[Investigation of reason and treatment methods for active, refractory and recurrence thyroid associated ophthalmopathy].

TAO is a common autoimmune orbital disease. 20% to 30% patients in active phase of moderate to severe TAO have no reponse to treatment or relapse after treatment, which become a difficult problem in clinical treatment. In this paper, the necessity of the treatment, etiology, treatment difficulties and treatment measures of TAO are discussed, in order to make great efforts to solve the clinical problems and to treat such patients. (Chin J Ophthalmol, 2017, 53: 408-412).

A rapid and high-quality method for total RNA isolation from Haematococcus pluvialis.

Haematococcus pluvialis, as the most potential natural source of astaxanthin, which is a powerful antioxidant with high economic value, has attracted more and more scientific attention in recent years. An in-depth understanding of the mechanism for how H. pluvialis produces astaxanthin requires the intensive investigations on its genetic information. In particular, many reported studies were based on a variety of RNA analyses. However, it is difficult to extract RNA with high quality and quantity from H. pluvialis, because of the blockage from its thick cell wall and contamination by a large quantity of pigments, polysaccharides, and lipids. Therefore, we proposed an optimized Trizol-based RNA extraction method for H. pluvialis by investigating the effect of cell wall broken ways, algal strains, and cell growth status on total RNA isolation. Using this rapid, convenient, and cost-saving method, isolated H. pluvialis RNA had high quantity and quality (with an RNA integrity number of 7.0 and a concentration of 1604.1 ng/µL) equivalent to that isolated by commercial kit, enabling its applications into downstream RNA analyses.

[A clinicopathological study of IgG4-related ocular disease and differentiated diagnosis with non-IgG4-related ocular disorders].

OBJECTIVE: To study the clinicopathologic features of IgG4-related ocular disease and differentiated diagnosis with non-IgG4-related ocular disorders. METHODS: Forty-three cases were selected from the Department of Pathology, West China Hospital, Sichuan University from December 2008 to January 2015 with retrospective series case study. The clinical manifestation, laboratory examination, imaging findings, treatment and histopathological characteristics were analyzed. According to the diagnostic criteria for IgG4-related ocular disease in 2014 and relevant literatures, our cases were divided into two groups: IgG4-related ocular disease (23 samples from 18patients) and non-IgG4-related ocular disease (31 samples from 25 patients). The male-to-female ratio, the periductal fibrosis and the orbital lesion area was analyzed by Pearson chi-square. The Immunohistochemistry of IgG4+/HPF, IgG+/HPF and IgG4+/IgG+ ratio and age was analyzed by two sample t test. The involved structural tissue and the infiltration of eosinophils and lymphocytes and plasma cells was analyzed by signed-rank test. RESULTS: The significant statistic differences were observed between these two groups in immunohistochemical staining of IgG4 (>40/HPF) (t=12.17, P<0.01), IgG (t=2.91, P<0.01) and IgG4+/IgG+ ratio (≥40%) (t=16.98, P<0.01), the lymphocytic and plasmacytic infiltration (F=3.62, P=0.03),the periductal fibrosis(F=4.71 , P=0.03), the involved region of orbit (t=1.11, P=0.01), but not in the mean age and male-to-female ratio (t=1.09, P=0.58). IgG4 ROD should be distinguished form the non-specific inflammation disorders, lymphoma and so on. CONCLUSIONS: IgG4-related ocular disease often arises from middle-old man, characterized by bilateral orbital painless swelling and abundant IgG4+ plasma cells infiltration. Nearly all orbital tissue could be involved by IgG4-related ocular disease, especially lacrimal gland. But the characteristic obliterating phlebitis were not seen. The immunohistochemical staining of IgG4 and IgG plays an important role in differentiating IgG4-related ocular disease from non-IgG4-related ocular disorders. Combining clinical manifestation, laboratory examination, imaging findings with histopathology is helpful for diagnosis and differential diagnosis. (Chin J Ophthalmol, 2016, 52: 256-262).

Expression profile of tumor suppressor gene RASSF1 in lacrimal gland carcinoma.

We examined the expression pattern of the tumor sup-pressor gene RAS association domain family gene 1 (RASSF1) in lacri-mal gland carcinoma and analyzed its relationship with the oncogenesis and progression of tumors. Sixty-two patients (30 males, 32 females, average age = 47 ± 3.5 years) admitted with lacrimal gland carcinoma to the Department of Ophthalmology of our hospital between January 2012 and January 2014 were enrolled in this study. Based on tumor ma-lignancy, patients were classified into a malignant group (N = 25) and benign group (N = 37). Healthy lacrimal gland resections from trauma surgery (N = 35) were recruited as a healthy control group. Expres-sion profiles of RASSF1 in all groups were quantified using reverse transcription-polymerase chain reaction and western blotting. Recur-rence of lacrimal gland carcinoma was surveyed through postopera-tive follow-up. Expression levels of RASSF1 in samples from the ma-lignant and benign groups were significantly lower than those in the healthy group (P < 0.05). Furthermore, the malignant group showed lower RASSF1 expression than the benign group (P < 0.05). Postopera-tive follow-up identified 22 cases of recurrence in the malignant group, with a recurrence rate of 88%, while 15 cases in the benign group had a recurrence rate of 40.5%. A direct relationship exists between RASSF1 expression levels and the malignancy grade of lacrimal gland carci-noma. Patients with lower RASSF1 expression showed a higher recur-rence probability, indicating unfavorable prognosis. Therefore, measur-ing RASSF1 expression can be used as a diagnostic method for lacrimal gland carcinoma.

Pharmacological investigation of a HPLC/MS standardized three herbal extracts containing formulae (Bu-Shen-Yi-Qi-Tang) on airway inflammation and hypothalamic-pituitary-adrenal axis activity in asthmatic mice.

Bu-Shen-Yi-Qi-Tang (BSYQT) which is prescribed on the basis of clinical experience is commonly used in clinic of traditional Chinese medicine (TCM) for asthma treatment. The components of BSYQT include Radix Astragali (RA), Herba Epimedii (HE) and Radix Rehmanniae (RR). The aim of this study was to screen extracts of BSYQT with best anti-inflammatory activity in asthmatic mice, and separate and identify the chemical compounds in them. Our results suggested that 60% ethanol extract of herbs (H60) and granules (G60) of BSYQT were the two extracts with best anti-inflammatory activity and effects of H60 were a little better than that of G60. High-performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry (HPLC-ESI-Q-TOF-MS/MS) analysis of the major chemical compounds of H60 and G60 revealed that 56 and 42 peaks were identified separately in H60 and G60. Further analysis revealed that 38 compounds were identified shared by H60 and G60, and 18 compounds were only in H60. There were 25 compounds in HE, 6 compounds in RR and 7 compounds in RA in the 38 compounds shared by G60 and H60. These 38 chemical components were tentatively considered the material basis of the anti-inflammatory activity of G60 and H60. The differences in the amount of the 38 chemical components as well as the 18 chemical components only in H60 were tentatively considered responsible for the activity differences between H60 and G60. In conclusion, these results suggested that extracts of BSYQT had inhibitory effects on airway inflammation in asthmatic mice, and H60 and G60 demonstrated the best anti-inflammatory activity. The 38 chemical compounds shared by H60 and G60 were responsible for their anti-inflammatory activity in asthmatic mice, and the differences in chemical compounds contents and amounts between H60 and G60 were responsible for this activity differences. This work would provide support for further pharmacodynamic material basis study of BSYQT.

Monoclonal antibody 55 (CD10) and complement used for purging autologous bone marrow in common acute lymphoblastic leukemia.

A CD10 monoclonal antibody 55 (McAb55) was intended for purging residual common acute lymphoblastic leukemia antigen (CALLA) positive leukemic cells from autotransplants of common acute lymphoblastic leukemia (C-ALL) patients. It was found that after two rounds of McAb55 and complement treatment, 4-5 logs of CALLA+ cells were removed from bone marrow detected by clonogenic assay. The standardization of separation, purgation and preservation of bone marrow for C-ALL patients' autotransplants was then set as follows: Following the carboxymethyl starch sedimentation and Ficoll-Hypaque gradient separation, the isolated mononuclear cells (MNCs) were treated with McAb55 and complement twice and kept in room temperature for 48-72 hours prior to infusion. This procedure resulted in the removal of more than 99% of CALLA+ cells, recovery of 10-30% MNCs, and leaving the hematopoiesis stem cells intact. After the intensive cytoreductive therapy, 4 patients with C-ALL received the purged autotransplants giving timely recovery of the hematopoietic function. The patients were all remaining in remission status for more than 40-250 days so far.

Orbital mucormycosis with retinal and ciliary artery occlusions.

A 61-year-old man presented with acute, painful loss of vision in the left eye due to a central retinal artery occlusion. Fluorescein angiography confirmed the central retinal artery occlusion and also identified a nasal posterior ciliary artery occlusion. CT scanning revealed a left medial orbital mass with adjacent ethmoid sinusitis. Transnasal ethmoid biopsy disclosed mucormycosis. A left external ethmoidectomy, maxillectomy, and orbital exploration were performed, after which the patient was treated with daily intravenous amphotericin B for six weeks. Coexistence of retinal and nasal posterior ciliary artery occlusion due to mucormycosis may relate to their common origin from the ophthalmic artery. Treatment without exenteration was successful.