Regulation of Intracellular Reactive Oxygen Species Levels after the Development of Phallus rubrovolvatus Rot Disease Due to Trichoderma koningii Mycoparasitism.

Phallus rubrovolvatus is a unique mushroom used for medicinal and dietary purposes in China. In recent years, however, the rot disease of P. rubrovolvatus has seriously affected its yield and quality, becoming an economically important threat. In this study, samples of symptomatic tissues were collected, isolated, and identified from five major P. rubrovolvatus production regions in Guizhou Province, China. Based on combined analyses of phylogenies (ITS and EF1-α), morphological characteristics and Koch's postulates, Trichoderma koningiopsis and Trichoderma koningii were identified as the pathogenic fungal species. Among these, T. koningii exhibited stronger pathogenicity than the other strains; thus, T. koningii was used as the test strain in the follow-up experiments. Upon co-culturing T. koningii with P. rubrovolvatus, the hyphae of the two species were intertwined, and the color of the P. rubrovolvatus hyphae changed from white to red. Moreover, T. koningii hyphae were wrapped around P. rubrovolvatus hyphae, leading to their shortening and convolution and ultimately inhibiting their growth due to wrinkling; T. koningii penetrated the entire basidiocarp tissue of P. rubrovolvatus, causing serious damage to the host basidiocarp cells. Further analyses revealed that T. koningii infection resulted in the swelling of basidiocarps and significantly enhanced the activity of defense-related enzymes, such as malondialdehyde, manganese peroxidase, and polyphenol oxidase. These findings offer theoretical support for further research on the infection mechanisms of pathogenic fungi and the prevention of diseases caused by them.

Six Additions to the Genus Periconia (Dothideomycetes: Periconiaceae) from Graminaceous Plants in China.

Periconia is a polyphyletic and asexual morphic genus within the family Periconiaceae (Pleosporales). The genus is characterized by a pale to dark brown stipe with an apical conidial head and ellipsoidal to oblong conidia. Species of Periconia are widely distributed throughout the world in various hosts, while most species are isolated from graminaceous plants. During our investigations of microfungal in Sichuan Province, China, 26 Periconia isolates were collected from a wide variety of graminaceous plants. These isolates corresponded to 11 species based on the examination of morphology and multi-locus phylogenetic analysis (SSU, ITS, LSU, TEF1, RPB2). This includes six new species (P. chengduensis, P. cynodontis, P. festucae, P. imperatae, P. penniseti, and P. spodiopogonis) and five new records (P. byssoides, P. chimonanthi, P. cookie, P. pseudobyssoides, and P. verrucosa). A comprehensive description and illustrations of the new species are provided and discussed with comparable taxa. These discoveries expand our knowledge of the species diversity of Periconia taxa in graminaceous plants in China.

Antiviral therapy inhibited HBV-reactivation and improved long-term outcomes in patients who underwent radiofrequency ablation for HBV-related hepatocellular carcinoma.

BACKGROUND: Hepatitis B virus (HBV) reactivation impact negatively the prognosis of patients with HBV-related hepatocellular carcinoma (HCC). This study aimed to observe the effect of antiviral therapy (AVT) on viral reactivation and long-term outcomes after percutaneous radiofrequency ablation (PRFA) for HBV-related HCC. METHODS: Data on 538 patients between 2009 and 2013 were reviewed. Propensity score matching (PSM) analysis was used to adjust for differences in baseline features between patients who received AVT (AVT group) and did not receive it (non-AVT group). Logistic regression was used to identify the independent factors for viral reactivation. The tumor recurrence and overall survival (OS) rates were analyzed using the Kaplan-Meier method. Recurrence patterns were also investigated. RESULTS: HBV reactivation developed in 10.8% (58/538) of patients after PRFA. AVT was associated independently with decreased viral reactivation (odd ratio: 0.061, 95% confidence interval: 0.018-0.200). In 215 pairs of patients obtained after PSM, the AVT group had lower 1-, 3-, and 5-year recurrence rates (24%, 55%, and 67% vs 33%, 75%, and 85%, respectively) and higher 1-, 3-, and 5-year OS rates (100%, 67%, and 59% vs 100%, 52%, and 42%, respectively) than non-AVT group (P < 0.001 for both). Additionally, the relapses in distant hepatic segments and the late recurrence after 2 years of PRFA were significantly reduced in the AVT group (78/215 vs 111/215 vs., P = 0.001; 39/109 vs. 61/91, P = 0.012, respectively). CONCLUSIONS: AVT reduced late and distal intrahepatic recurrence and improved OS in patients undergoing PRFA for HBV-related HCC by inhibiting viral reactivation.

Ophiocordycepsaphrophoridarum sp. nov., a new entomopathogenic species from Guizhou, China.

BACKGROUND: Ophiocordyceps is the largest genus in the family Ophiocordicipitaceae, including many entomopathogenic species. In recent years, many species have been described in this genus, with a wide range of host insects. Entomopathogenic fungi include ecologically, economically and medicinally important species, but a large portion of their diversity remains to be discovered and described. NEW INFORMATION: In this study, a new species, Ophiocordycepsaphrophoridarum sp. nov, parasitising Aphrophoridae sp. (Hemiptera) is proposed from China, based on evidence from morphology and molecular phylogenetic analyses. This species is characterised by fibrous, pigmented stromata, cylindrical asci and filiform ascospores. Compared to its closest relative, O.tricentri, the new species has wider perithecia and longer asci. Molecular phylogenetic analyses of a multilocus dataset (consisting of SSU, ITS, LSU, TEF1, RPB1 and RPB2) confirm its placement in Ophiocordyceps. Ophiocordycepsaphrophoridarum is morphologically described and illustrated with colour photographs. Morphological comparisons with closely-related species are also presented in tabulated format.

Microwave ablation for the treatment of hepatocellular carcinoma that met up-to-seven criteria: feasibility, local efficacy and long-term outcomes.

OBJECTIVES: This study aimed to evaluate the feasibility, local efficacy and long-term outcomes of microwave (MW) ablation for the treatment of hepatocellular carcinoma (HCC) that met up-to-seven criteria. METHODS: Between January 2007 and January 2012, 142 HCC patients with 294 nodules, which conformed to up-to-seven criteria, were enrolled into this retrospective study. All patients were followed up for more than 3 years after receiving MW ablation. Technical success, complications, local tumour progression (LTP) and distant recurrence (DR) were monitored. Recurrence-free survival (RFS), overall survival (OS) and prognostic factors were analysed. RESULTS: Primary technical efficacy was achieved in 95.2% (280/294) of the carcinomatous nodules, and major complications occurred in four (2.8%) patients. Among the 294 tumours, LTP was observed in 44 (15.0%) tumours. Among the 142 patients, DR was observed in 97 (68.3%) patients. The estimated OS rates after MW ablation at 1, 3 and 5 years were 97.2%, 75.4% and 50.6%, respectively; and the corresponding RFS rates were 76.1%, 33.1% and 19.5%, respectively. CONCLUSIONS: MW ablation is a safe and effective treatment with a high rate of primary technical efficacy for patients with HCC that met up-to-seven criteria. KEY POINTS: • The first study expanding MW ablation to HCC category beyond Milan criteria. • The first report that used up-to-seven criteria as indications for MW ablation. • HCC of up-to-seven criteria viewed as a subgroup of BCLC stage B. • MW ablation is safe and effective for treating HCC within up-to-seven criteria. • MW ablation is preferable in treating unresectable HCC within up-to-seven criteria.

Key Markers of Minimal Residual Disease in Childhood Acute Lymphoblastic Leukemia.

The aim of this study was to identify key markers of minimal residual disease (MRD) in childhood Acute Lymphoblastic Leukemia (ALL). Bone marrow samples were collected at presentation from 139 patients with newly diagnosed B-lineage ALL. On the basis of the expression of CD19, CD10, and CD34 antigens by bone marrow cells, combined with the terminal deoxynucleotide transferase (TdT), CD38, CD45, CD58, CD21, CD66c, CD22, and CD33 expression patterns characterized at diagnosis, leukemia-associated immunophenotypes (LAIPs) were identified. One hundred thirty-nine patients with a median age of 4.3 years were screened with 4-color flow cytometry MRD screening, and 119 of them exhibited 1 or more LAIP suitable for further monitoring, constituting a coverage rate of 85.6%. Only 20 of the 139 (14.4%) had no LAIP identified for follow-up. The most applicable antibody combination was TdT/CD10/CD34/CD19 (87/139, 62.6%), followed by CD38/CD10/CD34/CD19 (85/139, 61.2%) and CD45/CD10/CD34/CD19 (58/139, 41.7%). We have identified a relatively effective MRD panel, combined with TdT, CD38, and CD45 as key markers, that is applicable to the majority of newly diagnosed B-lineage ALL.

Real-time polymerase chain reaction for diagnosing infectious mononucleosis in pediatric patients: A systematic review and meta-analysis.

In this meta-analysis, we evaluated the diagnostic role of Epstein-Barr virus deoxyribonucleic acid detection and quantitation in the serum of pediatric and young adult patients with infectious mononucleosis. The primary outcome of this meta-analysis was the sensitivity and specificity of Epstein-Barr virus (EBV) deoxyribonucleic acid (DNA) detection and quantitation using polymerase chain reaction (PCR). A systematic review and meta-analysis was performed by searching for articles that were published through September 24, 2014 in the following databases: Medline, Cochrane, EMBASE, and Google Scholar. The following keywords were used for the search: "Epstein-Barr virus," "infectious mononucleosis," "children/young adults/infant/pediatric," and "polymerase chain reaction or PCR." Three were included in this analysis. We found that for detection by PCR, the pooled sensitivity for detecting EBV DNA was 77% (95%CI, 66-86%) and the pooled specificity for was 98% (95%CI, 93-100%). Our findings indicate that this PCR-based assay has high specificity and good sensitivity for detecting of EBV DNA, indicating it may useful for identifying patients with infectious mononucleosis. This assay may also be helpful to identify young athletic patients or highly physically active pediatric patients who are at risk for a splenic rupture due to acute infectious mononucleosis.

A single center clinical analysis of children with neuroblastoma.

In the present study, the cases of 59 children diagnosed with neuroblastoma (NB) were retrospectively analyzed to assess the association between the short-term efficacy of treatment and prognostic factors. In total, 59 patients with NB that were diagnosed between July 1, 2008 and June 30, 2013 at Shanghai Children's Hospital were enrolled in the present study. The follow-up was performed until December 31, 2013, and the data revealed that 43 patients (72.9%) achieved complete remission (CR) or partial remission (PR). The 3-year overall survival (OS) rate of patients with stage I, II, III, IV and IVs disease was 100, 100, 65.6, 34.8 and 85.7%, respectively (P=0.02). The 3-year OS and event-free survival rates were evidently increased in patients with favorable histology compared with the rates in the patients with unfavorable histology (P=0.046 and 0.030, respectively). Univariate statistical analysis revealed that the factors significantly associated with prognosis were patient age, tumor stage and risk group (P=0.004, 0.02 and 0.001, respectively). The present study identified that tumor stage, risk group and patient age are important prognostic factors for NB. An age of 18 months was also hypothesized to be the cut-off for the prognosis of patients.

Tumor-associated macrophages promote cancer stem cell-like properties via transforming growth factor-beta1-induced epithelial-mesenchymal transition in hepatocellular carcinoma.

Tumor-associated macrophages (TAMs), a crucial component of immune cells infiltrated in tumor microenvironment, have been found to be associated with progression and metastasis of hepatocellular carcinoma (HCC). In this study, we aimed to clarify the mechanism underlying the crosstalk between TAMs and cancer stem cells (CSCs) in HCC. Mouse macrophage cell line RAW264.7 cells were used to investigate the effects of TAMs on mouse hepatoma cell line Hepa1-6 cells in vivo and vitro. A total of 90 clinical samples had pathology-proven HCC were used to evaluate the distribution of TAMs and CSCs and analyze their value in predicting the prognosis. In the study, we have found that the number of TAMs has a positive correlation with the density of CSCs in the marginal of human HCC. Our results show that, cocultured with TAM-conditioned medium (CM) promoted CSC-like properties in Hepa1-6 cells, which underwent EMT and gained higher invasive capability. TAMs secreted more transforming growth factor- beta1 (TGF-beta1) than other phenotypes of macrophage. Furthermore, depletion of TGF-beta1 blocked acquisition of CSC-like properties by inhibition of TGF-beta1-induced EMT. High expression of CD68 in the EpCAM positive expression HCC tissues was strongly associated with both poor cancer-free survival and overall survival in patients. Our results indicate that the TAMs promote CSC-like properties via TGF-beta1-induced EMT and they may contribute to investigate the prognosis of HCC.

[Association between lymphangiogenesis and clinicopathological features and prognosis in laryngeal carcinoma].

OBJECTIVE: To detect lymphangiogenesis by labeling the lymphatic endothelial marker, lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), and study the prognostic relevance of lymphangiogenesis in laryngeal squamous carcinoma. METHODS: Clinical files and specimens of 78 patients with histologically diagnosed laryngeal carcinoma were stained with LYVE-1 as a specific lymphatic endothelial marker. The lymphatic vessel density (LVD) was measured, and the correlation between LVD and clinicopathological features of the tumor cases was analyzed. RESULTS: The mean LVD in laryngeal carcinoma (13.24 ± 5.09) was significantly higher than that in adult laryngeal papilloma (5.54 ± 3.15) and squamous dysplasia (6.76 ± 4.45, P < 0.05). The LVD of poorly differentiated tumors (15.74 ± 5.24) was significantly higher than that in the moderately differentiated tumors (13.84 ± 6.20), and the LVD in the moderately differentiated tumors was significantly higher than that in the well-differentiated tumors (11.68 ± 6.34). The LVD in stage 0 to stage II group (10.66 ± 5.70) was significantly lower than that in the stage III to IV group (17.01 ± 6.35). The lymph node metastasis group (17.25 ± 7.37) was significantly higher than non-lymph node metastasis group (8.60 ± 5.23, P < 0.05). There was no significant association between LVD and age, sex, primary site and distant metastasis. The overall survival in the patients with a LVD higher than the mean value was 33.5 month, and that of cases with a LVD lower than the mean value was 81.6 month (P < 0.05). The multivariate survival analysis showed that the clinical stage and LVD were independent prognostic factors of laryngeal cancer. CONCLUSIONS: The LYVE-1 staining histochemistry demonstrates that the lymphangiogenesis occurrs mainly at the edge of the tumors, and lymphangiogenesis plays an important role in the carcinogenesis, cancer progression and lymph node metastasis in laryngeal cancer. LVD may be an independent indicator of poor prognosis of laryngeal cancer.

[Clinical significance of sequential monitoring minimal residual disease in childhood B-cell acute lymphoblastic leukemia].

OBJECTIVE: To study the clinical significance of sequentially monitoring minimal residual disease (MRD) in childhood B-cell acute lymphoblastic leukemia (B-ALL). METHOD: Eighty one B-ALL cases were enrolled in the study from January 2004 to December 2009. Leukemia cell markers were detected by flow cytometry at diagnosis, then regularly followed-up. RESULTS: Of 81 cases, 80 achieved complete remission (CR) after induction therapy, 5-year event-free survival (EFS) was (76.80 ± 5.70)%. Among them, the EFS was (89.40 ± 5.90)% in standard risk group and (66.99 ± 13.60)% in intermediate risk group. Eight cases were screened for leukemia markers for MRD monitoring and identified in 68; and 5-year EFS was (79.10 ± 6.20)% and (62.50 ± 15.10)% (P > 0.05, respectively). MRD detection at day 35 in induction therapy showed that 52 of 68 cases were MRD negative (leukemia cells < 0.01%), the 5-year EFS being (88.50 ± 4.90)%, and 16 were MRD positive (leukemia cells ≥ 0.01%), the 5-year EFS being (42.10 ± 20.10)% (P > 0.05). Univariate analysis confirmed that there was a correlation between MRD monitoring and risk stratification. MRD detection at day 55 showed that among the 52 day 35 MRD negative cases, 51 were still negative, 1 positive, among 16 day 35 MRD positive cases, 14 (87.50%) turned negative, 2 still positive. Of the 68 cases, 9 were MRD positive within one year after CR (3 relapsed), 4 MRD positive after one year (2 relapsed) and 55 MRD negative (4 relapsed) (P > 0.05). CONCLUSIONS: Sequential monitoring MRD can find out treatment outcome and adjust therapy in time.

[Vocal fold augmentation by injection of autologous fascia and fat].

OBJECTIVE: To evaluate the effect of combination of autologous fascia and fat injection into vocal fold for the treatment of patients with unilateral vocal fold paralysis and to observe the long-term effectiveness of this procedure. METHODS: A total of 26 unilateral vocal fold paralysis patients underwent vocal fold injection under general anesthesia, meanwhile, the mucosa of the injected point was sutured through laryngoscope under direct vision. There were 6 patients underwent autologous fat injection into vocal fold (group A), and 20 patients underwent autologous anterior rectus sheath fascia and fat injection (group B). Therapeutic efficacy were evaluated by videostroboscopy, voice-related parameters analysis and voice evaluation before and after treatment. Clinical analysis of this procedure was retrospectively performed in this serial of patients. RESULTS: All patients were followed up for 24 months. On the third day after operation, there was an acute inflammatory reaction induced by the graft. This reaction disappeared three months later. In all 20 cases, videolaryngostroboscopy showed significant improvement of the glottic closure, the improvement in acoustical parameters was statistically significant (P < 0.01). Perceptual evaluation of GRBAS scale showed significant improvement of phonatory function on G, B, A scale. The results remained stable 6 - 24 months after operation and were not changed by the length of follow-up. And in the 6 cases, videolaryngostroboscopy showed significant improvement of the glottic closure at 3 months compared with preoperative observation, a little spindle-shaped disclosure. The improvement in acoustical parameters was significant statistically at 3, 6 and 24 months (P < 0.05 or < 0.01), the voice quality decreased significantly at 6 and 24 months compared with 3 months (P < 0.05 or < 0.01). The significant differences were not observed between 6 and 24 months (P > 0.05). No complications were observed in all patients perioperatively or during the follow-up period. Voice-related parameters jitter, normalized noise energy and maximum phonation time showed significant differences between Group A and Group B on 24 months (P < 0.05 or < 0.01). CONCLUSION: The combination of autologous fascia and fat vocal fold injection is an effective procedure for the treatment of unilateral vocal fold paralysis, and the stable results can be achieved during the follow-up period for 24 months.

[Tubed pectoralis major myocutaneous flap for reconstruction of circumference pharyngoesophageal defects].

OBJECTIVE: To investigate the feasibility and efficacy of tubed pectoralis major myocutaneous flap in the reconstruction of circumferential defects following resection for locally advanced hypopharyngeal and cervical esophageal carcinoma. METHODS: From Dec. 2004 to Oct. 2008, 30 patients underwent immediate reconstruction by tubed pectoralis major myocutaneous flap for circumferential defects following resection of primary tumours. Among them, 22 were hypopharyngeal carcinoma, 7 were cervical esophageal carcinoma and one was recurrent laryngeal carcinoma involved the hypopharyngeal lumen. Five of 30 patients had received previous radiotherapy and three had failed in the previous surgical procedure. In this series, 12 patients had total pharyngolaryngectomy and 18 had total pharyngolaryngectomy and partial cervical esophagectomy. RESULTS: Postoperative pharyngocutaneous fistula formation occurred in 4 patients, 2 of them with previous radiotherapy and 2 with diabetes, and the fistulae healed later. Two patients developed anastomotic strictures at the upper junction, but they had good responses to dilatation treatment and had satisfactory oral intake. The postoperative follow-up time ranged from 8 to 56 months. Median follow-up was 18 months. One-year survival rate was 71.4% and three-year survival rate was 42.5%. CONCLUSIONS: The tubed pectoralis major myocutaneous flap is a reliable procedure to reconstruct hypopharyngeal circumferential defects following resection of advanced hypopharyngeal and cervical esophageal carcinoma. This method may be the optimal choice for the reconstruction of hypopharyngeal circumferential defects following resection of recurrent carcinoma. The incidence of fistula and stenosis could be kept at an acceptable level.

Clinical studies of laser ablation in treatment of primary liver carcinoma-associated portal vein tumor thrombus.

OBJECTIVE: To explore the clinical values of laser ablation as a new treatment for portal vein tumor thrombus (PVTT). METHODS: A total of 43 male patients with primary liver carcinoma (PLC)-associated PVTT were randomized into a laser ablation group and a radiotherapy group. The laser ablation group of 21 cases received direct percutaneous laser ablation under ultrasonographic guide, followed by direct injection into the portal vein with chemotherapy drugs including mitomycin C (MMC) and fluorouracil (5-Fu). The radiotherapy group of 22 cases was treated with X-ray in three-dimensional conformal radiotherapy (3D CRT) once a day for a total of 10 days course. After the treatment, all patients were followed for postoperative adverse reactions, changes of tumor thrombus, and survival time. RESULTS: In comparison with the radiotherapy, laser ablation resulted in decreased postoperative complications such as limb fatigue, ascites, upper gastrointestinal ulcer, and bleeding. The overall remission rates (complete remission [CR] + partial remission [PR]) were 61.9% (13/21) and 31.8% (7/22) in the laser ablation and radiotherapy group, respectively, with a significant difference (P < .05). All cases in the laser group showed clinical improvement and the average survival time prolonged to greater than 19.5 months (P < .002). CONCLUSION: Percutaneous laser ablation to PVTT is an effective, safe, and simply procedure and has proven to offer significant clinical outcomes in the treatment of PLC-associated PVTT.

Treatment of portal vein tumor thrombus of hepatocellular carcinoma with percutaneous laser ablation.

BACKGROUND: Portal vein tumor thrombus (PVTT) is a common complication of hepatocellular carcinoma (HCC), and is associated with extremely poor prognosis. PATIENTS AND METHODS: In this retrospective study, we first evaluate the application of percutaneous laser ablation as a treatment for PVTT due to advanced hepatocellular carcinoma. 108 patients (2002.7-2005.12) that have adequate liver function and be in reasonably good general condition were enrolled at Eastern hepatobiliary surgery hospital. The thrombus was ablated via an optic fiber placed in the guide needle with the guiding of ultrasound. In the follow-ups, the serial imaging and laboratory routines were examined and the overall clinical progress was measured at regular intervals until time of death. In the clinical assessment, survival time and factors affecting survival time were analyzed. The changes of laboratory test (alanine transaminase and alpha fetoprotein) and clinical manifestation (ascites and diarrhea) of the PVTT patients before and after laser ablation were observed. RESULTS: Patency of the tumor-occluded portal vein branch is the only factor that affect the survival time, the longer the patency time, the longer the survival time. The long-term survivals of patients in our study are 55.56, 33.58 and 22.38% at 1, 2 and 3 years, respectively. Both laboratory test and clinical presentations were improved. Alphalpha fetoprotein in the positive patients decreased and alanine transaminase in the abnormal patients normalized at 1 month after the treatment. Ascites disappeared in 44.00% patients (11/25), and diarrhea ameliorated in 57.14% (12/21). CONCLUSION: Laser ablation might be a novel and effective treatment for PVTT associated with advanced HCC.

siRNA-mediated inhibition of hTERT enhances chemosensitivity of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. However, there is no effective treatment for HCC. It has been shown that sustained activation of telomerase is essential for the growth and progression of HCC, suggesting that telomerase is a rational target for HCC therapy. Here, we investigated the effects of siRNA-mediated knockdown of hTERT, the catalytic and rate-limiting subunit of telomerase, on the sensitivity of HCC cells to cisplatin. While silencing of hTERT and the resultant inhibition of telomerase activity by infection with the recombinant adenovirus expressing a hTERT siRNA (Ad-si/hTERT) alone did not affect the proliferation and viability of SMMC7721 and HepG2 HCC cells within five days, co-administration of Ad-si/hTERT, but not the empty adenovirus vector, with cisplatin caused much greater extent of apoptosis in vitro under the same conditions and induced significantly more robust inhibition of SMMC7721 and HepG2 tumors growth in a mouse tumor xenograft model than cisplatin monotherapy. Our results demonstrated the synergistic effect between hTERT siRNA and cisplatin in the suppression of HCC progression and indicated that the combination of hTERT-specific siRNA and cisplatin could be an effective therapy for HCC.

Methylation-related silencing of p14ARF gene correlates with telomerase activity and mRNA expression of human telomerase reverse transcriptase in hepatocellular carcinoma.

BACKGROUND AND OBJECTIVES: The p14(ARF) gene encodes a critical negative regulator of cell cycle progression. And upregulation of telomerase activity has been considered a critical step to tumors. We investigated promoter methylation of p14(ARF) and expression as well as their effect on hTERT expression and telomerase activity in HCC. METHODS: We studied p14(ARF) methylation and mRNA in 30 hepatocellular carcinomas and corresponding non-tumor liver tissues using methylation-specific PCR. Reverse transcription-PCR was performed for expression of p14(ARF) and hTERT. Telomerase activity was performed by TRAP assay. RESULTS: p14(ARF) hypermethylation was detected in 16 (53.3%) of HCC and 10 (33.3%) of non-tumor tissues, and 12 of 16 with methylation were tumor tissues around large size (> or =5 cm). p14(ARF) mRNA was observed in 25%, 80.0%, or 50.0% of tumors with hypermethylated, partially methylated or unmethylated p14(ARF) promoter, respectively. Among the 16 tumors with p14(ARF) hypermethylation, all 12 tumors lacking p14(ARF) mRNA were positive for hTERT expression, while only one of 4 tumors expressing p14(ARF) mRNA was positive for hTERT expression. CONCLUSIONS: p14(ARF) hypermethylation may contribute to silencing of p14(ARF) mRNA expression in HCC. Telomerase activity is association with inactivation of the p14(ARF)-p53 pathway induced by methylation in HCC.

[The expression and significance of urokinase-type plasminogen activator and its receptor in squamous cell carcinoma of hypopharynx].

OBJECTIVE: To investigate the expression of urokinase-type plasminogen activator (uPA) and its receptor (uPAR) in squamous cell carcinoma of hypopharynx and reveal the correlation of the major clinicopathological parameters and prognosis. METHOD: Samples of 48 hypopharyngeal carcinoma and 10 normal hypopharyngeal tissue were detected by immunohistochemistry method (SP method) for urokinase-type plasminogen activator (uPA) and its receptor (uPAR). The correlation between the expression of urokinase-type plasminogen activator and its receptor and the major clinicopathological parameters of hypopharyngeal carcinoma were analyzed by rank sum test and Spearman correlation analysis. Overall survival were analyzed according to Kaplan-Meier and log-rank statistics, the prognostic relevance of uPA and uPAR and conventional prognostic factors were analyzed by Cox analysis. RESULT: In 48 hypopharyngeal carcinoma specimens, positive expression rates of uPA and uPAR were 77.1% and 68.75% respectively, which were significantly higher than in normal tissues (P < 0.01). The uPA and uPAR positive expression was correlated with pathological grading, lymph node metastases and growth mode of hypopharyngeal carcinoma. The positive expression rate for uPA and uPAR in patients with lower pathological grading, lymph node metastases and invasion growth mode were significantly higher than in patients with higher pathological grading, non-lymph node metastases and non-invasion growth mode. Patients were followed-up postoperatively. The positive expression of uPA and uPAR were correlated with prognosis (P < 0.05 and P < 0.01). According to Log-rank statistics, patients with positive expression of uPA and uPAR had a significantly shorter survival time than those with negative expression of uPA and uPAR. Multivariate Cox analysis revealed that three independent prognostic factors for overall survival time were clinical stage, invasion growth mode and uPAR expression. CONCLUSION: The positive expression of uPA and uPAR in hypopharyngeal carcinoma were significantly higher than in normal tissues. uPAR is a new independent and strong biologically prognostic factors, which positive expression may be a powerful aid in evaluating metastatic potential and High-Risk patients in early stage of hypopharynx carcinoma ryngeal carcinoma.

[Percutaneous laser ablation in the treatment of portal vein tumor thrombus in 93 hepatocellular carcinoma patients].

OBJECTIVE: To evaluate the efficacy of percutaneous laser ablation (LA) in the treatment for portal vein tumor thrombus (PVTT) of hepatocellular carcinoma (HCC). METHODS: The PVTT of HCC patients were treated through percutaneous transhepatic laser ablation (PTLA). The survival rate, thrombus size, blood flow of embolized portal vein by thrombus, liver function, ascites and clinical presentation were observed. RESULTS: The 6-month, 1-year and 2-year survival rate of these 93 patients were 82.8%, 53.0% and 34.1%, respectively. In 11 patients with partially occluded portal vein by PVTT, the cut-surface of the PVTT diminished significantly 6 months after LA. The color blood stream signal was seen again one day after LA in all of the other 82 patients with totally occluded portal vein by thrombus, and it could still be seen in 67 of those one month later, 57 (of 71) 3 months later, 40 (of 57) 6 months later, 27 (of 32) 1 year and 4 (of 6) 2 years later after LA. In the 38 patients who survived over 1 year, PVTT was gradually atrophied and disappeared eventually in 14, PVTT was atrophied and the portal vein changed into honeycomb-like appearance in 14. In the remaining 10 patients, PVTT continued to grow and made the portal vein enlarged. It was also observed that liver function, clinical symptom and ascites were improved in various degree after LA. CONCLUSION: Percutaneous laser ablation might be an effective and safe treatment method for controlling portal vein tumor thrombus of hepatocellular carcinoma.

Expression of SSX-1 and NY-ESO-1 mRNA in tumor tissues and its corresponding peripheral blood expression in patients with hepatocellular carcinoma.

BACKGROUND: Cancer-testis antigen (CTA) is a family of the most noticeable tumor antigens which could be potential tumor markers for cancer diagnosis. In this research we aimed to investigate the expression of SSX-1 and NY-ESO-1 mRNA, two members of the CTA family, in tissue and peripheral blood of patients with hepatocellular carcinoma (HCC) to assess their feasibility for the immunotherapy and diagnosis of HCC and the association of their expression levels with diverse clinical indicators. METHODS: Thirty-six north Chinese patients with HCC and 30 normal controls were enrolled in this study. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of SSX-1 and NY-ESO-1 mRNA in tumor tissues and corresponding levels in peripheral blood of patients. RESULTS: The positive rates of SSX-1 and NY-ESO-1 mRNA expression were 61.1% (22/36) and 11.1% (4/36), respectively, in cancer tissues; 38.9% (14/36) and 5.6% (2/36), respectively, in the corresponding peripheral blood samples. No positive expression of either SSX-1 or NY-ESO-1 mRNA was detected in the samples of cancer-adjacent tissues, cirrhotic tissues, normal liver tissue or the peripheral blood of control patients. No significant relationship was found between the expression of these two genes and clinical indicators such as age, gender, tumor size, extent of differentiation, serum a-fetoprotein (AFP) level or infection with hepatitis B virus (P > 0.05). The short term recurrence rate was 46.2% (6/13) in patients whose peripheral blood expressed SSX-1 mRNA, while the recurrence rate in patients with negative SSX-1 mRNA was 28.6% (4/14). CONCLUSIONS: SSX-1 and NY-ESO-1 antigens might be new potentially promising targets for antigen-specific immunotherapy for HCC. High specific expression of SSX-1 and NY-ESO-1 mRNA suggested that we could apply them as tumor markers. The short term recurrence rate was significantly higher in patients whose peripheral blood expressed SSX-1 mRNA, suggesting that SSX-1 mRNA could be used as indicator for recurrence, metastasis and prognosis of HCC.