Risk stratification of gastric cancer screening in community population based on oral contrast-enhanced ultrasonography examination: A 3-year follow-up analysis report.

Objective: This study aimed to retrospectively investigate the use of oral contrast-enhanced ultrasonography (O-CEUS) in assessing the thickness of the gastric wall for gastric cancer (GC) screening and to establish screening strategies for GC with different risk stratifications based on the gastric wall thickness. Methods: From January 2015 to March 2020, people who underwent O-CEUS at the Physical Examination Center of our hospital with at least three years of follow-up were included in this study. The thickness of the gastric wall measured by O-CEUS was divided into three groups using 6 mm and 9 mm as cutoff values. The occurrence of GC in each group was observed. The imaging and clinical information of these populations were recorded and analyzed. Kaplan-Meier survival analysis and Cox's proportional hazards regression were performed to calculate the risk of GC occurrence. Results: A total of 4,047 people were finally included in this study. During the follow-up period, GC occurred in 7 individuals (incidence rate 0.17%). Among them, according to the thickness of the gastric wall, one case occurred in Group A (< 6 mm), two cases occurred in Group B (6-9 mm), and four cases occurred in Group C (>9mm). Based on Kaplan-Meier survival analysis, the curves of the three groups were significantly different (P < 0.01). The risk of GC occurrence in Group C and Group B were higher than that in Group A (4.76E+2-fold and 1.50E+2-fold). Conclusion: O-CEUS is a convenient, economical, safe, and noninvasive screening method for GC. Measuring the thickness of the gastric wall is helpful to predict the risk of GC occurrence according to our stratification screening system.

Efficacy and Safety of Microwave Ablation Assisted by Ultrasound Fusion Imaging for Primary and Secondary Liver Cancers with a Diameter of 3-7 Cm.

Purpose: To investigate the efficacy and safety of microwave ablation (MWA) assisted by ultrasound fusion imaging (FI) for primary and secondary liver cancers with a diameter of 3-7 cm. Patients and Methods: A retrospective analysis was conducted on patients with primary and secondary liver cancers (3-7 cm) who underwent MWA with ultrasound FI assistance in our hospital from April 2020 to May 2022. Technical success, technique efficacy, local tumor progression (LTP), major complication, intrahepatic distant recurrence (IDR), and overall survival (OS) were assessed during the follow-up period. In addition, the ablation results of tumors between the medium-sized group (3.1-5.0 cm) and large-sized group (5.1-7.0 cm) were compared. Results: 31 patients with 35 primary and secondary liver cancers were treated with MWA assisted by ultrasound FI. Complete ablation was achieved in 34 lesions with a technical success rate of 97.1%. Major complications occurred in 6.5% of patients (2/31), while no ablation-related deaths were reported. The median follow-up time of this study was 24 months (range:10 to 35 months). The technique efficacy rate was 97.1% (34/35), with LTP occurring in three lesions at a rate of 8.8% (3/34). The incidence of IDR was 38.7% (12/31) and the 2-year cumulative OS rate reached 96.7%. Moreover, there were no statistical differences in technique efficacy rate (p=0.286), LTP rate (p=0.328), major complication rate (p=0.503), IDR (p=0.857), and OS (p=0.118) between medium-sized group and large-sized group. Conclusion: Ultrasound FI-assisted MWA has the potential to be an effective and safe therapeutic strategy for primary and secondary liver cancers ranging from 3-7 cm in size.

Prediction of ablation zone disappearance after microwave ablation for the patients with papillary thyroid microcarcinoma using nomograph.

Objectives: To construct a prognostic nomogram to predict the ablation zone disappearance for patients with papillary thyroid microcarcinoma (PTMC) after microwave ablation (MWA). Materials and methods: From April 2020 to April 2022, patients with PTMC who underwent MWA treatment were collected retrospectively. Ultrasound (US) or contrast-enhanced ultrasound (CEUS) was performed at 1 day, 1, 3, 6, 12, 18 and 24 months after MWA to observe the curative effect after ablation. The volume, volume reduction rate (VRR) and complete disappearance rate of the ablation zone at each time point were calculated. Univariate and multivariate logistic regression analysis were used to determine the prognostic factors associated with the disappearance of the ablation zone after MWA, and the nomogram was established and validated. Results: 72 patients with PTMCs underwent MWA were enrolled into this study. After MWA, no tumor progression (residual, recurrence or lymph node metastasis) and major postoperative complications occurred. The ablation zone in 28 (38.89%) patients did not completely disappear after MWA in the follow-up period. Three variables, including age (odds ratio [OR]: 1.216), calcification type (OR: 12.283), initial maximum diameter (OR: 2.051) were found to be independent prognostic factors predicting ablation zone status after MWA by multivariate analysis. The above variables and outcomes were visualized by nomogram (C-index=0.847). Conclusions: MWA was a safe and effective treatment for PTMC. Older patients with macrocalcification and larger size PTMCs were more unlikely to obtain complete disappearance of ablation zones. Incomplete disappearance of ablation zone was not related to recurrence.

Radiofrequency Ablation of Hepatocellular Carcinomas Adjacent to the Gallbladder Without Isolation Under Contrast-Enhanced Ultrasound Monitoring: A Comparative Study with Long Term Follow-Up.

Objective: This study intends to compare the efficacy and safety between patients undergoing invasive isolation or monitoring measures and patients undergoing intra-operative contrast-enhanced ultrasound (CEUS) monitoring who underwent radiofrequency ablation (RFA) of hepatocellular carcinomas (HCC) adjacent to the gallbladder (GB). Methods: We retrospectively assessed patients with HCC adjacent to the GB who underwent ultrasound-guided RFA. They were divided into two groups: group A was monitored under intra-operative CEUS, while group B was assisted by invasive auxiliary means. The efficacy, complications and survival were followed up and compared. Results: Thirty-eight patients with 39 HCCs were enrolled into group A and 31 patients with 35 HCCs were enrolled into group B. The technique efficacy rates were both 100% in the two groups. There were no significant differences of the cumulative 1-, 3-, and 5-year local tumor progression, tumor-free survival and overall survival between the two groups (P = 0.851, 0.081 and 0.700, respectively). There were no significant differences of major and minor complications rates between the two groups (P = 1.000, 0.994, respectively). More importantly, no GB related complications occurred in group A. Conclusion: Intra-operative CEUS monitoring without protective isolation of the GB might be also a potentially safe and effective method for the RFA of HCC adjacent to the GB, when compared with those assisted with invasive auxiliary means.

Improving Ablation Safety for Hepatocellular Carcinoma Proximal to the Hilar Bile Ducts by Ultrasound-MR Fusion Imaging: A Preliminary Comparative Study.

AIM: To explore whether ablation safety could be improved by ultrasound (US)-magnetic resonance (MR) fusion imaging for hepatocellular carcinoma (HCC) proximal to the hilar bile ducts (HBDs) through a preliminary comparative study. METHODS: Between January 2014 and June 2019, 18 HCC nodules proximal to the HBDs were included in a US-MR fusion imaging-assisted radiofrequency ablation (RFA) group (study group), while 13 HCC nodules in a similar location were included as a control group. For the study group, the tumor and adjacent bile ducts were outlined on preprocedural MR images. Procedural ablation planning was conducted to assess the feasibility of ablating the tumors while avoiding biliary injury. Such tumors were then ablated under US-MR fusion imaging guidance. The control group nodules were ablated under conventional ultrasound guidance. Baseline characteristics and outcomes were compared between the groups. RESULTS: After preprocedural assessment, 14 of 18 patients with tumors that were feasible to ablate underwent US-MR fusion imaging-assisted RFA. No biliary complications were observed in these 14 patients; the complication rate was significantly lower in the study group than in the control group (30.8%, 4/13) (P = 0.041). There was no significant difference in the technique efficacy rates [92.9% (13/14) versus 100% (13/13), P = 1] or local progression rates [7.1% (1/14) versus 7.7% (1/13), P = 1] between the study and control groups. CONCLUSIONS: US-MR fusion imaging may be a non-invasive means for assisting RFA of HCC nodules proximal to the HBDs and ensuring ablation safety.

Radiofrequency ablation of liver cancers adjacent to the gallbladder without gallbladder isolation under contrast-enhanced ultrasound monitoring: a preliminary study.

PURPOSE: The purpose of this study was to investigate the feasibility, safety and efficacy of intra-procedural contrast-enhanced ultrasound (CEUS) monitoring of the radiofrequency ablation (RFA) of liver cancers adjacent to gallbladder (GB) without GB isolation. MATERIALS AND METHODS: From May 2016 to July 2017, patients with liver cancers adjacent to GB (≤10 mm) who intended to undergo ultrasound-guided RFA without GB isolation in our hospital were prospectively enrolled. During the RFA procedures, CEUS was employed to evaluate the therapeutic response and the perfusion of the intact GB wall. The outcomes of GB and liver cancers were followed up and recorded. RESULTS: 23 patients (18 male, 5 female) with 23 liver cancers (mean 18 mm, range 8-34 mm) adjacent to GB were enrolled. There were 12 tumors that abutted the GB while 11 tumors located within 10 mm of the GB. After the RFA procedures, intra-procedural CEUS evaluation demonstrated the perfusion of the GB wall was intact in all 23 patients and technical success rate of RFA was 100% (23/23). According to the contrast-enhanced CT/MR one month after RFA, the technical efficacy rate was 100% (23/23). During the follow-up period (range: 12-23 months, median: 17 months), no local tumor progression occurred and no major complications arised. Overall survival at 1-year was 100%. Thickening of GB wall was detected in 11 patients. The thickness of GB wall returned to the pre-ablation level in five patients. CONCLUSION: CEUS-monitored RFA of liver cancers adjacent to GB without GB isolation was feasible, safe and effective.

Evaluating histologic differentiation of hepatitis B virus-related hepatocellular carcinoma using intravoxel incoherent motion and AFP levels alone and in combination.

PURPOSE: To evaluate histologic differentiation of hepatitis B virus (HBV)-related hepatocellular carcinomas (HCCs) using apparent diffusion coefficient (ADC) and intravoxel incoherent motion (IVIM)-derived metrics and to compare findings with alpha-fetoprotein (AFP) levels alone and in combination. MATERIALS AND METHOD: One hundred and six chronic HBV-related HCC patients who underwent IVIM diffusion-weighted magnetic resonance imaging with eleven b values were enrolled. Mean ADC, diffusion coefficient (D), pseudodiffusion coefficient (D*), and perfusion fraction (f) values were determined for all detected lesions. The metrics and AFP levels of different histologically differentiated groups were compared. Spearman's rank correlation was used to assess the statistical dependence among the histologically differentiated HCCs. Receiver operating characteristic (ROC) analysis was performed to evaluate diagnostic performance of these metrics and AFP levels alone and in combination. RESULTS: ADC, D, and f values and AFP levels were significantly different among well-, moderately, and poorly differentiated HCCs. The four metrics were significantly correlated with histologic differentiation. The area under the ROC curve (AUC-ROC) of ADC, D, f, and AFP for diagnosing well-differentiated HCCs was 0.903, 0.84, 0.782, and 0.806, respectively, and the AUC-ROC of above metrics for diagnosing poorly differentiated HCCs was 0.787, 0.726, 0.624, and 0.633, respectively. The combination of ADC and AFP provided an AUC-ROC of 0.945 for well-differentiated HCC. However, this did not provide better performance for diagnosing poorly differentiated HCC. CONCLUSION: ADC, IVIM metrics, and AFP levels may be useful for evaluating histologic differentiation of HBV-related HCCs, and the combination of ADC and AFP provides better diagnostic performance for well-differentiated HCC.

The association of pretreatment thrombocytosis with prognosis and clinicopathological significance in cervical cancer: a systematic review and meta-analysis.

Previous studies reported inconsistent findings about the relationship between pretreatment thrombocytosis and survival in patients with cervical cancer. This study aimed to evaluate the prognostic significance of thrombocytosis in cervical cancer. We searched databases to identify relevant articles. Pooled hazard ratios (HRs), odds ratios (ORs), and 95% confidence intervals (CIs) were calculated. Fourteen studies including 3,394 patients were eligible for the meta-analysis. Overall, an elevated platelet count was significantly associated with inferior overall survival (OS, hazard ratio [HR]: 1.66, 95% confidence interval [CI]: 1.42-1.95, P < 0.001) and recurrence-free survival (RFS, HR: 1.67, 95% CI: 1.15-2.42, P = 0.007) but not progression-free survival (PFS, HR: 1.21, 95% CI: 0.89-1.64; P = 0.235). The results were similar for low stage patients treated with surgery alone. Moreover, a pretreatment thrombocytosis status was significantly associated with higher clinical stage (odd ratio [OR]: 2.39, 95% CI: 1.68-3.38, P < 0.001), positive pelvic node status (OR: 1.58, 95% CI: 1.01- 2.45, P = 0.044) and larger tumor size (OR: 2.32, 95% CI: 1.39-3.87, P = 0.001). Pretreatment thrombocytosis is an independent prognosis predictor in cervical cancer patients. It may be used as a readily available biomarker to refine clinical outcome prediction for cervical cancer patients.

In Vivo Bioluminescence Imaging of Transplanted Mesenchymal Stromal Cells and Their Rejection Mediated by Intrahepatic NK Cells.

PURPOSE: Mesenchymal stromal cells (MSCs) hold promise in the treatment of liver disease. However, short survival time of MSCs after intrahepatic transplantation limits their value; therefore, understanding the basis of MSCs survival and rejection may increase their utility. This study was aimed at determining the role of intrahepatic natural killer (NK) cells on MSCs survival and their retention in the liver shortly after transplant. PROCEDURES: Human MSCs were labeled with the Luc2-mKate2 dual-fusion reporter gene (MSCs-R), and the residence time and survival of MSCs-R xenografts after intrahepatic transplantation were evaluated by in vivo bioluminescence imaging (BLI). Coculture of MSCs and NK cells was performed to assess cytotoxicity. To evaluate the role of NK cells in rejection of the xenografted cells, the fates of transplanted MSCs-R were then assessed in vivo by BLI after activation of intrahepatic NK cells. RESULTS: We observed a linear correlation between luciferase activity from live MSCs-R and cell number in vitro (R 2 = 0.9956). In vivo, we observed a gradual decline in bioluminescent signals from transplanted MSCs-R over a region corresponding to the liver in both the control group and the NK-activated group. However, the survival time and retention of intrahepatic MSCs-R decreased more rapidly in the NK-activated group of mice compared to the control group. This indicated that activated NK cells accelerate the elimination of transplanted MSCs. Also, we found that the number of hepatic NK cells and the expression of NK activation markers significantly increased after intrahepatic delivery of MSCs. This suggested that resident NK cells, in a resting state, were activated by intrahepatic transplantation of human MSCs. Taken together, the data suggests that activated hepatic NK cells mediate, in part, rejection of the MSCs xenografts. Cytotoxicity assays showed that activated NK cells may inhibit the proliferation of MSCs and, to a certain extent, induce MSCs death. CONCLUSION: Human MSCs could be followed dynamically in vivo by BLI, and the role of murine hepatic NK cells, especially activated NK cells, could be inferred from the loss of signals from MSCs. This finding may have practical clinical implications in MSCs transplantation in treating liver disease.

Combined Diffusion Tensor Imaging and Arterial Spin Labeling as Markers of Early Parkinson's disease.

This study aimed to identify a PD-specific MRI pattern using combined diffusion tensor imaging (DTI) and arterial spin labeling (ASL) to discriminate patients with early PD from healthy subjects and evaluate disease status. Twenty-one early and 22 mid-late PD patients, and 22 healthy, age/gender-matched controls underwent 3-T MRI with apparent diffusion coefficient (ADC), fractional anisotropy (FA), fiber number (FN) and cerebral blood flow (CBF) measurements. We found that compared with healthy subjects, there was a profound reduction in FN passing through the SN in PD. FA in the SN and CBF in the caudate nucleus were inversely correlated with motor dysfunction. A negative correlation was observed between FA in the hippocampus (Hip) and the NMSS-Mood score, whereas CBF in the Hip and the prefrontal cortex(PFC) correlated with declined cognition. Stratified five-fold cross-validation identified FA in the SN(FA-SNAv), CBF in the PFC(CBF-PFCAv) and FA in the parietal white matter(FA-PWMAv), and the combination of these measurements offered relatively high accuracy (AUC 0.975, 90% sensitivity and 100% specificity) in distinguishing those with early PD from healthy subjects. We demonstrate that the decreased FNs through SN in combination with changes in FA-SNAv, CBF-PFCAv and FA-PWMAv values might serve as potential markers of early-stage PD.

[Preparation of magnetic near infrared fluorescent probe and targeted multimodal imaging of human mesenchymal stem cells in vitro].

OBJECTIVE: To prepare the magnetic near infrared fluorescent (NIRF) bifunctional molecular probe with human holo-transferrin (Tf) as a targeted ligand and detect human transferrin receptor (hTfR) actively. METHODS: Molecular probe Tf-cy5.5-IO was prepared and purified by conjugating Tf, superparamagnetic iron oxide (IO) and near infrared fluorescent dye (cy5.5). The particle size and morphology was determined by transmission electron microscopy (TEM), zeta potential and particle sizing analyzer. Human serum albumin (HSA) was used for conjugating with cy5.5 and IO as control. hMSCs and HeLa (as a positive control) were divided into 4 groups: A non-labeled, B Tf-cy5.5-IO, C HSA-cy5.5-IO and D competition assay to confirm the targeted connection. The fluorescent signals from intracellular probe were detected with laser scanning confocal microscope (LSCM) and flow cytometry. Intracellular iron was detected with iron concentration assay and TEM. MRI and NIRF imaging of 2×10(5) cells were performed respectively. Enhancements of R2 value and average intensity (AI) were analyzed qualitatively. RESULTS: The conjugation between IO, Tf and cy5.5 was confirmed with a molar ratio of 1: 2.89: 7.89. The hyperdense aqueous diameter of probe was 23.39 ± 2.42 nm. LSCM showed the fluorescence from Tf-cy5.5-IO and cy3-labeled monoclonal antibody against hTfR in cells and two markers were localized in intracellular compartments of similar appearance. After co-incubating with Tf-cy5.5-IO, the intracellular iron and average intensity were significantly higher than cells of other groups (P < 0.01). MRI and NIRF images showed that, after incubation, intracellular Tf-cy5.5-IO decreased the T2WI signal of human mesenchymal stem cells (hMSCs) and AI on NIRF image increased. Enhancements of R2 value and AI were higher in B group than those in other groups (P < 0.05). CONCLUSION: Tf-cy5.5-IO probe can recognize and conjugate with hTfR specifically. And targeted imaging in vitro of hTfR expressed in hMSCs may be performed by MRI and NIRF multimodal imaging.

Evaluation on therapeutic effects of orthotopic liver transplantation by megnetic resonance imaging in patients with portal hypertension.

BACKGROUND: Orthotopic liver transplantation (OLT) has become the therapeutic option of choice for end-stage liver disease. The aim of this study was to investigate the changes of splenic morphology, signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), apparent diffusion coefficient (ADC) values and explore their value in evaluating the therapeutic effects of orthotopic liver transplantation (OLT) on portal hypertension at 1.5 Tesla MRI. METHODS: Twenty patients with portal hypertension undergoing OLT were included in this study. Conventional MRI and diffusion-weighted image (DWI) (b value = 600 s/mm(2)) sequences were applied on each patient before and after OLT, and these patients were referred to as the preoperative and postoperative groups. Twenty healthy individuals were selected as the normal group. After image acquisition, the splenic width (W), thickness (T), length (L), the diameter of the portal vein (PD) and splenic vein (SD) were measured and the splenic volume (V) was calculated. The SNR and CNR were measured on T2WI. The ADC maps were calculated using the b600 in DWIs and the ADC values were measured. RESULTS: Compared with the preoperative group, the splenic V, PD and SD decreased significantly in the postoperative group (P < 0.05). All splenic morphological values were significantly different between preoperative and normal groups (P < 0.05). The splenic L and V were significantly different (P < 0.05) between postoperative and normal groups. The SNR and CNR values were 17.66 ± 4.62 and 13.18 ± 3.12, 11.50 ± 1.64 and 7.44 ± 4.32, 4.24 ± 1.24 and 3.03 ± 2.41 in the preoperative, postoperative and normal groups, respectively. Both SNR and CNR decreased after OLT, but they was still higher than the normal values. The SNR was significantly different between any two groups (P < 0.05). The CNR was significantly different (P < 0.05) between the preoperative and postoperative groups, preoperative and normal groups. The splenic ADC values were (1.339 ± 0.482) × 10(-3) mm(2)/s, (1.120 ± 0.254) × 10(-3) mm(2)/s and (0.997 ± 0.447) × 10(-3) mm(2)/s in the preoperative, postoperative and normal groups, respectively. The difference of ADC values were significant (P < 0.05) between the preoperative and postoperative groups, and the preoperative and normal groups. CONCLUSIONS: OLT is an effective method of treatment for portal hypertension. In addition to dramatically decreasing the splenic V, it can also decrease the splenic SNR, CNR and ADC values in patients with portal hypertension. The changes of splenic SNR, CNR and ADC after OLT may be helpful in providing noninvasive supplementary information in assessing the therapeutic effect of OLT on portal hypertension.

[Evaluation studies of hepatic artery complications after liver transplantation with 320-detector row computed tomography scanner for Surt-start imaging and dynamic volume scanning].

OBJECTIVE: To evaluate the clinical application comparison of Surt-start imaging and dynamic volume scanning acquired from 320-detector row computed tomography (CT) for the diagnosis of hepatic artery complications after liver transplantation. METHODS: Retrospective analysis was conducted for 20 patients undergoing 320-slice CT scanning. Aquilion One scanner after liver transplantation was performed from April 2009 to September 2011. Dynamic volume scanning group employed dynamic volume scanning pattern while Surt-start group used helical scanning pattern. The contrast medium (Ultravist, 370 mgI/ml iopromide) was infused at 6 ml/s with a total dose of 50 ml in dynamic volume scanning group versus 1.5 ml/kg in Surt-start group. Two groups were compared with regards to hepatic artery enhancing peak time, the best arterial phase time, maximum peak enhancement of artery, maximum absolute peak enhancement of artery and branches of hepatic artery. RESULTS: The hepatic artery enhancing peak time and the best arterial phase time was (21.5 ± 2.5) s and (22.4 ± 3.3) s in dynamic volume scanning group versus (19.5 ± 1.8) s in Surt-start group. And there was significant inter-group difference (P = 0.001, P = 0.000). The maximum peak enhancement of artery or the maximum absolute peak enhancement of artery and branches of hepatic artery had no inter-group difference. Two groups showed no difference in displaying hepatic artery anastomosis and variations. Dynamic volume scanning group was better than Surt-start group in displaying hepatic artery microscopic lesions. CONCLUSION: 320-detector CT dynamic volume scanning has the advantage of low-dose contrast medium. Capable of acquiring overall accurate imaging process of hepatic artery, it is effective for follow-ups of hepatic artery complications after liver transplantation.

[Clinical application of 320-row computed tomography 4D digital subtraction angiography in hepatocellular carcinoma].

OBJECTIVE: To explore the application value of 320-row computed tomography (CT) 4D digital subtraction angiography (DSA) for hepatocellular carcinoma (HCC). METHODS: A total of 40 HCC patients received 320-row CT contrast scans. The 4D DSA images were obtained on the basis of baseline data. The normal anatomy and anatomical variations of hepatic artery, tumor supplying arteries, tumor vessels, tumor staining were observed by comparing DSA (n = 20). RESULTS: 320-row CT 4D DSA could show 6-7 levels of intrahepatic arterial branch. Normal hepatic artery anatomy was found in 35 cases (87.5%, Michels I type) and variations in 5 cases (12.5%). The diagnose accordance rate was 100% between 4D DSA and DSA in showing the anatomy and variation of hepatic artery. Among them, 320-row CT 4D DSA showed tumor staining (n = 40), tumor vessels (n = 28), tumor supplying arteries (n = 26) and two hepatic supplying arteries (n = 3). The number of tumor supplying arteries observed by 4D DSA (n = 20) was 18 versus 19 by DSA. Compared with DSA, the accurate rate of 4D DSA was 94.7% (18/19) in detecting tumor supplying arteries. CONCLUSION: As a noninvasive vascular examination modality, 320-row CT 4D DSA can accurately visualize normal anatomy and variation of hepatic artery, dynamically display tumor staining and reproducibly delineate the three-dimension relationship between tumor and blood vessels. In consistency with DSA in detection blood supply of HCC, 320-row CT 4D DSA provides a rapid, DSA-like and non-invasive alternative.

[To evaluate the role of OLT on splenomegaly of portal hypertension by the radiological changes of splenic morphology and collaterals].

OBJECTIVE: To explore the effect of orthotopic liver transplantation (OLT) on portal hypertension by observing the radiological changes of splenic volume and collaterals before and after OLT. METHODS: In our hospital 56 patients performing OLT due to cirrhosis, portal hypertension and splenomegaly were classified into five groups according to their following-up time: A (≤3 months), B (>3-6 months), C (>6-12 months), D (>12-24 months), and E (>24 months). Twenty health people were chose as control group (F). The splenic width, thickness, length, volume, diameter of portal and splenic vein and collaterals were measured and observed in every patient of six groups before and after OLT respectively. RESULTS: After OLT, the splenic volume decreased by 25.4%, 27.8%, 21.9%, 25.2%, 27.7% in five groups respectively, which was still larger than the normal group (P<0.05). Gastroesophageal varices in 31 cases (81.6%, 31/36) became normal after OLT. The opened umbilical vein disappeared and the retroperitoneal varices persisted in five cases after OLT. CONCLUSIONS: Splenomegaly and opened collaterals can be relieved by OLT effectively. The splenic volume didn't change obviously until it decreased by 25% in the three months after OLT. Gastroesophageal varices can be removed in most of patients after OLT. The splenomegaly could last paralled with the splenic vein and retroperitoneal varices after OLT. After OLT, correct disposal of splenic and collateral changes could improve the success rate and the long-term treatment effect of OLT.

[Enhanced green fluorescence protein reporter gene labeling of mesenchymal stem cells mediated by lentivirus].

OBJECTIVE: To explore the effect of enhanced green fluorescence protein (EGFP) labeling mediated by lentivirus on the biophysical properties of mesenchymal stem cells (MSC), and whether the EGFP gene expression is permanent and stable. METHODS: MSC were infected with EGFP lentivirus at different virus multiplicity of infection (MOI). EGFP positive rate was measured with fluorescent-activated cell scanning (FACS) analysis, and EGFP expression in MSC was investigated under a fluorescence microscope. Cell viability, proliferation, apoptosis and cell cycle were detected with trypan blue stain, MTT colorimetric assay, Hoechst stain and FACS analysis respectively. To evaluate the stability of EGFP expression, EGFP lentivirus infected MSC were harvested after cultured continuously in vitro for 2, 4, 8 or 16 weeks, and EGFP positive rate and fluorescence strength were detected with FACS analysis. RESULTS: After infected with EGFP lentivirus (MOI = 20) for 96 h, EGFP positive rate of MSC was 97.39% +/- 0.68%. Cell viability, proliferation, apoptosis and cell cycle of MSC infected with EGFP lentivirus were unaffected, as compared with control MSC (P > 0.05). When cultured in vitro continuously for 2, 4, 8 or 16 weeks, EGFP positive rates of EGFP-MSC were 97.50% +/- 0.54%, 97.32% +/- 0.51%, 97.39% +/- 0.11%, and 97.48% +/- 0.13% respectively, while EGFP fluorescence strength were 440 +/- 13, 445 +/- 12, 458 +/- 13 and 456 +/- 16 respectively. Both EGFP positive rate and fluorescence strength kept in a stable level. CONCLUSION: EGFP lentivirus can efficiently label MSC and has no significant effect on the biophysical properties of MSC. EGFP gene expression in MSC is permanent and stable. EGFP-MSC can be used for further cell tracing research.

[Magnetic/luminescent quantum dots bifunctional nanoparticles labeling of rat bone mesenchymal stem cells].

OBJECTIVE: To evaluate the dual-labeling efficiency of magnetic and luminescent quantum dots bifunctional nanoparticles to rat bone mesenchymal stem cells (BMSC). METHODS: Rat BMSC were isolated, purified, and expanded. Magnetic/luminescent bifunctional nanoparticles (Fe(3)O(4)@CdTe@SiO(2)), were prepared by using silicon dioxide (SiO(2)) to encapsulate simultaneously Fe(3)O(4) and CdTe quantum dots. BMSC were incubated with the Fe(3)O(4)@CdTe@SiO(2) nanoparticles which iron concentration was 25 microg/ml. Fluorescence microscope was used to detect the fluorescence of the intracellular nanoparticles. The dual-labeled BMSC with various concentration underwent ex vivo MR imaging with T(1)WI, T(2)WI and T(2)(*)WI sequences. To show the intracellular iron of labeled cells, prussian blue stain was performed. Spectrophotometer was used to detect the iron concentration in the cells. RESULTS: Intracellular red fluorescence of Fe(3)O(4)@CdTe@SiO(2) can be observed via fluorescent microscopy. Rat BMSC could be effectively dual-labeled with approximately 90% efficiency. The MR images with T(2)WI and T(2)(*)WI sequences, especially with T(2)(*)WI sequence, showed that the signals of the dual-labeled BMSC were lower than those of the unlabeled cells. Cellular total iron is 14.05 + or - 4.15 pg per cell. Iron containing intracytoplasmic vesicles could be observed with Prussian blue staining. CONCLUSION: Rat BMSC can be dual-labeled successfully with Fe(3)O(4)@CdTe@SiO(2) magnetic/luminescent bifunctional nanoparticles successfully, and might serve as a tool for magnetic resonance imaging and in vivo optical imaging.

MR diffusion-weighed imaging of rabbit liver.

AIM: To study the techniques of MR diffusion-weighed imaging (DWI) for normal rabbit liver. METHODS: After 15 normal New Zealand white rabbits and one New Zealand white rabbit implanted with VX-2 tumor were anesthetized with 3% soluble pentobarbitone, DWI was performed respectively for different b values, repetition times (TR) or thicknesses, when other parameters were the same and magnetic resonance imaging (MRI) was performed respectively, or with different field of views (FOV) or coil when other parameters were the same. The distinction between groups was analyzed by SPSS10.0 with apparent diffusion coefficient (ADC), quality index (QI) or signal-noise ratio (SNR). RESULTS: As b value increased, liver ADC, QI and SNR of DWI became smaller and simultaneously (F = 292.87, 156.1, 88.23, P<0.01). QI of DWI was high, when b value was 10, 50 or 100 respectively, but the distinction between them was insignificant; when b value was 800, QI and SNR of DWI were low. QI and SNR of DWI had no significant difference between TR = 4 000, 6 000 and 8 000. QI of DWI with 2 mm thickness was bigger than that with 5 mm thickness (t = 3.04, P<0.01), but SNR of DWI with 2 mm thickness was significantly smaller (t = -17.86, P<0.01). SNR of MRI with knee joint coil was obviously bigger than that with cranium coil (t = -5.77 (T1WI) or -4.02 (T2WI), P<0.01), but QI of MRI was smaller on the contrary (t = 7.10 (T1WI) or 3.97 (T2WI), P<0.01). When FOV was enlarged gradually, SNR of MRI increased (F = 85.81 (T1WI) or 221.96 (T2WI), P<0.01), but QI firstly increased, then decreased (F = 68.67 (T1WI) or 69.46 (T2WI), P<0.01) and QI of MRI was the biggest when FOV was 20 cm x 15 cm. CONCLUSION: The scanning technique is very important in DWI of rabbit liver and the overall quality of DWI with b (100 s/mm2), thickness (2 mm), cranium coils and FOV (20 cm x 15 cm) was best in our study, when other parameters were the same.

[Production of polyclonal antibody against HBD-2 by immunization with recombinant GST-HBD-2 fusion protein].

For the purpose of detecting the HBD-2 expression at protein level, the recombinant prokaryotic expression vector pGEX-1lambdaT-HBD-2 was constructed and the E. coli-based product of GST-HBD-2 fusion protein was prepared. When rabbit was immunized with the fusion protein, the anti-serum against HBD-2 was produced. After caprylic acid and ammonium sulfate precipitation, high titer of specific polyclonal antibody against HBD-2, which was detected by ELISA and Western blot, was obtained. This result suggests that recombinant peptide fusion protein could be used instead of the conjugate of peptide-albumin or peptide-thyroid globulin to produce antibody. The obtained antibodies could be used for revealing the tissue distribution of HBD-2 and the regulation of its gene expression.

MR diffusion-weighted imaging of rabbit liver VX-2 tumor.

AIM: To investigate the implanting method of rabbit liver VX-2 tumor and its MR diffusion-weighted imaging (DWI) characteristics. METHODS: Thirty-five New Zealand rabbits were included in the study. VX-2 tumor was implanted subcutaneously in 14 rabbits and intrahepatically in 6 for pre-experiments. VX-2 tumor was implanted intrahepatically in 12 rabbits for experiment and three were used as the control group. DWI, T1- and T2-weighted of MRI were performed periodically in 15 rabbits for experiment before and after implantation. The distinction of VX-2 tumors on DWI was assessed by their apparent diffusion coefficient (ADC) values. The statistical significance was calculated by analysis of variance (ANOVA) of the randomized block design using SPSS10.0 software. RESULTS: The successful rate of subcutaneous implantation of VX-2 tumor was 29% (4/14) while that of intrahepatic implantation of it was 33% (2/6) in the pre-experiment. The successful rate of intrahepatic implantation of VX-2 tumor in the experiment was 83% (10/12) and 15 tumors grew in 10 successfully implanted rabbits. The DWI signal of VX-2 tumor was high and became lower when the b value increased step by step. The signal of VX-2 tumor on the map of ADC was low. When the b value was 100 or 300 s/mm2, the ADC value of normal group and VX-2 tumor group was respectively 2.57+/-0.26, 1.73+/-0.31, 1.87+/-0.25 and 1.57+/-0.23 mm2/s. Their distinction was significant (F = 43.26, P<0.01), the tumor ADC value between b values 100 and 300 s/mm2 was significant (Tukey HSP, P<0.05) and the ADC value between VX-2 tumor and normal liver was also significant (Tukey HSP, P<0.01). VX-2 tumor developed quickly and metastasized early to all body, especially to the lung, liver, lymph nodes of mediastinum, etc. CONCLUSION: The DWI signal of rabbit VX-2 tumor has its characteristics on MR DWI and DWI plays an important role in diagnosing and discovering VX-2 tumor.