The function and molecular mechanism of CEP55 in anaplastic thyroid cancer.

OBJECTIVE: The purpose of this study was to determine the role of centrosomal protein of 55 kDa (CEP55) in anaplastic thyroid cancer (ATC) and to further explore the mechanism, which might provide a new molecular marker for treatment of ATC. PATIENTS AND METHODS: The expression level of CEP55 in clinical cases was tested by fluorescence quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Also, qRT-PCR assay was performed in different TC cell lines. The relationship between CEP55 expression and clinicopathological characteristics was statistically analyzed. Kaplan-Meier curve and Cox's proportional hazards regression model were performed in survival analysis. Further, Western blot assay was used to analyze the protein expression changes in PI3K/Akt pathway. RESULTS: The expression level of CEP55 in TC tissues showed a noticeable upgrade, especially in ATC. In vitro, CEP55 expression was also increased in four kinds of TC cells, in which, the highest expression was found in ATC (TA-K) cells. The clinicopathological features, including lymph node metastasis, distant metastasis, and prognostic index were found to be correlated with the expression level of CEP55. Besides, the ATC patients with higher expression of CEP55 had a statistically worse overall survival (OS) time. In univariate analyses and multivariate analyses, the CEP55 level was an independent prognosis index of patients with ATC. In vitro study, CEP55 protein expression level was significantly reduced in si-CEP55-transfected TA-K cells. Notably, the downregulation of CEP55 could suppress the phosphorylation of PI3K and AKT. CONCLUSIONS: This study found that CEP55 could promote ATC progression, and PI3K/AKT pathway might be the downstream target of its action. These results provided a new therapeutic direction for the treatment of ATC.

First Report of Wilt of Eucalyptus Caused by Ceratocystis fimbriata in China.

Eucalyptus spp., most of which are native to Australia, are one of the world's most commonly planted forestry crops, and have been widely planted in the tropics and subtropics. Eucalyptus trees are distributed in more than 600 counties in 19 provinces/regions throughout China, especially in the southern regions of the country (1). From April of 2013 to May of 2014, eucalyptus trees were noted to be wilting and dying rapidly in Kunming, Yunnan Province (25°02' N, 102°42' E), southwest China. Symptomatic trees typically have many wilted or dead leaves hanging throughout the tree or on some major branches, sometimes followed by death of the whole tree. Dark brown to black discoloration can be seen in the woody xylem, and elongated cankers were also present on some affected trees. A fungus was consistently isolated from the leaves and cankers when symptomatic tissue was incubated between two slices of fresh carrot root. Spore masses were removed from the apices of perithecia, transferred to malt extract agar medium (MEA), and incubated at 25°C. Perithecia developed on the medium, and were black, globose, 212.5 to 242.4 × 207.5 to 254.2 µm, and possessed a long black neck (952.3 to 1,303.3 µm). Ascospores had the typical "hat" morphology and were 4.3 to 5.2 × 3.0 to 3.8 µm. Cylindrical endoconidia (11.2 to 22.2 × 3.9 to 6.1 µm) were found. Chlamydospores produced on media were ovoid and smooth, and were 7.8 to 9.7 × 9.9 to 12.8 µm. Chains of barrel-shaped conidia were not found. PCR amplification and sequencing of the ITS region of rDNA were carried out for one isolate, E2-2, using the procedures of Thorpe et al. (3). Analysis of ITS sequence data (GenBank Accession No. KJ511481) showed that the isolates were 99% homologous to the isolate of C. fimbriata from diseased Colocasia esculenta in Cuba, China, and Hawaii (AY526304 to 06) by BLAST analysis. Thus, the fungus was identified as C. fimbriata based on morphological and molecular characteristics. Pathogenicity tests were made on 1-year-old E. grandis seedlings as follows. A conidial suspension of each isolate was diluted to 106 conidia/ml, and 0.2 ml was injected into wounds on three petioles on each of five healthy plants of E. grandis, and control seedlings were injected with sterile water that had been placed on MEA plates. The seedlings were incubated at 25°C in moist chambers. After 3 days, all inoculated E. grandis plants showed dark brown to black discoloration in the leaf axils. After 5 days, leaf wilting was present. C. fimbriata was re-isolated from all inoculated symptomatic tissue. No symptoms were visible in the control plants and no fungus was isolated from them. Ceratocystis wilt was first observed in eucalyptus in 1997 in the state of Bahia. This was followed by a report of C. fimbriata causing wilt of E. grandis in the Republic of Congo, Uganda, and Uruguay (2). Chen et al. reported two species of Ceratocystis, C. acaciivora and a previously undescribed species C. chinaeucensis, from eucalyptus plantations in Guangdong Province in China (1). To our knowledge, this is the first report of C. fimbriata causing wilt of eucalyptus in China. References: (1) S. F. Chen et al. Fungal Diversity 58:267, 2011. (2) F. A. Ferreira et al. Fitopatol. Bras. 24:284, 1999. (3) D. J. Thorpe et al. Phytopathology 95:316, 2005.

Biological and genetic characteristics of two Trichinella isolates in China; comparison with European species.

The most recent taxonomic revision in Trichinella genus included 10 taxa. Trichinella spiralis (Ts) is a thoroughly studied species but most of the isolates came from the Europe or American continents. Few information is available from China about the Trichinella isolates and their diversity. In this report two Chinese isolates were characterized and compared with European strains. The in vitro release of newborn larvae (NBL) was determined for the two species. NBL is observed in supernatant of cell culture after adults purification on day 4 post infection (pi) with Ts, and on day 5 pi with Trichinella nativa (Tna). A new parameter was thus proposed to characterize Trichinella strains. Restriction fragment length polymorphism (RFLP) of genomic DNA and random amplified polymorphic cDNA (RAPD) were used to define genetic variability among different isolates. Species specific pattern could be obtained with RAPD but it was far more difficult to get geographical markers for Trichinella using these methods. A low genetic variability in Trichinella species (i.e. a strong "clonality") is suggested.

Reappraisal of diagnostic significance of a hepatoma-specific band of serum gamma-glutamyl transferase.

The hepatoma-specific band of serum gamma-glutamyl transferase II (GGT II) and other three markers were evaluated in 77 patients with primary hepatocellular carcinoma (PHC). The positive rate of GGT II (87%) was much higher than that of the increased alpha-fetoprotein (AFP greater than or equal to 400 ng/ml, 54.5%), the increased alpha-1-antitrypsin (AAT greater than or equal to 400 mg/dl, 64.9%) and alkaline phosphatase isoenzyme I (ALP I, 13.0%). In patients with AFP less than 400 ng/ml, the positive rate of GGT II was 95.2%, higher than that of ALP I (22.8%) and AAT (60.0%). The positive rate of GGT II was positively correlated to the volume of PHC (r = 0.324, P less than 0.05), but even in patients with small PHC (less than or equal to 65 cm3), the positive rate of GGT II (78.6%) was higher than that of AFP (50.0%) and AAT (28.6%). The ALP I positivity was only seen in patients with larger PHC. Follow-up study showed that GGT II, like AFP, might occur before liver tumor could be detected by B-mode ultrasonography and computerized tomography. Therefore, GGT II is a valuable marker of PHC, especially in patients whose AFP was negative or slightly increased; GGT II may be useful for relatively early diagnosis of PHC.

Serum alpha-1-antitrypsin and alpha-1-antichymotrypsin in the diagnosis of primary hepatocellular carcinoma.

Both alpha-1-antitrypsin (AAT) and alpha-1-antichymotrypsin (AAC) in serum were investigated in various liver diseases. Serum levels of AAT and AAC in normal controls were 293.3 +/- 37.9mg/dl and 119.6 +/- 18.9U/ml, respectively. Both AAT and AAC were significantly higher in 75 patients with primary hepatocellular carcinoma (PHC) than in patients with acute viral hepatitis, chronic hepatitis, liver cirrhosis as well as normal controls. In a cut-off value of more than 400 mg/dl AAT showed a sensitivity rate (SS) of 74.7%, specificity (SP) of 97.3%, positive predictive value (PV+) of 86.2%, negative predictive value (PV-) of 95.5% and a total accuracy of 93.3% for diagnosing PHC. In a cut-off value of more than 157 U/ml, the corresponding figures for AAC were 68.0%, 96.7%, 86.4%, 90.6% and 90.0%. If the increased levels in one or both tests are positive, the combined positive rate is 80.0% in PHC. In 11 PHC cases with serum alpha-fetoprotein (AFP) less than 50 ng/ml, the positive rate of AAT and AAC were 72.7% and 81.8%, respectively, compared to corresponding figures of 71.7%, and 65.2% in 46 cases of PHC with more than 50 ng/ml of AFP. It shows that combined assay of both serum AAT and AAC has a complemental value in the diagnosis of PHC, especially in patients without increased AFP.

[Coronary angiography in patients with U wave inversion during coronary artery spasm].

During ergonovine-induced vasospastic angina, U wave inversion without significant ST segment deviation on the precordial electrocardiograms was documented in four patients. Coronary angiography revealed incomplete spastic obstruction of the left anterior descending artery without delayed filling and runoff in three patients. In the remaining patient, the proximal left anterior descending artery was totally occluded and there were well-developed collaterals from the non-spastic artery. Thus, ergonovine-induced U wave inversion was related to the presence of coronary vasospasm, and angiography demonstrated less severe myocardial ischemia in such patients than in cases with ST segment elevation or depression, which is usually associated with subtotal or total obstruction of a major coronary artery without adequate collaterals. In their clinical courses, two patients had episodes of angina with ST segment elevations or depressions. It was suggested that vasospastic angina with U wave inversion alone is one aspect of a continuous spectrum of vasospastic myocardial ischemia.

[The significance of the kallikrein-kinin system in the pathogenesis of essential hypertension].

The levels of plasma kininase II (ACE), a high molecular weight-kininogen (HMW), alpha 2-mega-globular protein (alpha 2M), prekallikrein (PK), alpha 1-antitrypsin (alpha 1 AT) were determined in essential hypertension and control groups. The results showed that there is significant increase of ACE and PK activity and HMW content in patients with essential hypertension than in control group (P less than 0.001). No changes in alpha 2M and alpha 1AT were found. These results suggest that human plasma kallikrein system was inhibited in patients with essential hypertension.

The diagnostic value of a hepatoma-specific band of serum gamma-glutamyl transferase.

Using a vertical slab electrophoretic apparatus loaded with polyacrylamide stage gel plate, we observed a total of II bands of gamma-glutamyl transferase (EC 2.3.2.2,GGT) on a gel plate and designated these GGT I-XI by order of distance beginning at the positive pole. Positive GGT II was found in 81 (90%) of 90 cases of primary hepatic carcinoma (PHC), in 9(90%) of 10 cases of secondary hepatic carcinoma (SHC), and in only 4 (3.1%) of a total of 128 cases of acute and chronic liver disease. There was one case with positive GGT II among 45 cases of cancer of ampulla of Vater, but the possibility of hepatic metastasis could not be excluded. No GGT II was found in a total of 250 healthy persons and 21 pregnant women. PHC patients with negative alpha-fetoprotein (AFP) had a positive GGT II rate of 84.6%, whereas in those with negative GGT II the positive rate for AFP was only 44.4%.

[Clinical significance of serum "hepatoma-specific" gamma-glutamyl transferase and alkaline phosphatase].

By means of polyacrylamide gel stage electrophoresis designed by us, serum gamma-glutamyl transferase (GGT) and alkaline phosphatase (ALP) were separated into 9 (I to IX) and 7 (I to VII) isoenzyme bands. The GGT II and ALP I were found only in the sera of patients with primary hepatic carcinoma showing positive rates of 29.5% and 8.5%, respectively. The combined positive rate of the two kinds of isoenzymes was 36.2% which is lower than that of alpha-fetoprotein (AFP) in the liver cancer but it has a higher specificity compared to the latter test. Furthermore, positive GGT II and ALP I may also be present in patients with liver cancer who were negative for AFP. It is obvious that the simultaneous determinations of GGT II, ALP I and AFP are mutually supplementary in the diagnosis of liver cancer.