ultrasound and STE were eligible for enrollment from December 2020 to July 2021. The maximum and mean elastic values of masses, Emax and Emean, were determined. Shell function was used to measure the maximum and mean elastic values of tissues around masses in annular shells 0.5, 1.0, 1.5 and 2.0 mm wide, recorded as corresponding Emax-shell and Emean-shell. All parameters were analyzed and compared with histopathologic results. Receiver operating characteristic curves were constructed to assess diagnostic performance. Logistic regression analysis was conducted to determine the best diagnostic model. Collagen fiber content of tissues around breast lesions was evaluated using Masson staining and ImageJ software. Ninety women with breast masses were included in this study; 50 had benign (mean diameter 15.84 ± 4.39 mm) and 40 had malignant (mean diameter 17.40 ± 5.42 mm) masses. The diagnostic value of Emax-shell-2.0 was the highest (area under the curve = 0.930) with a sensitivity of 87.5% and specificity of 88%. According to stepwise logistic regression analysis, Emax-shell-2.0 and age were independent predictors of malignancy. Emax-shell-2.0 was also found to be highly correlated with the collagen fiber content of tissue in the malignant group (r = 0.877). Tissue stiffness around lesions measured by STE is a useful metric in identifying malignant breast masses by reflecting collagen fiber content, and Emax-shell-2.0 performs best.

Analysis of Risk Factors Associated With Central Lymph Node Metastasis in Papillary Thyroid Carcinoma With cT1N0 Stage.

Aim: Annual T1 stage papillary thyroid carcinoma (PTC) incidence rates continue to rise, yet the optimal treatment for this cancer type remains controversial. Central lymph node metastasis (CLNM) is a critical determinant in the context of treatment decision-making. While several prior studies have evaluated patients with clinica l T1a(cT1a) stage PTC, there have been fewer analyses of clinical T1b(cT1b) disease to date. The present study was thus formulated to explore predictors of CLNM in patients with cT1a and cT1b stage PTC. Methods: A retrospective analysis of data including clinicopathological characteristics and BRAFV600E mutation status was conducted for 452 PTC patients undergoing surgical treatment. Logistic univariate and multivariate analyses were performed to identify risk factors associated with CLNM in particular patients' characteristics and the accuracy of the established logistic regression models was evaluated using the R software platform. Results: Respective CLNM incidence rates in cT1a and cT1b disease were 39.39% and 67.21%. Factors associated with a higher risk of CLNM among PTC(cT1a) patients included male sex, young age, tumor size, contact with capsule, and multifocality as determined through comparisons of the area under the curve for logistic regression models. Whereas male sex and age were associated with CLNM risk in PTC(cT1b) patients in univariate and multivariate analyses, age was the only risk factor associated with CLNM incidence among women with PTC(cT1b). Conclusion: Predictors of CLNM differ between PTC patients with cT1a and cT1b stage disease, and a comprehensive assessment of these risk factors should thus be conducted when designing individualized treatment regimens for PTC patients.

Ectopic Schistosomiasis Involving Axillary Lymph Nodes.

Axillary lymph node schistosomiasis is a form of ectopic schistosomiasis, in which the Schistosoma adult worms or eggs are infected in the axillary lymph nodes. A 53-year-old woman visited our hospital with a small mass in the right armpit, which had been present for over 1 month. Histological examination of biopsy samples from the right axillary lymph nodes revealed granulomas containing calcified Schistosoma eggs. This is the first authentic case of ectopic schistosomiasis identified in the axillary lymph nodes.

Coiled-Coil Domain Containing 80 Suppresses Nonylphenol-Induced Colorectal Cancer Cell Proliferation by Inhibiting the Activation of ERK1/2.

Recently, the effect of endocrine-disrupting chemicals on the cancer procession has been a concern. Nonylphenol (NP) is a common environmental estrogen that has been shown to enhance the proliferation of colorectal cancer (CRC) cells in our previous studies; however, the underlying mechanism remains unclear. In this study, we confirmed the increased concentration of NP in the serum of patients with CRC. RNA sequencing was used to explore the differentially expressed genes after NP exposure. We found 16 upregulated genes and 12 downregulated genes in COLO205 cells after NP treatment. Among these differentially expressed genes, we found that coiled-coil domain containing 80 (CCDC80) was downregulated by NP treatment and was associated with CRC progression. Further experiments revealed that the overexpression of CCDC80 significantly suppressed NP-induced cell proliferation and recovered the reduced cell apoptosis. Meanwhile, the overexpression of CCDC80 significantly inhibited the activation of ERK1/2 induced by NP treatment. ERK1/2 inhibitor (PD98059) treatment also suppressed NP-induced CRC cell growth, but the overexpression of CCDC80 did not enhance the effect of ERK1/2 inhibitor. Taken together, NP treatment significantly inhibited the expression of CCDC80, and the overexpression of CCDC80 suppressed NP-induced CRC cell growth by inhibiting the activation of ERK1/2. These results suggest that NP could induce CRC cell growth by influencing the expression of multiple genes. CCDC80 and ERK1/2 inhibitors may be suitable therapeutic targets in NP-related CRC progression.

Exosomal miR-487a derived from m2 macrophage promotes the progression of gastric cancer.

Tumor-associated macrophages contribute to cell growth, development, and metastasis in various cancers. However, the underlying mechanisms of M2 macrophage that modulate the progression of gastric cancer (GC) remain largely unknown. In this study, we detected the ratio of macrophages in GC tissues and found that the proportion of M2 macrophages was increased in GC tissues. We then co-cultured GC cells with M1 and M2 macrophages, respectively, and then assessed cell proliferation and tumorigenicity of GC cells by MTT and colony formation assay. The results indicated that M2 macrophages promoted the proliferation of GC cells, but M1 not. Besides, GW4869, an exosomes inhibitor, reduced the effects induced by M2 macrophage. Then, we isolated and identified exosomes derived from M1 and M2 macrophage, and confirmed that the exosomes could be taken up by GC cells. We demonstrated that M2 macrophage-exosomes could induce the proliferation and tumorigenesis in vitro and in vivo. Moreover, miR-487a was enriched in M2 macrophage-exosomes and further determined that miR-487a exert the functions by targeting TIA1. In conclusion, exosomal miR-487a derived from M2 macrophage promotes the proliferation and tumorigenesis in gastric cancer, and the novel findings might be helpful to the development of novel diagnostic and therapeutic methods in GC.

Evaluation of the Perinodular Stiffness Potentially Predicts the Malignancy of Thyroid Nodules.

OBJECTIVES: To evaluate the surrounding tissue stiffness measured by sound touch elastography for differential diagnosis of thyroid nodules (TNs). METHODS: Thirty-nine benign and 90 malignant TNs were included in this study. The conventional ultrasound features, the maximum Young modulus value of the stiffness of the TNs (recorded as E), and the stiffness of the 0.5-, 1.0-, 1.5-, and 2.0-mm perinodular regions of the TNs (recorded as Eshell0.5 , Eshell1.0 , Eshell1.5 , and Eshell2.0 , respectively) were prospectively analyzed and compared to histopathologic results. The abundance of collagen fibers at various widths in the perinodular regions of the TNs was evaluated by Masson staining and ImageJ software (National Institutes of Health, Bethesda, MD). The fibrous structures in the perinodular regions of the TNs were classified. RESULTS: The various Eshell values of malignant TNs were significantly higher than those of benign TNs (P < .001 for all). Eshell0.5 correlated highly with E in the malignant TNs and in all samples (r = 0.722 and 0.772; P < .001 for both). Eshell2 yielded the highest area under the receiving operating characteristic curve value (0.96) for the differential diagnosis of TNs. The abundance of collagen fibers in the 2-mm perinodular region of the TNs was closely correlated with Eshell2 in the malignant TNs and in all samples (r =0.729 and 0.867; P < .001). The Eshell2 values for different levels of disorder of the tissue surrounding TNs were significantly different (P < .01 for all). CONCLUSIONS: Perinodular stiffness measured by sound touch elastography improved the diagnostic accuracy in TNs.

Hepatic angiomyolipoma: contrast patterns with SonoVue-enhanced real-time gray-scale ultrasonography.

This study was conducted to retrospectively evaluate the pattern of contrast enhancement with SonoVue on gray-scale ultrasonography of hepatic angiomyolipoma (HAML). Imaging features of 33 pathologically proven HAML lesions in 33 patients who underwent baseline ultrasound and contrast-enhanced ultrasonography (CEUS) were assessed retrospectively. All lesions were enhanced in the arterial phase and showed whole-tumor filling in. Thirty-two of 33 (97%) lesions showed early positive enhancement in the arterial phase. Twenty-three of these exhibited isoechoic or hyperechoic features in the portal phase. HAML demonstrate characteristic manifestations with SonoVue-enhanced real-time gray-scale ultrasonography.

[Relationship between TIM-4 polymorphism and childhood asthma].

OBJECTIVE: To elucidate whether the polymorphism of asthma immune regulator gene TIM-4 is associated with the risk of childhood allergic asthma in the southwest region of China. METHODS: TIM-4 gene promoter region RS6882076 and intron RS4704727 were studied. PCR-RFLP was used to test the genotypes of two polymorphism loci among 579 cases (average 7.2 years old) of asthma and 524 controls (average 7.6 years old) in a case-control study. RESULTS: There were significant differences in the frequency of gene types at RS4704727 site between the asthma and the control groups (P<0.01). The results of PCR-RFLP showed that the polyporphisms of RS6882076 and RS4704727 in TIM-4 gene were present in this study population. The frequency of T allele at the RS4704727 site in the asthma group was significantly lower than that in the control group (OR=1.603; 95%CI 1.304-1.971; P<0.01). There were no significant differences in the frequencies of gene types and allele at RS6882076 site between the two groups (P>0.05). CONCLUSIONS: RS4704727 polymorphism of TIM-4 gene may be associated with childhood asthma, providing a better understanding of the pathogenesis of childhood asthma in the Southwest region of China.

Antioxidant, antiproliferative, and pro-apoptotic activities of a saponin extract derived from the roots of Panax notoginseng (Burk.) F.H. Chen.

Dietary and medicinal uses of Panax notoginseng have been associated with reduced risk of cancer. This study was designed to investigate the profiles of P. notoginseng saponin extract (PNSE), the major bioactive ingredients in P. notoginseng (Burk.) F.H. Chen, by high-performance liquid chromatography, and, for the first time, the anticancer effect of PNSE in the human colon cancer cell line LoVo was further evaluated. The major saponins present in PNSE were ginsenosides Rg1 (31.1%) and Rb1 (34.4%), and the total content of the eight saponins identified (notoginsenoside R1, ginsenosides Rg1, Re, Rb1, Rc, and Rd, and isomeric ginsenosides Rb2 and Rb3) was 81.7%, indicating that it was a highly purified standardized saponin extract. Furthermore, PNSE was found to have a markedly cytotoxic effect and antiproliferative activity against the LoVo cell line in a dose- and time-dependent manner. Flow cytometry analysis demonstrated that PNSE caused cell cycle arrest at S phase. Moreover, PNSE was found to possess antioxidative capacities in the 1,1-diphenyl-2-picrylhydrazyl free radical scavenging assay and hydroxyl radical scavenging assay in vitro. Taken together, the present results suggest that naturally occurring PNSE may provide significant natural defense against human colon cancer.

Two clinically relevant pressures of carbon dioxide pneumoperitoneum cause hepatic injury in a rabbit model.

AIM: To observe the hepatic injury induced by carbon dioxide pneumoperitoneum (CDP) in rabbits, compare the effects of low- and high-pressure pneumoperitoneum, and to determine the degree of hepatic injury induced by these two clinically relevant CDP pressures. METHODS: Thirty healthy male New Zealand rabbits weighing 3.0 to 3.5 kg were randomly divided into three groups (n = 10 for each group) and subjected to the following to CDP pressures: no gas control, 10 mmHg, or 15 mmHg. Histological changes in liver tissues were observed with hematoxylin and eosin staining and transmission electron microscopy. Liver function was evaluated using an automatic biochemical analyzer. Adenine nucleotide translocator (ANT) activity in liver tissue was detected with the atractyloside-inhibitor stop technique. Bax and Bcl-2 expression levels were detected by western blotting. RESULTS: Liver functions in the 10 mmHg and 15 mmHg experimental groups were significantly disturbed compared with the control group. After CDP, the levels of alanine transaminase and aspartate transaminase were 77.3 ± 14.5 IU/L and 60.1 ± 11.4 IU/L, respectively, in the 10 mmHg experimental group and 165.1 ± 19.4 IU/L and 103.8 ± 12.3 IU/L, respectively, in the 15 mmHg experimental group, which were all higher than those of the control group (P < 0.05). There was no difference in pre-albumin concentration between the 10 mmHg experimental group and the control group, but the pre-albumin level of the 15 mmHg experimental group was significantly lower than that of the control group (P < 0.05). No significant differences were observed in the levels of total bilirubin or albumin among the three groups. After 30 and 60 min of CDP, pH was reduced (P < 0.05) and PaCO2 was elevated (P < 0.05) in the 10 mmHg group compared with controls, and these changes were more pronounced in the 15 mmHg group. Hematoxylin and eosin staining showed no significant change in liver morphology, except for mild hyperemia in the two experimental groups. Transmission electron microscopy showed mild mitochondrial swelling in hepatocytes of the 10 mmHg group, and this was more pronounced in the 15 mmHg group. No significant difference in ANT levels was found between the control and 10 mmHg groups. However, ANT concentration was significantly lower in the 15 mmHg group compared with the control group. The expression of hepatic Bax was significantly increased in the two experimental groups compared with the controls, but there were no differences in Bcl-2 levels among the three groups. Twelve hours after CDP induction, the expression of hepatic Bax was more significant in the 15 mmHg group than in the 10 mmHg group. CONCLUSION: A CDP pressure of 15 mmHg caused more substantial hepatic injury, such as increased levels of acidosis, mitochondrial damage, and apoptosis; therefore, 10 mmHg CDP is preferable for laparoscopic operations.

Hepatic injury induced by carbon dioxide pneumoperitoneum in experimental rats.

AIM: To observe the hepatic injury induced by carbon dioxide pneumoperitoneum in rats and to explore its potential mechanism. METHODS: Thirty healthy male SD rats were randomly divided into control group (n = 10), 0 h experimental group (n = 10) and 1 h experimental group (n = 10) after sham operation with carbon dioxide pneumoperitoneum. Histological changes in liver tissue were observed with hematoxylin-eosin staining. Liver function was assayed with an automatic biochemical analyzer. Concentration of malonyldialdehyde (MDA) and activity of superoxide dismutase (SOD) were assayed by colorimetry. Activity of adenine nucleotide translocator in liver tissue was detected with the atractyloside-inhibitor stop technique. Expression of hypoxia inducible factor-1 (HIF-1) mRNA in liver tissue was detected with in situ hybridization. RESULTS: Carbon dioxide pneumoperitoneum for 60 min could induce liver injury in rats. Alanine aminotransferase and aspartate aminotransferase were 95.7 +/- 7.8 U/L and 86.8 +/- 6.9 U/L in 0 h experimental group, and 101.4 +/- 9.3 U/L and 106.6 +/- 8.7 U/L in 1 h experimental group. However, no significant difference was found in total billirubin, albumin, and pre-albumin in the three groups. In 0 h experimental group, the concentration of MDA was 9.83 +/- 2.53 micromol/g in liver homogenate and 7.64 +/- 2.19 micromol/g in serum respectively, the activity of SOD was 67.58 +/- 9.75 nu/mg in liver and 64.47 +/- 10.23 nu/mg in serum respectively. In 1 h experimental group, the concentration of MDA was 16.57 +/- 3.45 micromol/g in liver tissue and 12.49 +/- 4.21 micromol/g in serum respectively, the activity of SOD was 54.29 +/- 7.96 nu/mg in liver tissue and 56.31 +/- 9.85 nu/mg in serum, respectively. The activity of ANT in liver tissue was 9.52 +/- 1.56 in control group, 6.37 +/- 1.33 in 0 h experimental group and 7.28 +/- 1.45 (10(-9) mol/min per gram protein) in 1 h experimental group, respectively. The expression of HIF-1 mRNA in liver tissue was not detected in control group, and its optical density difference value was 6.14 +/- 1.03 in 0 h experimental group and 9.51 +/- 1.74 in 1 h experimental group, respectively. CONCLUSION: Carbon dioxide pneumoperitoneum during the sham operation can induce hepatic injury in rats. The probable mechanisms of liver injury include anoxia, ischemia reperfusion and oxidative stress. Liver injury should be avoided during clinical laparoscopic operation with carbon dioxide pneumoperitoneum.

[Changes of rabbit IL-1 and TNF-alpha, etc cytokines in response to acute normovolemic hemodilution with HAES-balanced solution as diluting agent].

AIM: To observe effect of acute normovolemic hemodilution(ANH) with HAES-balanced solution as diluting agent on levels of cytokines including IL-1, IL-2, IL-6 and TNF-alpha in rabbit serum so as to provide theoretical basis for clinical application. METHODS: A total of 20 healthy adult rabbits were enrolled in the study and randomly divided into two groups (10 rabbits per group), i.e., control group (Group C) and HAES group (Group H). Under anesthesia of the rabbits, we performed incision of trachea, high-frequency jet ventilation and liberation of femoral artery and femoral veins. Group C was free from hemodilution. Group H was injected with dilution (2-fold of blood letting volume) via femoral veins during blood letting of the femoral artery. 6% HAES-steril plus compound solution of sodium lactate, with crystal/gel ratio of 2:1, blood letting volume = TBV x (Ho-Hf)/Hav. All blood was transfused back 60-120 min after blood letting. Venous blood was collected before blood letting (T0) and 30 min (T1), 60 min (T2), 120 min (T3) and 24 h(T4) after blood letting to detect Hb and Hct and measure level of IL-1, IL-2, IL-6 and TNF-alpha in serum. RESULTS: In Group H, levels of IL-1, IL-2, IL-6 and TNF-alpha in serum were increased from T1 after ANH, reached peak at T3 but showed decrease at T4, with significant difference compared with Group C at T1, T2, T3 and T4 (P < 0.01) and significant difference compared with those before ANH (P <0.01). In Group C, there was no significant difference upon IL-1, IL-2, IL-6 and TNF-alpha in serum at different time points. CONCLUSION: ANH with HAES-balanced solution as diluting agent can up-regulate the levels of cytokines IL-1, IL-2, IL-6 and TNF-alpha in rabbit serum. In the meantime, ANH may arouse eustress with low intensity and short action time, which exerts effect of enhancing immune function of the organisms.

[Human fetal liver nonparenchymal mesenchymal stem cells differentiate into functional hepatocyte-like cells in vitro].

OBJECTIVE: To induce nonparenchymal mesenchymal stem cells (NPMSCs) differentiating into functional hepatocyte-like cells in vitro, and to identify the molecular biology and functional characteristics of those hepatocyte-like cells. METHODS: Human NPMSCs were isolated and cultured with cell culture technique. NPMSCs were induced (on 1% Matrigel as a matrix and then submitted to 2.5 mmol/L AZA pretreatment for 10-12 h), by adding HGF 10 microg/L + FGF4 10microg/L + HGM into the culture medium. The characteristics of proliferation and growth of human NPMSCs were studied with methyl thiazolyl tetrazolium (MTT). The phenotypes of NPMSCs were identified by flow cytometry, immunohistochemistry and RT-PCR. Albumin (Alb) levels in culture supernatants were determined with ELISA. Staining for glycogen of undifferentiated NPMSCs and NPMSCs derivated hepatocyte-like cells was conducted with periodic acid-Schiff (PAS) test. RESULTS: Growth and division of adherent cells obtained from fetal livers were good and the amount of NPMSCs resourced from each fetus could be amplified to 109 cells after 10 serial subcultivations. The phenotype of NPMSCs was CD166 positive and CD34 negative. The shape of NPMSCs plated on Matrigel with FGF4 and HGF changed from long fusiform to polygonal or round on days 21-28. The rate of cell rounding was 40% and the ratio of dikaryocytes was 5%. Immunohistochemical and RT-PCR detection showed that undifferentiated NPMSCs expressed few alpha fetoprotein (AFP) and AFP mRNA, and did not express any of the liver-specific transcription factors or cytoplasmic markers. Many cells in early induction expressed GATA4, AFP and CK18 proteins and their mRNAs, and their expressions were reduced at the late induction, but the expressions of Alb, CK18, GST-and hepatocyte transcription factor HNF1increased gradually. The ratio of Alb and CK18 positive cells was 60%. Undifferentiated NPMSCs did not produce Alb. Alb production by induced NPMSCs increased in a time-dependent manner. Glycogen storage was first seen on day 14, and maximum levels were seen after day 28. CONCLUSIONS: There are MSCs among nonparenchymal cells of fetal livers. A high ratio of hepatocyte-like cells was obtained under our induction condition. NPMSCs differentiate firstly into hepatocyte precursors, and then differentiate into mature hepatocytes and hepatocyte-like cells with positive hepatocyte markers. The induced NPMSCs have hepatocyte specific functional features.

[Isolation, cultivation and biological identification of human fetal marrow mesenchymal stem cells].

OBJECTIVE: Noting the morphological and cytobiology characteristics and phenotypes of MMSCs, to establish an isolation and culture method for fetal MMSCs in order to provide a source of marrow mesenchymal stem cells (MMSCs). METHODS: Fetal MMSCs were isolated and cultured with in vitro cell culture technique; the characteristics of the proliferating and growing fetal MMSCs were studied with MTT and image analysis; the phenotypes of MMSCs were identified by flow cytometry and immunohistochemistry. RESULTS: Bone marrow of 12 fetuses was isolated within 0.5-2 h, and about 300+/-80 adherent cells were obtained at 24 h. Colonies with more than 5 cells were 15+/-6, growth detention period of culture cell was at 1-3 d after planting, log phase growth period was at day 4, and the amount of disintegration phase cells was reduced significantly. Original culture and serial subcultivations showed that cells divided prosperiously; unequal divisions special for stem cells were observed, and the amount of MMSCs harvested from each fetus was as much as 10(11)-10(12) cells after 10 serial subcultivations. The phenotype of MMSCs was CD166 positive and CD34 negative. Serial subcultivated MMSCs expressed a microamount of AFP and did not express albumine or CK18. CONCLUSION: Fetal MMSCs are easily isolated and proliferate prosperouly. Serial subcultivated MMSCs did not differentiate into hepatocyte-like cells under common culture condition and are feasibile as seed cells for tissue engineering reconstruction.

Imaging of focal liver lesions: low-mechanical-index real-time ultrasonography with SonoVue.

OBJECTIVE: The purpose of this study was to evaluate the usefulness of a contrast-enhanced contrast-specific ultrasonographic technique with a low mechanical index for characterization of focal liver lesions. METHODS: Contrast-specific ultrasonography was used to assess 144 patients with 147 focal liver lesions: 87 primary liver carcinomas, 27 hemangiomas, 16 focal nodular hyperplasias, 5 hepatic abscesses, 3 inflammatory pseudotumors of the liver, and 9 metastases. A sulfur hexafluoride gas-based contrast agent was used with a mechanical index of 0.08 to 0.11. RESULTS: On contrast-enhanced ultrasonography, the typical hemodynamic pattern of primary liver carcinoma was the whole-lesion enhancement or mosaic enhancement in the arterial phase with an enhancement defect in the late phase (sensitivity, 92.0%; specificity, 86.7%). The most common enhancement pattern of hemangioma was that enhancement appeared in the periphery first and progressively filled into the lesion center (sensitivity, 96.3%; specificity, 97.5%). The enhancement pattern of focal nodular hyperplasia was that the whole lesion enhanced early and rapidly in the arterial phase with a centrifugal radiating configuration and appeared isoechoic or hyperechoic until the late phase (sensitivity, 87.6%; specificity, 94.5%). The central scar was detected in 31.3% of cases in the late phase. The specific enhancement of a hepatic abscess was the honeycomblike enhancement in all phases (sensitivity, 80.0%; specificity, 100%). No enhancement of a lesion in all phases was specific for an inflammatory pseudotumor of the liver. CONCLUSIONS: Contrast-enhanced real-time ultrasonography is a promising approach in the noninvasive characterization of focal liver lesions and can be useful as a first-line imaging technique clinically when a focal liver lesion is detectable on ultrasonography.

[Clinical and pathological features of borderline ovarian tumors].

OBJECTIVE: To study the relationship between clinical pathological factors and prognosis. METHODS: Thirty-four cases with borderline ovarian tumors (BOT) and 30 cases with stage I ovarian epithelial cancer admitted in our hospital from Jan.1973 to Dec.2000 were studied retrospectively. All the cases were diagnosed according to the International Histological Classification and Staging of Ovarian Tumors (WHO, 1999). RESULTS: Thirty-seven cases were finally diagnosed of BOT, 6 cases with "microinvasive" had been misinterpreted as stage I ovarian cancer and one had "non-invasive peritoneal implants". Serous (38%) and mucinous (51%) tumors were dominant type of BOT and 95% of tumors were at stage I (International Federation of Gynecology and Obstetrics, FIGO). All patients were operated, 11 cases given conservative surgery with a recurrence rate of 9%, 26 cases had adjuvant chemotherapy, cyclophosphamide + adriamycin + cis-platinum used mostly. 5-year and 10 year survival rates were both 100%. The prognosis of BOT is related to pathology and adjuvant therapy. CONCLUSION: Surgery is the main treatment choice of BOT, and the criteria of chemotherapy must be adequately used.