Global trends in colorectal cancer and metabolic syndrome research: a bibliometric and visualization analysis.

Numerous studies have demonstrated a robust correlation between metabolic syndrome (MetS) and colorectal cancer (CRC). Nonetheless, no systematic analysis or visualization of relevant publications has been conducted via bibliometrics. This research, centred on 616 publications obtainable through the Web of Science Core Collection (WoSCC), employed CiteSpace software and VOSviewer software for correlation analyses of authors, journals, institutions, countries, keywords, and citations. The findings indicate that the Public Library of Science had the highest number of publications, while the United States, China and South Korea were the most contributory nations. Recent years have seen the mechanisms linking Metabolic Syndrome with Colorectal Cancer, including diet, obesity, insulin resistance and intestinal flora, remain a burgeoning research area. Furthermore, bariatric surgery appears to be a promising new area of study. This paper presents the initial bibliometric and visualization analysis of research literature concerning CRC and MetS which examines research trends and hotspots.

Role of antibiotics in Milligan-Morgan hemorrhoidectomy for Grade III to IV Hemorrhoids: A randomized clinical trial.

PURPOSE: This study was mainly to compare the safety and long-term clinical efficacy of using intravenous antibiotics in Milligan Morgan hemorrhoidectomy for Grade III to IV Prolapsing Hemorrhoids. METHODS: This was a parallel group, 3-arm, randomized clinical trial to evaluate the efficacy of intravenous prophylactic antibiotics. A total of 150 consecutive patients undergoing Milligan Morgan hemorrhoidectomy (MMH) in a tertiary hospital for grade III/IV hemorrhoids from January 2020 to August 2022 were enrolled. Patients were randomly assigned to three groups using a computer-generated table. Group A did not receive any prophylactic antibiotic, group B received 2 g I/V Cefoxitin Sodium before the induction of anesthesia, and group C received 2 g I/V Cefoxitin Sodium before the induction of anesthesia and 6 h after operation. RESULTS: There was no significant difference in measured VAS values on the 1st day,3rd day and 7th day after surgery (p> 0.05). Compared with VAS values on the 1st day postoperatively, these values got decreased on the 3rd day and 7th day after surgery (p< 0.05). In addition, there was no significant difference among the first defecation time, wound edema, bleeding, urinary retention after surgery (p> 0.05). There was no significant difference in the outcome comparison between all 3 groups' basal and the 3rd day postoperatively no matter in WBC, NUET% or CRP (p> 0.05). However, compared with basal, the WBC, NUET%,CRP(p< 0.05) of group A and group B on the 3rd day postoperatively got rised, the rate of recurrence of hemorrhoids follow-up for 1 year was 1.4%. CONCLUSIONS: Our results suggest that there is no efficacy on intravenous prophylactic antibiotics in Milligan Morgan hemorrhoidectomy.

Gentianine alleviates dextran sulfate sodium-induced ulcerative colitis via inhibition of TLR4/NLRP3-mediated pyroptosis.

OBJECTIVES: Ulcerative colitis (UC) is a common inflammatory bowel disorder. Gentiana scabra Bunge is a traditional medicinal plant that is used to treat a variety of diseases. Studies have shown that gentianine (GTN) from Gentiana scabra inhibits the development of inflammatory diseases. The purpose of this study was to investigate the effect and possible mechanism of action of GTN on UC in mice. METHODS: An animal model of UC was established using dextran sulfate sodium (DSS). Mice were administered intraperitoneally with GTN (12.5, 25, or 50 mg/kg/day) for seven days. Body weight and disease activity index (DAI) were monitored daily during GTN administration. Colon length, pathological changes, and myeloperoxidase (MPO) activity were measured following GTN administration. The signalling pathways regulated by GTN were analysed using machine learning. HT-29 cells were used to verify the effect and mechanism of action of GTN on UC in vitro. RESULTS: GTN suppressed weight loss, shortened colon length, alleviated colon injury, and reduced the DAI score and MPO activity of mice with UC in a dose-dependent manner. Further analysis showed that GTN inhibited the NOD-like receptor (NLR) signalling pathway. GTN markedly decreased the levels of NLR signalling pathway-related proteins. Moreover, GTN decreased the levels of pyroptosis-related proteins, IL-1ß and IL-18. The in vitro data were consistent with those of animal experiments. Furthermore, TLR4 and NLRP3 overexpression eliminated the protective effects of GTN in HT-29 cells. CONCLUSION: Gentianine alleviated DSS-induced UC by inhibiting TLR4/NLRP3-mediated pyroptosis.

Intra and inter: Alterations in functional brain resting-state networks in patients with functional constipation.

Background: Functional constipation (FCon), is a symptom-based functional gastrointestinal disorder without an organic etiology and altering brain structure and function. However, previous studies mainly focused on isolated brain regions involved in brain plasticity. Therefore, little is known about the altered large-scale interaction of brain networks in FCon. Methods: For this study, we recruited 20 patients with FCon and 20 healthy controls. We used group independent component analysis to identify resting-state networks (RSNs) and documented intra- and inter-network alterations in the RSNs of the patients with FCon. Results: We found 14 independent RSNs. Differences in the intra-networks included decreased activities in the bilateral caudate of RSN 3 (strongly related to emotional and autonomic processes) and decreased activities in the left precuneus of RSN 10 (default mode network). Notably, the patients with FCon exhibited significantly decreased interactive connectivity between RSNs, mostly involving the connections to the visual perception network (RSN 7-9). Conclusion: Compared with healthy controls, patients with FCon had extensive brain plastic changes within and across related RSNs. Furthermore, the macroscopic brain alterations in FCon were associated with interoceptive abilities, emotion processing, and sensorimotor control. These insights could therefore lead to the development of new treatment strategies for FCon.

[Effect of warm-needle moxibustion intervention on immune function and intestinal flora in patients after colorectal cancer radical operation].

OBJECTIVE: To explore the effect of warm-needle moxibustion (WNM) on the levels of T cell subgroups and serum inflammatory factors, intestinal microecological balance and postoperative adverse reactions in patients with colorectal cancer. METHODS: Eighty-four patients who underwent elective radical resection of colorectal cancer were randomly and equally divided into control (medication) group (23 men and 19 women) and WNM group (24 men and 18 women). Patients of the control group received conventional medication treatment (such as postoperative anti-infection and fluid supplementation), and those of the WNM group received conventional medication plus WNM stimulation (the acupuncture needle handle warmed by ignited moxa stick) of bilateral Zusanli(ST36), Sanyinjiao(SP6), Yinlingquan(SP9), Shangjuxu(ST37), and Zhaohai(KI6). The acupuncture needles were retained for 45 minutes every time, starting on the first day after surgery, once a day for 15 days. The number of T cell subsets (CD3+, CD4+, CD8+) positive cells was counted under fluorescence microscope, and the contents of serum tumor necrosis factor α(TNF-α) and interleukin-6 (IL-6) were detected by using ELISA, and the level of C-reactive protein (CRP) was detected by using immunoturbidimetry. The levels (logarithm of colony-forming units per gram of wet fecal weight) of Bifidobacterium, Lactobacillus, Escherichia coli and Enterococcus were determined. The adverse reactions (leukocyte decline, nausea and vomiting, peripheral phlebitis, cold stimulation sensitivity) were recorded after surgery. RESULTS: Before treatment, there were no significant differences between the two groups in the number of T cell subgroups, TNF-α and IL-6 contents, and intestinal flora numbers (P>0.05). After the treatment, self-comparison showed that the numbers of CD3+ and CD4+ positive cells, the ratio of CD4+/CD8+ and the intestinal Bifidobacterium and Lactobacillus levels in the WNM group were significantly increased (P<0.05), whereas the number of CD8+positive cells, intestinal Escherichia coli and Enterococcus levels in the WNM group, and the levels of TNF-α, IL-6 and CRP in both groups were obviously decreased in comparison with their own pretreatment (P<0.05), but no significant changes were found in the levels of CD3+ and CD4+ positive cells, CD4+/CD8+ and intestinal Bifidobacterium, Lactobacillus, Escherichia coli and Enterococcus in the control group (P>0.05). Comparison between two groups displayed that after the treatment, the numbers of CD3+ and CD4+ positive cells, the ratio of CD4+/CD8+, as well as the levels of Bifidobacterium and Lactobacillus were significantly higher in the WNM group than in the control group (P<0.05), whereas the number of CD8+ positive cells， TNF-a, IL-6 and CRP, and the levels of Escherichia coli and Enterococcus were obviously lower in the WNM group than in the control group (P<0.05). The incidence of adverse reactions including leukopenia, nausea and vomiting, peripheral phlebitis, and sensitivity to cold stimulation in the WNM group were markedly lower than those of the control group (P<0.05). CONCLUSION: WNM intervention can significantly improve the immune function, reduce the level of inflammatory factors, regulate the level of beneficial intestinal flora, and also reduce the incidence of postoperative adverse reactions in patients experiencing radical resection of colorectal cancer.

Interleukin-12B gene rs6887695 and rs2288831 polymorphisms are associated with an increased risk of ulcerative colitis development in Chinese Han population: A case-control study.

BACKGROUND: The association of the interleukin (IL)-12B gene rs6887695 and rs2288831 polymorphisms with ulcerative colitis (UC) risk has been extensively investigated, but results are conflicting. In this study, we investigated potential link between the IL-12B gene rs6887695 and rs2288831 polymorphisms and UC development in Chinese Han population. MATERIAL AND METHODS: Genotyping was performed in 367 patients and 456 controls through polymerase chain reaction-restriction fragment length polymorphism analysis. Plasma levels of IL-12B were tested using an enzyme-linked immunosorbent assay. RESULTS: We found that the IL-12B gene rs6887695 and rs2288831 polymorphisms were related to a significantly increased risk of UC. Subgroup analyses revealed significant associations of the IL-12B gene rs6887695 and rs2288831 polymorphisms with UC risk among females, consumers of alcohol, and those aged <40 years. Additionally, the rs6887695 and rs2288831 polymorphisms were associated with lesion location and UC treatment. Last, we found that these two polymorphisms were associated with IL-12B levels. CONCLUSIONS: The IL-12B gene rs6887695 and rs2288831 polymorphisms were associated with a higher risk, and the clinical characteristics, of UC.

Anti-tumor activity and mechanism of oligoclonal hepatocellular carcinoma tumor-infiltrating lymphocytes in vivo and in vitro.

Objective: To explore a method for culturing hepatocellular carcinoma and tumor-infiltrating lymphocytes (HCC-TIL) and investigate the mechanism of TIL in killing tumors. Methods: The distribution of regulatory T cells (Treg) in HCC was detected by immunohistochemistry. Conventional TIL and oligoclonal TIL were isolated by the traditional method of enzyme digestion combined with mechanical treatment for whole HCC and micro HCC tissue block culturing method. MTT was used to compare the killing activity of TIL. Flow cytometry was used to analyze the proportion of CD8+ T cells and Treg cells in TIL. Tumor-bearing mice were established, and TIL adoptive immunotherapy was performed. Results: Treg cells were mainly distributed in the stroma of HCC. In vitro experiments showed oligoclonal TIL had higher cytotoxicity to tumor cells which negatively correlated with the proportion of Treg cells. In vivo experiments showed oligoclonal TIL had a higher anti-tumor effect. IFN-γ in peripheral blood and the positive rate of intratumoral lymphocytic infiltration in oligoclonal TIL group were both higher. TGF-ß and IL-10 in peripheral blood and the positive rate of intratumoral FoxP3 and IL-17 were both lower than those in conventional TIL group. Conclusion: The oligoclonal TIL culture method could obtain TIL with higher purity, and cytotoxicity to tumor cells was associated with Treg cells. The oligoclonal TIL had cytotoxicity to autologous HCC cells and significant inhibitory effect on the growth of transplanted tumors. The mechanism might be associated with the inhibition of Treg cells proliferation, increase of IFN-γ secretion, and decrease of TGF-ß, IL-10, and IL-17 secretion.

Aqueous Extracts of Herba Cistanche Promoted Intestinal Motility in Loperamide-Induced Constipation Rats by Ameliorating the Interstitial Cells of Cajal.

Traditional Chinese medicine was reported to have good effects in treating functional constipation. This work attempted to prove the effects of aqueous extracts of Herba Cistanche (AEHC) on STC treatment and to determine the possible mechanisms by a loperamide-induced slow transit constipation (STC) model. HPLC was performed for identification and confirmation of the bioactive components in the AEHC. It was found that AEHC attenuated STC responses based on increased fecal quantity, moisture content, and intestinal transit rate, as well as serum levels of GAS, MTL, SS, and CGRP. The protein and mRNA levels of c-kit, a labeling of interstitial cells of Cajal (ICC), also increased. Meanwhile, only the protein level of SCF, a ligand of c-kit, increased. The analysis of our data suggested that AEHC could obviously improve the function of ICC via a signaling pathway involving PI3K, SCF, and c-kit and enhance colonic motility indices such as GAS, MTL, SS, and CGRP. It is interesting to note that AEHC appeared to be effective on constipation, so further experiments are necessary to clarify the exact mechanisms involved.

[Effect on Postoperative Complications of Mixed Hemorrhoids Treated with Electroacupuncture at Chengshan(BL 57), Dachangshu(BL 25)and Erbai(EX-UE 2)].

OBJECTIVE: To evaluate the effect on postoperative complications of mixed hemorrhoids treated with electroacupuncture(EA) at Chengshan(BL 57)，Dachangshu(BL 25)and Erbai(EX-UE 2). METHODS: A total of 60 patients with mixed hemorrhoids were randomly divided into control and EA groups，30 cases in each group. In addition to basic treatment, EA was applied at BL 57, BL 25 and EX-UE 2 for 30 min (2 Hz/15 Hz，0.5－2 mA) each day for 3 days after surgery in the EA group, while the control group received basic treatment after surgery. Postoperative complications after treatment were evaluated each day. RESULTS: Following surgery, anal pain, anal pendant expansion and hematochezia were observed in both the EA group and control group. On the first day after surgery, symptom improvement was not statistically significantly different between the two groups (P>0.05)．On the 2nd and 3rd days after surgery, anal pain, anal pendant expansion and hematochezia were markedly improved in the EA group compared with those in the control group (P<0.05). CONCLUSIONS: Electroacupuncture can reduce anal pain, anal pendant expansion and hematochezia after surgery in patients with mixed hemorrhoids.

Improved method increases sensitivity for circulating hepatocellular carcinoma cells.

AIM: To improve an asialoglycoprotein receptor (ASGPR)-based enrichment method for detection of circulating tumor cells (CTCs) of hepatocellular carcinoma (HCC). METHODS: Peripheral blood samples were collected from healthy subjects, patients with HCC or various other cancers, and patients with hepatic lesions or hepatitis. CTCs were enriched from whole blood by extracting CD45-expressing leukocytes with monoclonal antibody coated-beads following density gradient centrifugation. The remaining cells were cytocentrifuged on polylysine-coated slides. Isolated cells were treated by triple immunofluorescence staining with CD45 antibody and a combination of antibodies against ASGPR and carbamoyl phosphate synthetase 1 (CPS1), used as liver-specific markers, and costained with DAPI. The cell slide was imaged and stained tumor cells that met preset criteria were counted. Recovery, sensitivity and specificity of the detection methods were determined and compared by spiking experiments with various types of cultured human tumor cell lines. Expression of ASGPR and CPS1 in cultured tumor cells and tumor tissue specimens was analyzed by flow cytometry and triple immunofluorescence staining, respectively. RESULTS: CD45 depletion of leukocytes resulted in a significantly greater recovery of multiple amounts of spiked HCC cells than the ASGPR(+) selection (Ps < 0.05). The expression rates of either ASGPR or CPS1 were different in various liver cancer cell lines, ranging between 18% and 99% for ASGPR and between 9% and 98% for CPS1. In both human HCC tissues and liver cancer cell lines, there were a few HCC cells that did not stain positive for ASGPR or CPS1. The mixture of monoclonal antibodies against ASGPR and CPS1 identified more HCC cells than either antibody alone. However, these antibodies did not detect any tumor cells in blood samples spiked with the human breast cancer cell line MCF-7 and the human renal cancer cell line A498. ASGPR(+) or/and CPS1(+) CTCs were detected in 29/32 (91%) patients with HCC, but not in patients with any other kind of cancer or any of the other test subjects. Furthermore, the improved method detected a higher CTC count in all patients examined than did the previous method (P = 0.001), and consistently achieved 12%-21% higher sensitivity of CTC detection in all seven HCC patients with more than 40 CTCs. CONCLUSION: Negative depletion enrichment combined with identification using a mixture of antibodies against ASGPR and CPS1 improves sensitivity and specificity for detecting circulating HCC cells.

CXCL8, overexpressed in colorectal cancer, enhances the resistance of colorectal cancer cells to anoikis.

Anoikis is a form of apoptosis which occurs when anchorage-dependent cells either show loss of adhesion or inappropriate adhesion. Only a few cancer cells that detach from the primary site of the tumor acquire the ability to resist anoikis and form metastasis. The mechanism underlying the resistance of colorectal cancer (CRC) cells to anoikis remains unclear. Interleukin-8 (alternatively known as CXCL8) is associated with CRC angiogenesis and progression. Here, we found that a high abundance of CXCL8 or TOPK strongly correlated with poor overall and disease-free survival of 186 patients with CRC. A combination of high CXCL8 and high TOPK expressions had the worst prognosis. We showed that CXCL8 expression was negatively correlated with anoikis in CRC cells. CXCL8 treatment enhanced the resistance of CRC cells to apoptosis, which was accompanied by the increase of TOPK, and the activation of AKT and ERK. Moreover, we demonstrated that the inhibition of either ERK or AKT by specific chemical inhibitors attenuated the CXCL8-mediated resistance to anoikis. Treatment with AKT inhibitor abolished the effects of CXCL8 on TOPK expression, suggesting that TOPK was downstream of AKT in the process of anoikis. Taken together, we demonstrated that CXCL8 is strongly implicated in the resistance of CRC cells to anoikis, and that the AKT, TOPK and ERK pathway may be a potential therapeutic target for CRC.

Prognostic value of TTF-1 expression in patients with non-small cell lung cancer: A meta-analysis.

BACKGROUND: Observational studies on the prognostic role of thyroid transcription factor 1 (TTF-1) in non-small-cell lung cancer (NSCLC) are controversial. METHODS: To clarify the impact of TTF-1 in NSCLC survival, we performed this meta-analysis that included eligible studies. The combined hazard ratios and their corresponding 95% confidence intervals were calculated in terms of overall survival. RESULTS: A total of 17 studies with 2235 patients were evaluable for this meta-analysis. The studies were categorized by histology, disease stage and patient race. Our results suggested that TTF-1 overexpression had a favorable impact on survival of patients with NSCLC, the HR (95% CI) was 0.49 (0.42 to 0.55) overall, 0.46 (0.38-0.54) in Asian patients, 0.52 (0.42-0.63) in non-Asian patients, 0.45 (0.38-0.52) in adenocarcinoma, 0.63 (0.39-0.86) in stage I NSCLC, and 0.43 (0.33-0.53) in stage IIIb-IV NSCLC. The data collected were not sufficient to determine the prognostic value of VEGF in patients with squamous cell lung carcinomas. But there was a high heterogeneity between the studies. CONCLUSION: TTF-1 overexpression indicates a favorable prognosis for patients with NSCLC, this effect appears also significant when the analysis is restricted in lung AC patients, stage I and stage IIIb-IV NSCLC.

Icaritin ameliorates carbon tetrachloride-induced acute liver injury mainly because of the antioxidative function through estrogen-like effects.

To investigate the effects of icaritin, an active ingredient extracted from Epimedium Sagittatum (Sieb. et Zucc.), on CCl4-induced liver injury and its possible mechanisms. Hepatocytes isolated from Sprague-Dawley male rats were treated with 3 mmol/L CCl4 for 24 h to induce acute liver cell injury, then icaritin (0.1, 1, 10, 100 µmol/L, respectively) was administrated to the cells, and estrogen receptor antagonist ICI182,780 (1 µmol/L) was co-treated with 10 µmol/L icaritin. Biochemical parameters (alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), and superoxide dismutase (SOD)) and cell apoptosis were detected to evaluate the injury degree. Protein expressions of Bax, Bcl-2, liver fatty acid-binding protein (L-FABP), and peroxisome proliferator-activated receptor-α (PPAR-α) as well as reactive oxygen species (ROS) generation were determined by western blot. Icaritin alleviated CCl4-induced liver cell injury in a concentration-dependent manner and 10 µmol/L was the optimal concentration. Icaritin (10 µmol/L) significantly reduced activities of ALT, AST in cell culture medium and MDA level of the impaired liver cells, but increased the intercellular SOD activity. The apoptotic rate of the impaired liver cells was also decreased by icaritin (10 µmol/L) treatment. Icaritin might exert antioxidative and anti-apoptotic functions via estrogen-like effect, as the ratio of Bcl-2/Bax was significantly increased, while protein expressions of L-FABP and PPAR-α were markedly increased, and this function was blocked by the estrogen receptor antagonist ICI182,780 efficiently. Icaritin may be a promising drug candidate for acute liver injury benefiting from the antioxidative and anti-apoptotic functions via estrogen-like effect.

CCL20 and CXCL8 synergize to promote progression and poor survival outcome in patients with colorectal cancer by collaborative induction of the epithelial-mesenchymal transition.

Liver metastases represent the major cause of death in patients with colorectal cancer (CRC). Recent studies have suggested that the chemotactic responses of tumor cells are necessary for metastatic spread to the liver, and CCL20 and CXCL8 have a strong association with CRC metastasis. The aim of our study was to identify the mechanisms by which CCL20 and CXCL8 synergize to promote metastatic progression and evaluated their potential as prognostic markers for CRC patients. The abilities of CCL20 and CXCL8 to promote CRC cell progression and epithelial-mesenchymal transition(EMT)phenotype were analyzed in vitro. Possible signaling pathways were investigated with specific pathway inhibitors and small interfering RNA (siRNA). 213 Patients with CRC who underwent surgery were enrolled for analysis of CCL20, CXCL8 and E-cadherin expressions in tumor tissues. Prognostic factors were then identified. CCL20 or CXCL8 alone was not sufficient to induce complete EMT in CRC cells, but both of them could coordinately induce EMT-like phenotype that was required to maintain CRC cell proliferation, migration and invasion. PI3K/AKT-ERK1/2 pathway crosstalk was demonstrated to be responsible for this process. Coexpression of CCL20 and CXCL8 was negatively correlated with E-cadherin expression in human CRC tissues. CRC patients with coexpression of CCL20 and CXCL8 were more likely to develop liver metastases and both coexpression was an independent high-risk factor for a most poor prognosis. CCL20 and CXCL8 synergize to promote CRC metastatic progression by coordinated induction of EMT via PI3K/AKT-ERK1/2 signaling axis. Detection of both coexpressions can be used to predict clinical outcomes in CRC patients.

Changes of serum alpha-fetoprotein and alpha-fetoprotein-L3 after hepatectomy for hepatocellular carcinoma: prognostic significance.

BACKGROUND: Alpha-fetoprotein (AFP) is the most established tumor marker of hepatocellular carcinoma (HCC), but one of its limitations is non-specificity. Many studies have demonstrated that alpha-fetoprotein-L3 (AFP-L3) is more specific than AFP in the early diagnosis and prognosis of HCC. However, there is a lack of knowledge about the post-hepatectomy profiles of serum AFP and AFP-L3 values in HCC patients. To identify the profiles after surgical resection of HCC, we analyzed the correlation between the profiles and postoperative HCC recurrence or survival, and evaluated their utility in predicting postoperative therapeutic efficacy and prognosis. METHODS: From August 2003 to December 2004, 318 patients with positive serum AFP who had received surgical resections were enrolled in this study. Preoperative and postoperative serum AFP and AFP-L3 levels were measured simultaneously and regularly, and their postoperative profiles during a long-term follow-up were recorded and summarized. RESULTS: A high ratio of AFP-L3 to total AFP was shown to correlate with pathologic features of aggressiveness. The overall 1-, 3-, and 5-year recurrence rates of the whole series were 28%, 57%, and 84%, and the overall survival rates were 86%, 61%, and 33%, respectively. The changes of serum AFP and AFP-L3 after hepatectomy for HCC were classified into 3 groups (group A: AFP-L3 undetectable; group B: AFP-L3 <10%; and group C: AFP-L3 >10%). Patients with positive postoperative AFP-L3 had significantly earlier recurrence than those with negative results. The overall survival was significantly shorter in the positive groups than in the groups negative for postoperative AFP-L3. CONCLUSION: Post-hepatectomy changes in serum AFP and AFP-L3 levels occurred in three distinct patterns, which were closely correlated with HCC recurrence and patient survival with different prognostic values.

Isolation of circulating tumor cells in patients with hepatocellular carcinoma using a novel cell separation strategy.

PURPOSE: To establish a sensitive and specific isolation and enumeration system for circulating tumor cells (CTC) in patients with hepatocellular carcinoma (HCC). EXPERIMENTAL DESIGN: HCC cells were bound by biotinylated asialofetuin, a ligand of asialoglycoprotein receptor, and subsequently magnetically labeled by antibiotin antibody-coated magnetic beads, followed by magnetic separation. Isolated HCC cells were identified by immunofluorescence staining using Hep Par 1 antibody. The system was used to detect CTCs in 5 mL blood. Blood samples spiked with Hep3B cells (ranging from 10 to 810 cells) were used to determine recovery and sensitivity. Prevalence of CTCs was examined in samples from HCC patients, healthy volunteers, and patients with benign liver diseases or non-HCC cancers. CTC samples were also analyzed by FISH. RESULTS: The average recovery was 61% or more at each spiking level. No healthy, benign liver disease or non-HCC cancer subjects had CTCs detected. CTCs were identified in 69 of 85 (81%) HCC patients, with an average of 19 ± 24 CTCs per 5 mL. Both the positivity rate and the number of CTCs were significantly correlated with tumor size, portal vein tumor thrombus, differentiation status, and the disease extent as classified by the TNM (tumor-node-metastasis) classification and the Milan criteria. HER-2 gene amplification and TP53 gene deletion were detected in CTCs. CONCLUSION: Our system provides a new tool allowing for highly sensitive and specific detection and genetic analysis of CTCs in HCC patients. It is likely clinically useful in diagnosis and monitoring of HCC and may have a role in clinical decision making.

Lens culinaris agglutinin-reactive fraction of alpha-fetoprotein as a marker of prognosis and a monitor of recurrence of hepatocellular carcinoma after curative liver resection.

BACKGROUND: The aim of this study was to determine the role of Lens culinaris agglutinin-reactive fraction of alpha-fetoprotein (AFP-L3) as a prognostic marker and a monitor marker of recurrence after curative resection of hepatocellular carcinoma (HCC). METHODS: From December 2002 to May 2004, 395 consecutive patients with HCC who underwent curative partial hepatectomy were included in the study. The tumor characteristics and clinical outcomes of patients with positive preoperative and postoperative AFP-L3 were compared with those with negative results. RESULTS: A high ratio of AFP-L3 to total AFP was an indicator of pathologic aggressiveness. Patients with positive preoperative AFP-L3 had significantly earlier recurrence (median time to recurrence 22.0 ± 2.4 months vs 45.0 ± 6.9 months, P < .001) when compared with those with negative preoperative results. Significantly more patients with continuously positive or negative-turn-positive AFP-L3 results after surgery developed recurrence, particularly distant metastases, when compared with patients with continuously negative AFP-L3 results. The overall and disease-free survivals were significantly shorter in the positive than the negative preoperative AFP-L3 group. The overall and disease-free survivals were significantly shorter in the continuously positive and the negative-turn-positive than the continuously negative postoperative AFP-L3 group. CONCLUSION: Positive preoperative AFP-L3 and continuously positive or negative-turn-positive AFP-L3 results after surgery predicted a more aggressive tumor behavior, higher tumor recurrence, and poorer clinical outcomes. HCC patients with an increased proportion of AFP-L3 to total AFP should be more aggressively treated and closely followed-up.

[Clinical observation on the effect of acupuncture-assisted anesthesia for 40 cases of mixed hemorrhoids surgery].

OBJECTIVE: To observe the efficacy of comined administration of acupuncture and anesthetics for mixed hemorrhoids surgery and its postoperative complications. METHODS: A total of 40 cases of mixed hemorrhoid patients were randomized into acupuncture-assisted anesthesia (AAA) group and narcotic drug group (control), with 20 cases in each. For patients of AAA group, electroacupuncture (EA, 2 Hz/100 Hz, 9 mA) was applied to Changqiang (GV 1) and Yaoshu (GV 2) for 20 min, and 1% lidocaine (5 mL) was injected into GV 1. For patients of control group, 20 mL and 10 mL of 1% lidocaine were injected into GV 2 and GV 1, separately. The patients' pain response, anus muscular relaxion state, and postoperative complications were observed to evaluate their clinical efficacy. RESULTS: Of the 20 cases in AAA and control groups, 16 and 18 cases had a striking analgesic effect, 4 and 2 were effective, respectively. No significant difference was found between two groups in the analgesic efficacy for surgery (P>0.05). Postoperative urinary retention, pain severity I and II and the cases with intoxic response in AAA and control groups were 1 and 8, 16 and 6, 4 and 14, and 1 and 9, separately. The effects of AAA group were significantly superior to those of control group in lessening urinary retention and toxication, and in inducing analgesia (P<0.05). CONCLUSION: Acupuncture combined with narcotic drugs is superior to simple anesthetics for mixed hemorrhoids surgery in reducing postoperative complications.

Troglitazone, a peroxisome proliferator-activated receptor gamma ligand, induces growth inhibition and apoptosis of HepG2 human liver cancer cells.

AIM: To examine the effect of troglitazone, a peroxisome proliferator-activated receptor gamma (PPAR gamma) ligand, on the proliferation and apoptosis of human liver cancer cells. METHODS: Liver cancer cell line HepG2 was cultured and treated with troglitazone. Cell proliferation was detected by 3-(4-,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay; apoptosis was detected by flow cytometry and terminal deoxynucleotidyl transferase-mediated nick end labeling of DNA fragmentation sites (TUNEL) assay; and apoptosis-related protein was detected by immunocytochemistry and Western blotting. RESULTS: Troglitazone inhibited growth and induced apoptosis of HepG2 cells in a dose-dependent manner, and induced activation of caspase-3 expression. Troglitazone not only drove apoptosis-inhibiting factor survivin to translocate incompletely from the nucleus to the cytoplasm, but also inhibited expression of survivin, while it did not affect expression of apoptosis-promoting factor Bax. CONCLUSION: PPAR gamma ligands inhibit growth and induce apoptosis of liver cancer cells, and may have applications for the prevention and treatment of liver cancer.

[The specific cytotoxic effect of tumor infiltrating lymphocytes transfected with chimeric T cell receptor on cells which express KDR].

OBJECTIVE: To investigate the specific cytotoxity of tumor infiltrating lymphocytes (TIL) transfected with chimeric T cell receptor (CTCR) on cells which express KDR. METHODS: A recombinant retroviral plasmid (pMSCVneo-Vhgamma) was constructed by cloning VEGF121-hinger-FcRgamma (Vhgamma) into retroviral vector pMSCVneo. After packaging by PT67, the virus with high titer was used to infect TIL isolated from liver cancer tissues, and then MSCVneo-Vhgamma-TIL was generated. TIL infected with MSCVneo was used as a control. The cytotoxicty of the transgenic TIL on NIH3T3 and HepG2 expressing no KDR and on ECV304 and A375 expressing KDR was detected with MTT colorimetric assay. RESULTS: The sequences of VEGF121 and hinger-FcRgamma were different from those reported, but the deduced amino sequences were identical to the reported ones. The cytotoxity of TIL infected with MSCVneo on target cell was similar to that of the control TIL; both only had mild cytotoxity on cancer cell line. No significant cytotoxity was found in TIL infected with MSCVneo-cTCR on NIH3T3, but its cytotoxity on ECV304 was significant. The cytotoxity on HepG2 was similar to that of MSCVneo-TIL and uninfected TIL, but cytotoxity on A375 was significantly higher. CONCLUSION: Chimeric T cell receptor permanently grafts TIL cell with predefined new specificity. TIL expressing Vhgamma can selectively recognize and kill vascular endothelial cell and tumor cells which express VEGF receptor KDR.