Preliminary study on the resection of parapharyngeal and lateral skull base tumors by using transoral endoscopy with 3D visualization and navigation technologies.

OBJECTIVES: With the assistance of 3D visualization and real-time navigation technologies, the tumors in the parapharyngeal and lateral skull base should be removed through oral the approach with endoscopy. METHODS: The preoperative CT data of eight patients with parapharyngeal or lateral skull base soft tissue tumors were modeled, and the anatomical position relationship between the tumor and surrounding blood vessels and other important structures was reconstructed using 3D visualization technology, and preoperative design was performed. The intraoperative oral approach and real-time navigation guidance were adopted in the endoscopic resection of soft tissue tumors in the parapharyngeal and lateral skull base, and the clinical application value of this method was evaluated. RESULTS: The blood loss during the operation was controlled within 150 mL, and the average blood loss was approximately 125 mL. The incidence of postoperative complications was low, and patients could recover well through functional training. The oral approach did not leave any wounds nor scars on the patient's facial skin after the operation and had no effect on the patient's appearance. CONCLUSIONS: The combination of 3D visualization technology, intraoperative real-time navigation, and endoscopy provides a beautiful, safe, and minimally invasive surgical method for patients with parapharyngeal or lateral skull base tumors.

Isolation and Identification of Culturable Bacteria from South China Seawater and Preliminary Screening of Marine Biocontrol Bacteria.

Marine microorganisms have evolved special metabolic pathways to produce numerous bioactive substances with novel structures and unique functions. This study analyzed the diversity of culturable bacteria in marine water samples from the South China Sea and screened the isolated bacteria with pathogenic fungi. A total of 200 culturable strains of 72 different bacteria were obtained from 56 water samples from the South China Sea. They belonged to three phyla and four classes, namely Gammaproteobacteria, Alphaproteobacteria, Bacilli and Actinomycetia. Bacilli was the dominant class, comprising up to 59.72%, followed by Gammaproteobacteria (20.83%). Bacillus, Pseudomonas, Paenibacillus and Rhizobium were the most dominant genera. Among these strains, HY-88 and HY-91 encoding BamC, FenB and PKSI genes were selected and identified as Bacillus subtilis. The respective inhibition rates of the HY-88 caused by plate confrontation against Magnaporthe grisea, Fusarium oxysporum, Botrytis cinerea, anthrax and Botrytis cinerea were 90.91%, 54.29%, 52.17% and 51.72%, in comparison with HY-91 86.36%, 48.57%, 47.83% and 34.48%. In addition, the supernatant of HY-88 showed a lesion inhibition rate of 74.5%, which was significantly higher than HY-91 (60.55%). In addition, HY-88 and HY-91 showed strong antifungal activity to Colletotrichum viniferum on detached Shine Muscat grapes. Tolerance tests showed that the HY-88 and HY-91 grew at 10-40 °C, 7-10% NaCl and pH 3-11. HY-88 and HY-91 could inhibit various fungal plant diseases, which lays a foundation for the development of new biopesticides.

Isolation and genomic characterization of a cypovirus from Clanis bilineata.

Clanis bilineata Walker, soybean hawkmoth, belongs to the subfamily Ambulicinae (Sphingidae, Lepidoptera) and is an edible insect that usually grows on soybean leaves. In this study, we isolated a new cypovirus from naturally diseased Clanis bilineata larvae (named CbCPV), scanned its structure, sequenced its genome, and studied its phylogenetic relationship to other cypoviruses. Microscopy showed that CbCPV polyhedral occlusion bodies were about 1.878 µm on average and contained many virions in the ultrathin sections. The complete genome sequence of CbCPV is 22,812 bp comprising 10 segmented double-stranded RNAs. Apart from segment 1 containing one open reading frame (ORF) and one sub-ORF, the other nine segments all contain one open reading frame and encoded one putative protein. The non-coding regions contained conserved sequences at 5' termini (AGUCAAA) and 3' termini (AGC), except segment 4 containing a different 5' termini (AUGUUUA). The whole sequence of the polyhedrin gene in CbCPV contained 892 nucleotides, encoding a protein of 246 amino acids. Based on amino acid sequences of polyhedrin or RNA dependent RNA polymerase (RdRp), the phylogenetic analysis indicated that CbCPV was closely related to DnCPV-23. The putative function of all segments differed from each other, but the most closely related species of segments were DnCPV-23 with 98.2-99.8% nucleotide identity. Overall, the evidence of morphology, protein analysis and nucleic acids (genomic pattern) showed that CbCPV is a new isolate in the cypovirus-23 type and can be termed Clanis bilineata cypovirus type 23 (CbCPV-23).

Talaromyces sp. Ethyl Acetate Crude Extract as Potential Mosquitocide to Control Culex pipiens quinquefasciatus.

Vector control is considered an effective approach to controlling diseases spread by mosquito bites. Entomopathogenic fungi are widely used in agriculture to control insect pests, and fungal metabolites can potentially be developed as effective mosquitocides. In this study, a high-throughput screening method was used to search for potential mosquitocides in the Global Fungal Extract Library (GFEL). We tested the larvicidal activity of 264 fungal ethyl acetate crude extracts against Culex pipiens quinquefasciatus. Nine fungal extracts caused moderate to high mortality rates (>50%), with two fungal extracts (58A7 and 101H12) causing a 100% mortality rate. The lethal concentrations for 50% of the population (LC50) were 44.27 mg/L and 31.90 mg/L, respectively. Fraction 14 had a high mortality rate, with an LC50 value of 12.13 mg/L, and was isolated from 58A7 (Fractions 1-11) and 101H12 (Fractions 12-15). Further analyses showed that Fraction 14 was made up of vermistatin and dihydrovermistatin. In a Cx. p. quinquefasciatus larvicidal bioassay, vermistatin (LC50 = 28.13 mg/L) was more toxic than dihydrovermistatin (LC50 = 83.87 mg/L). Our findings suggested that the active fungal extract 101H12 from Talaromyces sp. and its compound vermistatin could be developed as mosquitocides.

LKB1 alleviates high glucose­ and high fat­induced inflammation and the expression of GnRH and sexual precocity­related genes, in mouse hypothalamic cells by activating the AMPK/FOXO1 signaling pathway.

Precocious puberty (PP) is a developmental disorder. Hypothalamic cells can produce gonadotropin­releasing hormone (GnRH), the final output of neuroendocrine regulation that occurs during puberty. The aim of the present study was to investigate the role of live kinase B1 (LKB1), also known as serine/threonine kinase, in the progression of PP and identify the underlying mechanisms. First, the levels of LKB1 in peripheral blood and peripheral blood mononuclear cells of children with PP were detected by reverse transcription­quantitative (RT­q) PCR or western blotting. After the GT1­7 mouse hypothalamus cell line was treated with high glucose (HG) and high fat (HF), the expression of LKB1 and GnRH was tested. LKB1 was overexpressed by transfection with a pcDNA3.1 plasmid and the levels of inflammatory factors, GnRH, PP­related factors and proteins in the AMP­activated protein kinase (AMPK)/forkhead box protein O1 (FOXO1) pathway were determined using RT­qPCR or western blot analysis. Subsequently, Compound C, an inhibitor of AMPK/FOXO1 signaling, was used to clarify whether the effects of LKB1 on PP were mediated by the regulation of this pathway. Results indicated that children with PP exhibited a lower LKB1 expression. In addition, HG and HF culture resulted in an enhanced GnRH expression and a reduced LKB1 expression in GT1­7 cells. LKB1 overexpression inhibited the contents of TNF­α, IL­6 and GnRH in in GT1­7 cells exposed to HG and HF and reduced the expression of PP­related proteins, including estrogen receptor­ß, cluster of differentiation 36 and G­protein­coupled receptor. In addition, the expression of phosphorylated (p)­AMPK and p­FOXO1 was markedly downregulated following LKBI overexpression. Furthermore, compound C intervention partially diminished the inhibitory effects of LKB1­mediated upregulation on the levels of inflammation and PP­related factors. In conclusion, these results demonstrated that LKB1 alleviated HG­ and HF­induced inflammation, as well as the expression of GnRH and sexual precocity­related genes, in GT1­7 cells by activating the AMPK/FOXO1 signaling pathway.

Transcriptional responses of Daphnis nerii larval midgut to oral infection by Daphnis nerii cypovirus-23.

BACKGROUND: Daphnis nerii cypovirus-23 (DnCPV-23) is a new type of cypovirus and has a lethal effect on the oleander hawk moth, Daphnis nerii which feeds on leave of Oleander and Catharanthus et al. After DnCPV-23 infection, the change of Daphnis nerii responses has not been reported. METHODS: To better understand the pathogenic mechanism of DnCPV-23 infection, 3rd-instar Daphnis nerii larvae were orally infected with DnCPV-23 occlusion bodies and the transcriptional responses of the Daphnis nerii midgut were analyzed 72 h post-infection using RNA-seq. RESULTS: The results showed that 1979 differentially expressed Daphnis nerii transcripts in the infected midgut had been identified. KEGG analysis showed that protein digestion and absorption, Toll and Imd signaling pathway were down-regulated. Based on the result, we speculated that food digestion and absorption in insect midgut might be impaired after virus infection. In addition, the down-regulation of the immune response may make D. nerii more susceptible to bacterial infections. Glycerophospholipid metabolism and xenobiotics metabolism were up-regulated. These two types of pathways may affect the viral replication and xenobiotic detoxification of insect, respectively. CONCLUSION: These results may facilitate a better understanding of the changes in Daphnis nerii metabolism during cypovirus infection and serve as a basis for future research on the molecular mechanism of DnCPV-23 invasion.

Discovery of mosquitocides from fungal extracts through a high-throughput cytotoxicity-screening approach.

BACKGROUND: Mosquitoes transmit a variety of diseases. Due to widespread insecticide resistance, new effective pesticides are urgently needed. Entomopathogenic fungi are widely utilized to control pest insects in agriculture. We hypothesized that certain fungal metabolites may be effective insecticides against mosquitoes. METHODS: A high-throughput cytotoxicity-based screening approach was developed to search for insecticidal compounds in our newly established global fungal extract library. We first determined cell survival rates after adding various fungal extracts. Candidate insecticides were further analyzed using traditional larval and adult survival bioassays. RESULTS: Twelve ethyl acetate extracts from a total of 192 fungal extracts displayed > 85% inhibition of cabbage looper ovary cell proliferation. Ten of these 12 candidates were confirmed to be toxic to Anopheles gambiae Sua5B cell line, and six showed > 85% inhibition of Anopheles mosquito cell growth. Further bioassays determined a LC50, the lethal concentration that kills 50% of larval or adult mosquitoes, of 122 µg/mL and 1.7 µg/mosquito, respectively, after 24 h for extract 76F6 from Penicillium toxicarium. CONCLUSIONS: We established a high-throughput MTT-based cytotoxicity screening approach for the discovery of new mosquitocides from fungal extracts. We discovered a candidate extract from P. toxicarium that exhibited high toxicity to mosquito larvae and adults, and thus were able to demonstrate the value of our recently developed approach. The active fungal extracts discovered here are ideal candidates for further development as mosquitocides.

Isolation and genomic characterization of a cypovirus from the oleander hawk moth, Daphnis nerii.

The oleander hawk moth, Daphnis nerii, is a serious pest of plants belonging to the family Apocynaceae. Thus far, pathogen infection has not been reported in D. nerii. In this study, a new cytoplasmic polyhedrosis virus (cypovirus; CPV) was isolated from naturally diseased D. nerii larvae and named DnCPV-23. Virions were observed in ultrathin sections of DnCPV polyhedral bodies. Electrophoretic analysis revealed that the DnCPV genome consisted of 10 segments of double-stranded RNA (dsRNA). cDNA copies of these dsRNA segments were amplified using the method of full-length amplification of cDNAs (FLAC), cloned, and sequenced. Sequencing results showed that all segments contained one open reading frame (ORF); They shared the conserved terminal sequences AGUCAAA and AGC at 5' and 3' ends respectively, except segment 4, which is different from previously reported 22 cypoviruses. Phylogenetic analysis based on amino acid sequences of polyhedrin (encoded by segment 10) indicated that this CPV was closely related to CPV type 19. Altogether, DnCPV-23 is a new type of cypovirus.

Layer Caused an Anisotropic Terahertz Response of a 3D-printed Simulative Shale Core.

Energy demands have motivated the development of shale formations as significant unconventional reservoirs. The anisotropy of shales plays a significant role in both the mechanical behavior and engineering activities. Alternating layers presented in shales affect the propagation of waves, causing anisotropy at various frequencies. Simplifying the complicated interior structures of shales is conducive to characterize the anisotropic properties. Therefore, simulative shale core samples were designed and fabricated using additive manufacturing processes, and layer-caused dielectric anisotropy was investigated by terahertz (THz) time-domain spectroscopy. On the basis of effective medium theory, the change of the optical length caused by refraction of rays was discussed and modeled. It is believed that the refraction of rays at the interfaces is the source of THz propagation anisotropy in the multilayered structure, and the anisotropy degree is mainly influenced by the layer thickness as well as the refractive index.

[Isolation and Identification of a Novel Cypovirus from Daphnis nerii].

In order to develop a novel effective biological insecticide for controlling oleander hawk moth, a new pathogen was isolated from naturally diseased Daphnis nerii. Based on scanning electron microscopy, full-length amplification of cDNAs (FLAC), and phylogenetic analysis of genome segments 2and 10,the virus was identified as a new type of cypovirus (Da phnis nerii cypovirus [DnCPV]). Electrophoresis analysis showed that DnCPV had a genome comprising 10double-stranded RNA (dsRNA) segments, ranging from 892 to 4160bp.Using FLAC, the cDNAs from the 10 dsRNA segments of the new CPV were cloned and genome segments 2and 10 were sequenced. Sequencing results showed that segment 2 encoded RNA-dependent-RNA-polymerases (RdRps) and segment 10 encoded polyhedrin. These two segments shared conserved terminal sequences of AGUCAAA and AGC at the 5'and 3'ends,respectively.These conserved terminal sequences were not consistent with any of the known CPV types.Phylogenetic analysis of the RdRp and polyhedrin indicated that this CPV was more closely related to CPV type 19 and type 5than other CPV types. Based on the unique conserved terminal sequences and the electrophoresis pattern of the new virus, we tentatively named it DnCPV Nanchang isolate: DnCPV-NC.

Association of MDR1 gene C3435T polymorphism with childhood intractable epilepsy: a meta-analysis.

Drug-resistant epilepsy is also referred to as intractable, medically refractory, or pharmacoresistant epilepsy. Approximately, one-third of patients with epilepsy have recurrent seizures despite therapy. Multidrug resistance 1 (MDR1) gene may play a role in drug-resistance in epilepsy. To assess the association between MDR1 C3435T polymorphism and the response to anticonvulsants in childhood intractable epilepsy, we conducted a systematic review and meta-analysis. Studies were obtained from the electronic database of PubMed, Medline, Embase and CNKI up to September 2013. All the case-control association researches evaluating the role of MDR1 C3435T polymorphism in childhood epilepsy to antiepileptic drugs were identified. The odds ratios (ORs) with 95% confidence intervals (CIs) were calculated for comparisons of the alleles and genotypes with co-dominant (C/C vs. T/T, C/T vs. T/T), dominant (C/C + C/T vs. T/T), and recessive (C/C vs. C/T + T/T) models in overall and in ethnicity subgroups to measure the strength of genetic association. A total of 8 related studies, including 634 drug-resistant patients, 615 drug-responsive patients and 1,052 healthy controls were pooled in this meta-analysis. The allelic association of MDR1 C3435T with risk of drug-resistance was not significant (OR 1.03, 95% CI 0.87-1.22, P = 0.73; OR 1.00, 95% CI 0.86-1.16, P = 0.98) in overall and in the subgroup analysis by ethnicity (Asian: OR 0.95, 95% CI 0.77-1.18, P = 0.67; Caucasian: OR 1.18, 95% CI 0.89-1.57, P = 0.25). Neither association was found in other genetic models. Our results did not show a significant association between MDR1 C3435T polymorphism and response to anticonvulsant drugs, suggesting that this polymorphism may not be a risk factor to childhood intractable epilepsy.

[Diagnosis and treatment of the complex fracture of the maxillary and nasal bone].

OBJECTIVE: To explore the effect of surgical treatment for compound fracture of nasal bone and frontal process of maxilla with endoscopy and small local incision. METHOD: Twenty-two cases with compound fracture of nasal bone and frontal process of maxilla were treated with surgery, and the postoperative effect were analysed. RESULT: Twenty-two cases healed well. CONCLUSION: CT scan and 3D reconstruction before operation can help the diagnosis. Corresponding incision and assisted endoscopy can make good surgical field and achieve surgical success. Rigid internal fixation is also the key for surgical success.

[Repair and functional reconstruction of oropharyngeal defects after resection of advanced-stage tonsillar cancer].

OBJECTIVE: To study the repair and functional reconstruction of oropharyngeal defects after resection of advanced-stage tonsillar cancer, and to select the donor site of appropriate flap. METHODS: Between October 2000 and February 2010, 13 patients with advanced-stage tonsillar cancer were treated, including 5 cases of high differentiation squamous cell carcinomas and 8 cases of medium differentiation squamous cell carcinomas. There were 11 males and 2 females, with an average age of 53.6 years (range, 39-67 years). According to Union for International Cancer Control (UICC) 1997 standards of oropharyngeal cancer, 1 case was classified as T1N1M0, 2 as T2NIM0, 2 as T2N2M0, 3 as T3N1M0, 2 as T3N2M0, 2 as T4N1M0, and 1 as T4N2M0. The disease duration was 1-8 months with an average of 4.3 months. The tumor invaded lateral wall of nasopharyngeal in 1 case, lateral wall of hypopharynx in 3 cases, epiglottis in 1 case, soft palate in 4 cases, and tongue root in 3 cases. The tumor infiltrating range was from 2 cm x 2 cm to 12 cm x 6 cm. All the 13 cases underwent integrated methods of surgery and postoperative radiotherapy. After resection of tumor by combined neck-mandible-oral cavity approach, pectoralis major myocutaneous flaps were transplanted in 5 cases, forearm free skin flaps in 5 cases, and anterolateral thigh free skin flaps in 3 cases. RESULTS: The postoperative pathological results showed 10 cases of cervical lymph node metastasis; 2 cases had local recurrence and 3 cases had cervical lymph node metastasis after postoperative radiotherapy. Neck infection occurred at 5 days after operation in 1 case undergoing transplantation of pectoralis major myocutaneous flap, and vascular crisis occurred at 12 hours after operation in 1 case undergoing transplantation of forearm free skin flap, which were cured after correspondent treatments. The other flaps survived with incision healing by first intention. Second suture was carried out in 1 case undergoing anterolateral thigh free skin flap transplantation because of wound disruption at the donor site. All the patients were followed up 1 to 6 years, with an average of 3.6 years. In 5 cases undergoing pectoralis major myocutaneous flap transplantation, swallowing obstruction and stomatolalia occurred. In 8 cases undergoing free skin flaps transplantation, the appearances of the flaps and the functions of swallowing or speaking were satisfactory, with no dysfunction at the donor site. All the patients returned to normal occlusion, facial appearance and function were normal. According to the direct calculation method, the three-year survival rate was 60.0% (6/10), and five-year survival rate was 37.5% (3/8). CONCLUSION: For the patients with advanced-stage tonsillar cancer, forearm free skin flaps, or anterolateral thigh free skin flaps is the first choice for repairing defect. However, it is better to choose pectoralis major myocutaneous flaps in patients who need large flap and fail to radiotherapy.

Free jejunal graft for reconstruction of defects in the hypopharynx and cervical esophagus following the cancer resections.

OBJECTIVES: The reconstruction of esophagus defects after hypopharyngeal and cervical esophageal carcinoma resection is an ongoing problem. The objective of this article was to investigate the techniques of the free jejunal graft for the reconstruction of hypopharyngeal and cervical esophagus and discuss the outcome related to the procedures. SUBJECTS AND METHODS: From July of 2005 to December 2007, seven patients with hypopharyngeal and cervical esophageal cancer underwent free jejunal graft reconstruction of the hypopharyngeal and cervical esophagus. Their clinical data were retrospectively analyzed. All patients received postoperative radiotherapy and were followed up for 7-24 months. RESULTS: Despite the multistep and time-consuming procedure, free jejunal graft survival was 100%. Operation-induced complications did not occur in six patients. One patient developed pharyngeal fistula. CONCLUSION: The present experience supports the use of free jejunal grafts in reconstruction of the hypopharyngeal and cervical esophagus defects after exenteration of the central compartment of the neck. A high successful rate with low incidence of complications in reconstruction of the hypopharyngeal and cervical esophagus was obtained in this study.