An injectable signal-amplifying device elicits a specific immune response against malignant glioblastoma.

Despite exciting achievements with some malignancies, immunotherapy for hypoimmunogenic cancers, especially glioblastoma (GBM), remains a formidable clinical challenge. Poor immunogenicity and deficient immune infiltrates are two major limitations to an effective cancer-specific immune response. Herein, we propose that an injectable signal-amplifying nanocomposite/hydrogel system consisting of granulocyte-macrophage colony-stimulating factor and imiquimod-loaded antigen-capturing nanoparticles can simultaneously amplify the chemotactic signal of antigen-presenting cells and the "danger" signal of GBM. We demonstrated the feasibility of this strategy in two scenarios of GBM. In the first scenario, we showed that this simultaneous amplification system, in conjunction with local chemotherapy, enhanced both the immunogenicity and immune infiltrates in a recurrent GBM model; thus, ultimately making a cold GBM hot and suppressing postoperative relapse. Encouraged by excellent efficacy, we further exploited this signal-amplifying system to improve the efficiency of vaccine lysate in the treatment of refractory multiple GBM, a disease with limited clinical treatment options. In general, this biomaterial-based immune signal amplification system represents a unique approach to restore GBM-specific immunity and may provide a beneficial preliminary treatment for other clinically refractory malignancies.

Case Report: Identification of a novel LYN::LINC01900 transcript with promyelocytic phenotype and TP53 mutation in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a malignant disease of myeloid hematopoietic stem/progenitor cells characterized by the abnormal proliferation of primitive and naive random cells in the bone marrow and peripheral blood. Acute promyelocytic leukemia (APL) is a type (AML-M3) of AML. Most patients with APL have the characteristic chromosomal translocation t(15; 17)(q22; q12), forming PML::RARA fusion. The occurrence and progression of AML are often accompanied by the emergence of gene fusions such as PML::RARA, CBFß::MYH11, and RUNX1::RUNX1T1, among others. Gene fusions are the main molecular biological abnormalities in acute leukemia, and all fusion genes act as crucial oncogenic factors in leukemia. Herein, we report the first case of LYN::LINC01900 fusion transcript in AML with a promyelocytic phenotype and TP53 mutation. Further studies should address whether new protein products may result from this fusion, as well as the biological function of these new products in disease occurrence and progression.

[Study on the levels of main inflammatory factors in septic shock and their clinical significance].

OBJECTIVE: To explore the correlation between major inflammatory factors and septic shock in intensive care unit (ICU) patients, and to provide a basis for the diagnosis and treatment of septic shock. METHODS: The patients admitted to ICU of the Third People's Hospital of Datong from March 2017 to August 2018 were selected as the research objects. According to the diagnostic criteria of septic shock, the patients were divided into septic shock group and non-septic group. The basic information and inflammatory factors levels of the two groups, including age, gender, body mass index (BMI), course of disease, acute physiology and chronic health evaluation II (APACHE II), infection site and pathogenic; and C-reactive protein (CRP), procalcitonin (PCT), neutrophil lymphocyte ratio (NLR), N-terminal pro-B-type natriuretic peptide (NT-proBNP), tumor necrosis factor-α (TNF-α), γ-interferon (IFN-γ), interleukins (IL-1ß, IL-2, IL-6, IL-8) at 8 hours after diagnosis, were compared. Logistic regression was used to analyze the influencing factors of septic shock in ICU patients. RESULTS: A total of 154 ICU patients were selected, of whom 74 had septic shock. The APACHE II score of septic shock group was significantly higher than that of non-sepsis group (23.42±3.64 vs. 15.67±2.26, P < 0.05). There was no significant difference in other baseline data between the two groups. The levels of CRP, NT-proBNP, TNF-α, IFN-γ, PCT, IL-6, IL-8 in the septic shock group were significantly higher than those in the non-septic group [CRP (mg/L): 164.3±22.6 vs. 52.3±16.2, NT-proBNP (ng/L): 426.3±288.9 vs. 167.3±80.6, TNF-α (ng/L): 193.4±39.3 vs. 88.1±20.3, IFN-γ (ng/L): 133.3±52.0 vs. 97.0±56.1, PCT (ng/L): 27.6±10.2 vs. 7.3±4.1, IL-6 (ng/L): 83.0±17.6 vs. 20.9±6.4, IL-8 (ng/L): 445.8±34.0 vs. 84.0±25.7, all P < 0.05]. It was shown by Logistic regression analysis that CRP, NT-proBNP, TNF-α, PCT, IL-6 were independent risk factors for septic shock [CRP: odds ratio (OR) = 1.662, 95% confidence interval (95%CI) = 1.132-2.567; NT-proBNP: OR = 14.688, 95%CI = 3.580-20.238; TNF-α: OR = 1.093, 95%CI = 1.043-1.343; PCT: OR = 6.378, 95%CI = 4.556-12.243; IL-6: OR = 9.641, 95%CI = 2.242-13.786; all P < 0.05]. CONCLUSIONS: The levels of inflammatory factors CRP, NT-proBNP, TNF-α, PCT and IL-6 were significantly increased, which were important factors for early diagnosis of septic shock.

The development of low-calorie sugar and functional jujube food using biological transformation and fermentation coupling technology.

Jujube juice has been used as ingredient in a range of foods and dietary supplements. In this study, an enzyme transformation and fermentation coupling technology was applied to increase the nutritional value of concentrated/extracted Jinsi jujube juice. Two enzymes, D-glucose isomerase (GI) and D-allulose 3-epimerase (DAE), were employed to convert the glucose and fructose to a low-calorie sweeter D-allulose with a concentration of 110 g/L in jujube juice. Furthermore, the mixed cultures of Pediococcus pentosaceus PC-5 and Lactobacillus plantarum M were employed to increase the content of nutrition components related to bioactivities and flavor volatiles in jujube juice. Accordingly, this fermentation accumulated 100 mg/L gamma-aminobutyric acid (GABA), which has neurotransmission, hypotension, diuretic, and tranquilizer effects, and increased the content of branched-chain amino acids (BCAAs) and many free amino acids (Asp, Glu, Gly, and Ala) at different level. The fermentation not only maintained the concentration of native functional components such as cyclic adenosine monophosphate (cAMP) and minerals, but also increased the content of iron (Fe2+) and zinc (Zn2+), which have blood and eyesight tonic function. The value-added jujube juice might serve as a low-calorie and probiotic functional beverage and show high application potential in food industry.

Biosynthesis of dendroketose from different carbon sources using in vitro and in vivo metabolic engineering strategies.

BACKGROUND: Asymmetric aldol-type C-C bond formation with ketones used as electrophilic receptor remains a challenging reaction for aldolases as biocatalysts. To date, only one kind of dihydroxyacetone phosphate (DHAP)-dependent aldolases has been discovered and applied to synthesize branched-chain sugars directly using DHAP and dihydroxyacetone (DHA) as substrate. However, the unstable and high-cost properties of DHAP limit large-scale application. Therefore, biosynthesis of branched-chain sugar from low-cost and abundant carbon sources is essential. RESULTS: The detailed catalytic property of l-rhamnulose-1-phosphate aldolase (RhaD) and l-fuculose-1-phosphate aldolase (FucA) from Escherichia coli in catalyzing the aldol reactions with DHA as electrophilic receptors was characterized. Furthermore, we calculated the Bürgi-Dunitz trajectory using molecular dynamics simulations, thereby revealing the original sources of the catalytic efficiency of RhaD and FucA. A multi-enzyme reaction system composed of formolase, DHA kinase, RhaD, fructose-1-phosphatase, and polyphosphate kinase was constructed to in vitro produce dendroketose, a branched-chain sugar, from one-carbon formaldehyde. The conversion rate reached 86% through employing a one-pot, two-stage reaction process. Moreover, we constructed two artificial pathways in Corynebacterium glutamicum to obtain this product in vivo starting from glucose or glycerol. Fermentation with glycerol as feedstock produced 6.4 g/L dendroketose with a yield of 0.45 mol/mol glycerol, representing 90% of the maximum theoretical value. Additionally, the dendroketose production reached 36.3 g/L with a yield of 0.46 mol/mol glucose when glucose served as the sole carbon resource. CONCLUSIONS: The detailed enzyme kinetics data of the two DHAP-dependent aldolases with DHA as electrophilic receptors were presented in this study. In addition, insights into this catalytic property were given via in silico simulations. Moreover, the cost-effective synthesis of dendroketose starting from one-, three-, and six-carbon resources was achieved through in vivo and in vitro metabolic engineering strategies. This rare branched-chain ketohexose may serve as precursor to prepare 4-hydroxymethylfurfural and branched-chain alkanes using chemical method.

One-Pot Synthesis of Ginsenoside Rh2 and Bioactive Unnatural Ginsenoside by Coupling Promiscuous Glycosyltransferase from Bacillus subtilis 168 to Sucrose Synthase.

Ginsenosides, the major effective ingredients of Panax ginseng, exhibit various biological properties. UDP-glycosyltransferase (UGT)-mediated glycosylation is the last biosynthetic step of ginsenosides and contributes to their immense structural and functional diversity. In this study, UGT Bs-YjiC from Bacillus subtilis 168 was demonstrated to transfer a glucosyl moiety to the free C3-OH and C12-OH of protopanaxadiol (PPD) and PPD-type ginsenosides to synthesize natural and unnatural ginsenosides. In vitro assays showed that unnatural ginsenoside F12 (3- O-ß-d-glucopyranosyl-12- O-ß-d-glucopyranosyl-20( S)-protopanaxadiol) exhibited remarkable activity against diverse human cancer cell lines. A one-pot reaction by coupling Bs-YjiC to sucrose synthase (SuSy) was performed to regenerate UDP-glucose from sucrose and UDP. With PPD as the aglycon, an unprecedented high yield of ginsenosides F12 (3.98 g L-1) and Rh2 (0.20 g L-1) was obtained by optimizing the conversion conditions. This study provides an efficient approach for the biosynthesis of ginsenosides using a UGT-SuSy cascade reaction.

Adipose tissue-derived mesenchymal stem cells differentiated into hepatocyte-like cells in vivo and in vitro.

Cell­based therapy is a potential alternative to liver transplantation. The goal of the present study was to examine the in vivo and in vitro hepatic differentiation potential of adipose tissue­derived mesenchymal stem cells (AT­MSCs) and to explore its therapeutic use. AT­MSCs were isolated and cultured with hepatic differentiation medium. Bioactivity assays were used to study the properties of AT­MSCs. The morphology of differentiated AT­MSCs in serum­free hepatic differentiation medium changed into polygonal epithelial cells, while the morphology of AT­MSCs in a similar medium containing 2% fetal bovine serum remained unchanged. The differentiated cells cultured without serum showed hepatocyte­like cell morphology and hepatocyte­specific markers, including albumin (ALB) and α­fetoprotein. The bioactivity assays revealed that hepatocyte­like cells could take up low­density lipoprotein (LDL) and store glycogen. Furthermore, trichostatin A (TSA) enhanced ALB production and LDL uptake by the hepatocyte­like cells, analogous to the functions of human liver cells. ALB was detected in the livers of the CCl4­injured mice one month post­transplantation. This suggested that transplantation of the human AT­MSCs could relieve the impairment of acute CCl4­injured livers in nude mice. This therefore implied that adipose tissue was a source of multipotent stem cells which had the potential to differentiate into mature, transplantable hepatocyte­like cells in vivo and in vitro. In addition, the present study determined that TSA was essential to promoting differentiation of human MSC towards functional hepatocyte­like cells. The relief of liver injury following treatment with AT­MSCs suggested their potential as a novel therapeutic method for liver disorders or injury.

Alteration of UCP2 and ZO-1 expression in trabecular meshwork of neovascular glaucoma patients.

PURPOSE: The aim of this study was to investigate the expression of uncoupling protein 2 (UCP2) and zonula occludens-1 (ZO-1) in the trabecular meshwork (TM) of neovascular glaucoma (NVG) patients treated with trabeculectomy. MATERIALS AND METHODS: Six eyes with NVG underwent trabeculectomy for therapeutic purposes. The data consisted of patient demographics, presurgical and postsurgical visual acuity, intraocular pressure, gonioscopy, and neovascularization of iris and/or the anterior chamber angle. TM samples were obtained from the NVG eyes that had undergone surgery. Immunofluorescence and confocal laser scanning microscopy were carried out to determine the expression of UCP2 and ZO-1 in the TM cells. RESULTS: The baseline median visual acuity was light perception, and the mean intraocular pressure (standard error) was 52.5 (8.3) mm Hg. All eyes displayed neovascularization of the iris and the anterior chamber angle. The expression of UCP2 was significantly decreased in TM cells of NVG compared with the control (P=0.000), whereas increase in ZO-1 expression was detected in staining cells with NVG in comparison with the control (P=0.000). The necrotic cells in the TM were increased (P=0.000), whereas the viable cells were reduced (P=0.000) in comparison with the control. CONCLUSIONS: The decreased UCP2 expression and increased ZO-1 expression suggest that the oxidative stress-induced mitochondrial dysfunction and tight junction formation may play pivotal roles in the progress of NVG.

[Discussion on the electromagnetic compatibility testing and evaluation of radio frequency ablation catheter].

With the enforcement of YY 0505-2012, the testing items and evaluation points of radio frequency ablation catheter in electromagnetic compatibility field should be studied and discussed. Based on the requirements of relevant standards, this paper discusses on the testing items that should be applied and the evaluation points that should be focused on by analyzing the intended use and the structure of radio frequency ablation catheter, when it intends to apply registration individually with the basic knowledge of electromagnetic compatibility field.

Overexpression of CDKN1B inhibits fibroblast proliferation in a rabbit model of experimental glaucoma filtration surgery.

PURPOSE: To investigate the potential antiproliferative effect of cyclin-dependent kinase inhibitor 1B (CDKN1B) overexpression in a rabbit model of glaucoma filtration surgery (GFS). METHODS: The recombinant adenovector expressing exogenous CDKN1B was delivered to Tenon's capsule by subconjunctival injection during unilateral filtration surgery. The time course of CDKN1B expression was monitored by immunohistochemistry and Western blot analysis. Evaluation of proliferating activity was performed by proliferating cell nuclear antigen (PCNA), argyrophilic nucleolar organizing region (AgNOR) staining, and fibroblast-specific protein 1 (FSP-1). Cyclin-dependent kinase 2 (Cdk2) and Cdk4 expression were detected with immunohistochemical analysis. RESULTS: The overexpression of CDKN1B in Tenon's capsule was monitored throughout the experimental period. Immunoreactivity to CDKN1B was mainly observed in the nucleus of fibroblasts. The increased expression of CDKN1B in sclera was detected up to 21 days after viral infection, whereas the level of CDKN1B protein in corneal stroma was not significantly increased. The overexpression of CDKN1B induced a significant decrease in AgNOR number/nucleus and area/nucleus, PCNA staining, FSP-1 positive cells, and the decreased expressions of Cdk2 and Cdk4, as evidenced by nuclear and cytoplasmic immunoreactivity to Cdk2 and Cdk4 antibodies in positive fibroblasts. CONCLUSIONS: The persistent overexpression of CDKN1B mediated by the recombinant adenovector expressing exogenous CDKN1B in Tenon's fibroblasts after GFS may lead to the inhibition of fibroblast proliferation and the downregulation of Cdk2 and Cdk4 activity, thereby reducing the severity of scar formation and the surgical outcome.

[Pro-angiogenic effects of four Chinese medicines and three herbal prescription in chicken chorioallantoic membrane model].

OBJECTIVE: To observe the pro-angiogenic effect of four Chinese medicines and three herbal prescriptions, screen the effective components from them. METHODS: Chicken chorioallantoic membrane (CAM) model was employed to observe the pro-angiogenic activities of Angelicae Sinensis Radix, Salviae Miltiorrhizae Radix, Notoginseng Radix, Astragali Radix, Xuefuzhuyu decoction, Dangguibuxue decoction and Taohongsiwu decoction, all of them were claimed to promote angiogenesis. The effective components were screened from the extracts. RESULTS: Compared with negative control group, the blood vessel densities in Angelicae Sinensis Radix and Notoginseng Radix groups were not increased significantly (P > 0.05). However, blood vessel densities in Astragali Radix group, Salviae Miltiorrhizae Radix group, Xuefuzhuyu decoction group, Dangguibuxue decoction group and Taohongsiwu decoction group were notably enhanced (P < 0.05). Dangguibuxue decoction showed a more than 90% of increase in blood vessel densities as compared with the negative control group (P < 0.01), and components contained ferulic acid and astragaloside from Dangguibuxue decoction displayed significantly pro-angiogenic effect (P < 0.05). CONCLUSION: Dangguibuxue decoction and its extract, components contained ferulic acid and astragaloside, can improve angiogenesis in CAM model significantly.

Clinical evaluation of traumatic ciliochoroidal detachment with surgical treatment.

PURPOSE: To determine the clinical features of traumatic ciliochoroidal detachment (CCD), and to evaluate the surgical outcomes. METHODS: We retrospectively reviewed the records of 37 consecutive patients with traumatic CCD who underwent surgical procedures, including ciliary body suturing, transscleral cyclophotocoagulation, and cyclocryopexy. A complete ocular examination was performed at pre-surgery and at periodical post-surgery follow-ups. We compared visual acuity (VA), intraocular pressure (IOP), and morphologic changes with UBM among the different surgical procedures at the pre-surgery and periodical follow-ups. RESULTS: The mean IOP was 6.62 mmHg, and the median VA was 20/200 at baseline. The mean final IOP was 11.03 mmHg, and the final median VA improved to 20/50. IOPs were significantly different in post-surgery compared with those at baseline (P = 0.000) among the ciliary body suturing, cyclophotocoagulation, and cyclocryopexy groups. However, no significant differences were noted at each follow-up among the 3 groups (P > 0.05). The post-surgical morphological figures consisted of complete reattachment, partial reattachment, and the complete detachment. Cyclocryopexy (71.4%), suturing (68.4%), and cyclophotocoagulation (63.6%) produced similar surgical outcomes of the complete reattachment based on UBM images. CONCLUSION: Prompt treatment and periodic follow-ups are necessary after traumatic CCD, based on accurate dimensions and configuration by UBM. The appropriate choice of surgical procedures is pivotal for an optimal outcome.

Histological and ultrastructural features for proliferation inhibition by delivery of exogenous p27Kip1 to rabbit models after glaucoma filtration surgery.

PURPOSE: To investigate postoperative inhibition of cell proliferation in animals treated with exogenous p27Kip1 during an experimental glaucoma filtration surgery (GFS) by histological and ultrastructural examinations. METHODS: The primer was designed according to p27Kip1 gene sequence of GenBank. The p27Kip1-expressing adenovirus-mediated Ad-p27 was constructed according to standard techniques. Gene therapy was performed by subconjunctival delivery of Ad-p27, mitomycin C (MMC) and PBS served as controls. Histological and ultastructural changes at surgical sites were observed for 28 days postoperatively. RESULTS: Histologically, evident cellular proliferation induced by Ad-p27 was observed on day 7, with appreciable filtering cavity. However, thin conjunctival layers were noted, and the number of fibroblasts decreased in the Tenon's capsule on day 14. On day 28, the filtering cavity partially disappeared. The histological features of the animals with MMC delivery were similar to those treated by Ad-p27. In contrast, in the PBS treatment group, thick conjunctival layers were observed, and the number of goblet cells and fibroblasts, increased markedly. The filtering cavity disappeared at postoperative day 21. Twenty eight day postoperatively, ultrastructural findings showed that most conjunctival epithelial cells in the Ad-p27 treatment group were under static state, and that organelles were inactive. The number of goblet cells, and the secretion of mucin, were decreased. The amount of fibroblasts also decreased with partial apoptotic cells. The ultrastructural features presented by rabbits with the MMC delivery resembled those treated with Ad-p27. In the PBS treatment group, the number of conjunctival epithelial cells increased; the amount of goblet cells increased as the secretion of mucin was strengthened, and a substantial amount of fibroblasts were observed with active and productive organelles. CONCLUSION: Ad-p27 inhibits the proliferation of Tenon's capsule fibroblasts at surgical sites, and suppresses the formation of scars, thereby promoting surgical efficacy.

Activation of Notch signal pathway is associated with a poorer prognosis in acute myeloid leukemia.

OBJECTIVE: Notch signal pathway plays a fundamental role in regulating haematopoietic development. It is also an important mediator of growth and survival in several cancer types, with Notch pathway genes functioning as oncogenes or tumor suppressors in different cancers. However, the clinical role of Notch signal pathway in acute myeloid leukemia (AML) remains unclear. METHODS: To address this problem, we investigated the gene expression levels of Notch signal pathway members (Notch1, Jagged1 and Delta1) in bone marrow mononuclear cells by real-time quantitative PCR in a cohort of 100 patients with newly diagnosed de novo AML and normal marrow donors. The prognostic values of the three molecules in AML were also analyzed. RESULTS: Comparing with the normal controls, we show the transcriptional up-regulation of Notch1, Jagged1 and Delta1 in the bone marrow of AML patients with statistic differences (P = 0.008, 0.01 and 0.01, respectively). In addition, univariate analysis of factors associated with relapse-free survival and overall survival showed a significantly shorter survival in the patients with unfavorable karyotype, higher Notch1 expression, higher Jagged1 expression, or higher Delta1 expression. Moreover, Cox proportional hazards multivariate analysis of the univariate predictors identified karyotype and gene expression levels of Notch1, Jagged1 and Delta1 as independent prognostic factors for relapse-free survival and overall survival. Furthermore, the prognostic significance of Notch1, Jagged1 and Delta1 expression was more obvious in the subgroup of patients with intermediate-risk cytogenetics. CONCLUSION: Taken together, our data suggest for the first time that the activation of Notch pathway may indicate a poor prognosis in AML. Especially, Notch1, Jagged1 and Delta1 expression may be relevant prognostic markers in intermediate risk AML.

Surgical treatment for 42 patients with traumatic annular ciliochoroidal detachment.

AIM: To analyze the clinical features of traumatic annular ciliochoroidal detachment (CCD) with ultrasound biomicroscopy (UBM) images, to investigate the surgical outcomes of ciliary body suturing and the prognostic factors. METHODS: Forty-two patients with traumatic annular CCD who had undergone ciliary body suturing were enrolled for complete ocular examinations, including visual acuity (VA), slitlamp microscopy, tonometer, indirect ophthalscopy and UBM. Comparisons of clinical features were performed among baseline and follow-ups, and the morphologic alterations on UBM images were analyzed between pre- and post-surgery. RESULTS: The mean intraocular pressure (IOP) was 5.54mmHg, and the median VA was 0.1 in traumatic eyes at baseline. The pre-surgical morphological features on UBM images consisted of supraciliochoroidal effusion (33.33%), multilayer splits (40.48%) and CCD with cyclodialysis cleft (26.19%). After surgery, the median VA was 0.4 at the final follow-up. IOPs were significantly increased, which the mean final IOP was to 10.36mmHg (P<0.01). UBM images displayed complete reattachment in 40.48% of patients, partial reattachment in 50.00% of patients and 360-degree detachment in 9.52% of patients. Analyzing the prognostic factors, the significant factors were duration, VA at baseline, ocular laterality (P<0.01), gender, age and the presence of hypotonous maculopathy (P<0.05). CONCLUSION: Ciliary body suturing is the optimized procedure for traumatic annular CCD. UBM is a useful equipment for diagnosis and monitoring post-surgical morphological changes. The periodical detection of IOP and UBM is necessary for the observation of surgical outcomes.

Adenovirus-mediated delivery of p27(KIP1) to prevent wound healing after experimental glaucoma filtration surgery.

AIM: The aim of the study was to evaluate the outcome of adenovirus-mediated p27(KIP1) (Ad-p27) expression on wound healing after filtration surgery and to investigate the inhibition of cell proliferation induced by Ad-p27. METHODS: We constructed the adenovirus recombinant vector Ad-p27 and administered it to a rabbit model of glaucoma filtration surgery by subconjunctival injection; phosphate-buffered saline (PBS) and mitomycin C (MMC) were used as controls. Intraocular pressure (IOP), bleb scores, and anterior chamber depths were observed during a 28-d period. Histological examinations, fluorescence observations and Western blot analyses were evaluated. RESULTS: Ad-p27 enhanced the surgical outcome and inhibited cell proliferation when compared with PBS. Bleb scores in the Ad-p27-treated eyes were higher than those in the PBS-treated eyes on d 7 (P<0.01), 14 (P<0.01) and 21 (P<0.05). On d 28, IOP remained significantly decreased in the Ad-p27 group compared with the PBS group (P<0.05). However, no differences in bleb scores or IOPs were observed between the Ad-p27 and MMC groups. Histological analysis showed that total cell numbers were markedly reduced, and less scar tissue was observed at the surgical site in eyes treated with Ad-p27. The number of fibroblasts was decreased in Tenon's capsule in Ad-p27-treated eyes; however, a marked and diffuse signal from the green fluorescent protein (GFP) was observed in fibroblasts. Western blot analysis revealed a high level of p27(KIP1) expression in conjunctival epithelium (P<0.01), relatively high expression in superficial scleral stroma (P<0.01), and low expression in corneal epithelium in the Ad-p27 group. CONCLUSIONS: Ad-p27 administration significantly improves the outcome of filtration surgery and inhibits postoperative proliferation in rabbit eyes. These findings suggest that p27(KIP1) is a potential adjunctive agent for inhibition of wound healing after filtration surgery.

Subconjunctival sustained release 5-fluorouracil for glaucoma filtration surgery.

AIM: To determine the release characteristics of a 5-fluorouracil-loaded poly (lactic acid) disc (5-FU-PLA-DS) and the effect of sustained drug delivery on the success of glaucoma filtration surgery in rabbit eyes. METHODS: A method of microspheres accumulated by excessive carriers was used in the preparation of the 5-FU-PLA-DS. The disc was characterized for drug loading, entrapment efficiency, in vitro release, and external morphology. It was then implanted subconjunctivally into rabbit eyes with trabeculectomy. Intraocular pressure, ocular inflammatory reaction, filtration bleb appearance, and persistence were evaluated up to postoperative d 90. A quantitative analysis of 5-fluorouracil (5-FU) was performed in the aqueous humor. Ultrasound biomicroscopy was used to assess the appearance of the filtering fistula. RESULTS: The 5-FU-PLA-DS was produced with the drug-loading of 3.07+/-0.08 mg (mean+/-SD). 5-FU was released for 91 d with suppressive concentrations. The decrease in intraocular pressure from baseline was significantly more marked in the 5-FU-PLA-DSimplanted eyes during postoperative d 3-90, and the persistence of bleb and filtration fistula was longer than the control eyes (P<0.05). Corneal toxicity and hyperemia triggered by 5-FU was lower in the 5-FU-PLA-DS-implanted eyes than those exposed to 5-FU intraoperatively. The 5-FU concentration in the aqueous humor was insufficient for corneal endothelial damage. No evidence of toxic reaction was found in the conjunctival biopsy. CONCLUSION: 5-FU-PLADS displaying sustained intraocular release of 5-FU, reduced intraocular pressure, and prolonged bleb persistence, while significantly reducing 5-FU toxicity.

[Reconstruction of eyelid with hard palate mucosa autograft after resection of Malignant tumor of eyelid].

OBJECTIVE: To evaluate the clinical effect of the eyelid reconstruction using hard palate mucosa graft after resection of malignant tumor of eyelid. METHODS: From January 1998 to October 2003, 18 cases of malignant tumor of eyelid underwent the eyelid reconstruction with local flap grafting and hard palate mucosa autotransplantation. Of 18 cases, there were 10 males and 8 females, aging from 35 to 67 years. The defect was caused by basal cell carcinoma of eyelid in 10 cases, by carcinoma of meibomian glands in 6 cases and by squamous cell carcinoma of eyelid in 2 cases, including 12 cases of complete eyelid defect and 6 cases of 2/3 eyelid defect. RESULTS: The appearance and function of the eyelid in all cases were almost normal after the eyelid reconstruction with hard palate mucosa autotransplantation in replacing posterior layer of eyelid. The complete eyelid closure and no entropion were observed in all cases. The infection and contracture of the graft, immunologic rejection were not observed in all cases during the follow-up period of 6 to 48 months. CONCLUSION: The eyelid reconstruction using autotransplantation with hard palate mucosa in replacing posterior layer of eyelid was easy-to-operate and has satisfactory clinical effect. The hard palate mucosa as substitute of tarsus is superior to the traditional substitute.