A combined pre- and intra-operative nomogram in evaluation of degrees of liver cirrhosis predicts post-hepatectomy liver failure: a multicenter prospective study.

Background: Adequate evaluation of degrees of liver cirrhosis is essential in surgical treatment of hepatocellular carcinoma (HCC) patients. The impact of the degrees of cirrhosis on prediction of post-hepatectomy liver failure (PHLF) remains poorly defined. This study aimed to construct and validate a combined pre- and intra-operative nomogram based on the degrees of cirrhosis in predicting PHLF in HCC patients using prospective multi-center's data. Methods: Consecutive HCC patients who underwent hepatectomy between May 18, 2019 and Dec 19, 2020 were enrolled at five tertiary hospitals. Preoperative cirrhotic severity scoring (CSS) and intra-operative direct liver stiffness measurement (DSM) were performed to correlate with the Laennec histopathological grading system. The performances of the pre-operative nomogram and combined pre- and intra-operative nomogram in predicting PHLF were compared with conventional predictive models of PHLF. Results: For 327 patients in this study, histopathological studies showed the rates of HCC patients with no, mild, moderate, and severe cirrhosis were 41.9%, 29.1%, 22.9%, and 6.1%, respectively. Either CSS or DSM was closely correlated with histopathological stages of cirrhosis. Thirty-three (10.1%) patients developed PHLF. The 30- and 90-day mortality rates were 0.9%. Multivariate regression analysis showed four pre-operative variables [HBV-DNA level, ICG-R15, prothrombin time (PT), and CSS], and one intra-operative variable (DSM) to be independent risk factors of PHLF. The pre-operative nomogram was constructed based on these four pre-operative variables together with total bilirubin. The combined pre- and intra-operative nomogram was constructed by adding the intra-operative DSM. The pre-operative nomogram was better than the conventional models in predicting PHLF. The prediction was further improved with the combined pre- and intra-operative nomogram. Conclusions: The combined pre- and intra-operative nomogram further improved prediction of PHLF when compared with the pre-operative nomogram. Trial Registration: Clinicaltrials.gov Identifier: NCT04076631.

Unexpected discovery of intrahepatic cholangiocarcinoma during follow-up in two cases of liver cysts: case report.

Background: The liver cyst is commonly treated by hepatobiliary surgery. Generally, most patients show no apparent symptoms and often get diagnosed accidentally during the imaging examinations. In addition, most patients with liver cysts follow a benign course, with fewer severe complications and rare occurrences of malignant changes. Therefore, based on disease characteristics and healthcare costs, long-term regular follow-up of liver cysts are rarely performed clinically. Case Description: Here, we reported two previously treated or observed cases for liver cysts, where intrahepatic neoplastic lesions were found unexpectedly at the liver cyst during follow-up. These two patients' clinical manifestations and laboratory examinations lacked specificity with unclear pre-operative diagnosis, whereas the post-operative pathology confirmed cholangiocarcinoma. One of the patients was a 64-year-old female with right upper abdominal distension. She underwent cyst fenestration for a liver cyst 3 years ago. In the latest admission, imaging examination revealed a tumor in the left inner lobe of the liver. The tumor was located in the exact fenestration location, and the pathological diagnosis of cholangiocarcinoma was made after surgical resection. The patient received Lenvatinib post-operatively and had no recurrence during the follow-up. Another patient, a 68-year-old woman, was asymptomatic, but the liver margin was palpable under the ribs on her physical examination. She had a previous diagnosis of liver cysts and was on regular yearly follow-up. In the last follow-up, a tumor was found close to a cyst. It was diagnosed as intrahepatic cystadenocarcinoma before surgery; however, the pathological features after surgical resection were more consistent with the cholangiocarcinoma. The patient had lung metastases 2 months after the surgery, but her condition improved after receiving targeted therapy and immunotherapy. Moreover, she is alive to this day. Conclusions: We reported 2 cases of intrahepatic cholangiocarcinoma discovered accidentally during the follow-up of hepatic cysts. The location of the malignant tumor coincided with the location of the cyst, making the clinical differential diagnosis problematic. Therefore, it is necessary to be vigilant about the possibility of combined malignant tumors for the follow-up of complex cysts, as early detection and treatment may help improve the prognosis of these patients. After surgery, multimodal therapy, including chemotherapy, immunotherapy, and targeted therapy, is helpful.

SV40T reprograms Schwann cells into stem-like cells that can re-differentiate into terminal nerve cells.

BACKGROUND: The specific effect of SV40T on neurocytes has seldom been investigated by the researchers. We transfected Schwann cells (SCs) that did not have differentiation ability with MPH 86 plasmid containing SV40T, in order to explore the effects of SV40T on Schwann cells. METHODS: SCs were transfected with MPH 86 plasmid carrying the SV40T gene and cultured in different media, and also co-cultured with neural stem cells (NSCs). In our study, SCs overexpressing SV40T were defined as SV40T-SCs. The proliferation of these cells was detected by WST-1, and the expression of different biomarkers was analyzed by qPCR and immunohistochemistry. RESULTS: SV40T induced the characteristics of NSCs, such as the ability to grow in suspension, form spheroid colonies and proliferate rapidly, in the SCs, which were reversed by knocking out SV40T by the Flip-adenovirus. In addition, SV40T up-regulated the expressions of neural crest-associated markers Nestin, Pax3 and Slug, and down-regulated S100b as well as the markers of mature SCs MBP, GFAP and Olig1/2. These cells also expressed NSC markers like Nestin, Sox2, CD133 and SSEA-1, as well as early development markers of embryonic stem cells (ESCs) like BMP4, c-Myc, OCT4 and Gbx2. Co-culturing with NSCs induced differentiation of the SV40T-SCs into neuronal and glial cells. CONCLUSIONS: SV40T reprograms Schwann cells to stem-like cells at the stage of neural crest cells (NCCs) that can differentiate to neurocytes.

BMP9 Can Induce Schwann Cells Expressing Simian Virus 40 T Antigen to Differentiate into Fat and Bone In Vivo and In Vitro.

In our previous study, we constructed Schwann cells (SCs) that stably express Simian virus 40 T antigen (SV40T-SCs). SV40T-SCs functions and markers are similar to those of neural crest cells. There we used bone morphogenetic protein 9 (BMP9) to induce SV40T-SCs differentiation in vitro and in vivo and study possible related mechanism. SV40T-SCs differentiation was induced by BMP9 conditioned medium. The lipogenic differentiation of SV40T-SCs was assessed by Oil Red O staining. Alizarin red and Alcian blue staining, and alkaline phosphatase (ALP) assays were used to evaluate the SV40T-SCs osteogenic differentiation. The expression of adipocyte differentiation (c/EBPα and c/EBPß) and osteoblast differentiation markers (OSX and RUNX2) were detected by quantitative polymerase chain reaction (qPCR). To study possible mechanism related to SV40T-SCs differentiation, the P53 and E2F1 activity were assessed by luciferase reporter plasmid, and Slug and E-cadherin expression by qPCR. In vivo, SV40T-SCs infected by Ad-BMP9 or Ad-GFP were injected under the skin of nude mice. After 4-6 W, the mice were euthanized and subcutaneously mass formed at injecting sites was collected for pathological analysis. After SV40T-SCs were cultured in BMP9 conditioned medium, lipid droplets were formed in the cytoplasm of these cells. Alizarin red and Alcian blue staining were positive, and ALP activity of SV40T-SCs increased significantly. The expression of adipocyte differentiation (c/EBPα and c/EBPß) and osteoblast differentiation markers (OSX and RUNX2) in SV40T-SCs was upregulated by BMP9. SV40T significantly increased Slug expression and decreased E-cadherin expression. SV40T-SCs infected with Ad-BMP9 were able to differentiate into adipose tissue and form a small bone matrix under the nude mice skin. SV40T-SCs have the ability to differentiate into adipocytes and osteoblasts in vivo and in vitro. SV40T can upregulate the Slug expression and downregulate the E-cadherin expression to produce endothelial-to-mesenchymal transition (EMT). The multidirectional differentiation ability of SV40T-SCs may be related to EMT.

Ethylene and salicylic acid synergistically accelerate leaf senescence in Arabidopsis.

The phytohormones ethylene and salicylic acid (SA) have long been known to promote senescence, but their interplay during this process remains elusive. Here we report the synergistic effects of ethylene and SA on promoting leaf senescence in Arabidopsis. EIN3, a key transcription factor of ethylene signaling, physically interacted with the core SA signaling regulator NPR1 in senescing leaves. EIN3 and NPR1 synergistically promoted the expression of the senescence-associated genes ORE1 and SAG29. The senescence phenotype was more delayed for the ein3eil1npr1 triple mutant than ein3eil1 or npr1 with ethylene or/and SA treatment. NPR1-promoted leaf senescence may depend on functional EIN3/EIL1.

Isorhamnetin: A review of pharmacological effects.

Isorhamnetin is one of the most important active ingredients in the fruits of Hippophae rhamnoides L. and the leaves of Ginkgo biloba L., which possesses extensive pharmacological activities. At present, there have been numerous investigations on isorhamnetin, which has the effects of cardiovascular and cerebrovascular protection, anti-tumor, anti-inflammatory, anti-oxidation, organ protection, prevention of obesity, etc. The related mechanisms involve the regulation of PI3K/AKT/PKB, NF-κB, MAPK and other signaling pathways as well as the expression of related cytokines and kinases. Isorhamnetin has a high value of development and application. However, the investigations on its mechanism of action are limited and lack of detailed scientific validation. The manuscript reviewed the pharmacological effects of isorhamnetin and related mechanisms of action for the development of its medicinal properties further.

Effect of CLIC1 gene silencing on proliferation, migration, invasion and apoptosis of human gallbladder cancer cells.

This study aimed to explore the effects of CLIC1 gene silencing on proliferation, migration, invasion and apoptosis of human gallbladder cancer (GBC). GBC and normal gallbladder tissues were extracted for the detection of mRNA and protein expressions of CLIC1. GBC-SD and NOZ cells in the logarithmic growth phase were selected to conduct the experiment. Three different siRNA recombined expression vectors were established using CLIC1 as a target at different sites. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting were, respectively, used to detect the CLIC1 mRNA and protein expressions. MTT assay was performed to detect the cell proliferation. Flow cytometry was applied to measure the cell apoptosis and cell cycle distribution. The variations of cell migration and invasion were evaluated using Transwell assay. GBC tissues showed higher CLIC1 mRNA and protein expressions than normal gallbladder tissues. The CLIC1 mRNA and protein expressions in the CLIC1 siRNA group were significantly lower than those in the NC and blank groups. Compared with the NC and blank groups, the CLIC1 siRNA group showed a significant decrease in cell proliferation but an obvious increase in apoptosis rate in GBC cells. Besides, in the CLIC1 siRNA group, cell percentage in G0/G1 and G2/M phase was gradually increased but decreased in S phases. The migration and invasion abilities in GBC cells were significantly lower than those in the NC and blank groups. Our study demonstrates that CLIC1 gene silencing could promote apoptosis and inhibit proliferation migration and invasion of GBC cells.

Potentially optimal body size to adjust tube current for individualized radiation dose control in retrospective ECG-triggered 256-slice CT coronary angiography.

INTRODUCTION: We aimed to determine a potentially optimal body size index for adjusting the tube current in retrospective ECG-triggered helical 256-slice CT coronary angiography (CTCA) for individualized radiation dose control. METHODS: Consecutive patients (n=102) with suspected coronary artery disease underwent retrospective ECG-triggered CTCA with a 256-slice CT scanner. Body mass index (BMI), nipple level (NL) bust and six anthropometric body size indexes, including thoracic anteroposterior diameter at NL, chest circumference (CC) at NL, left main (LM) and right coronary artery (RCA) origin level, chest area (CAr) and chest attenuation (CAt) at RCA origin level were measured, and their correlation with image noise in the aorta was evaluated using linear regression. Pearson correlation analysis was performed respectively to determine the body size index that correlated best with the other body size indexes. An equation was derived to use the best correlated body size index for adjusting tube current. RESULTS: Linear regression demonstrated that CCRCA had the best correlation with image noise. Pearson correlation analysis showed that CCNL, CCLM and CArRCA had the closest linear relationship with CCRCA. The equations connecting CCRCA and tube current for males and females were XmA = 662 × (0.055 × CCRCA - 28.594) / 302 and XmA = 662 × (0.049 × CCRCA - 21.584) / 302, respectively, for a fixed image noise value of 30 HU. CONCLUSIONS: Tube current selection is different for males and females, particularly in patients with a small chest circumference. CCRCA is an ideal body index for appropriately adjusting tube current in CTCA for individualized radiation dose control.

Suppression of angiogenesis and tumor growth in vitro and in vivo using an anti-angiopoietin-2 single-chain antibody.

Hepatocellular carcinomas (HCCs) are tumors with a highly developed vascular architecture. HCC cells require access to blood vessels for growth and metastasis; therefore, the inhibition of angiogenesis represents a potential therapeutic target for HCC that may reduce the mortality and morbidity from HCC. Various attempts to develop an anti-angiogenic therapy have been made in past decades; however, modest results have been achieved in clinical trials and the challenge of HCC treatment remains. Single-chain antibodies (scFv) are characterized by low molecular weight, low immunogenicity, high penetration and a short half-life, and are easy to produce on a large scale by genetic engineering. Accordingly, an scFv against a specific angiogenic regulator, such as angiopoietin (Ang), may be a promising anti-angiogenic therapy for HCC. Our previous study indicated that an imbalanced expression of angiopoietin-2 (Ang-2) vs. angiopoietin-1 (Ang-1) in HCCs contributes to initiation of neovascularization and promotes the angiogenesis and progression of HCCs. Therefore, we suggest that specific Ang-2-targeting interventions may be valuable in the treatment of HCC via remodeling the neovascular network and changing the tumor microenvironment. In this study, a prokaryotic expression vector of Ang-2 was constructed and purified human Ang-2 protein was isolated. An scFv against human Ang-2 (scFv-Ang2) was identified and purified via phage display technology, and the effects of scFv-Ang2 in vitro and in vivo on HCC in nude mice were evaluated. The results show that scFv-Ang2 inhibits vascular endothelial growth factor (VEGF) and Ang-2 induces the proliferation, migration and tubule formation of human umbilical vein endothelial cells (HUVECs) in vitro. In the in vivo assay, statistical indices, including tumor weight and volume, metastases to lungs, CD31 expression and the microvessel density (MVD) count in the scFv-Ang2-treated group of mice were significantly lower than those in the control group (P<0.05). In conclusion, the successfully generated scFv-Ang2 showed significant inhibitory effects on the angiogenesis and tumor growth of human HCC in vitro and in vivo.

[CT and MRI features of Castleman's disease of the abdomen and pelvis].

OBJECTIVE: To analyze the CT/MRI features of Castleman's disease of the abdomen and pelvis. METHODS: CT/MRI images of 6 cases of pathologically confirmed Castleman's disease of the abdomen and pelvis were retrospectively reviewed. All the patients received plain CT scan and dynamic enhanced scan, and one had an additional MR scan. RESULTS: One case was identified as the disseminated type with multicentric enlarged lymph nodes and hepatosplenomegaly, and 5 cases were found to have localized type, of which 3 had retroperitoneal, 1 had mesentery and 1 had pelvic lesions. On CT scan, all the 5 cases with localized lesions showed single, round or ellipse soft tissue masses, with intra-tumoral calcification in 2 cases, fascial thickening around the mass in 3 cases, and satellite nodules in 4 cases. Enhanced scanning revealed obvious enhancement in the arterial phase and continuous enhancement in the portal vein and delayed phase in all the lesions, with an attenuation pattern similar to that of large vessels; enlarged blood vessels within or around the mass were displayed in each case. In 4 cases, the intra-tumoral radial or fissured non-enhanced areas in early stage of enhancement were gradually filled up as the scan time was delayed. The patient receiving MRI showed hypo-intensity on T(1)WI and hyper-intensity on T(2)WI, presenting with an enhancement feature similar to that of CT. CONCLUSION: Castleman's disease in the abdomen and pelvis is rare and liable to misdiagnosis, but its characteristic imaging features can help in the diagnosis and differential diagnosis.

[Computed tomography and magnetic resonance imaging features of desmoid-type fibromatosis: comparison with the pathological findings].

OBJECTIVE: To explore the computed tomography (CT) and magnetic resonance imaging (MRI) features of desmoid-type fibromatosis, and improve the diagnostic accuracy and understanding of the disease. METHODS: The CT and MRI features of 18 cases of surgically and pathologically confirmed desmoid-type fibromatosis were reviewed retrospectively. Among the patients, 10 received CT pre- and post-contrast scanning, and 8 patients had MRI pre- and post-contrast scanning. The CT and MRI features were analyzed in comparison with the pathological findings. RESULTS: In the extra abdominal cases, the tumors occurred in the head and neck in 3, in the dorsal part of the chest in 2, in the abdominal wall and groin area in 9, and in the peritoneal cavity in 4; concomitant Gardner syndrome was found in 1 case. In 4 cases the tumor occurred within 1 to 3 years after abdominal surgeries. Pathologically, the lesion was hard and composed of fusiform fibroblasts and myofibroblast. The cells showed no obvious heteromorphism with few karyokinesis, growing invasively and recurrent locally but without distant metastasis. Immunohistochemically, the fibroblasts and myofibroblasts expressed vimentin, and the myofibroblasts were positive for SMA. On CT and MRI, the lesion appeared benign with malignant growth pattern, and caused compression of the adjacent organs and vessels or encasement of the vessels; the border was unclear without encapsulation, and necrosis and calcification was scarce. The density and signal of the tumor were well distributed. Twelve patients displayed obvious enhancement and 5 showed uneven enhancement. CONCLUSION: The CT and MRI features of desmoid-type fibromatosis are characteristic, and CT and MRI are valuable modalities for the diagnosis and differential diagnosis of the tumor.

[Association of vascular endothelial growth factor 936C/T polymorphism and the susceptibility to colorectal adenoma].

OBJECTIVE: To examine the association between polymorphism of vascular endothelial growth factor(VEGF)1498 C/T,936 C/T and colorectal adenoma genetic susceptibility. METHODS: A case-control study of 224 colorectal adenomas and 200 controls was conducted and VEGF genotypes were determined based on TaqMan-probe assay. The epidemiological factors were collected through questionnaire. Accordingly, the clinicopathological data of each sample were also investigated. RESULTS: The carriage of 936 CT and CT+TT genotypes had significantly higher risk of colorectal adenoma (CT vs. CC, OR=2.00, 95% CI: 1.23-3.25, P=0.006; CT+TT vs. CC, OR=2.04, 95% CI:1.28-3.26, P=0.003). 936-T allele carriage had increased risk of colorectal adenoma (OR=1.91, 95% CI:1.25-2.91, P=0.003). The genotypes of 1498 C/T and the frequency of C/T allele showed no differences between healthy persons and patients (P>0.05). In patients with 936 CT+TT and 936-T allele implied a tendency of villous adenoma category (CT+TT vs. CC, OR=2.54, 95% CI:1.12-5.75, P=0.040; T allele vs. C allele, OR=3.08, 95% CI, 1.64-5.80, P=0.001). CONCLUSION: VEGF 936 C/T polymorphism can influence susceptibility to colorectal adenoma.

[Clinical study of transcatheter arterial chemoembolization plus radiofrequency ablation in hepatocellular carcinoma by magnetic resonance imaging and functional diffusion-weighted imaging].

OBJECTIVE: To evaluate the apparent diffusion coefficient (ADC) value features of the lesions after transcatheter arterial chemoembolization (TACE) plus radiofrequency ablation in hepatocellular carcinoma (HCC) with 3.0T magnetic resonance imaging (MRI) and diffusion-weight imaging (DWI) and analyze the value of 3.0T DWI in detecting the pathological lesion features of post-TACE plus radiofrequency ablation in HCC. METHODS: Twenty-eight HCC patients were enrolled to receive TACE firstly. Then all viable tumors around the lesions underwent radiofrequency ablation. At 1-4 months after radiofrequency ablation, 3.0T MRI and DWI (b = 600 sec/mm(2)) were performed to measure the ADC values of different lesions of post-TACE plus radiofrequency ablation. The features of MRI and ADC values of different lesions, the difference of contrast enhancement sequence and DWI in evaluating the lesions of post-TACE plus radiofrequency ablation were analyzed. RESULTS: Viable tumors occurred in 14 of 28 HCC patients after TACE plus radiofrequency ablation. The ADC values of necrotic tissues with lipiodol, necrotic tissues without lipiodol, viable tumors and normal liver tissues were 1.905 ± 0.487, 0.726 ± 0.116, 1.449 ± 0.054 and 1.777 ± 0.094 (10(-3) mm(2)/sec) respectively. There was no significant difference of ADC values between necrotic tissues with lipiodol and normal tissues (P = 0.115). But there were significant differences of ADC values among necrotic tissues with lipiodol, necrotic tissues without lipiodol and viable tumors (P < 0.05). The viable tumor tissues after TACE plus radiofrequency ablation appeared as nodular lesions with slightly heightened signal intensities around the necrotic tissues, the lesions with heterogeneous enhancement during arterial phase, portal vein phase and parenchymal phase. Necrotic tissues without lipiodol occurred outside necrotic tissues without lipiodol, around normal liver tissues, with low signal intensities on T2WI, without enhancement during arterial phase, portal vein phase and parenchymal phase. There were no significant difference between contrast enhancement and DWI sequence in detecting viable tumors after TACE plus radiofrequency ablation (P > 0.05). CONCLUSION: The ADC values of 3.0T MR DWI may be used to distinguish the viable residue or recurrent tumor tissues, necrotic tissues in HCC after TACE plus radiofrequency ablation.

[Preoperative evaluation of the 64-slice spiral CT three dimensional angiography for vascular invasion in gastric cancer].

OBJECTIVE: To evaluate the preoperative diagnosis value of 64-slice spiral CT three dimensional angiography (3D CTA) for the vascular invasion in gastric cancer. METHODS: CT images of 40 patients diagnosed as gastric cancer by endoscope,who proceeded to surgical exploration from August 2006 to December 2007,were collected. These images were rebuilt by 3D CTA to judge vascular invasion by gastric cancer in comparison with the surgical finding as standard reference. RESULTS: Successful 3D CTA reconstructions were performed for all these 40 patient images. Out of 40 cases, 14 cases presented vascular invasion in the 3D CTA, and 12 of 14 cases were proved to have vascular invasion in the surgery. For assessing vascular invasion with CTA, the sensitivity was 98.1% and the specificity was 96.4% respectively (Chi Square chi(2)=0.0099,P>0.05). There was no significant differences regarding vascular invasion in gastric cancer between preoperative 3D CTA assessment and surgical finding. CONCLUSION: Sixty-four-slice spiral CT 3D angiography is effective in assessing vascular invasion in gastric cancer and is also valuable in clinical application.

Crohn disease of small bowel: multidetector row CT with CT enteroclysis, dynamic contrast enhancement, CT angiography, and 3D imaging.

BACKGROUND: CT could be used to evaluate abnormalities in the bowel wall, mesentery, adjacent structures, vasculature, and even the activity of Crohn disease (CD). To our knowledge, few direct comparisons of CD characterization using multidetector row CT with dynamic contrast enhancement, 3D imaging, CT angiography (CTA), and CT-enteroclysis (CT-E) on the same cohort of patients. The purpose of this study was to evaluate the diagnostic value of CD using multidetector helical CT with CT-E, dynamic contrast enhancement, 3D imaging, and CTA. METHODS: Twenty-eight patients known or suspected CD underwent CT-E, dynamic contract enhancement, CTA, and 3D imaging. The multidetector CT series images were performed on eight-slice CT scanner. All the examinations were performed when water was used as an oral contrast starting 25 s after 140 mL of intravenous contrast agent was administered, followed by portal venous phase (60 s), and a 60-70 s delay, then sending 1.25-mm slices to the 3D workstation, CT angiograms and 3D images were reconstructed. All the images were reviewed to detect abnormalities of CD. The abnormalities of the bowel wall, mucosal and submucosal ulceration, prominent perienteric vasculature, sinus tracts or fistulae, abscess were evaluated. RESULTS: Crohn disease was diagnosed in 28 patients by CT images, and 54 inflammatory segments were revealed. In active inflammatory cases, the diseased bowel wall thickened and the enhancement of diseased bowel wall increased significantly in 34 inflammatory segments of 22 cases, the enhancement of diseased bowel wall increased significantly but without the wall thickened in three patients. Prominent vasculature was found in CTA and 3D images in 21 patients with active diseases. In 16 patients, the sharp interface between bowel and mesentery was lost and the attenuation of fat increased. Sinus tracts or fistulae were observed in eight patients, four of 28 patients demonstrated abscesses, all were active inflammatory patients. In three chronic inflammatory patients, normal bowel, bowel lumen stricture, and the normal enhancement of the wall were displayed. CONCLUSION: The abnormalities of CD and its complications can be identified by multidetector CT with CT-E, dynamic enhancement, CTA, and 3D imaging, and they are important methods in diagnosing CD. Complications of CD can be shown better when CT-E is performed.

Expression of angiopoietins, Tie2 and vascular endothelial growth factor in angiogenesis and progression of hepatocellular carcinoma.

AIM: To investigate the significance of angiopoietins, Tie2 and vascular endothelial growth factor (VEGF) expression in the angiogenesis and progress of hepatocellular carcinoma (HCC). METHODS: Fresh surgically resected specimens of HCC and noncancerous liver (NCL) tissue from 38 patients with HCC were obtained, and expression of angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2), Tie2, and VEGF messenger RNA (mRNA) was examined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Expression pattern of each gene in HCC and NCL tissue specimens was compared and the potential role and interaction in angiogenesis of HCC were analyzed. Genes' expression level and its relationship with tumor's clinicopathological parameters were also investigated. Immunohistochemical staining of CD34 was performed to determine the microvessel density (MVD) and Ang-2/Ang-1 ratio was calculated. Relationships between Ang-2/Ang-1 ratio, VEGF and MVD and clinicopathological features were also tested so as to evaluate their significance in the progression of HCC. RESULTS: Ang-2 and VEGF mRNAs in HCC were significantly higher than those in NCL tissue (P < 0.05), whereas the Ang-1 and Tie2 mRNAs showed no statistical significance (P > 0.05), though slightly lower level of Ang-1 mRNA in HCC was observed. Ang-2/Ang-1 ratio and VEGF were both positively correlated to MVD. The Ang-2/ Ang-1 ratio, Ang-2 and VEGF were all associated with tumor's clinicopathological parameters (P < 0.05) except for histological grades (P > 0.05). Ang-1 and Tie2 levels in different clinicopathological groups were not significantly different (P > 0.05). CONCLUSION: Dominant Ang-2 expression against Ang-1 through Tie2 receptor in the presence of VEGF plays a critical role in initiating early neovascularization and transformation of noncancerous liver to hepatocellular carcinoma. Its consequently constant operation in formed HCC induces further angiogenesis and progression of HCC.

[Anti-tumor effect of thalidomide and paclitaxel on hepatocellular carcinoma in nude mice].

BACKGROUND: Thalidomide is reviving for its antiangiogenic effect on corneal neovascularization models. Recently, it has been employed in tumor research in several types of solid carcinomas. However, its effect on hepatocellular carcinoma (HCC) has not yet been clarified. METHODS: A total of 48 nude mice bearing human HCC with a high metastatic potential were randomly divided into 4 groups. Thalidomide (200 mg/kg), paclitaxel (13 mg/kg), or their combination, which was dissolved in 0.5% sodium carboxyl methyl cellulose (CMC) suspension, was intraperitoneally injected in each group since the second day of the establishment of animal model. The group simply administered with 0.5% CMC was set as placebo-control. The mice were sacrificed on the 30th day, for the measurement of tumor size, weight and metastasis in the lungs. The levels of CD34 and endothelial growth factor (VEGF) mRNA in tumor tissues were detected by immunohistochemistry and semiquantitative RT-PCR, respectively, and microvessel density (MVD) was evaluated. RESULTS: No statistical difference was found in tumor weight and volume between the thalidomide group and control (P>0.05). Paclitaxel showed a growth-inhibiting effect on tumors (P<0.05). The value of MVD and VEGF mRNA and metastases to the lungs in each group were lower than those in the placebo-control group (P<0.05); such difference in the combination group was statistically significant (P<0.05). CONCLUSIONS: Paclitaxel, but not thalidomide, has significant growth inhibitory effect on tumors, but both significantly inhibit angiogenesis and metastasis of human HCC in nude mice, such effects of paclitaxel can be amplified by thalidomide.

Effects of thalidomide on angiogenesis and tumor growth and metastasis of human hepatocellular carcinoma in nude mice.

AIM: To investigate the effects of thalidomide on angiogenesis, tumor growth and metastasis of hepatocellular carcinoma in nude mice. METHODS: Twenty-four nude mice were randomly divided into therapy group and control group, 12 mice in each group. Thalidomide dissolved in 0.5% sodium carboxyl methyl cellulose (CMC) suspension was administered intraperitoneally once a day at the dose of 200 mg/kg in therapy group, and an equivalent volume of 0.5% CMC in control group. Mice were sacrificed on the 30th d, tumor size and weight and metastases in liver and lungs were measured. CD34 and VEGF mRNA in tumor tissue were detected by immunohistochemistry and semi-quantitative RT-PCR respectively and microvessel density (MVD) was counted. Serum concentrations of TNF-alpha and ALT and AFP were also tested. RESULTS: MVD and VEGF mRNA in therapy group were less than those in control group (31.08+/-16.23 vessels/HP vs 80.00+/-26.27 vessels/HP, 0.0538+/-0.0165 vs 0.7373+/-0.1297, respectively, P<0.05). No statistical difference was observed in tumor size and weight and metastases in liver and lungs. TNF-alpha was significantly lower in therapy group than in control group (28.64+/-4.64 ng/L vs 42.69+/-6.99 ng/L, P<0.05). No statistical difference in ALT and AFP was observed between groups. CONCLUSION: Thalidomide can significantly inhibit angiogenesis and metastasis of hepatocellular carcinoma. It also has inhibitory effects on circulating TNF-alpha.

Chloroplast transformation in oilseed rape.

The chloroplast transformation vector pNRAB carries two expression cassettes for the spectinomycin resistance gene aadA and the insect resistance gene cry1Aa10. The two cassettes are sited between the rps7 and ndhB targeting fragments. Biolistic delivery of the vector DNA, followed by spectinomycin selection, yielded chloroplast transformants at a frequency of four in 1000 bombarded cotyledon petioles. PCR analysis and Southern blot of PCR products confirmed the site-specific integration of aadA and cry1Aa10 into the chloroplast genomes of transgenic oilseed rape. When transgenic oilseed rape leaves were fed to second instar Plutella xylostera larvae, 47% mortality was observed against this insect and the surviving larvae had significantly lower weight than the control. This is the first report of chloroplast transformation in oilseed rape and the introduction of novel genes between the rps7 and ndhB genes in the chloroplast genome. This offers an opportunity for improvement of oilseed rape by chloroplast genetic engineering.

Transplastomic Plants Homoplasmic for Foreign Transgenes.

Scientists pay more and more attention to the research on plastid engineering for its following advantages foreign genes can be integrated site-specifically into the plastid genome (plastome) there are no position effects as experienced with random insertion of transgenes in nuclear transformation pollen-mediated dispersion of transgenes can be avoided because chloroplasts are maternally transmitted gene silencing does not occur in plastids and therefore transgene expression is stable in progeny of transplastomic plants and the high ploidy level of the plastome in leaf cells makes high levels of transgene expression feasible. At the same time, however, the highly polyploid plastome makes it difficult to get transplastomic plants homoplasmic for foreign transgenes. In this work, chloroplast transforming vector pTRCH205, which carries two psbA5'-nifH-psbA3'and Prrn-aadA-TpsbA cassettes flanked by plastid DNA sequence to target their insertion between psbA and trnK operons, was constructed. Plastid transformation of Nicotiana tabacum was carried out by the biolistic delivery of transforming plasmid pTRCH205 DNA into leaf cells. Integration of nifH and aadA by two homologous recombination events via the flanking ptDNA sequences, and selective amplification of the transplastomes on MS medium with high concentration of spectinomycin, yielded resistant cell lines. All the independent transplastomic lines were subjected to three additional rounds of regeneration and were subcultured for 6--10 times through stem sections on MS medium containing 500 mg/L spectinomycin, to obtain homoplasmic tissues. The results of PCR assay and Southern blot hybridization, probed with 0.9 kb BglII/SnaBI homologous fragment, indicated that foreign genes had been integrated into the plastomes of transgenic plants, which finally became homoplasmic for foreign transgenes.