Analysis of Key Genes for Slow Transit Constipation Based on RNA Sequencing.

Purpose: This study aims to identify key genes in slow transit constipation (STC). We also sought to explore the potential link between STC and colorectal cancer. Patients and Methods: mRNA expression profiles were obtained by RNA sequencing, and differentially expressed genes were identified. Functional enrichment analysis and a protein-protein interaction (PPI) network was explored, and differentially expressed genes common to STC and colorectal cancer were examined. Analysis of the effect of constipation and colorectal cancer common genes on the overall survival of colorectal cancer patients based on GEPIA database. Results: Functional enrichment showed that significantly different genes are related to lymphocyte chemotaxis, positive regulation of inflammatory response, cellular response to tumor necrosis factor, extracellular region, extracellular space and chemokine activity. The hub gene for STC was found in the PPI network. In addition, AQP8 and CFD were common differential genes for STC and colorectal cancer. AQP8 affects overall survival in patients with colorectal cancer. Conclusion: Our findings will contribute to understanding the pathology of STC at the molecular level, with the first discovery that AQP8 may be a hub gene in the transition from STC to colorectal cancer.

Anatomic patterns and clinical significance of gastrocolic trunk of Henlé in laparoscopic right colectomy for colon cancer: Results of the HeLaRC trial.

BACKGROUND: Recent interest in laparoscopic right colectomy with D3 lymphadenectomy for right colon cancer, has raised renewed attention to the anatomic variations of the gastrocolic trunk of Henlé (GTH). Understanding the vascular structure of the GTH region for individual patients should improve surgical outcomes. The goal of this nationwide multicenter study (Anatomical Classification of Henlé's Trunk in Laparoscopic Right Hemi-colectomy (HeLaRC) trial) was to study the anatomic patterns of the GTH region, to clarify the implications of GTH in laparoscopic right colectomy with D3 lymphadenectomy (D3-RC) and analyze their clinical significance. METHODS: We enrolled 583 patients from 26 centers across China who underwent D3-RC. The number of tributaries, length and types of GTH constitutions and their influence on intra-operative data were investigated. A nomogram score (based on the length of GTH, body mass index (BMI), tumor location, T stage and type of GTH (type I vs. non-type I) was established to assess the potential hazard of bleeding. RESULTS: The GTH was found in 567 patients (97.3%). The distribution of GTH types was 0 (14.1%, n = 80), I (53.3%, n = 302), II (27.0%, n = 153), III (5.6%, n = 32). Of note, the type I GTH, T1 stage and tumor location at ileocecal or ascending colon were correlated with shorter exposure time of the GTH region (P < 0.0001). Short length of GTH (P = 0.002) and tumor location (transverse colon vs. non transverse colon) (P = 0.003) were correlated with the amount of GTH bleeding during the surgery. Nomogram discrimination was good (C-index: 0.72 (95% CI: 0.64, 0.80)). The dissection plane was better in patients with type I GTH than with other types (P = 0.023). CONCLUSION: GTH pattern variations may affect surgical outcomes in patients undergoing D3-RC. Better recognition of GTH anatomy might lead to a safer operation with better oncologic quality.

Short-term outcomes of complete mesocolic excision versus D2 dissection in patients undergoing laparoscopic colectomy for right colon cancer (RELARC): a randomised, controlled, phase 3, superiority trial.

BACKGROUND: Whether extended lymphadenectomy for right colon cancer leads to increased perioperative complications or improves survival is still controversial. This trial aimed to compare the efficacy and safety of complete mesocolic excision (CME) versus D2 dissection in laparoscopic right hemicolectomy for patients with right colon cancer. This article reports the early safety results from the trial. METHODS: This randomised, controlled, phase 3, superiority, trial was done at 17 hospitals in nine provinces of China. Eligible patients were aged 18-75 years with histologically confirmed primary adenocarcinoma located between the caecum and the right third of the transverse colon, without evidence of distant metastases. Central randomisation was done by means of the Clinical Information Management-Central Randomisation System via block randomisation (block size of four). Patients were randomly assigned (1:1) to CME or D2 dissection during laparoscopic right colectomy. Central lymph nodes were dissected in the CME but not in the D2 procedure. Neither investigators nor patients were masked to their group assignment but the quality control committee were masked to group assignment. The primary endpoint was 3-year disease-free survival, but the data for this endpoint are not yet mature; thus, only the secondary outcomes-intraoperative surgical complications and postoperative complications within 30 days of surgery, graded according to the Clavien-Dindo classification, mortality (death from any cause within 30 days of surgery), and central lymph node metastasis rate in the CME group only-are reported in this Article. This early analysis of safety was preplanned. The outcomes were analysed according to a modified intention-to-treat principle (excluding patients who no longer met inclusion criteria after surgery or who did not have surgery). This study is registered with ClinicalTrials.gov, NCT02619942. Study recruitment is complete, and follow-up is ongoing. FINDINGS: Between Jan 11, 2016, and Dec 26, 2019, 1072 patients were enrolled and randomly assigned. After exclusion of 77 patients, 995 patients were included in the modified intention-to-treat population (495 in the CME group and 500 in the D2 dissection group). The postoperative surgical complication rate was 20% (97 of 495 patients) in the CME group versus 22% (109 of 500 patients) in the D2 group (difference, -2·2% [95% CI -7·2 to 2·8]; p=0·39); the frequency of Clavien-Dindo grade I-II complications were similar between groups (91 [18%] vs 92 [18%], difference, -0·0% [95% CI -4·8 to 4·8]; p=1·0) but Clavien-Dindo grade III-IV complications were significantly less frequent in the CME group than in the D2 group (six [1%] vs 17 [3%], -2·2% [-4·1 to -0·3]; p=0·022); no deaths occurred in either group. Of the intraoperative complications, vascular injury was significantly more common in the CME group than in the D2 group (15 [3%] vs six [1%], difference, 1·8 [95% CI 0·04 to 3·6]; p=0·045). Metastases in the central lymph nodes were detected in 13 (3%) of 394 patients who underwent central lymph node biopsy in the CME group; no patient had isolated metastases to central lymph nodes. INTERPRETATION: Although the CME procedure might increase the risk of intraoperative vascular injury, it generally seems to be safe and feasible for experienced surgeons. FUNDING: The Capital Characteristic Clinical Project of Beijing and the Chinese Academy of Medical Sciences.

Targeting CLK3 inhibits the progression of cholangiocarcinoma by reprogramming nucleotide metabolism.

CDC-like kinase 3 (CLK3) is a dual specificity kinase that functions on substrates containing serine/threonine and tyrosine. But its role in human cancer remains unknown. Herein, we demonstrated that CLK3 was significantly up-regulated in cholangiocarcinoma (CCA) and identified a recurrent Q607R somatic substitution that represented a gain-of-function mutation in the CLK3 kinase domain. Gene ontology term enrichment suggested that high CLK3 expression in CCA patients mainly was associated with nucleotide metabolism reprogramming, which was further confirmed by comparing metabolic profiling of CCA cells. CLK3 directly phosphorylated USP13 at Y708, which promoted its binding to c-Myc, thereby preventing Fbxl14-mediated c-Myc ubiquitination and activating the transcription of purine metabolic genes. Notably, the CCA-associated CLK3-Q607R mutant induced USP13-Y708 phosphorylation and enhanced the activity of c-Myc. In turn, c-Myc transcriptionally up-regulated CLK3. Finally, we identified tacrine hydrochloride as a potential drug to inhibit aberrant CLK3-induced CCA. These findings demonstrate that CLK3 plays a crucial role in CCA purine metabolism, suggesting a potential therapeutic utility.

Anatomical characteristics and classifications of gastrocolic trunk of Henle in laparoscopic right colectomy: preliminary results of multicenter observational study.

BACKGROUND: As a key landmark during laparoscopic right colectomy, the classification and variation of the gastrocolic trunk of Henle (GTH) remains to be clarified. The aim of this nationwide multicenter study was to describe the characteristics of the GTH intra-operatively during laparoscopic right colectomies. METHODS: Three hundred seventy-one patients who underwent laparoscopic right colectomies from January 2018 to March 2019 in 25 hospitals across China were enrolled in the study. The length of the GTH, the classification with a precise description of confluent tributaries, and other variations were analyzed. RESULTS: Of the 371 patients, 363 had a GTH. The proportion of type-0, type-I, type-II, and type-III was 15.2% (n = 55), 54.8% (n = 199), 25.3% (n = 92), and 4.7% (n = 17), respectively. The average length of the GTH was 8.5 mm, ranging from 2 to 30 mm. CONCLUSIONS: This is the first multicenter study with a large sample by which the GTH was classified based on laparoscopic intraoperative observation. Variations in the GTH were classified into four types based on the number of colic drainage veins (right colic, superior right colic, middle colic, accessory middle colic, and ileocolic veins), among which the right colic vein was the most common. The length of the GTH was relatively short, and thus might carry a risk of bleeding. Further clinical data should be correlated with the characteristics of the GTH.

LncRNA DLEU1 contributes to colorectal cancer progression via activation of KPNA3.

BACKGROUND: Accumulating evidences show that long noncoding RNAs (lncRNA) play essential roles in the development and progression of various malignancies. However, their functions remains poorly understood and many lncRNAs have not been defined in colorectal cancer (CRC). In this study, we investigated the role of DLEU1 in CRC. METHODS: Quantitative real-time PCR was used to detect the expression of DLEU1 and survival analysis was adopted to explore the association between DLEU1 expression and the prognosis of CRC patients. CRC cells were stably transfected with lentivirus approach and cell proliferation, migration, invasion and cell apoptosis, as well as tumorigenesis in nude mice were performed to assess the effects of DLEU1 in BCa. Biotin-coupled probe pull down assay, RNA immunoprecipitation and Fluorescence in situ hybridization assays were conducted to confirm the relationship between DLEU1 and SMARCA1. RESULTS: Here we revealed that DLEU1 was crucial for activation of KPNA3 by recruiting SMARCA1, an essential subunit of the NURF chromatin remodeling complex, in CRC. DLEU1 was indispensible for the deposition of SMARCA1 at the promoter of KPNA3 gene. Increased expression of DLEU1 and KPNA3 was observed in human CRC tissues. And higher expression of DLEU1 or KPNA3 in patients indicates lower survival rate and poorer prognosis. DLEU1 knockdown remarkably inhibited CRC cell proliferation, migration and invasion in vitro and in vivo while overexpressing KPNA3 in the meantime reversed it. CONCLUSIONS: Our results identify DLEU1 as a key regulator by a novel DLEU1/SMARCA1/KPNA3 axis in CRC development and progression, which may provide a potential biomarker and therapeutic target for the management of CRC.

Comprehensive analysis of differential expression profiles of mRNAs and lncRNAs and identification of a 14-lncRNA prognostic signature for patients with colon adenocarcinoma.

The objective of this study was to identify potentially significant genes and long non-coding RNAs (lncRNAs) in colon cancer for a panel of lncRNA signatures that could be used as prognostic markers for colon adenocarcinoma (COAD) based on the data from The Cancer Genome Atlas (TCGA). RNA-seq V2 exon data of COAD were downloaded from the TCGA data portal for 285 tumor samples and 41 normal tissue samples adjacent to tumors. Differentially expressed mRNAs and lncRNAs were identified. A functional enrichment analysis of differentially expressed mRNAs was performed, followed by protein-protein interaction (PPI) network construction and significant module selection. Additionally, the regulatory relationships in differentially expressed mRNAs and lncRNAs were assessed, and an lncRNA-lncRNA co-regulation and functional synergistic analysis were performed. Furthermore, the risk score model and Cox regression analysis based on the expression levels of lncRNAs were used to develop a prognostic lncRNA signature. A total of 976 differentially expressed mRNAs and 169 differentially expressed lncRNAs were identified. MDFI and MEOX2 were the PPI network hubs. We found these lncRNAs to be mainly involved in vascular smooth muscle contraction and the cGMP-PKG signaling pathway. Several lncRNA-lncRNA pairs had co-regulatory relationships or functional synergistic effects, including BVES-AS1/MYLK-AS1, ADAMTS9-AS1/MYLK-AS1 and FENDRR/MYLK-AS1. The differential expression profile analysis of four candidate lncRNAs (MYLK-AS1, BVES-AS1, ADAMTS9-AS1, and FENDRR) in COAD tumors were confirmed by reverse transcription-quantitative PCR. Moreover, this study identified a 14-lncRNA signature that could predict the survival for COAD patients.

GSK-3ß phosphorylation-dependent degradation of ZNF281 by ß-TrCP2 suppresses colorectal cancer progression.

Zinc finger protein 281 (ZNF281) has been recently shown to be critical for CRC progression. However, the immediate upstream regulators of ZNF281 remain unclear. Here we reported that the E3 ligase the ß-transducin repeat-containing protein 2 (ß-TrCP2) governs the ubiquitination and degradation of ZNF281. In human CRC specimens, endogenous ß-TrCP2 were inversely correlated with ZNF281. Beta-TrCP2 reversed the phenotype of CRC cell with overexpressed ZNF281. Moreover, we found that glycogen synthase kinase 3ß (GSK-3ß), not GSK-α, could bind to and phosphorylate ZNF281 at one consensus motif (TSGEHS; phosphorylation site is shown in italics), which promotes the interaction of ZNF281 with ß-TrCP2, not ß-TrCP1, and leads to the subsequent ubiquitination and degradation of phosphorylated ZNF281. A mutant of ZNF281 (ZNF281-S638A) is much more stable than wild-type ZNF281 because ZNF281-S638A mutant abolishes the phosphorylation by GSK-3ß and can not be ubiquitinated and degraded by ß-TrCP2. Conversely, ZNF281 transcriptionally repressed the expression of ß-TrCP2, indicating a negative feedback loop between ZNF281 and ß-TrCP2 in CRC cells. These findings suggest that the turnover of ZNF281 by ß-TrCP2 might provide a potentially novel treatment for patients with CRC.

[Pathogenesis and treatment of slow transit constipation].

Slow transit constipation (STC) is generally considered as a complex idiopathic disease affected by multiple factors synergistically. Primarily caused by the condition of gut dysmotility, the transit of intestinal contents turned so slow that the moisture absorption increases, defecation frequency decreases, bowel movement is weakened or even disappeared with or without abdominal distension, dry and hard stool. Its etiology and pathogenesis remains unclear.Recently some researches reported the pathogenesis may be associated with the changes of the enteric nervous system (ENS), such as the change or degeneration of intestinal nerve cells, gut glial cell damage and neurotransmitter changes. Besides, intestinal myopathy, ICC reduction, immune factors, endocrine factors, laxative, mental psychological factors, diet and exercise habits may also be associated with the occurrence and aggravation of STC. The current understanding of STC mechanism can not meet the needs of clinical diagnosis and treatment. Conservative treatment is the main treatment of STC nowadays. For those receiving normative medical treatment but with little effect, surgery is necessary. "Jingling procedure" and "antiperistaltic anastomosis" can both get good efficacy. Treatment aiming at causes of disease will be uncovered as the development of the researches on the pathogenesis and treatment of slow transit constipation.

Changes in T lymphocyte subsets in mice with CT26 colon tumors after treatment with donor lymphocyte infusion.

The objective of this study was to detect changes in T lymphocyte subpopulations in mice with CT26 subcutaneous colon cancer after treatment with donor lymphocyte infusion (DLI) and cyclophosphamide (CP) chemotherapy. A colon cancer model was established by subcutaneous injection of CT26 carcinoma cells into BALB/C mice. The mice were randomized into different treatment groups. We recorded survival times, tumor growth inhibition rates, histopathological changes, and T lymphocyte subsets in peripheral blood of the mice. Mice treated with DLI and CP survived 33.5 ± 5.02 days, which was significantly longer than the survival time of untreated control mice (16.7 ± 2.98 days, P < 0.01). In addition, the tumor inhibitory rate was higher in mice treated with DLI and CP (89.3 %) than that in mice treated with CP or DLI alone (67.1 and 34.5 %, respectively). There were higher levels of T lymphocytes that were CD3(+) and CD4(+) in mice treated with DLI alone or the combination of CP and DLI (P < 0.05), and the ratio of CD4(+)/CD8(+) cells was significantly improved in these mice (P < 0.05). DLI combined with chemotherapy significantly prolonged survival and inhibited tumor growth in mice with CT26 colon cancer. This treatment might also improve immune function in these mice. Donor spleen cells that include high numbers of allogeneic lymphocytes and a few stem cells could induce a graft-versus-tumor effect, leading to elimination of residual cancer cells. This indicates that it is potentially a feasible adoptive cellular immunotherapy strategy for the management of solid tumors.

Gravity line strategy may reduce risks of intraoperative injury during laparoscopic surgery.

BACKGROUND: "Tilt" of surgical view was commonly shown on the monitor due to unintentional rotation of camera along its long axis by camera driver. Surgeons may be influenced on identification of anatomical structures by the tilt vision. We aimed to analyze the surgical records and videos of laparoscopic surgery, and to reveal the correlation between intraoperative complications and tilt view. METHODS: A series of 425 consecutive patients who received laparoscopic low anterior resection and abdominoperineal resection were studied, and 398 surgery videos were reviewed. Still pictures showing intraoperative injury were selected. A method was established to measure tilt angle in the still pictures according to the reference line based on several anatomic landmarks. The patients were grouped with two methods according to different study purposes. Incidence of intraoperative complication and tilt angle were calculated, and statistical analysis was performed. RESULTS: The incidence of intraoperative complications was 8.3%. Tilt of the surgical field at different degrees (<15°, 15°-30° and >30°) was found in a relatively high rate in these surgery videos (31.4%). Compared with controls, comparatively bigger tilt angles were found in all cases of complication group. It is interesting to note that intraoperative complications happened more often when the tilt angle was in the range of 15°-30° (72.7%) than >30° (18.2%). We also noted a high incidence of complication (72.7%), while tilt angle was over 15° (26%) in the first 100 cases; comparatively a steady declining low rate of complication occurrence (5-7%) and also tilt angle over 15° (9-11%) were noted in the later 298 cases. CONCLUSIONS: Rotation of camera is common during laparoscopic procedures. The tilt view increased the risk of laparoscopic procedures. Tilt angle at 15-30° is the most dangerous rotation for laparoscopic surgeries. Therefore, we propose the "Gravity Line Strategy" principle as one of the basic operating criteria to correct the tilt angle.

Clinical implications of AEG-1 in liver metastasis of colorectal cancer.

The aim of the study was to investigate the role of AEG-1 in colorectal cancer and the relationship between AEG-1 protein expression and pathological characteristics and prognosis in colorectal cancer. The AEG-1 mRNA and protein expression level between the tumor cells from the cases with liver metastasis and those from the ones without live metastasis were detected by RT-PCR and immunohistochemistry staining. The relationship between AEG-1 and clinicopathological parameters of colorectal cancer was determined. AEG-1 expressed positively in 218 (41.92%) of the 520 cases examined. It showed that both mRNA and protein level AEG-1 were over-expressed in the cases with liver metastasis compared with those without liver metastasis (P=0.01). After immunohistochemistry analysis, the expression of AEG-1 protein was related to age, Duke's stage and distant metastasis (P=0.001, 0.001 and 0.016, respectively). After survival analysis, the cases with highly expressed AEG-1 protein attained a significantly poorer postoperative disease-specific survival than those with none/low-expressed AEG-1 protein (P=0.001). In the Cox regression test, histological grade, Duke's stage and AEG-1 were detected as the independent prognostic factors (P=0.035, 0.001 and 0.010, respectively). AEG-1 protein may be a potential new early liver metastasis biomarker of colorectal cancer.

Clinical implications of activated leukocyte cell adhesion molecule expression in breast cancer.

In the study, we enrolled 150 breast cancer cases to investigate the expression status of activated leukocyte cell adhesion molecule (ALCAM), and the relationships between ALCAM expression and clinical-pathological characteristics and prognosis of breast cancer. It was observed that ALCAM was expressed at higher levels in breast cancer tissue compared to levels observed for tumor-adjacent tissue. Compared to cancers with low membranous ALCAM expression, cancers with high membranous ALCAM expression were prone to lymph node metastasis (χ2=15.910, P=0.010) and metastasis in general (χ2=5.211, P=0.029). High cytoplasmic ALCAM expression was noticeably correlated with local recurrence (χ2=7.379, P=0.012), especially for short-term recurrence (interval<2 years) (χ2=5.562, P=0.037), while not associated to long-term local recurrence (interval>2 years). The content of ALCAM protein is closely associated with the expression of estrogen receptor (ER) (P=0.024). The disease-free survival of patients with high cytoplasmic ALCAM expression was significantly shorter compared to the cases with low cytoplasmic ALCAM expression (P=0.036). In conclusion, ALCAM expressed at high levels in breast cancer. High membranous expression of ALCAM probably resulted in weakened adherent ability and metastasis. In addition, high cytoplasmic ALCAM expression strengthened invasive ability of malignant cells and then promoted tumor development.

Formation of microchimerism in rat small bowel transplantation by splenocyte infusion.

AIM: To investigate the effect of donor splenocyte infusion combined with cyclosporine A (CsA) on rejection of rat small bowel transplantation (SBT). METHODS: Male Sprague-Dawley (SD) rats and female Wistar rats weighing 230-270 g were used as donors and recipients respectively in the study. Heterotopic small bowel transplantation was performed. The rats were divided into three groups: group one receiving allotransplantation (SD rarr Wistar), group two receiving allotransplantation (SD rarr Wistar) + donor splenocyte infusion, group three receiving allotransplantation (SD rarr Wistar) + donor splenocyte infusion + CsA followed by CsA 10 mg/kg per day after transplantation, in which recipient Wistar rats were injected with 2 x 10(8) SD splenocytes 28 d before transplantation, and treated with CsA after transplantation. Finally, the specific DNA fragment of donor Y chromosome was detected in recipient peripheral blood and skin by PCR. The survival time after small bowel transplantation was observed. Gross and histopathological examinations were performed. RESULTS: The survival time after small bowel trans-plantation was 7.1 +/- 1.2 d in group 1, 18.4 +/- 3.6 d in group 2 and 31.5 +/- 3.1 d in group 3. The survival time was significant longer (P < 0.01) in group 3 than in groups 1 and 2. The gross and histopathological examination showed that the rejection degree in group 3 was lower than that in groups 1 and 2. CONCLUSION: Donor splenocyte infusion combined with CsA decreases remarkably the rejection and prolongs the survival time after rat small bowel transplantation.

[The study on the morphology character of blood-spleen barrier].

OBJECTIVE: To study the morphology and functional character of blood-spleen barrier (BSB) and establish the concept of BSB. METHODS: Thirty healthy Wistar rats were studied. Ten rats were injected with 1.5 ml mixed fluid of India ink and physiological saline through the tail vein. Histological changes of the spleen in all animals were observed with light and electron microscopy, including HE, Foot, Masson staining and immunohistochemistry of CD68 and CD34. RESULTS: Most of the carbon particles were within the splenic sinuses in marginal zone but not in the white pulp after 6 h. There was a characteristic distribution of the macrophagocytes, vessel endothelial cell, reticular tissue and collagen fiber in the BSB. CONCLUSIONS: BSB, surrounding the white pulp, is composed of macrophagocytes, marginal-sinus-endothelial cells and their basement membrane, the reticular tissue (reticular cells and reticular fibers) and collagen fibers. The role of BSB is to keep the microenvironment of white pulp stable. It becomes mature while the formation of germinal center of the white pulp. The permeability of BSB changes during its development.

[The effect and mechanism of arsenic trioxide on hepatocellular carcinoma].

OBJECTIVE: To investigate the effect and mechanism of arsenic trioxide on hepatoma cell line BEL-7402 and observe the effect and best administration method of arsenic trioxide on hepatocellular carcinoma patients who were not suitable for operation. METHODS: The cell activity and morphologic changes were studied after being treated with arsenic trioxide in different concentrations. The apoptosis was detected by flow cytometry assay and DNA fragmentation assay. The caspase-3 level of mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and fluoro-spectrophotometer. The growth inhibition of implant tumor was observed in nude mice treated with arsenic trioxide in different concentrations. Arsenic trioxide was used in hepatocellular carcinoma patients by intravenous dropping and continuous regional infusion through hepatic artery. RESULTS: The effect of arsenic trioxide on hepatoma cell lines was dependent on the time and concentration obviously. The decrease in cell number was preceded by morphological changes in the treated BEL-7402 cells that were characteristic of apoptosis, including membrane blebbing, shrunken cytoplasm, nuclear condensation and loss of adhesion. Flow cytometry assay showed an arrestment at G(2)/M phase and sub-G(1) cell peak. DNA fragmentation assay showed a marked DNA ladder. The mRNA level of caspase-3 was no change in RT-PCR whereas the protein of caspase-3 was increased after added As(2)O(3) 1 - 36 h. Caspase-3 activity began to increase after 2 h and reached a maximal level after 12 h in a linear fashion. Then, the level of caspase-3 was decreased, but still in a high level. The growth inhibition of implant tumors was obviously in nude mice. The intravenous usage of arsenic trioxide could improve the quality of life with low toxicity in hepatocellular carcinoma patients not suitable for operation. The tumor size decreased in 30 patients and AFP value decreased in 19 patients by continuous regional infusion through hepatic artery. CONCLUSIONS: Arsenic trioxide can obviously inhibit the growth of hepatoma cell line BEL-7402 through inducing hepatoma cell apoptosis. The activation and increase of Caspase-3 is the possible mechanism of apoptosis, and the acting point is in pro-enzyme level. The best result of arsenic trioxide on non-operative patients should be gotten in continuous infusion through hepatic artery.

Liver regional continuous chemotherapy: use of femoral or subclavian artery for percutaneous implantation of catheter-port systems.

AIM: To evaluate the feasibility and safety of the intraarterial chemotherapy of the liver cancer by an interventional method, catheter-port system. METHODS: Thirty-two catheter-port systems were implanted percutaneously via the femoral artery or subclavian artery. Chemotherapies were performed 0-5 d after the implantation of the catheter-port systems. The mean interval between two sequential chemotherapies was 4 wk. The occurrence of side effects of the implantation was examined clinically. RESULTS: Implantation of the catheter-port was successful in all patients. Mean patency period was 210 d. One occlusion (3.1%) of the catheter was observed. Displacement of the catheter was observed in one case (3.1%). One patient rated a hematoma in the chest wall as important. Mild hematoma was reported in 8 cases (25%). In 3 of 32 cases (9.4%), mild pain was reported initially, and dysesthesia was reported in seven (21.9%). No patient rated overall discomfort as mild, severe, or important. CONCLUSION: Percutaneous placement is feasible and safe for liver regional continuous chemotherapy. Compared with surgical placement, the overall complication rate is comparable or less.

Expression of cell cycle/growth regulator genes in human hepatocellular carcinoma and adjacent normal liver tissues.

We investigated the gene expression of the cell cycle/growth regulators in hepatocellular carcinoma (HCC) through the usage of Atlas human cancer array membranes, semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Northern blot. Hybridization of cDNA array membrane was performed with alpha-32P-labeled cDNA probes synthesized from RNA, isolated from HCC and adjacent non-cirrhotic normal liver. RT-PCR of 24 paired specimens and Northern blot of 4 paired specimens were used to confirm the expression patterns of the cell cycle/growth regulator genes identified by Atlas array hybridization. Among 79 genes related to cell cycle/growth regulators, transcription factor DP2 (TFDP-2) and E2F-3 were up-regulated, whereas dual-specificity mitogen-activated protein kinase kinase 1 (MAPKK1) and cell division protein kinase 3 (CDK3) were down-regulated in HCC. RT-PCR of TFDP-2 gave result consistent with Atlas human cancer cDNA array findings. Northern blot analysis of TFDP-2 and E2F-3 of 4 paired specimens all showed up-regulation in HCC compared to normal liver tissues. The results obtained from Atlas microarray provided for the first time a liver cancer-specific expression profile, which identified the gene expressions comprehensively and systematically. The findings may lead to better understanding of the mechanism of onset and progression of HCC. The rapid and high-throughout method of profiling gene expression by cDNA array provides an overview of the key factors that may be involved in HCC. Some genes are reported for the first time in HCC.

Gene expression profiles of hepatoma cell line BEL-7402.

BACKGROUND/AIMS: To investigate the gene expression of cancer related genes in hepatoma cell line BEL-7402 through the usage of Atlas Human Cancer Array with 588 well-characterized human genes related with cancer biology. METHODOLOGY: Hybridization of cDNA membrane was performed with 32P-labeled cDNA probes synthesized from RNA isolated from Human hepatoma cell line BEL-7402 and non-cirrhotic normal liver which was liver transplantation donor. Bioinformatics was used to analyze the result. The reverse transcription polymerase chain reaction of 24 pairs of specimens and northern blot of 4 pairs of specimens were used to confirm the expression pattern of some genes identified by Atlas arrays hybridization. RESULTS: The differential expression cell cycle/growth regulator in hepatoma cell line BEL-7402 showed a stronger tendency toward cell proliferation with up-regulation of E2F-3 and TFDP-2. The anti-apoptotic factors such as Akt-1 were up-regulated. Whereas the promotive genes of apoptosis were down-regulated, such as BAK and caspase-3. Besides this, some genes were up-regulated, such as Integrin beta 8, DNA-PK, CSPCP, cyclin C etc. A number of genes were down-regulated, which included LAR, ABL2, SKY, TDGF1 etc. In general, expression of the cancer progression genes were up-regulated, while expression of anti-cancer progression genes were down-regulated. The results of reverse transcription polymerase chain reaction of 24 pairs of specimens and Northern Blot of 4 pairs of specimens were consistent with the expression pattern of some genes identified by Atals arrays hybridization. CONCLUSIONS: Investigation of gene expression profile of hepatoma cell line BEL-7402 should help to disclose the molecular mechanism of the onset, progression and prognosis of hepatocellular carcinoma. The quick and high-throughout method of profiling gene expression by cDNA array provides us with an overview of key factors that may be involved in hepatocellular carcinoma, and may find the clue to the study of hepatocellular carcinoma carcinogenesis and molecular targets of diagnosis and therapy.

K-ras gene mutation in the diagnosis of ultrasound guided fine-needle biopsy of pancreatic masses.

AIM: To investigate the utility of K-ras mutation analysis of ultrasound guided fine-needle aspirate biopsy of pancreatic masses. METHODS: Sixty-six ultrasound guided fine-needle biopsies were evaluated by cytology, histology and k-ras mutation. The mutation at codon 12 of the k-ras oncogene was detected by artificial restriction fragment length polymorphisms using Bst NI approach. RESULTS: The presence of malignant cells was reported in 40 of 54 pancreatic carcinomas and K-ras mutations were detected in 45 of the 54 FNABs of pancreatic carcinomas. The sensitivity of cytology and k-ras mutation were 74 % and 83 %, respectively. The specialty of cytology and k-ras mutation were both 100 %. The sensitivity and specialty of k-ras mutation combined with cytology were 83 % and 100 %, respectively. CONCLUSION: High diagnostic accuracy with acceptable discomfort of FNAB make it useful in diagnosis of pancreatic carcinoma. Ultrasound guided fine-needle biopsy is a safe and feasible method for diagnosing pancreatic cancer. Pancreatic carcinoma has the highest K-ras mutation rate among all solid tumors. The mutation rate of k-ras is about 80-100 %. The usage of mutation of codon 12 of k-ras oncogene combined with cytology is a good alternative for evaluation of pancreatic masses.