Population genomics of Agrotis segetum provide insights into the local adaptive evolution of agricultural pests.

BACKGROUND: The adaptive mechanisms of agricultural pests are the key to understanding the evolution of the pests and to developing new control strategies. However, there are few studies on the genetic basis of adaptations of agricultural pests. The turnip moth, Agrotis segetum (Lepidoptera: Noctuidae) is an important underground pest that affects a wide range of host plants and has a strong capacity to adapt to new environments. It is thus a good model for studying the adaptive evolution of pest species. RESULTS: We assembled a high-quality reference genome of A. segetum using PacBio reads. Then, we constructed a variation map of A. segetum by resequencing 98 individuals collected from six natural populations in China. The analysis of the population structure showed that all individuals were divided into four well-differentiated populations, corresponding to their geographical distribution. Selective sweep analysis and environmental association studies showed that candidate genes associated with local adaptation were functionally correlated with detoxification metabolism and glucose metabolism. CONCLUSIONS: Our study of A. segetum has provided insights into the genetic mechanisms of local adaptation and evolution; it has also produced genetic resources for developing new pest management strategies.

MiR-29a-3p inhibits high-grade transformation and epithelial-mesenchymal transition of lacrimal gland adenoid cystic carcinoma by targeting Quaking.

BACKGROUND: Lacrimal adenoid cystic carcinoma (LACC) is the most common orbital malignant epithelial neoplasm. LACC with high-grade transformation (LACC-HGT) has higher rates of recurrence, metastasis, and mortality than LACC without HGT. This study investigated the effects of microRNA-29a-3p (miR-29a-3p) in the pathogenesis of LACC-HGT. METHODS: An Agilent human miRNA microarray was used to screen the differentially expressed miRNAs (DEMs) in LACC and LACC-HGT tumor tissues. Then, the primary cells obtained in previous studies were used to determine the effect of miR-29a-3p. RESULTS: The expression of miR-29a-3p was abnormally lower in LACC-HGT than in LACC. miR-29a-3p can specifically target the 3' UTR of Quaking mRNA and down-regulate Quaking expression, thereby inhibiting the proliferation, migration, and epithelial-mesenchymal transition of LACC cells. CONCLUSIONS: This study illustrated that miR-29a-3p functions as a tumor suppressor by down-regulating the expression of Quaking to inhibit the tumorigenesis of LACC cells. This study may also reveal the pathogenesis of HGT in LACC cells and provide a reference for LACC-HGT targeted diagnosis.

Eye-Preserving Therapies for Advanced Retinoblastoma: A Multicenter Cohort of 1678 Patients in China.

PURPOSE: This study attempted to estimate the impact of eye-preserving therapies for the long-term prognosis of patients with advanced retinoblastoma with regard to overall survival and ocular salvage. DESIGN: Retrospective cohort study covering all 31 provinces (38 retinoblastoma treating centers) of mainland China. PARTICIPANTS: One thousand six hundred seventy-eight patients diagnosed with group D or E retinoblastoma from January 2006 through May 2016. METHODS: Chart review was performed. The patients were divided into primary enucleation and eye-preserving groups, and they were followed up for survival status. The impact of initial treatment on survival was evaluated by Cox analyses. MAIN OUTCOME MEASURES: Overall survival and final eye preservation. RESULTS: After a median follow-up of 43.9 months, 196 patients (12%) died, and the 5-year overall survival was 86%. In total, the eyeball preservation rate was 48%. In this cohort, 1172 patients (70%) had unilateral retinoblastoma, whereas 506 patients (30%) had bilateral disease. For patients with unilateral disease, 570 eyes (49%) underwent primary enucleation, and 602 patients (51%) received eye-preserving therapies initially. During the follow-up (median, 45.6 months), 59 patients (10%) from the primary enucleation group and 56 patients (9.3%) from the eye-preserving group died. Multivariate Cox analyses indicated no significant difference in overall survival between the 2 groups (hazard ratio [HR], 1.25; 95% confidence interval [CI], 0.85-1.84; P = 0.250). For patients with bilateral disease, 95 eyes (19%) underwent primary enucleation, and 411 patients (81%) received eye-preserving therapies initially. During the follow-up (median, 40.1 months), 12 patients (13%) from the primary enucleation group and 69 patients (17%) from the eye-preserving group died. For bilateral retinoblastoma with the worse eye classified as group E, patients undergoing primary enucleation exhibited better overall survival (HR, 2.35; 95% CI, 1.10-5.01; P = 0.027); however, this survival advantage was not evident until passing 22.6 months after initial diagnosis. CONCLUSIONS: Eye-preserving therapies have been used widely for advanced retinoblastoma in China. Patients with bilateral disease whose worse eye was classified as group E and who initially underwent eye-preserving therapies exhibited a worse overall survival. The choice of primary treatment for advanced retinoblastoma should be weighed carefully.

Establishment of a human meibomian gland carcinoma cell model and analysis of differently expressed genes.

Meibomian gland carcinoma (MGC) is a malignant eyelid tumor with a high malignancy degree and poor prognosis. However, the lack of suitable cell and animal models has limited the study of MGC pathogenesis. In the present study, we established and identified one human MGC cell and one meibomian gland (MG) cell model by fresh surgical resection tissue block primary culture and differentially expressed gene assays. The outgrowth of MGC and MG cells was periodically observed after primary culture, and the first passage of MGC cells proceeded on the 14th day, whereas that for MG cells after three weeks. Cell ultrastructures were observed by transmission electron microscopy (TEM). Immunofluorescence staining showed that MGC and MG cells were both positive for cytokeratin (CK) and androgen receptor (AR). Orange granules were observed in the cytoplasm of MGC and MG cells using Oil red O staining, but they were stronger for MG cells than for MGC. CCK-8 detection demonstrated that the proliferation ability of MGC cells was stronger than that of MG cells. Moreover, during RNA sequence analyses, 3023 differential expressed genes were detected between MGC and MG cells. These genes were involved in biological processes such as cell division and positive regulation of cell migration; the signaling pathways mainly covered cell cycle and DNA replication. Further, the tumorigenic potential of MGC cells was examined by inoculating them subcutaneously into the right abdomen of three severely immunodeficient NOD -SCID mice. Transplanted tumors formed on day 11 after inoculation. The xenograft mouse tissues retained the same histological characteristics as the human MGC original tumor and MGC primary cells. Altogether, these results showed that the MGC and MG models were successfully cultured and established, and differentially expressed genes were successfully detected. We provided a useful model and molecular basis for studying the biological characteristics and pathogenesis of human MGC.

Clinical outcome observation of the embolization of orbital vascular malformation with medical glue under direct intra-operative view.

BACKGROUND: Orbital vascular malformation often encircles normal tissue with ill-defined borders. It is easy to bleed during resection operation, making surgical treatment difficult and lesions hard to be removed completely. In this study we aimed to summarize the treatment outcomes by embolizing orbital vascular malformation with intraoperative intracavitary injection of medical glue . METHODS: A retrospective observational and cross-sectional case series study enrolled 31 patients (male = 9, female = 22) with orbital vascular malformations, who were treated from March 2008 to September 2017 at our institution. The clinical features, operation records, pathological reports and follow-up data were analyzed. RESULTS: The location of vascular malformations involved intraorbital (14 cases), superficial area of eyelid and/or face (7 cases), both intraorbital and superficial area (10 cases). Imaging examination showed a solitary mass with regular shape in 8 cases and a space occupying lesion with irregular shape and ill-defined margins in 23 cases. There were 9 cases had optic nerve involved. Surgical debulkling were performed via skin incision on the mass surface (5 cases), lateral orbitotomy (2 cases), and anterior orbitotomy (24 cases). During the operation, lesions were partly exposed and injected with medical glue. The amount of injected glue was 0.25 ml to 2.5 ml in divided doses. The lesions and remnant glue were removed after the glue had turned hard. The whole procedure caused less bleeding and was easier performing than usual. Topical skin aseptic inflammation took place on the same side of the superficial eyelid lesions in 3 cases. One patient suffered from sudden central retinal artery embolism on the third day post operation. With timely rescue and appropriate procedure, visual acuity recovered to 20/32. There were no recurrences in 29 cases. CONCLUSIONS: Embolization of orbital vascular malformation with medical glue intraoperatively made it easy to control hemorrhage. Surgeons should be careful with glue application methods in order to avoid complications.

The evaluation of ICHD-3 beta diagnostic criteria for Tolosa-Hunt syndrome: a study of 22 cases of Tolosa-Hunt syndrome.

The objective of the study was to evaluate the amended International Classification of Headache Disorders (third edition, beta version, ICHD-3 beta) with a retrospective analysis. A total of 22 patients diagnosed with painful ophthalmoplegia and Tolosa-Hunt syndrome (THS) in our hospital were retrospectively studied. The following clinical data were collected: symptoms, signs, location of inflammatory tissue, time interval of paresis following the onset of pain, pain and signs of resolution, follow-up and relapse. Pain and diplopia were found in 22 (100 %) and 20 cases (91 %). The sympathetic nerve was involved in 6 cases (27 %). Paresis followed the pain for an average of 8 ± 5.87 days. Serial magnetic resonance imaging (MRI) revealed granulomatous lesion that was visible in 20 patients (91 %). 19 patients (86 %) demonstrated the lesions located in the cavernous sinus, orbital apex or superior orbital fissure. One lesion extended to the intracranial structure. Pain was relieved in 20 cases (91 %) within 72 h and no patient had complete relief from paresis. According to our study, we think the time course of relief should be undefined. Headache location is hard to describe accurately. Normal MRI should be involved in THS diagnoses. The lesion of THS can extend beyond the cavernous sinus and the orbit. The time interval between headache and paresis can exceed 2 weeks.

[The cultivation and identification of lacrimal gland adenoid cystic cancer stem cells].

OBJECTIVE: To isolate and cultivate the Lacrimal gland Adenoid Cystic Carcinoma cells line, study Cancer Stem Cells properties. METHOD: Experimental study. Lacrimal gland adenoid cystic carcinoma cancer stem cells were cultivated in serum-free suspension culture and the morphological changes were observed. Cells were divided into two groups, the LACC-CSC experimental group and the LACC control group. The flow cytometry instrument was used to detect the expression of classical stem cell markers CD133 and ABCG2. Transwell chamber was used to detect the cancer stem cell aggressivity and differentiated into the vascular endothelial cells. The tumorigenic force in vitro xenotransplantation were applied. RESULT: LACC cells can grow suspensively after vaccinated in serum free medium and form tumor microspheres after 10-12 days. Flow cytometry experiments showed that the expression ratio of stem cell markers CD133 in LACC-CSC was (35.67 ± 6.86)%, significantly different to LACC with (0.46 ± 0.48)%, (t = 8.867, P < 0.05). Similarly, the expression ratio of stem cell marker ABCG2 in LACC-CSC was (39.99 ± 4.54)%, significantly different to LACC with (6.75 ± 1.34)%, (t = -9.932, P < 0.05). In vitro experiment of Matrigel invasion, LACC-CSC went through the matrigel basement membrane averagely (32.60 ± 8.79)/HP contrary to LACC with average (10.20 ± 2.77)/HP after 24 hours, showing statistically significance (t = 5.433, P < 0.05) between the two groups. After training for 48 hours, the difference between two groups was still obvious (t = 5.779, P < 0.05) with LACC-CSC average (62.60 ± 4.83)/HP to LACC (44.00 ± 5.34)/HP. When induced by serum medium containing VEGF and bFGF, LACC-CSC grew adherent gradually and cell morphological changes occurred after continuous induction to long spindle cells. When cultured into three-dimensional matrix structure they formed vessel samples and expressed vascular endothelial marker CD31 and CD34. Transplanted tumor in vitro experiment, mice of LACC-CSC group grew tumors in 9 days with 100% tumorigenic rate, whereas LACC group 12 days with 100% tumorigenic rate. CONCLUSIONS: LACC-CSC can be obtained through serum-free culture method. LACC-CSC grew suspensively and expressed classical stem cell markers. LACC-CSC were identified as cancer stem cells with stronger migration and invasion. LACC-CSC have tumorigenic force and multi-directional differentiation potential with general characteristics of the stem cell.

Primary orbital melanoma combined with giant divided nevus of the eyelid.

The authors report a rare case of primary orbital melanoma (POM) combined with giant divided nevus of the eyelid. An 8-year-old Chinese girl is referred for evaluation of 2-month duration of exophthalmos with decreased vision, epiphora, and pain on her right eye. His presentation, imaging, biopsy, histopathology, and management are presented. The possible cellular origin of the POM and the relationship of POM and divided nevus are discussed. We presume that divided nevus may be one of rarely preexisting lesions of POM.

[Isolation and identification of cancer stem cells from human LACC cell line].

OBJECTIVE: To find ways to isolate and culture lacrimal adenoid cystic carcinoma (LACC) stem cells from human LACC cell line and to identify their biological properties. METHODS: Experimental research. LACC cells were digested, centrifuged and suspended in specific serum free medium (SFM) to get LACC cancer stem cell spheres. Limiting dilution analysis and monoclonal formation assay were performed to determine the proportion of cancer stem cells in LACC cell line. MTT assay was used to determine the proliferation and chemotherapeutic drug resistance of stem cell spheres. The expressions of cell surface markers CD44⁺/CD24⁻ were detected by flow cytometry. Stem cell spheres differentiation were induced by dropping serum medium into SFM and the morphologic changes were observed. The IC50 and UV optical density of two kinds of cells were tested by Student's t-test. RESULTS: About (0.92 ± 0.02) % cells were clonogenic in LACC cell line. They could survive and proliferate to form free-floating cell spheres in SFM, which could stably passaged. The IC50 values of cancer stem cells and LACC cell line were 22.53 mg/L and 11.06 mg/L (t = 37.94, P < 0.05), which suggested that cancer stem cells were more resistant to cis-platinum than LACC cell line. In flow cytometry assay, 84.25% stem cell spheres were CD44⁺/CD24⁻ cells, comparing with 23.77% in LACC cell line. Stem cell spheres could differentiate into normal adenoid cystic carcinoma cells in medium with serum. CONCLUSION: LACC stem cell spheres, containing a large number of cancer stem cells, could be obtained by serum free suspension culture. This provide us an ideal model system for cancer stem cell research.

[Heterotransplantation of human lacrimal adenoid cystic carcinoma cell line into nude mouse].

OBJECTIVE: To investigate characterization of human lacrimal adenoid cystic carcinoma line LACC and establish a model of human LACC. METHODS: Experimental research. Human LACC cells were injected into subcutaneous tissue of nude mice. The length (L) and width (W) of the tumor were measured every day after injection of LACC cells, and the tumor volume was calculated as L×W(2)×1/2. Two mice bearing LACC tumor were randomly killed, removed the tumors anatomy and their lungs, livers, and lymph nodes of oxter, 14, 28, 35, 42, 49 days after injection of LACC cells. Histologic examination, immunohistochemical analysis for Keratin,Vimentin,α-SMA, S-100, Desmin, PCR analysis for human ß-actin and electron microscopy were performed. Whether tumor metastasis to lungs, livers and lymph nodes of oxter were observed by hematoxylin-cosin staining method. RESULTS: Heterotransplanted tumors were observed in all 10 mice after injection of LACC cells. The results of HE staining and transmission electron microscope indicated that heterotransplanted tumor possesses typical histological characterization of epithelial carcinoma. The results of immunohistochemistry showed that keratin, S-100, Vimentin, α-SMA expression in tumor tissue were positive, but Desmin expression were negative. RT-PCR analysis revealed that tumors expressed human ß-actin, indicating their human origin. Histologic examination show that tumors didn't metastasize to lung, liver and lymph node. CONCLUSIONS: Human LACC cell line possesses characterization of malignant carcinoma and also possesses characterization of adenoid cystic carcinoma. It is found that the heterotransplanted LACC tumor has histologic features similar to the original LACC of the patient. This model with lacrimal cystic adenoid carcinoma in nude mice is relatively easy, quick, and especially with high successful rate. So it can be considered as an ideal animal model for study on lacrimal cystic adenoid carcinoma in vivo.

Stroke rehabilitation: from diagnosis to therapy.

Stroke remains a significant global health burden, necessitating comprehensive and innovative approaches in rehabilitation to optimize recovery outcomes. This paper provides a thorough exploration of rehabilitation strategies in stroke management, focusing on diagnostic methods, acute management, and diverse modalities encompassing physical, occupational, speech, and cognitive therapies. Emphasizing the importance of early identification of rehabilitation needs and leveraging technological advancements, including neurostimulation techniques and assistive technologies, this manuscript highlights the challenges and opportunities in stroke rehabilitation. Additionally, it discusses future directions, such as personalized rehabilitation approaches, neuroplasticity concepts, and advancements in assistive technologies, which hold promise in reshaping the landscape of stroke rehabilitation. By delineating these multifaceted aspects, this manuscript aims to provide insights and directions for optimizing stroke rehabilitation practices and enhancing the quality of life for stroke survivors.

METTL3 promotes osteogenic differentiation of human umbilical cord mesenchymal stem cells by up-regulating m6A modification of circCTTN.

BACKGROUND: Human umbilical cord mesenchymal stem cells (hUCMSCs) are promising seed cells in bone tissue engineering. circRNA and N6-methyladenosine (m6A) RNA methylation play important roles in osteogenic differentiation. Here, we investigated the potential relevance of a critical circRNA, hsa_circ_0003376 (circCTTN), and methyltransferase-like 3 (METTL3) in osteogenic differentiation of hUCMSCs. METHODS: Expression of circCTTN after hUCMSC osteogenic induction was detected by qRT-PCR. Three databases (RMBase v2.0, BERMP, and SRAMP) were used to predict m6A sites of circCTTN. RNA was enriched by methylated RNA immunoprecipitation (MeRIP), followed by quantitative real-time polymerase chain reaction to detect m6A level of circCTTN after METTL3 overexpression and osteogenic induction. RNA pull-down, Western blotting, and protein mass spectrometry were performed to investigate the potential mechanisms by which METTL3 promoted m6A modification of circCTTN. Bioinformatic analyses based on database (STRING) search and co-immunoprecipitation were used to analyze the proteins that interacted with METTL3. RESULTS: Overexpression of METTL3 promoted osteogenic differentiation of hUCMSCs and increased m6A level of circCTTN. Two potential m6A modification sites of circCTTN were predicted. No direct interaction between METTL3 and circCTTN was observed. Thirty-one proteins were pulled down by probes specific for circCTTN, including NOP2, and two m6A reading proteins, EIF3A and SND1. Bioinformatics analysis and co-immunoprecipitation showed that METTL3 interacted with EIF3A indirectly through NOP2. CONCLUSIONS: METTL3 promotes the osteogenic differentiation of hUCMSCs by increasing the m6A level of circCTTN. However, METTL3 does not bind directly to circCTTN. METTL3 interacts with circCTTN indirectly through NOP2 and EIF3A.

Dicyanopyridine derivatives: One-pot preparation, ACQ-to-AIE transformation, light-conversion quality and photostability.

Low cost and strong fluorescence emission are two important guarantees for luminogens used as light conversion agents. By one-pot multicomponent approach and inexpensive starting materials, three dicyanopyridine (DP) derivatives named as DCP (2-amino-6-methoxy-4-phenylpyridine-3,5-dicarbonitrile), DCO (2-amino-6-methoxy-4-(4-methoxyphenyl) pyridine-3,5-dicarbonitrile) and DCC (2-amino-4-(4-cyanophenyl)-6-methoxypyridine-3,5-dicarbonitrile) were designed and synthesized. Meanwhile, the ACQ-to-AIE transformation was successfully realized by altering substituent groups rather than traditional rotor-stator theory. Based on crystal analysis and theoretical calculations, the ACQ-to-AIE transformation is attributed to the tunable stacking modes and intermolecular weak interactions. Owing to matched fluorescence emission, low lost, high yield, and AIE activity, DCC is used as light conversion agents and doped in EVA matrix. The light conversion quality confirms that DCC can not only convert ultraviolet light, but also significantly improve the transmittance of 25 %/40 % EVA, whose photosynthetic photon flux density at 400-500 nm and 600-700 nm increased to 30.67 %/30.21 % and 25.37 %/37.82 % of the blank film, respectively. After 20 h of UV irradiation (365 nm, 40 W), the fluorescence intensities of DCC films can maintain 92 % of the initial values, indicating good photostability in the doping films. This work not only provides an excellent and low-cost light conversion agent, but also has important significance for ACQ-to-AIE transformation of luminogens.

[The advance of ß-blockers in the treatment of infantile hemangiomas].

Infantile hemangiomas (IH) is the most common benign tumour in early infancy.IH often grows under skins and subcutaneous tissues around head and neck, but it is not rare in ophthalmologic clinical work.IH in the eyelids, orbit and other organs, characterized by their very rapid growth, can cause severe complications, including ptosis and translocation of eyeball. Periocular IH might cause serious visual loss through induction of strabismic, deprivational or anisometropic astigmatism.Visceral organ involvement may become life-threatening. Patients need to be treated immediately in these scenarios. Conventional treatments for IH include the use of corticosteroids, sclerosing agents, interferon, vincristine, etc., which had some side effects and also not effective to all IH.Recently, many specialized clinics reported the impressive effect of ß-blockers, mainly propranolol, in the treatment of IH. Although IH is not among the approved indications for ß-blockers, propranolol is still recommended as the first line treatment for IH. They acted as vasoconstrictors, regulating angiogenic pathways and inducing apoptosis of vascular endothelial cells. This review covered the current understanding of the indications, mechanism of action, dose regimen, administration route, treatment duration, clinical response, and adverse effects of ß-blockers in the treatment of infantile hemangiomas.

[Primary culture and morphological characteristics of human lacrimal adenoid cystic carcinoma cells].

OBJECTIVE: To explore a method of primary culturing human adenoid cystic carcinoma cells of lacrimal gland. METHODS: Experiment research. Tumor tissue was obtained from the surgical material of a patient diagnosed as lacrimal adenoid cystic carcinoma in Tianjin Medical University Eye Hospital during May 16th to June 1st.We gained primary cells via tissue culture techniques. Mixed cells were removed through several ways.Observed cell morphological characteristics by phase contrast microscope, scanning electron microscope and transmission electron microscope. Cyto-immunochemical staining was applied to detect the expressions of vimentin, desmin, S-100, cytokeratin, pan and CD34. Their expressions were also detected in the tumor tissue except CD34. Made cell growth curve and calculated cell doubling time. RESULTS: The outgrowth of cells was observed by day 5 after seeding tissues, and then cells generated slowly. The first passage proceeded by day 32, and the classical epithelioid cell colonies was observed by day 69 after inoculation. Purified cells of human lacrimal adenoid cystic carcinoma were obtained after the removal of mixed cells through several ways, which have been successfully subcultured for more than 100 passages. The 25th passage LACC cells appeared to be typically epithelioid cells, they showed contact inhibition as the density high enough.SEM and TEM showed the 25th passage LACC cells were malignant tumor cells poorly differentiated. They showed positive reaction with vimentin, cytokeratin (pan) as well as S-100, but negative reaction with desmin and CD34, which were consistent with the tumor tissue. The cell growth curve turned like a sigmoid one, and the cell doubling time was 37.1 h. CONCLUSIONS: We gained purified LACC cells, and understood the morphological characteristics, laying the foundation for the establishment of a human lacrimal adenoid cystic carcinoma cell line.

Breaking the bottleneck of organic light conversion agents: Preparation, performance evaluation and intrinsic mechanism.

Poor photostability has become a major obstacle of organic fluorescent dyes (OFD) used as light conversion agent. To explore the intrinsic mechanisms of photodegradation and highly efficient means to enhance photostability, here, three s-triazine dyes and two 1,3,5-triphenylbenzene luminescent agents were designed and synthesized. Further, the relationships of photostability, intramolecular charge transfer effect, energy gap between singlet and triplet, and active oxygen generating capacity are analyzed and discussed. AIE activity, solid-state fluorescence emission, light conversion quality, and photostability combined with thermostability show TPT-DB (2,4,6-tris(4-(3,6-ditertbutyl-9H-carbazol-9-yl)phenyl)-1,3,5-triazine) is the best light conversion agent among of the dyes, whose photosynthetic photon flux density at 400-500 nm and 600-700 nm in doping film increased successively to 6.20% and 25.78% of the blank film, emission intensity can maintain 93.4% of the initial value after intensified UV radiation of 20 h (365 nm, 40 w), and has good thermal stability, Td up to 374 °C. Furthermore, oxygen-free environment was confirmed to be the most effective measure to enhance the photostability of OFD, thereby a simple and efficient method is adopted to block the diffusion of oxygen and significantly enhance the photostability of OFD by amphiphilic ethylene-vinyl alcohol copolymer. The work not only provides an excellent light conversion agent, but also clears the obstacles for the large-scale application of OFD as light conversion agents.

Comparison of biological behavior of lacrimal gland adenoid cystic carcinoma with high-grade transformation cells.

AIM: To evaluate the differences between human lacrimal gland adenoid cystic carcinoma with high-grade transformation (LACC-HGT) primary cells cultured by high-grade transformation tissue and non-high-grade transformation (non-HGT) primary cells cultured by non-high-grade transformation tissue in proliferation, metastasis, drug susceptibility, and genes. METHODS: LACC-HGT primary cells were established by tissue block culture, and the 4th to 10th generation primary cells were selected as research objects. The cells were preliminarily identified by immunofluorescent staining. The differences between non-HGT and LACC-HGT primary cells in terms of proliferation, metastasis, and drug susceptibility were compared by cell counting kit-8 (CCK-8) assay, wound healing, and drug sensitivity experiments. Differentially expressed genes were screened using mRNA array. Gene expression was analyzed using real-time quantitative polymerase chain reaction (RT-qPCR). RESULTS: LACC-HGT primary cells were successfully cultured by tissue block culture. Immunofluorescence staining results showed that cytokeratin (CK) and CK7 expression levels were positive in LACC-HGT primary cells. CCK-8 results showed that the proliferation ability of LACC-HGT cells was significantly higher than that of non-HGT cells. Wound healing experiment showed that the migration ability of LACC-HGT cells was significantly higher than that of non-HGT cells. LACC-HGT cells were also less sensitive to cisplatin and paclitaxel than non-HGT cells. Compared with non-HGT cells, 9566 differentially expressed genes were found in LACC-HGT primary cells, of which 5162 were up-regulated and 4404 were down-regulated. The expression of N-acetylneuraminate pyruvate lyase (NPL), MARVEL domain containing 3 (MARVELD3), syntabulin (SYBU), and allograft inflammatory factor 1 (AIF1) was higher in LACC-HGT cells than in non-HGT cells, whereas that of periostin (POSTN) was lower. CONCLUSION: LACC-HGT primary cells have faster proliferation, stronger migration ability, and poorer sensitivity to chemotherapy drugs than non-HGT primary cells. The expression of mRNAs in non-HGT and LACC-HGT primary cells are significantly different. These features are speculated to be the reasons why high-grade transformation tissues exhibit higher malignant degree and poorer prognosis than their counterparts.

[The application of serum free medium in the research of isolating cancer stem cells].

The isolation and identification of cancer stem cells are the key points in exploring characteristics of cancer stem cells at present. Several species of cancer stem cells, for instance, retinoblastoma tumor stem cells, cancer stem cells of melanoma of choroid, breast cancer stem cells, lung cancer stem cells, colon cancer stem cells, etc., have been isolated and cultured successfully by serum free medium while their biological functions and characteristics are acquired. This review focus on the application of serum free medium in the research of isolating cancer stem cells, both ocular and general, in terms of providing foundation for further research on ocular cancer stem cells.

[Overexpression of the receptor tyrosine kinase EphA2 in choroidal melanoma: correlation with vesculogenic mimicry and prognosis].

OBJECTIVE: To study the correlation of EphA2 protein expression with vesculogenic mimicry (VM), clinicopathological characteristics and prognosis in choroidal melanoma (CM). METHODS: It was a retrospective case series study. Between January 1992 and December 2005, 56 cases of human CM with clinicopathologic data from the Second Hospital of Tianjin Medical University were studied. HE stainings were performed to observe the microcirculation patterns in tumor tissue specimens. VM was found in 26 of the 56 cases using CD31/periodic acid-Schiff (PAS) double staining and transelectron microscopy. All cases were divided into two groups: VM-positive and VM-negative. Immunohistochemical staining was performed on paraffin sections of the 56 cases of CM specimens to investigate the expression of EphA2. According to tumor cells positive rate and staining intensity of the results of evaluation, the specimens were divided into low expression and high expression groups.χ(2)-test and t-test were used to analyzed the enumeration data and measurement data, respectively. Survival analysis was used to further elucidate its correlation with clinicopathological characteristics, VM and prognosis. Cox proportional hazard model was used to analyzed the influence factors of prognosis. RESULTS: VM channels were found in 26 of the 56 CM cases and VM-negative 30 cases. VM-positivity was related to cell type, tumor size and recurrence and metastasis, and the differences were statistically significant (χ(2) = 4.612, 5.346, 5.213; P = 0.036, 0.021, 0.027). The results showed that EphA2 was up-regulated in the VM-positive group compared with the group of VM-negative group. The positive rates of EphA2 expression in the VM-positive group and VM-negative group were 92.3% (25/26) and 70.0% (21/30), respectively, with a significant difference between the two groups (t = 2.247, P = 0.009). The EphA2 protein was expressed in epithelioid (10/12), mixed (11/15) and spindle (41.40%) cell types, with a significant difference among these histological types (χ(2) = 6.513, P = 0.010). The expression rate of EphA2 protein were significantly higher in large (54.55%, 18/33) than small (45.45%, 15/33) tumors, and the expression of EphA2 in metastatic and recurrence patients (10/11) were significantly higher compared with controls (31.11%, 14/45) (χ(2) = 4.556, 8.211;P = 0.016, 0.005). Kaplan-Meier survival analysis showed the presence of VM resulted in a poor prognosis (t = 9.263, P = 0.000). The Cox proportional hazards model indicated that the EphA2 overexpression and the presence of VM were independent predictors of a poor prognosis (χ(2) = 12.041, P = 0.001). Moreover, there was a significant positive correlation between them (r = 0.412, P < 0.05). CONCLUSION: The results of this study demonstrate that EphA2 may play a critical role in the formation process of VM in CM, implicating EphA2 as a valuable marker for the prediction of recurrence, metastasis and prognosis in CM patients.

Comparison of early osseointegration of non-thermal atmospheric plasma-functionalized/ SLActive titanium implant surfaces in beagle dogs.

Background: Hydrophilic dental implants are gaining increasing interest for their ability to accelerate bone formation. However, commercially available hydrophilic implants, such as SLActive™, have some major limitations due to their time-dependent biological aging and lower cost-effectiveness. The non-thermal atmospheric plasma (NTAP) treatment is a reliable way to gain a hydrophilic surface and enhance osseointegration. However, a few studies have been carried out to compare the osseointegration of NTAP-functionalized titanium implants and commercially available hydrophilic implants. Purpose: In this study, we compare the osseointegration abilities of the NTAP-functionalized titanium implant and Straumann SLActive. Material and methods: The NTAP effectiveness was examined using in vitro cell experiments. Then, six beagle dogs were included in the in vivo experiment. Straumann SLActive implants, SLA implants, and SLA implants treated with NTAP were implanted in the mandibular premolar area of dogs. After 2 w, 4 w, and 8 w, the animals were sacrificed and specimens were collected. Radiographic and histological analyses were used to measure osseointegration. Results: NTAP treatment accelerated the initial attachment and differentiation of MC3T3-E1 cells. In the in vivo experiment, bone parameters (e.g., BIC value and BV/TV) and volume of new bone of NTAP groups were close to those of the SLActive group. Additionally, although there was no statistical difference, the osseointegration of SLActive and NTAP groups was evidently superior to that of the SLA group. Conclusion: NTAP-functionalized implants enhanced cell interaction with material and subsequent bone formation. The osseointegration of the NTAP-functionalized implant was comparable to that of the SLActive implant at the early osseointegration stage.