An endoparasitoid uses its egg surface proteins to regulate its host immune response.

With proteomic analysis, we identified 379 egg surface proteins from an endoparasitoid, Cotesia chilonis. Proteins containing conserved enzymatic domains constitute a large proportion of egg surface components. Some proteins, such as superoxidase dismutase, homolog of C. rubecula 32-kDa protein, and immunoevasive protein-2A, are classical parasitism factors that have known functions in host immunity regulation. Melanization assays revealed that a novel egg surface protein, C. chilonis egg surface serpin domain-containing protein had the same function as a C. chilonis venom serpin, as both suppressed host melanization in a dose-dependent manner. C. chilonis egg surface serpin domain-containing protein is mainly transcribed in C. chilonis oocytes with follicular cells, and it is located on both the anterior and posterior sides of the mature egg surface. Additionally, we used LC-MS/MS to identify 586 binding proteins sourced from C. suppressalis plasma located on the eggshell surface of C. chilonis, which included some immunity-related proteins. These results not only indicate that C. chilonis uses its egg surface proteins to reduce the immune response of its host but also imply that endoparasitoid egg surface proteins might be a new parasitism factor involved in host immune regulation.

Review of the Chinese Leucospidae (Hymenoptera, Chalcidoidea).

The Chinese fauna of the family Leucospidae (Hymenoptera, Chalcidoidea) is reviewed and illustrated for the first time. Twelve species of Leucospis Fabricius, 1775 are recorded; of which two species are new to science: Leucospis aequidentatasp. n. and Leucospis shaanxiensissp. n. and one species is reported new for China: Leucospis intermedia Illiger, 1807. An identification key to Chinese species is included. A lectotype is designated for Leucospis aurantiaca Shestakov, 1923.

Venom of Parasitoid Pteromalus puparum Impairs Host Humoral Antimicrobial Activity by Decreasing Host Cecropin and Lysozyme Gene Expression.

Insect host/parasitoid interactions are co-evolved systems in which host defenses are balanced by parasitoid mechanisms to disable or hide from host immune effectors. Here, we report that Pteromalus puparum venom impairs the antimicrobial activity of its host Pieris rapae. Inhibition zone results showed that bead injection induced the antimicrobial activity of the host hemolymph but that venom inhibited it. The cDNAs encoding cecropin and lysozyme were screened. Relative quantitative PCR results indicated that all of the microorganisms and bead injections up-regulated the transcript levels of the two genes but that venom down-regulated them. At 8 h post bead challenge, there was a peak in the transcript level of the cecropin gene, whereas the peak of lysozyme gene occurred at 24 h. The transcripts levels of the two genes were higher in the granulocytes and fat body than in other tissues. RNA interference decreased the transcript levels of the two genes and the antimicrobial activity of the pupal hemolymph. Venom injections similarly silenced the expression of the two genes during the first 8 h post-treatment in time- and dose-dependent manners, after which the silence effects abated. Additionally, recombinant cecropin and lysozyme had no significant effect on the emergence rate of pupae that were parasitized by P. puparum females. These findings suggest one mechanism of impairing host antimicrobial activity by parasitoid venom.

A Venom Gland Extracellular Chitin-Binding-Like Protein from Pupal Endoparasitoid Wasps, Pteromalus Puparum, Selectively Binds Chitin.

Chitin-binding proteins (CBPs) are present in many species and they act in a variety of biological processes. We analyzed a Pteromalus puparum venom apparatus proteome and transcriptome and identified a partial gene encoding a possible CBP. Here, we report cloning a full-length cDNA of a sequence encoding a chitin-binding-like protein (PpCBP) from P. puparum, a pupal endoparasitoid of Pieris rapae. The cDNA encoded a 96-amino-acid protein, including a secretory signal peptide and a chitin-binding peritrophin-A domain. Phylogenetic analysis of chitin binding domains (CBDs) of cuticle proteins and peritrophic matrix proteins in selected insects revealed that the CBD of PpCBP clustered with the CBD of Nasonia vitripennis. The PpCBP is specifically expressed in the venom apparatus of P. puparum, mostly in the venom gland. PpCBP expression was highest at day one after adult eclosion and much lower for the following five days. We produced a recombinant PpCBP and binding assays showed the recombinant protein selectively binds chitin but not cellulose in vitro. We infer that PpCBP serves a structural role in the venom reservoir, or may be injected into the host to help wound healing of the host exoskeleton.

Involvement of acid-sensing ion channel 1a in functions of cultured human retinal pigment epithelial cells.

In the retina, pH fluctuations may play an important role in adapting retinal responses to different light intensities and are involved in the fine tuning of visual perception. Acidosis occurs in the subretinal space (SRS) under pathological conditions such as age-related macular degeneration (AMD). Although it is well known that many transporters in the retinal pigment epithelium (RPE) cells can maintain pH homeostasis efficiently, other receptors in RPE may also be involved in sensing acidosis, such as acid-sensing ion channels (ASICs). In this study, we investigated whether ASIC1a was expressed in the RPE cells and whether it was involved in the function of these cells. Real-time RT-PCR and Western blotting were used to analyze the ASIC1a expression in ARPE-19 cells during oxidative stress induced by hydrogen peroxide (H(2)O(2)). Furthermore, inhibition or over-expression of ASIC1a in RPE cells was obtained using inhibitors (amiloride and PCTx1) or by the transfection of cDNA encoding hASIC1a. Cell viability was determined by using the MTT assay. The real-time RT-PCR and Western blotting results showed that both the mRNA and protein of ASIC1a were expressed in RPE cells. Inhibition of ASICs by amiloride in normal RPE cells resulted in cell death, indicating that ASICs play an important physiological role in RPE cells. Furthermore, over-expression of ASIC1a in RPE cells prolonged cell survival under oxidative stress induced by H(2)O(2). In conclusion, ASIC1a is functionally expressed in RPE cells and may play an important role in the physiological function of RPE cells by protecting them from oxidative stress.

[Frontalis muscle fascial flap passing through the pulley of orbital septum for correction of severe blepharoptosis].

OBJECTIVE: To evaluate the clinical result of the frontalis muscle fascial flap passing through the pulley of orbital septum for correction of severe blepharoptosis. METHODS: 57 eyes in 52 cases with congenital severe blepharoptosis were treated in recent two years. After the frontalis muscle fascial flap was prepared beneath the orbicularis oculi muscle, the pulley was created by two parallel horizontal incision on the orbital septum at the upper orbital rim and 1 cm under the upper rim. The frontalis muscle fascial flap was then pulled down behind the pulley and out to be attached to the upper margin of tarsal plate. RESULTS: The following-up period was 3-6 months. Satisfactory cosmetic result was achieved in 52 eyes. Three eyes had ptosis relapse and 2 eyes had unnatural contour of the palpebral margin which required another corrective operation. No other complication was observed. CONCLUSIONS: The pulley created by the orbital septum makes the traction lines of the frontalis muscle fascial flap in a similar direction as the natural movement of levator muscle. So both the postoperative static and dynamic appearance of the upper lid is more natural. The technique is very practicable in correction of blepharoptosis.

[Reconstruction of orbital deformity after radiotherapy with transcranial orbitotomy advancement combining free flap].

OBJECTIVE: To study clinic therapeutic effect about reconstruction of severe orbital and cul-de-sac deformity after the radiotherapy with transcranial orbitotomy advancement combining cascade free flap both dorsum pedis flap and anterior tibial fascial flap. METHODS: Five cases was subjected to orbital and cut-de-sac severe deformities after both operation and radiotherapy because of retinoblastoma. The technique included transcranial orbital advancement by anterior orbital osteotomy and rigid fixed with titanic plate by coronal incision, and meanwhile incising the cul-de-sac which would be extended circumference around the central incision separation, and then designing extent of cascade flap consisted of dorsum pedis flap and anterior tibial fascial flap according to the size of cul-de-sac defect and extent of temporal depression. Then, the aforementioned two parts of cascade flap were transplanted into cul-de-sac and temple respectively. There is either the superficial temporal artery and vein or facial artery and jugular vein to chose vascular anastomosis. RESULTS: All flaps survived. After 3 to 6 months following up, the results showed satisfactory orbital contour and temporal depression improved significantly in all cases. After the conjunctival sac were fixiformed with prefabricated eye prosthesis mode about 3 months. 3 cases have good appearance with wearing eye prosthesis and the other 2 cases' appearance is poor. One of the poor appearance cases, with depressed eye socket, have orbital implant underlying conjunctival sac in secondary operation. The other one, with swallowed inferior fornix, is transplanted autogenous hard palatal mucosa into inferior fornix in secondary operation. In addition, delayed healing in donor site of dorsum pedis occurred in one of the 4 cases. CONCLUSIONS: It is a reliable procedure about reconstruction of severe orbital and cul-de-sac deformity after both the operation and radiotherapy with transcranial orbitotomy advancement combining cascade free flap both dorsum pedis flap and anterior tibial fascial flap. All deformity was corrected by one staged procedure which lessen patient's suffering and shorten patient's hospital stay and spare patient's costs.

[Modified transcranial orbitotomy for treating exophthalmic intraorbital tumor].

OBJECTIVE: To study the feasibility of the modified osteotomy of transcranial orbitotomy in the treatment of intraorbital tumor. METHODS: We treated 8 patients with intraorbital tumor during six years. By the bicoronary incision, all cases underwent double bone flap osteotomy on the frontal bone: the superior orbital rim bone flap and roof flap instead of single fronto-orbital bone flap in the conventional transcranial orbitotomy. After removal of bone flaps, intracranial and intraorbital operation was performed. Then, two bone flaps were reduced respectively and fixated with titanic micro-plates and nails. At last, the scalp flap was sutured. RESULTS: The operative field was very well exposed. It was found that the retrobulbar tumor was located at the superolateral, median and superonasal area respectively, which was coincided with the preoperative CT and MRI. The tumor included adenocarcinoma of the lacrimal gland, neurinoma, meningioma, and cavernous hemangioma. Four patients had blood transfusion during the operation. No other postoperative complications happened except 3 cases of diplopia and one case of blindness. After 3 to 6 months follow up, diplopia of the 3 cases gradually disappeared. Of the 8 cases, 6 reached the same visual acuity as the preoperative state. One decreased visual acuity and one lost light perception. There was well-balanced fronto-orbital appearance and eyeball position compared with the healthy side in all cases. CONCLUSIONS: This surgical method for the treatment of intraorbital tumor is safe with well-exposed operative field. It has advantages not only in simpleness and less trauma, but also in keeping orbital roof and anterior fossa intact and decreasing complications.