Selective cyclooxygenase-2 inhibitor NS-398 attenuates myocardial fibrosis in mice after myocardial infarction via Snail signaling pathway.

OBJECTIVE: The role of NS-398 in Snail pathway of myocardial cells in mice after myocardial infarction and its effect on myocardial fibrosis were investigated in this study. MATERIALS AND METHODS: C57BL/6 mice were selected to establish mouse models of myocardial infarction with permanent ligation of anterior descending branch and sham-operation models without ligation. After successful establishment of models, 30 mice were randomly divided into sham-operation group, myocardial infarction group and drug intervention group. The drug intervention group was treated with intraperitoneal injection of NS-398 (5 mg/kg) at 1 week after modeling for 3 weeks. The survival status of mice after operation was monitored, the cardiac function was detected via echocardiography, the collagen levels in heart tissue pathological sections were detected via Masson staining and Sirius red staining. Moreover, the expressions of Snail and type I collagen levels were detected via immunohistochemistry, and the Snail protein expression level and the activity and expression level of E-cadherin protein were detected via Western blotting. RESULTS: At 4 weeks after establishment of myocardial infarction model, the fibrosis reaction was obvious, and the cardiac function was decreased, accompanied with Snail activation. The administration of NS-398 for 3 weeks inhibited the Snail activity expression and significantly improved the fibrosis degree after infarction. However, it did not improve the cardiac function. Inhibiting Snail improved the fibrosis reaction after infarction, in which Snail/E-cadherin signaling pathway was involved. CONCLUSIONS: NS-398 improves the myocardial fibrosis in mice after myocardial infarction through inhibiting the Snail signaling pathway.

First Report of Aspergillus niger Causing Root Rot of Peanut in China.

Peanut (Arachis hypogaea) is one of the most important oil crops and food legumes worldwide. China sows approximately 3.5 million hectares each year and produces 40% of the world's peanuts. Fungal diseases are among the main biotic stresses affecting peanut production. Root rot is a serious disease caused by several fungi. Pythium spp., Fusarium spp., and Rhizopus spp. are some of the root rot fungi that have been reported in China. In 2012 and 2013, root rot symptoms were observed in several fields in Laixi District, Qingdao City, Shandong Province, China. The first symptoms appeared in July. Initial symptoms of the disease were brown spots on the stem base and root. Affected plants were stunted, with leaf chlorosis, reduced growth, or sudden wilting. As disease progressed, the infected tissues showed brown discoloration and rot, and abundant dark brown and black powdery spores were visible on the surfaces of affected parts. Eventually, affected plants collapsed and died. To isolate the causal organism, roots and stems were cut into sections, which were surface-disinfected with 70% ethanol solution (v/v) for 20 s, soaked in 0.1% mercuric chloride solution for 50 s, rinsed with sterilized water three times, dried, placed on Czapek's Dox agar supplemented with chloramphenicol (100 µg/ml), and incubated at 28°C for 7 days. Fungal colonies were white initially and then covered with a dense layer of dark brown or black conidial heads. The conidial head was radiate; vesicles were nearly spherical and covered with irregular metulae and phialides. Conidia were globose or subglobose (3.0 to 5.5 µm in diameter), dark brown to black, with rough cell walls. Total genomic DNA was extracted from mycelia using the EasyPure Genomic DNA Kit (TransGEN, Beijing, China). The rDNA-ITS region was amplified using PCR with the universal fungal primers ITS1 and ITS4 (2). The purified products were separately sequenced in both directions using the same primer pair. The sequences (GenBank Accession No. KJ848716) obtained were 99% similar to the ITS sequence of isolates of Aspergillus niger. This, together with the morphological characters (1) described above, suggested that the microorganism we had isolated was A. niger. Koch's postulates were completed in the laboratory by inoculating peanut. Thirty Huayu20 peanut seeds were placed in a 500-ml sterile pot with 300 g of autoclaved soil. Twenty days after seedling emergence, 15 peanut plants were wounded with a needle and inoculated with 5 ml of conidia suspension (106 ml-1). The same number of peanuts were similarly wounded and inoculated with 5 ml of sterile distilled water to serve as controls in the same pot. All peanuts were kept in a randomized complete block design at 30°C under a 12-h photoperiod. After 7 days, disease symptoms similar to those observed in the field appeared in all inoculated but not in non-inoculated peanuts. The tests were repeated three times in the greenhouse. Koch's postulates were satisfied after re-isolating the A. niger from inoculated peanuts using the method described above. To our knowledge, this is the first report of A. niger causing root rot in peanut in China. References: (1) M. A. Klich. Page 12 in: Identification of Common Aspergillus Species. Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, 2002. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Potential modulation on P-glycoprotein and CYP3A by soymilk and miso: in vivo and ex-vivo studies.

P-glycoprotein (P-gp) and CYP3A4 both play very important roles in drug bioavailability, resistance and interactions. Our in vitro studies indicated that P-gp function was activated by many isoflavones. This study investigated the in vivo effects of soymilk and miso, isoflavone-rich soy foods, on P-gp and CYP3A by tracing the pharmacokinetics of cyclosporine (CSP), a probe drug of P-gp. Rats were orally administered CSP with and without soymilk or miso. A specific monoclonal fluorescence polarisation immunoassay was used to determine the blood concentration of CSP. The results showed that soymilk and miso significantly decreased the C(max) of CSP by 64.5% and 78.3%, and reduced the AUC(0-540) by 64.9% and 78.3%, respectively. Mechanism studies revealed that the activities of P-gp and CYP3A4 were induced by soymilk and miso. In conclusion, ingestion of soymilk and miso significantly activated the functions of P-gp and CYP3A.

Rhizoctonia solani Identified as the Disease Causing Agent of Peanut Leaf Rot in China.

Peanut (Arachis hypogaea L.) is one of the most economically important oil crops in the world. Since the 1990s, the peanut industry has developed rapidly in China. However, because of the use of high-yield varieties and increased plant density, a peanut leaf rot disease occurred in Laixi Experimental Fields in Shandong Province, China in 2007. Leaves had nearly circular, brown lesions that enlarged quickly developing yellow-brown halos at the edges of the lesions. High relative humidity under field conditions led to complete necrosis of the leaves with cotton wool-like mycelia observed followed by the development of sclerotia on the leaf surface. Symptomatic plants were observed between 2007 and 2010, and symptomatic leaf tissue was collected from the Laixi Experimental Fields. An isolate (designated YF-1) from symptomatic peanut leaves was isolated and purified on potato dextrose agar (PDA) and water agar (WA) medium. On PDA, the colony appeared initially as colorless and grew to the diameter of a 9-cm petri dish within 3 days. As the mycelium aged, the colony color gradually became light brown, and sclerotia developed on the surface of the colony. YF-1 was identified as Rhizoctonia solani Kühn based on the number of nuclei per cell ranging from 4 to 13 (average 6.1), hyphal diameter being 7.5 to 12.9 µm (average 8.3 µm), branching at right angles, a septum was present near each hyphal branch with a slight constriction, and no clamp connection structures or conidia were ever observed (4). To further confirm the identity of isolate YF-1, genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Shanghai Leifeng Biotechnol. Co., Ltd.), and the complete internal transcribed spacer (ITS) region of ribosomal DNA was amplified and sequenced with a pair of primers ITS1/ITS4 (2). A GenBank BLAST search produced an exact match for the sequences of R. solani (AY154301), with 100% sequence similarity. To estimate the mode of anastomosis, YF-1 was paired on WA medium with each reference strain belonging to anastomosis groups (AGs) 1 through 8 (provided by Shandong Agriculture University) (1,3). The results indicated that YF-1 belonged to group AG-1, subgroup AG-1-IA of R. solani. Pathogenicity tests were conducted by inoculating 10 peanut leaves using a colonized paper disc method (filter paper 1 cm in diameter suspended in the mycelia suspension). Ten control leaves received paper discs without mycelium. Inoculated and non-inoculated plants were kept in humid chambers for 24 h at 25°C. Three days after inoculation, the leaves developed typical brown lesions that were similar to those of naturally diseased plants. Koch's postulates were fulfilled by reisolation of R. solani from symptomatic leaves. No symptoms were observed on control leaves. To our knowledge, this is the first report of peanut leaf rot caused by R. solani. Occurrence of the disease in China is a new threat to the health of peanut. References: (1) Y. X. Chen et al. Acta Phytopathol. Sin. 3:139, 1985. (2) T. Misawa and S. Kuninaga. J. Gen. Plant Pathol. 76:310, 2010. (3) A. Ogoshi. Ann. Phytopathol. Soc. Jpn. 38:117, 1972. (4) J. R. Jr. Pameter and H. S. Whitmey. UC Press. 135, 1970.

Enhancement of anaerobic biohydrogen/methane production from cellulose using heat-treated activated sludge.

Anaerobic digestion is an effective technology to convert cellulosic wastes to methane and hydrogen. Heat-treatment is a well known method to inhibit hydrogen-consuming bacteria in using anaerobic mixed cultures for seeding. This study aims to investigate the effects of heat-treatment temperature and time on activated sludge for fermentative hydrogen production from alpha-cellulose by response surface methodology. Hydrogen and methane production was evaluated based on the production rate and yield (the ability of converting cellulose into hydrogen and methane) with heat-treated sludge as the seed at various temperatures (60-97 degrees C) and times (20-60 min). Batch experiments were conducted at 55 degrees C and initial pH of 8.0. The results indicate that hydrogen and methane production yields peaked at 4.3 mmol H2/g cellulose and 11.6 mmol CH4/g cellulose using the seed activated sludge that was thermally treated at 60 degrees C for 40 min. These parameter values are higher than those of no-treatment seed (HY 3.6 mmol H2/g cellulose and MY 10.4 mmol CH4/g cellulose). The maximum hydrogen production rate of 26.0 mmol H2/L/d and methane production rate of 23.2 mmol CH4/L/d were obtained for the seed activated sludge that was thermally treated at 70 degrees C for 50 min and 60 degrees C for 40 min, respectively.

Molecular analysis of the coat protein gene of peanut stripe virus from China.

Peanut stripe virus (PStV) is one of the most common viruses infecting peanut that causes great economic losses every year. The 3ʹ-terminal 1082 bp of 74 PStV isolates collected from 12 districts of Shandong province, China were sequenced. Their coat protein (CP) genes were 864 bp in length and shared identities of 98.0%~100% and 98.3% ~100% at nt and aa levels. The identities between the CP genes of these isolates and other 36 isolates from the GenBank were 93.5%~100% and 92.0%~100% at nt and aa levels, respectively. PStV isolates can be clustered into two phylogenetic groups. The isolates from United States, mainland China, and Indonesia formed group I and those from Viet Nam, Thailand, and Taiwan formed group II. The PStV isolates in group I can be further classified to two subgroups. The gene flow of PStV populations within a country was frequent, but that between countries was infrequent.

A novel macrocyclic spermine alkaloid from Incarvillea sinensis.

A novel macrocyclic spermine alkaloid incasine C' (1), along with a known compound incasine C (2), were isolated from the whole plants of Incarvillea sinensis, and their structures were elucidated on the basis of chemical and spectroscopic evidence.

Parental concerns for the child with febrile convulsion: long-term effects of educational interventions.

OBJECTIVE: To evaluate long-term effects of educational interventions on parental concerns for children with febrile convulsions (FC). SUBJECTS AND METHODS: 209 FC parents were recruited from southern Taiwan, and assigned into program or pamphlet groups by their choices. A questionnaire was used to examine the parental concerns at 4 time points: prior to the interventions, 3 months, 1 year, and 2 years after the interventions. RESULTS: The concern scores were high before the interventions. Parents concerned that their child is apt to get a fever, has further seizures in the night, that FC results in potential brain damage and is life threatening. After interventions the concern scores were significantly lower for the program group than for the pamphlet group. The educational program was the most significant factor that influenced the change in parental concerns. The recurrence of FC did not significantly influence the change in the concern trajectory in the program group but showed a significant difference in the pamphlet group. CONCLUSION: FC parental concerns decreased as time passed particularly in the program group. The educational program was the significant factor that influenced the change of concern trajectories, and the recurrent/non-recurrent factor influenced the change of concern only in pamphlet group. FC parental concerns should be assessed, and information given combined with emotional support.

Risk factors for a first febrile convulsion in children: a population study in southern Taiwan.

PURPOSE: To identify risk factors for a first febrile convulsion among 3-year-old children by a matched case-control population study. METHODS: All 11,714 neonatal survivors born in Tainan City between October 1989 and September 1991 were enrolled. At age 3, 10,460 children were available for telephone survey for febrile convulsions, and were confirmed by home visit interviews. Those without history of seizure were randomly matched to each febrile convulsion case by age, gender, and residence district. RESULTS: Two hundred fifty six children had febrile convulsions, and 218 of them and their matched controls were available for analysis. The febrile convulsion cases had significantly more febrile episodes (four or more) per year (33.0 vs. 22.5%; p = 0.021), and cases had a higher percentage of developmental delay (3.7 vs. 0.4%; p = 0.046) and a higher percentage of febrile convulsions in their siblings (12 vs. 0.4%; p = 0.011) than controls. The other sociodemographic, environmental, and biologic variables showed no differences between cases and controls. Step-wise logistic regression showed a highly significant independent association between febrile convulsions and history of febrile convulsions in the siblings, and a moderate one between febrile convulsions and the number of febrile episodes per year. CONCLUSIONS: The presence of febrile convulsions in the siblings and the number of fever episodes per year were the independent and significant predictors of febrile convulsion for an individual case in our population-based sample.

Prevention of chronic lung disease in preterm infants by early postnatal dexamethasone therapy.

Recent studies suggest that early dexamethasone therapy may lessen the pulmonary inflammation in preterm infants with respiratory distress syndrome (RDS). To investigate whether early (<12 hr) postnatal dexamethasone therapy would reduce the incidence of chronic lung disease (CLD), a randomized, double-blind, controlled trial was conducted in 40 infants (birth weights from 500 to 1,999 gm) who had severe RDS and required mechanical ventilation within 6 hr of birth. All infants received one dose of Survanta before they were randomly assigned to control (saline placebo) or dexamethasone-treated groups (0.5 mg/kg/d for 1 week, then tapered over 3 weeks). Sequential analysis was performed with the end point of assessment being the presence or absence of CLD on postnatal Day 28. Statistical significance favoring dexamethasone was reached when 12 consecutive pairs in which one infant had CLD and the other did not have CLD showed that ten pairs favored dexamethasone and two pairs favored control treatment. Among the survivors, 12/15 were extubated in the dexamethasone group and 9/16 in the control group at the end of study. Infants in the treated group had transient hyperglycemia and hypertension. There was no difference between the groups in mortality and in incidence of sepsis or intraventricular hemorrhage. We conclude that early postnatal dexamethasone therapy is potentially effective in the lessening of CLD in preterm infants. To substantiate our result, large randomized controlled trials are needed and warranted.

Potential value of urinary intercellular adhesion molecule-1 determination in patients with bladder cancer.

OBJECTIVE: Intercellular adhesion molecule-1 (ICAM-1) is known to play a role in immunity against bladder cancer and can be detected in the supernatants of cultured bladder cancer cells that constitutively express ICAM-1. This study was performed to examine the relevance of the ICAM-1 urine test in patients with bladder cancer. METHODS: A total of 53 patients with bladder carcinoma, 35 with history of bladder cancer, and 30 normal control subjects were included in this analysis. Urinary ICAM-1 (ulCAM-1) levels were measured by immunoassay and corrected for hydration status. RESULTS: Levels of ulCAM-1 were significantly elevated in patients with bladder cancer or those at tumor-free status compared with normal control subjects (P=0.001). However, there was no apparent difference between the two groups of urothelial disorders (P >0.1). ulCAM-1 did not correlate with clinicopathologic variables of bladder cancer or patient outcome (P >0.1). Six patients at tumor-free status had multiple ulCAM-1 determinations during the study period. Three of these 6 patients had elevated ulCAM-1 levels and proved to have recurrent tumors; 3 of the 6 had stable ulCAM-1 levels and were still free of disease. CONCLUSIONS: Our results suggest that urinary excretion of ICAM-1 is elevated in the early stage of bladder carcinogenesis, but is independent of biologic properties of bladder cancer. Serial monitoring of ulCAM-1 may be helpful in selecting patients who are at risk of tumor recurrence.

Urinary excretion of transforming growth factor-alpha in patients with transitional cell carcinoma.

This study was performed to examine the involvement of transforming growth factor-alpha (TGF-alpha) in urothelial tumorigenesis. TGF-alpha urine levels were measured in patients with urothelial carcinoma (n = 68), patients who were tumor-free (n = 58), patients with non-neoplastic inflammatory disease (n = 20), and normal controls (n = 39). Both inflammatory and neoplastic urologic diseases had elevated TGF-alpha urine levels (169.5 ng/gm and 116.7 ng/gm, respectively) as compared to normal controls (39.1 ng/gm) (P = 0.0001). For patients with active cancer, TGF-alpha levels were positively associated with histologic grading (P = 0.009), nodular shape, expression of epidermal growth factor receptor in primary tumor (P = 0.03, respectively). But, there was no important relationship with staging classification, number and size of tumor (P > 0.1, respectively). TGF-alpha urine levels did not correlate with the serum content (n = 26; P > 0.5), or the immunohistochemical expression of TGF-alpha (n = 60) in corresponding tumor (P < 0.05, 0.1). Significant factors in predicting patient survival were clinical staging, nodular shape and size of tumor (P < 0.05, respectively). Our data implies that interaction of urinary TGF-alpha/urothelial epidermal growth factor receptor may play a positive role in the carcinogenesis of human urothelium.

Unusual lesions associated with avian poxvirus infection in rosy-faced lovebirds (Agapornis roseicollis).

An epornitic of avian pox occurred in rosy-faced lovebirds (Agapomis roseicollis). The infected birds showed a variety of lesions including cutaneous, diphtheritic, systemic and oncogenic entities. Proliferative changes with cytoplasmic inclusion bodies in the cornea, bursa of Fabricius, and cranial and nasal bones which were found in the present cases have not been described previously. Electron microscopic examination of the skin, cornea, and cranial and nasal bones revealed poxvirus virions in the inclusions. Secondary infection of candidiasis was very common in cutaneous pox lesions.

[Study of effects of Chinese herbal prescription combined with copper, iron on malignancy of cancer cells].

BALB/c mice bearing ascitic liver cancer were used, and Chinese herbal prescriptions combined with Cu and Fe (CHCF) was given by gavage continuously for 10 days, and some cell biological parameters were measured; furthermore, the ascitic cancer cells (control and treated group) were removed, and retransplanted to another mice and observed. The results showed that in CHCF treated group, DNA content of the cancer cells was decreased, and the proliferation index (PI) was reduced (control: 83.4 +/- 2.6, CHCF group: 78.8 +/- 1.5; or control: 67.2 +/- 1.3, CHCF group: 64.2 +/- 1.6, P < 0.05). The number of the cancer cells in G1 phase increased obviously, but, those of S+G2M phases decreased considerably (P < 0.05-0.01); on the DNA histogram, the diploid peak became higher and bigger, but multiploid peaks became smaller. Furthermore, retransplanted experiments showed that in 2/10 animals, the tumor did not grow, the growth inhibition rate was 71.7%-88.3%; and the survival period of retransplanted animals was prolonged significantly (from 26.1 +/- 11.8 days to 38.1 +/- 9.6 days, or to 39.6 +/- 7.2 days, P < 0.01); the increase in life span was 46%-52% respectively. These results suggested that CHCF could decrease the malignancy of liver cancer cells in mice.

Expression of c-erbB-2 protein in normal and neoplastic urothelium: lack of adverse prognostic effect in human urinary bladder cancer.

The clinical significance of c-erb B-2 expression in urinary bladder cancer remains controversial. We performed an immunohistochemical study to examine the expression of c-erb B-2 in non-neoplastic urothelium (n = 12) and transitional cell carcinoma of the urinary bladder (n = 82). c-erb B-2 protein was localized in superficial and some intermediate cells of non-neoplastic urothelium. A total of 29 out of 82 (35%) tumors were positive for c-erb B-2 over-expression. There was no significant association of c-erbB-2 expression with tumor grade (p = 0.12), stage (p = 0.93), DNA ploidy status (p = 0.56) and the sex of patients (p = 0.5). Expression of epidermal growth factor receptor and Ki-67 index was available in 33 cases. Both parameters showed no apparent association with c-erbB-2 expression (p = 0.53 and 0.58 respectively). Factors correlated with poor patient survival by univariate analysis were tumor stage (p = 0.0001), tumor grade (p = 0.001), development of second recurrence (p = 0.002) and negative expression of c-erbB-2 (p = 0.017). Important indicators associated with first recurrence were tumor stage (p = 0.028), and c-erbB-2 expression with the risk of second recurrence (p = 0.031). Multivariate survival analysis revealed that tumor stage was among the most important prognostic factors (p = 0.029), followed by tumors without c-erbB-2 expression (p = 0.031) with a median follow-up at 46 months. The age of patients and c-erbB-2 expression were significantly associated with developing second recurrence (p = 0.031 and 0.046 respectively). The results indicate that expression of c-erbB-2 is independent of the stage and grade of bladder cancer. Although c-erbB-2 status can discriminate subpopulations with a high risk of recurrence, evaluation of its expression in paraffin-embedded tumors does not indicate poor prognosis for patients with urinary bladder cancer. To address this discrepancy a better understanding of the regulatory mechanism and physiological properties of c-erbB-2 protein in urothelium is required.

Study on late effects of radioiodine on rats.

This paper describes the results observed on 850 male Wistar rats for two years after intraperitoneal injection with 131I, 132I or 125I of different radioactivities. The incidence of thyroid malignant tumor was 45.8% when the dose absorbed by the thyroid gland was 7.8 Gy in the 131I groups, and even tumors of lung and soft tissue, and malfunction of kidney and immune organs could be elicited when the radioiodine was overdosed. Both 132I and 125I have their optimal carcinogenic dose ranges and optimal doses of carcinogenesis. As the dosage decreases to a certain level, a low-limit dose incapable of inducing cancers may be present. 131I induced mainly papillary and mixed type carcinomas, 132I chiefly follicular and undifferentiated type carcinomas, while 125I induced follicular and medullary carcinomas. The paper also discusses cytochemical DNA quantitative assays of different histological types of thyroid gland carcinomas, holding that reference to such data as decreasing serum T4, elevating serum TSH and CIC may be helpful for early diagnosis of thyroid carcinomas.