Use of the supraclavicular artery island flap for reconstruction of maxillofacial defects: a case report and literature review.

BACKGROUND: Free flaps are widely used in maxillofacial reconstruction; however, this approach was not feasible in the current case. It was not possible because the free flap method requires microvascular anastomosis expertise, which is difficult, time-consuming and costly. CASE PRESENTATION: An 86-year-old woman suffered squamous cell carcinoma on the right side of her face, which resulted in a large soft-tissue defect. Here, we present a case of facial reconstruction from the inferior margin of the jaw to the top of the head. The size of the defect was 18.5 cm × 7.5 cm, which is rare for a patient of this age in the maxillofacial area. We used the supraclavicular artery island flap (SCAIFP) which measured 19.3 cm × 8.3 cm to repair the defect. After the operation, the flap survived without complications. Then, the patient was followed for 10 months and was satisfied with the aesthetic and functional results at the donor and recipient sites following the tumour resection. The tumour did not recur, and facial nerve function was preserved. CONCLUSION: Our results provide a new choice for the reconstruction of large defects of the head and face, and expand the potential applications of the SCAIFP.

First Report of Root Rot on Atractylodes macrocephala (Largehead Atractylodes Rhizome) Caused by Ceratobasidium sp. in China.

Atractylodes macrocephala is a perennial herbaceous plant (family Asteraceae) native to China. The biennial root, Largehead Atractylodes Rhizome (LAR), is the most commonly used Chinese herbal medicine to prevent early pregnancy loss due to miscarriage. From summer 2010 to spring 2012, symptoms of root rot were observed on LAR in Xianfeng county, Enshi city, Hubei Province, China. White mold on the root of LAR could be observed at an early growth stage in the field and the white mold spread over the entire plant after 10 days, which differs from root rot of LAR caused by Fusarium oxysporum and Rhizoctonia solani, neither of which are characterized as having mycelium spreading over the whole plant (4). Where root rot symptoms were present, rhizome yield was reduced by 15% on average, with up to 40% yield loss in some fields. Under humid conditions in mid-June, the disease in the field spread quickly and the rhizomes of LAR were completely rotted. After rainfall and increasing temperature from 16 to 35°C, white mycelium appeared and plants withered within a few weeks. In April 2011 and 2012, a fungus was consistently recovered from symptomatic rhizome samples after they were surface sterilized with 0.1% mercuric chloride solution and plated onto potato dextrose agar (PDA). Pale gray colonies with short aerial mycelia and brown sclerotia formed on PDA after 7 days incubation at 28°C. Binucleate cells were observed using light microscopy and the characteristics were matched with morphological characteristics of a Ceratobasidium sp (3). Genomic DNA of the culture was extracted, and the rDNA-internal transcribed spacer sequence (GenBank Accession No. JQ926741) showed 99% identity to Ceratobasidium sp (GenBank No. H269825.1). Mycelial plugs of the culture taken from PDA were inoculated onto 40 rhizomes of 1-year-old seedlings and plants were incubated with a 16-h photoperiod at 28°C and 90% relative humidity in an artificial climate chamber where they developed typical disease symptoms after 2 days. Ten rhizomes of 1-year-old seedlings and were treated with PDA plugs only. All seedlings inoculated with the pathogen were withered and the rhizomes were completely covered with gray mycelium 2 days after inoculation, which was similar to the symptoms observed in the field. After 7 days, the symptoms were more severe than those observed in the field, with seedlings rotted completely. The main stalk of all inoculated plants was covered with gray mycelia in 4 days, and the stalk became withered, which was similar to the symptoms observed in the field. No symptoms were observed on control seedlings and plants. Koch's postulates were fulfilled by successful reisolation of Ceratobasidium sp. from diseased seedlings. The pathogenicity tests were carried out twice. Ceratobasidium sp. has been reported to cause root rot of canola in Washington (2). It has also been observed on Rehmannia in China (1). To our knowledge, this is the first report of Ceratobasidium sp. causing root rot on LAR. References: (1) B. B. Chen et al. Chin. J. Chin. Material Medica (In Chinese) 9:1137, 2011. (2) K. L. Schroeder et al. Plant Dis. 96:591, 2012. (3) B. Sneh et al. Page 39 in: Identification of Rhizoctonia Species. The American Phytopathological Society, 1991. (4) S. X. Zang et al. J. Agric. Univ. Hebei (In Chinese) 28:73, 2005.

PEDF induces p53-mediated apoptosis through PPAR gamma signaling in human umbilical vein endothelial cells.

OBJECTIVE: Pigment epithelial-derived factor (PEDF) is a potent anti-angiogenic factor whose effects are partially mediated through the induction of endothelial cell apoptosis. The pathway mediating endothelial cell apoptosis has not been fully established. Here we investigated the participation of peroxisome proliferator-activated receptor gamma (PPARgamma) and p53 in the apoptosis of human umbilical vein endothelial cells (HUVECs). METHODS AND RESULTS: HUVECs pretreated with either PPARgamma antagonist or PPARgamma small interfering RNA (siRNA) suppressed PEDF-induced apoptosis as determined by TUNEL assay, annexin V-FITC/PI staining, and cleavage of procaspase-8, -9, -3. PEDF sequentially induced PPARgamma and p53 expression as observed in immunoblotting and immunofluoresence assays. PEDF also increased the transcriptional activity of PPARgamma as evident from electromobility shift assays, and p53 as determined by the phosphorylation and acetylation of p53 and the induction of Bax. The induction of p53 by PEDF was abolished by either PPARgamma antagonist or PPARgamma siRNA. PEDF-mediated HUVEC apoptosis and cleavage of procaspases were significantly attenuated by p53 siRNA. CONCLUSIONS: Our observations indicate that PEDF induces HUVECs apoptosis through the sequential induction of PPARgamma and p53 overexpression. With the growing interest in anti-angiogenesis as a novel approach to cancer therapy, defining the mechanism of PEDF-mediated HUVEC apoptosis may facilitate the development of new therapeutics.

Metoprolol and mephenytoin oxidation polymorphisms in Far Eastern Oriental subjects: Japanese versus mainland Chinese.

We examined genetically determined oxidation polymorphisms of metoprolol and mephenytoin in 200 unrelated, healthy Japanese subjects and in 98 mainland Chinese subjects simultaneously. This examination was done according to the respective reported phenotyping criteria by use of the urinary metabolic ratio of metoprolol and of the percentage of excretion of 4-hydroxymephenytoin 8 hours after dose administration. The frequencies of occurrence of poor metabolizers (PMs) in the Japanese versus the Chinese subjects were 0.5% versus 0% for metoprolol and 22.5% versus 17.4% for mephenytoin, respectively. There were no statistically significant differences in these frequencies between the two Oriental populations. However, Chinese extensive metabolizers (EMs) showed a significantly lower excretion of alpha-hydroxymetoprolol (p less than 0.01) and 4-hydroxymephenytoin (p less than 0.001) than that of Japanese EMs, and the mode of the distribution histogram of the Chinese EMs for the two test probes was skewed compared with that of the Japanese EMs. The findings indicate that the two Far Eastern Oriental subject groups have a lower frequency of PM phenotype of debrisoquin/sparteine-type oxidation and a greater incidence of PM phenotype of mephenytoin oxidation compared with the respective frequencies reported from white subjects. However, the explanation for the observation that the metabolic capacities of the test drugs differed between the EMs of the two populations who had a similar ethnic origin and who resided in the same geographic area remains obscure.

A highly attenuated strain of Japanese encephalitis virus induces a protective immune response in mice.

A pair of virulent (RP-9) and attenuated (RP-2ms) mutants of Japanese encephalitis virus (JEV) were generated from a Taiwanese isolate NT109. The mutants differed in several aspects in vitro and in vivo. RP-2ms exhibited smaller plaque than RP-9 on BHK-21 cells, and when intracerebrally injected, RP-2ms was much less neurovirulent than RP-9. As peripherally inoculated, RP-2ms lost neuroinvasiveness while RP-9 penetrated blood-brain barrier, replicated in mouse brain, and killed all the mice. Single RP-2ms immunization completely protected C3H and ICR mice from a lethal challenge with RP-9; the sera from such mice contained antibodies against JEV envelope and nonstructural 1 proteins, indicating RP-2ms had replicated in the mice Neutralizing activity against NT109 in such sera was further demonstrated by plaque reduction neutralization test. In addition, significant lymphoproliferation was detected in spleen cells from the RP-2ms-immunized mice, and cytotoxic activity in these cells specific for the MHC-matched, JEV-infected cells, but not mock cells, was also observed. Altogether, these results demonstrate that RP-2ms, a highly attenuated JEV strain, can induce a protective immunity in mice.

Regulation of cell growth by selective COX-2 inhibitors in oral carcinoma cell lines.

Evidence indicates that NSAIDs that inhibit prostaglandin (PG) synthesis can reduce the incidence of colorectal cancers and that inhibition of cyclooxygenase-2 (COX-2) may be the underlying mechanism. The objective of this study was to investigate this putative mechanism by examining the effect of selective COX-2 inhibitors (Celebrex, DFU, NS-398) and COX-1 inhibitors (Aspirin) on the growth of two human oral carcinoma cell lines (OEC-M1 and KB) and one normal fibroblast cell line (NF). We found that the growth of OEC-M1 cells could be significantly inhibited by DFU concentrations above 30 microM (31%) after 4 days, and above 50 microM (35%) after 2 days in culture; by Celebrex at concentrations above 20 microM (52%) after 6 days, above 30 microM (36%) after 5 days, and above 40 microM (33%) after 4 days in culture; and by NS-398 above 1 microM (30%) after 6 days, and above 10 microM (35%) after 5 days in culture. The growth of KB cells could be significantly inhibited by DFU concentrations above 10 microM (33%) after 6 days, above 20 microM (35%) after 4 days in culture; and by Celebrex at concentrations above 10 microM (33%) after 5 days, and above 50 microM (30%) after 4 days in culture; and by NS-398 above 1 microM (45%) after 5 days, above 20 microM (36%) after 4 days in culture. The growth of NF cells could be significantly inhibited by DFU above 30 microM (45%) after 6 days, and above 40 microM (32%) after 3 days in culture, and by Celebrex at concentrations above 10 microM (42%) after 6 days, above 30 microM (31%) after 4 days, above 50 microM (32%) after 3 days in culture, and by NS-398 above 0.1 microM (35%) after 4 days, and above 1 microM (32%) after 3 days in culture. The growth-inhibitory concentration (IC50) values for DFU on OEC-M1, KB, and NF cells were about 39.1, 14.8, and 42.9 microM at 144 h, respectively, and on KB was about 45.2 microM at 120 h. The IC50 values for Celebrex on OEC-M1, KB, and NF cells were about 19.1, 8.6, and 15.8 microM at 144 h, respectively, and on KB and NF were about 27.7 and 35.3 microM, respectively, at 120 h. The IC50 values for NS-398 on OEC-M1, KB, and NF were about 18.9, 0.7 and 1 microM, respectively, at 144 h; on KB and NF values were about 10.8 and 1.4 microM, respectively, at 120 h and on KB and NF were about 26.6 and 4.1 microM, respectively, at 96 h. The results show that the growth of these cell lines is inhibited by three COX-2 selective inhibitors but not by any COX-1 selective inhibitors. These findings suggest that COX-2 may play an important role in the generation of biochemical mediators that stimulate the growth of human oral cancer and normal fibroblast cell lines.

Use of the phenylalanine:tyrosine ratio to test newborns for phenylketonuria in a large public health screening programme.

OBJECTIVE: To assess the benefits of using the phenylalanine:tyrosine ratio to screen newborns for phenylketonuria (PKU). SETTING: Data were collected from all newborns in California during a ten month period (n = 404,381). METHODS: Dried blood spot specimens were analysed at nine laboratories. To assure that the results reported from multiple sites were matched accurately, an automated methodology was chosen that included sample processing, analysis, telecommunications, reporting, and information technology. Phenylalanine and tyrosine concentrations were measured independently by continuous flow fluorometry, for which precision, recovery, detection limits, carryover, chemical specificity, reportable range, and number of repeats are reported. RESULTS: In this study, 37% of the newborns were tested at less than 24 hours of age. For this population, using a phenylalanine only cut off of 200 mumol/l, there were 48 recalled infants per case of classic PKU. Using the phenylalanine:tyrosine ratio with a cut off of 1.50, screen positives could be reported with phenylalanine as low as 150 mumol/l and with only 12 recalls per case. CONCLUSIONS: The phenylalanine:tyrosine ratio can be measured accurately at multiple laboratories using two channel chemical analyses. Having applied the methods to the routine clinical screening of a large population, it was confirmed that the clinical sensitivity and specificity of the PKU screening test are higher when the phenylalanine:tyrosine ratio is incorporated into the cut off than when the cut off is based on the phenylalanine concentration alone.

Using a defective-interfering RNA system to express the HE protein of mouse hepatitis virus for studying viral pathogenesis.

We have developed a defective-interfering (DI) RNA of mouse hepatitis virus (MHV) as a vector for expressing a variety of cellular and viral genes including the chloramphenicol acetyltransferase (CAT), hemagglutinin' esterase (HE), and gamma interferon. Here, we used the HE-expressing DI RNA for examining the role of HE protein in viral pathogenesis. The pseudorecombinant virus containing an expressed HE protein was generated by infecting cells with MHV-A59, which does not express, HE, and transfecting the in vitro-transcribed DI RNA containing the HE gene. These pseudorecombinant viruses (DE-HE A59) were then inoculated intracerebrally into mice. Viruses recovered from cells infected with A59 and transfected with DI RNA expressing the CAT gene (DE-CAT A59) were used as a control. At various time points after inoculation, mice were observed for clinical symptoms. Tissues (brains and livers) were obtained for determining the replication of DI RNA by RT-PCR, virus replication by plaque assay, antigen expression by immunohistochemistry, and pathological changes. Results showed that all mice infected with DE-CAT A59 succumbed to infection by 9 days postinfection (d p.i). These data are identical to the pathogenesis of the parental A59 virus, demonstrating that inclusion of the DI RNA did not by itself alter pathogenesis. In contrast, only 40% of mice infected with DE-HE A59 succumbed to infection. The subgenomic mRNAs transcribed from the DI vector were detected at 1 and 2 d p.i. but not at subsequent time points, indicating that the genes in the DI vector were expressed only at an early stage of viral infection. No significant difference in virus replication in the brains was detected between these two groups of mice, suggesting that virus replication in brains was not affected by the expression of the HE. Histopathological examination showed only a small increase in the extent of inflammatory cell infiltration and reduced viral antigen in the mice infected with DE-HE A59. There was no difference in virus replication in the livers at 2 and 4 d p.i., but a 3 log10 reduction was detected in the livers of mice infected with DE-HE A59 at 6 d p.i. Histological examination showed a significant reduction in viral antigen, inflammation and necrosis in mice infected with DE-HE A59. These results indicate that the expression of HE from the DI vector altered the viral pathogenesis. This study thus demonstrates the usefulness of this system in studying the role of viral or cellular genes expressed locally at the sites of viral infection in viral pathogenesis.

Staged operation for Graves' ophthalmopathy.

A four-staged operation is currently the most effective treatment available in the management of stable Graves' ophthalmopathy. This report describes the results of four-staged therapy in 51 patients with Graves' ophthalmopathy. Stage I, orbital decompression: 58 orbits in 32 patients underwent orbital decompression, with an average retroplacement effect of 4.2 +/- 2.2 mm (mean +/- SD). This procedure is effective to restore vision and correct the proptosis due to Graves' ophthalmopathy. Careful intraoperative titration of the retroplacement effect during orbital decompression is very important to achieve successful results. Stage II, strabismus surgery: 24 patients underwent strabismus surgery, including 11 with previous decompression surgery and six with previous simultaneous decompression and strabismus surgery. The overall success rate was 87% and previous decompression or strabismus surgery had no influence on the final results. Stage III, fissure width adjustment: 28 patients (45 eyes) received fissure width adjustment. Various procedures were performed and we found Müllerectomy with levator muscle stripping to be the most useful procedure for fissure width adjustment. The average improvement of fissure height was 3.1 +/- 1.8 mm. The rate of overall satisfactory results was 89%. There were five patients who received Staged IV cosmetic procedures with satisfactory results. Graves' ophthalmopathy is a chronic disease that needs thorough cooperation between doctor and patient. Careful evaluation of clinical parameters and individualized surgical goals are the keys to success.

Ultrasonic detection of persistent small unilateral hematocolpos in two girls.

Two girls with unilateral hematocolpos are reported. In both cases, a small amount of blood which had accumulated in the partially obstructed hemivagina was detected by real-time high resolution ultrasonography, and was confirmed by magnetic resonance imaging. Both patients were asymptomatic, and were regularly followed up at an outpatient clinic. To date, the hematocolpos persists but continues to be small.