Bombyx mori cell division cycle protein 37 promotes the proliferation of BmNPV.

Cell division cycle protein 37 (Cdc37) is a molecular chaperone that actively participates in many intracellular physiological and biochemical processes as well as pathogen infection. However, the function of Cdc37 in silkworm cells under Bombyx mori nucleopolyhedrovirus (BmNPV) infection is unknown. We cloned and identified BmCdc37, a Cdc37 gene from B. mori, which is highly conserved among other species. After BmNPV infection, the expression level of the BmCdc37 gene was up-regulated and showed an expression pattern similar to the BmHsp90 gene, which relies on Cdc37 to stabilize and activate specific protein kinases. The immunofluorescence, bimolecular fluorescence complementation (BiFC), and co-immunoprecipitation (Co-IP) assays all indicated that BmCdc37 interacts with BmHsp90 in silkworm cells. Both BmCdc37 and BmHsp90 promote the reproduction of BmNPV. Co-expression of BmCdc37 and BmHsp90 was better at promoting virus proliferation than overexpression alone. These findings all indicate that BmCdc37 plays an active role in the proliferation of BmNPV.

Bombyx mori protein BmREEPa and BmPtchd could form a complex with BmNPV envelope protein GP64.

Our previous studies have indicated that Bombyx mori receptor expression enhancing protein a (BmREEPa) could participate in BV invasion in vivo and in vitro, however, the mechanism is still unclear. In this study, we screened BmREEPa interacting protein through co-immunoprecipitation and finally identified a membrane protein, Bombyx mori patched domain containing protein (BmPtchd, KR338939), which contains receptor activity. Further studies showed that BmPtchd, BmREEPa and Glycoprotein 64 could form a protein complex and the expression level of BmREEPa and BmPtchd could be affected by cellular cholesterol level. These findings may provide an important basis for explaining the invasion mechanism of Bombyx mori Nucleopolyhedrovirus budded virus.

BmNHR96 participate BV entry of BmN-SWU1 cells via affecting the cellular cholesterol level.

B.mori nucleopolyhedrovirus (BmNPV), which produces BV and ODV two virion phenotypes in its life cycle, caused the amount of economic loss in sericulture. But the mechanism of its infection was still unclear. In this study we characterized B.mori nuclear hormone receptor 96 (BmNHR96) as a NHR96 family member, which was localized in the nucleus. We also found BmNHR96 over-expression could enhance the entry of BV as well as cellular cholesterol level. Furthermore, we validated that BmNHR96 increased membrane fusion mediated by GP64, which could probably promote BV-infection. In summary, our study suggested that BmNHR96 plays an important role in BV infection and this function probably actualized by affecting cellular cholesterol level, and our results provided insights to the mechanisms of BV-infection of B.mori.

Transgenic RNAi of BmREEPa in silkworms can enhance the resistance of silkworm to Bombyxmori Nucleopolyhedrovirus.

Our previous study has identified a gene, BmREEPa, which affects BmNPV invasion in silkworm cells. In this study, we interfered with BmREEPa in silkworm larvae through transgenic technology and screened BmREEPa-RNAi silkworm strains (RP). We found the mortality in RP was lower than that in Dazao, when silkworm larvae were infected with BmNPV via oral and injection routes. And the expression level of VP39 was lower in RP than in Dazao in the group infected via injection. In the oral infection group, VP39 expression level showed significant reduction at 48 h post-infection. These results revealed that the anti-BmNPV activity was enhanced in RP, and this enhancement probably presents itself during secondary infection via BVs.

In vivo RNA interference of BmNHR96 enhances the resistance of transgenic silkworm to BmNPV.

We previously identified a nuclear hormone receptor gene, BmNHR96, which promotes Bombyx mori nucleopolyhedrovirus (BmNPV) entry into silkworm cells. In an attempt to create an antiviral silkworm strain for better silk production, we used RNAi to downregulate BmNHR96 in silkworm larvae. We screened the resulting BmNHR96-RNAi silkworm strain (NHR) and also explored the antiviral mechanism in vivo. We found that the survival rate of the NHR strain was higher than that of the Dazao strain, when silkworm larvae were infected with BmNPV via oral ODV infection and BV injection. More importantly, the economic characteristics (silk yield) of the transgenic line remained unchanged. These findings reveal that RNAi of BmNHR96 could be an effective way to enhance the tolerance of B. mori to BmNPV infection.

Serum soluble ST2 is associated with ER-positive breast cancer.

BACKGROUND: ST2, a member of the interleukin (IL)-1receptor family, regulates Th1/Th2 immune responses in autoimmune and inflammatory conditions. However, the role of ST2 signaling in tumor growth and metastasis of breast cancers has not been investigated. This study investigated the possible role of soluble ST2 (sST2) in breast cancer. METHODS: The serum levels of IL-33, sST2, and vascular endothelial growth factor (VEGF) in 150 breast cancer patients and 90 healthy women were measured by enzyme-linked immunosorbent assay. Estrogen receptor(ER), progesterone receptor, human epithelial receptor (HER)-2, and cell cycle regulated protein Ki-67 were measured. Clinical stage, tumor size, lymph node metastasis, and histological type were also recorded. RESULTS: The serum levels of sST2, IL-33, and VEGF were significantly higher in breast cancer patients than in the control group (P < 0.05, each). Serum sST2 levels in ER-positive breast cancer patients were significantly associated with age, histological type, clinical stage, tumor size, and Ki-67 status (P < 0.05, each). Moreover, the serum levels of IL-33 and sST2 in breast cancers significantly correlated with VEGF levels (IL-33: r = 0.375, P < 0.0001; sST2: r = 0.164, P = 0.045). Serum levels of sST2, IL-33, and VEGF decreased after modified radical mastectomy in ER-positive breast cancers. Serum levels of IL-33, sST2, and VEGF and clinicopathological factors were not significantly correlated with disease-free survival and overall survival of ER-positive breast cancer women during follow-up. CONCLUSION: Serum sST2 levels in ER-positive breast cancer patients are significantly associated with factors that indicate poor prognosis.

Complete mitochondrial genome of Saldoida armata Horváth, 1911 (Heteroptera: Saldidae) and phylogenetic analysis.

The complete mitochondrial genome of Saldoida armata (Heteroptera: Saldidae) is 16,049 bp in length, comprising 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs) and a control region. All the PCGs are initially encoded by ATN, TTG or GTG, and terminated coding with TAA or a single T. With the exception of trnS(AGN), all tRNAs exhibit a typical cloverleaf secondary structure. Phylogenetic analysis reveals the sister relationship of S. armata with other Saldidae members. The complete mitogenome of S. armata will provide useful genetic information for species identification, phylogenetic analysis and conservation of this species.

Complete mitochondrial genome data and phylogenetic analysis of Papilio macilentus Janson, 1877 (Lepidoptera: Papilionoidea: Papilionidae).

In the present study, the complete mitochondrial genome (mitogenome) of the Papilio macilentus (Lepidoptera: Papilionoidea: Papilionidae) was sequenced by next-generation sequencing method. The mitochondrial genome is a circular DNA molecule of 15,264 bp in size with 80.7% AT content, including 37 genes (13 protein-coding genes, 2 rRNA genes, and 22 tRNA genes), and a long non-coding region (Control region). All protein-coding genes are initiated by ATN codons, and terminated with TAA, TAG, or single T. All tRNAs can be folded into common clover leaf secondary structure, except trn-S1. Phylogenetic analyses based on 13 protein-coding genes and 2 rRNA genes using maximum likelihood and Bayesian inference confirmed that P. macilentus and Papilio memnon are clustered into a clade, and revealed the relationships between Papilionini, Troidini, Teinopaippini and Leptocircini.

Transient receptor potential vanilloid 4 promotes the growth of non-small cell lung cancer by regulating Foxp3.

OBJECTIVE(S): Transient receptor potential vanilloid 4 (TRPV4) participates in malignant tumor. However, the role of TRPV4 in non-small cell lung cancer (NSCLC) remains unclear. In this study, we demonstrated TRPV4 was upregulated in NSCLC tissues and NSCLC cell lines. MATERIALS AND METHODS: TRPV4 level in the NSCLC patients and cell lines were detected, and its function was studied both in vivo and vitro. RESULTS: The level of TRPV4 showed a positive correlation with tumor size of NSCLC patients. Activation TRPV4 by agonist GSK1016790A promoted cell proliferation and decreased apoptosis in A549 cells, and these effects were enhanced when the cells have overexpressed TRPV4. Moreover, GSK1016790A induced inhibitory effects on apoptosis of A549 cells was impaired when GSK1016790A used together with TRPV4 selective antagonist HC-067047, or impaired when the cells have already downregulated TRPV4 expression by TRPV4 siRNA. In vivo study, pharmacological inhibition of TRPV4 prevented A549 cells transplanted tumor growth. It was showed Foxp3 level was significantly increased in the NSCLC tissues, and showed a positive correlation with the level of TRPV4. Deactivation of TRPV4 using TRPV4 siRNA or HC-067047 significantly reduced expression of Foxp3 in GSK1016790A treated NSCLC cells. Moreover, downregulation Foxp3 by transfection of Foxp3 siRNA significantly impaired TRPV4 induced NSCLC cells proliferations in vitro. CONCLUSIONS: Antitumor effects caused by TRPV4 inhibition in NSCLC might be attributed to the suppression of Foxp3 which induced subsequent cell apoptosis. Thus, pharmacological inhibition of TRPV4 may be a promising option for NSCLC treatment.

[Effects of chitosan oligosaccharide on alveolar bone resorption, Th17/Treg balance and OPG/RANKL/RANK pathway in periodontitis rats].

PURPOSE: To investigate the effect of chitosan oligosaccharide on alveolar bone resorption, Th17/Treg balance and OPG/RANKL/RANK pathway in rats with periodontitis. METHODS: Rat model of periodontitis was established, and the periodontitis rats were randomly divided into model group, low-dose chitosan oligosaccharide group, middle-dose chitosan oligosaccharide group, high-dose chitosan oligosaccharide group and metronidazole group, with 12 rats in each group, another 12 rats were set as control group. After treatment, gingival index and alveolar bone absorption were evaluated. H-E staining was used to observe the pathological changes of periodontal tissues. The ratio of Th17/Treg cells in peripheral blood was detected by flow cytometry, the levels of serum IL-17, TGF-ß, RANKL and OPG were detected by ELISA, and the expressions of OPG and RANKL mRNA in periodontal tissues of rats in each group were detected by real-time fluorescent quantitative PCR(qRT-PCR). SPSS 24.0 software package was used to analyze the data. RESULTS: Compared with the control group, the periodontal tissue of the model group showed periodontal membrane fiber bundle rupture, disordered arrangement, capillary expansion, proliferation, inflammatory cell infiltration and other pathological damage. Gingival index, alveolar bone resorption value, Th17/Treg ratio, serum RANKL and IL-17 levels, and periodontal RANKL mRNA level were significantly increased(P<0.05), while the levels of serum OPG, TGF-ß and OPG mRNA in periodontal tissues were significantly decreased (P<0.05). Compared with the model group, the pathological damage of periodontal tissue in the low-middle-and high-dose chitosan oligosaccharide groups and metronidazole group was reduced; gingival index, alveolar bone resorption value, Th17/Treg ratio, serum RANKL and IL-17 levels, and periodontal RANKL mRNA level were significantly decreased(P<0.05), while the levels of serum OPG, TGF-ß and OPG mRNA in periodontal tissues were significantly increased(P<0.05); there was a dose-dependent relationship between the chitosan oligosaccharide groups, and there was no significant difference between the high-dose chitosan oligosaccharide group and metronidazole group(P>0.05). CONCLUSIONS: Chitosan oligosaccharide can promote Th17/Treg balance to return to normal, up-regulate OPG expression, down-regulate RANKL expression, inhibit alveolar bone resorption in periodontitis rats and improve their clinical symptoms.

The Phylogenetic Implications of the Mitochondrial Genomes of Macropsis notata and Oncopsis nigrofasciata.

Macropsinae are forest pests that feed on woody plants. They can damage the growth of trees and crops, and some species can also spread plant pathogens. Due to their widespread effects, these leafhoppers are of great economic significance, which is why there is a need to study their genomes. To fill the gap in the mitochondrial genomic data of the subfamily Macropsinae, we sequenced the complete mitochondrial genomes of Macropsis notata and Oncopsis nigrofasciata (which were 16,323 and 15,927 bp long, respectively). These two species are representative species of the leafhoppers group (Cicadellidae); the mitochondrial genomes of these species range from a length of 15,131 bp (Trocnadella arisana) to 16,811 bp (Parocerus laurifoliae). Both mitogenomes contained 37 typical insect mitochondrial genes and a control region; there were no long non-coding sequences. The genes within the mitogenome were very compact. The mitogenomes from both species contained two kinds of parallel repeat units in the control region. The whole mitogenomes of Macropsinae showed a heavy AT nucleotide bias (M. notata 76.8% and O. nigrofasciata 79.0%), a positive AT Skew (0.15 and 0.12), and a negative GC Skew (-0.14 and -0.08). Upon comparative ML and BI analysis, some clade relationships were consistent among the six trees. Most subfamilies were reconstructed into monophyletic groups with strong support in all analyses, with the exception of Evacanthinae and Cicadellinae. Unlike the results of previous research, it was shown that although all Deltocephalinae species are grouped into one clade, they were not the sister group to all other leafhoppers. Further, Cicadellinae and Evacanthinae were occasionally reconstructed as a polyphyletic and a paraphyletic group, respectively, possibly due to the limited numbers of samples and sequences. This mitogenome information for M. notata and O. nigrofasciata could facilitate future studies on the mitogenomic diversity and evolution of the related Membracoidea, and eventually help to control their effects on plants for the betterment of society at large.

Path-folded infrared spectrometer consisting of 10 sub-gratings and a two-dimensional InGaAs detector.

A new compact infrared spectrometer without any mechanical moving elements has been designed and constructed using a two-dimensional InGaAs array detector and 10 sub-gratings. The instrument is compact, with a double-folded optical path configuration. The spectra are densely 10-folded to achieve 0.07-nm spectral resolution and a 2-ms data acquisition time in the 1450- to 1650-nm wavelength region, making the instrument useful for real-time spectroscopic data analyses in optical communication and many other fields.

[Relationship between serosal invasion types and lymph node metastasis after total gastrectomy in gastric cancer and its significance in selection of rational dissection].

OBJECTIVE: To evaluate the relationship between serosal invasion types and lymph node metastasis after total gastrectomy in gastric cancer patients, and explore its significance in planning practice rational dissection based on the serosa types of gastric cancer during surgery. METHODS: A total of 73 gastric cancer patients, who underwent total gastrectomy and lymph node dissection, were included in this study, and their clinicopathological data were analyzed. The serosa of gastric cancer was divided into five types: normal, reactive, nodular, tendonoid, and color-diffused, then they were combined into 3 groups: group 1: normal and reactive, group 2: nodular (including protruding nodular and flat nodular), and group 3: tendonoid and color-diffused. The lymph node metastasis ratios in the 3 groups were compared. The lymph nodes in each of the 3 groups were divided into 16 subgroups and the lymph node metastasis ratios of each subgroup in the 3 groups were compared and analyzed. RESULTS: The lymph node metastasis ratio of the gastric cancer with normal and reactive type serosa was 5.3% (26/492), the nodular was 37.1% (250/673), the tendonoid and color-diffused was 50.0% (486/972). The lymph node metastasis ratio of normal and reactive type groups was the lowest, that of the tendonoid and color-diffused groups was the highest, and the nodular type in between, showing a statistically significant difference (P<0.01). The results of comparing the lymph node metastasis ratios from the 1st to 16th subgroup in the 3 groups showed the same trend (P<0.05). CONCLUSION: Among all serosa types of gastric cancer, the lymph node metastasis ratio of the tendonoid and color-diffused is the highest, the normal and reactive type is the lowest, and the nodular in between. The extent of rational dissection should be carried out on the basis of serosa types of gastric cancer during surgery. An extended dissection including D2 and D3 lymphadenectomy should be performed for the patients with tendonoid and color-diffused serosa, a rational decreased operation including D1-D1+ lymphadenectomy should be performed for those with a normal and reactive type serosa, and for the patients with nodular type serosa, we suggest performing standard D2 dissection.

[Features of metastasis in different lymph node groups and their significance in lymph node dissection in total gastrectomy for gastric cancer].

OBJECTIVE: To summarize the features of metastasis in different lymph node groups (from 1 to 16 groups) in gastric cancer patients treated by total gastrectomy, and evaluate their clinical significance in lymph node dissection. METHODS: The data of 73 gastric cancer patients with total gastrectomy and lymph node dissection from January 2004 to April 2006 were analyzed retrospectively. The lymph nodes were divided into 16 groups according to the 13(th) edition of gastric cancer treatment guideline of JGCA (The Japan Gastric Cancer Association). The metastatic rate and degree of dissected lymph nodes in these patients were compared. RESULTS: The metastatic rates of lymph node groups in these patients from lower to higher were as follows: group 15, 13/16, 14v, 12, 10, 9, 11, 8, 2, 6/7, 5, 1, 4, 3. The lowest was the 15(th) group lymph nodes (1.4%), the highest was the 3rd group (65.8%), with a statistically significant difference between those two groups (P < 0.01). The metastatic degrees of the lymph node groups from lower to higher were as follows: 13, 16, 1, 7, 6, 5, 12, 4, 11, 8, 2, 15, 9, 3, 10, 14v. There was a statistically significant difference between the lowest group of lymph node (13(th) group, 10.7%) and the highest (14v(th), 56.3%, P < 0.01). CONCLUSION: In the radical total gastrectomy for patients with gastric cancer, it is suggested that the regional lymph nodes with higher metastatic rate should be resected necessarily, and the group with a higher metastatic degree should be dissected completely. If the result of sentinel lymph node biopsy in the 3(rd) or 14v(th) group is negative, the operation extent can be reduced. If positive, it should be extended. When the biopsy result in the 13(th) or 16(th) is positive, palliative operation may be indicated. However, if the biopsy result is negative in the 13(th) or 16(th), but positive in the 14v(th) group, extended operation is indicated.

Three new species of the leafhopper subfamily Iassinae (Hemiptera: Cicadellidae) from China.

Three new species of leafhoppers: Siniassus hamulus sp. nov., Decliviassus sagittatus sp. nov., Krisna quadrimaculosus sp. nov. are described and illustrated from China. An updated checklist and key to 22 of the 24 known Chinese species of these three genera (Siniassus, Decliviassus, Krisna) are provided.

Identification and characterization of the BmCyclin L1-BmCDK11A/B complex in relation to cell cycle regulation.

Cyclin proteins are the key regulatory and activity partner of cyclin-dependent kinases (CDKs), which play pivotal regulatory roles in cell cycle progression. In the present study, we identified a Cyclin L1 and 2 CDK11 2 CDK11 splice variants, CDK11A and CDK11B, from silkworm, Bombyx mori. We determined that both Cyclin L1 and CDK11A/B are nuclear proteins, and further investigations were conducted to elucidate their spatiofunctional features. Cyclin L1 forms a complex with CDK11A/B and were co-localized to the nucleus. Moreover, the dimerization of CDK11A and CDK11B and the effects of Cyclin L1 and CDK11A/B on cell cycle regulation were also investigated. Using overexpression or RNA interference experiments, we demonstrated that the abnormal expression of Cyclin L1 and CDK11A/B leads to cell cycle arrest and cell proliferation suppression. Together, these findings indicate that CDK11A/B interacts with Cyclin L1 to regulate the cell cycle.

A newly discovered member of the Atlastin family, BmAtlastin-n, has an antiviral effect against BmNPV in Bombyx mori.

Atlastin is a member of the dynamin protein superfamily and it can mediate homotypic fusion of endoplasmic reticulum (ER) membranes, which is required for many biological processes. In this study, a new Atlastin homologous protein, BmAtlastin-n, was characterized in silkworms and was found to contain an N-terminal conserved GTPase domain and a coiled-coil middle domain. BmAtlastin-n is localized in the cytoplasm and enriched in silkworm midgut. Results also showed that overexpression of BmAtlastin-n in BmN-SWU1 cells could enhance resistance to BmNPV. To better confirm its antiviral effect, microRNA was used to knock down the expression of BmAtlastin-n in BmE-SWU1 cells with inducing the reproduction of BmNPV. A transgenic expression vector of BmAtlastin-n was constructed and introduced to silkworm embryos by microinjection. The transgenic silkworm also showed considerable antiviral capacity. In conclusion, these findings demonstrate that BmAtlastin-n plays an important role in BmNPV defense. More importantly, the current study may provide a new clue for Atlastin research.

[Evaluation of the correlations of cell proliferation activity with lymphatic and vascular invasion and prognosis in gastric carcinoma].

OBJECTIVE: To evaluate bromodeoxyuridine/DNA (BrdUrd/DNA) double-parametric flow cytometric analysis in detection of gastric carcinoma and to study the correlations of cellular BrdUrd labeling index (LI), G(2)/M phase fraction (G(2)/MPF) and DNA content, with lymphatic and vascular invasion and prognosis. METHODS: Sixty cases of fresh tumor samples were examined by BrdUrd/DNA double-parametric flow cytometry. RESULTS: BrdUrd LI and G(2)/MPF values were both significantly higher in patients with lymphatic invasion than those without invasion (P < 0.01). There was statistically significant difference between the 5-year survival rates in cases with and without lymphatic invasion (P < 0.01). Both BrdUrd LI and G(2)/MPF values were significantly higher in patients with lymph node metastasis than those in cases without metastasis (P < 0.01). There was a significant difference in 5-year survival rates between patients with and without lymph node metastases. The incidence of lymph node metastasis was significantly higher in aneuploid carcinoma (P < 0.05), and the patients with aneuploidy had significantly poor prognosis. BrdUrd LI was significantly higher in patients with more than 5 metastatic lymph nodes than those with 1-4 metastatic lymph nodes (P < 0.05) and no metastasis (P < 0.01). G(2)/MPF values in cases with more than 5 and 1-4 metastatic lymph nodes were higher than that in cases with no metastasis (P < 0.01 and P < 0.05, respectively). There were significant differences in 5-year survival rates among patients with no lymph node metastasis, 1-4 nodes metastasis and more than 5 nodes metastasis. G(2)/MPF values were significantly higher in patients with and without vascular invasion (P < 0.01). CONCLUSION: Correlations exist among cellular BrdUrd LI, G(2)/MPF, DNA content and lymphatic involvement, vascular invasions and prognosis in gastric carcinoma.

A new picrotoxane sesquiterpene from the berries of Baccaurea ramiflora with antifungal activity against Colletotrichum gloeosporioides.

Three picrotoxane sesquiterpenes including one new glycoside and two known constituents, sapidolide A (2) and picrotoximaesin (3), were isolated from the berries of Baccaurea ramiflora. The structure of the new sesquiterpene glycoside, ramifloside (1), was elucidated as 2-one-6α-hydroxy-13-nor-11-picrotoxen-3(15ß)-olide 10-O-ß-D-glucopyranoside on the basis of extensive spectroscopic analysis. Compounds 1-3 exhibited antifungal activity against Colletotrichum gloeosporioides with MICs of 12.5, 12.5 and 50 µg/mL.

BmREEPa Is a Novel Gene that Facilitates BmNPV Entry into Silkworm Cells.

We previously established two silkworm cell lines, BmN-SWU1 and BmN-SWU2, from Bombyx mori ovaries. BmN-SWU1 cells are susceptible while BmN-SWU2 cells are highly resistant to BmNPV infection. Interestingly, we found that the entry of BmNPV into BmN-SWU2 cells was largely inhibited. To explore the mechanism of this inhibition, in this study we used isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative protein expression profiling and identified 629 differentially expressed proteins between the two cell lines. Among them, we identified a new membrane protein termed BmREEPa. The gene encoding BmREEPa transcribes two splice variants; a 573 bp long BmREEPa-L encoding a protein with 190 amino acids and a 501 bp long BmREEPa-S encoding a protein with 166 amino acids. BmREEPa contains a conserved TB2/DP, HVA22 domain and three transmembrane domains. It is localized in the plasma membrane with a cytoplasmic C-terminus and an extracellular N-terminus. We found that limiting the expression of BmREEPa in BmN-SWU1 cells inhibited BmNPV entry, whereas over-expression of BmREEPa in BmN-SWU2 cells promoted BmNPV entry. Our results also indicated that BmREEPa can interact with GP64, which is the key envelope fusion protein for BmNPV entry. Taken together, the findings of our study revealed that BmREEPa is required for BmNPV to gain entry into silkworm cells, and may provide insights for the identification of BmNPV receptors.