Ubenimex combined with Albendazole for the treatment of Echinococcus multilocularis-induced alveolar echinococcosis in mice.

Introduction: Alveolar echinococcosis (AE) is a parasitic disease caused by E. multilocularis metacestodes and it is highly prevalent in the northern hemisphere. We have previously found that vaccination with E. multilocularis-Leucine aminopeptidase (EM-LAP) could inhibit the growth and invasion of E. multilocularis in host liver, and Ubenimex, a broad-spectrum inhibitor of LAP, could also inhibit E. multilocularis invasion but had a limited effect on the growth and development of E. multilocularis. Methods: In this study, the therapeutic effect of Ubenimex combined with Albendazole on AE was evaluated. Mice were intraperitoneally injected with protoscoleces and imaging examination was performed at week 8 and week 16 to detect cyst change. During this period, mice were intraperitoneally injected with Ubenimex and intragastrically administered with Albendazole suspension. At last, the therapeutic effect was evaluated by morphological and pathological examination and liver function. Results: The results revealed that the combined treatment could inhibit the growth and infiltration of cysts in BALB/c mice infected with E. multilocularis protoscoleces. The weight, number, invasion and fibrosis of cysts were reduced in mice treated with Ubenimex in combination with Albendazole. The same effect was achieved by the single Ubenimex treatment because of its inhibitory effect on LAP activity, but it was less effective in inhibiting the growth of cysts. The levels of ALT, AST, TBIL, DBIL, ALP, and γ-GT were reduced after the combined treatment, indicating that treatment with both Ubenimex and Albendazole could alleviate liver damage. Discussion: This study suggests that the combined treatment with Ubenimex and Albendazole could be a potential therapeutic strategy for E. multilocularis infections.

Enantioselective Oxidative Stress and DNA Damage Induced by Rac- and S-metolachlor on the Earthworm Eisenia fetida.

Metolachlor is a widely used chiral herbicide. However, information on its enantioselective toxicity to earthworms, an important soil organism, remains limited. Herein, the effects of Rac- and S-metolachlor on oxidative stress and DNA damage in Eisenia fetida were investigated and compared. Moreover, the degradation of both herbicides in the soil was also determined. The results showed that reactive oxygen species (ROS) in E. fetida were more easily induced by Rac-metolachlor than S-metolachlor at a higher concentration (above 16 µg/g). Similarly, the effects of Rac-metolachlor on superoxide dismutase (SOD) activity and DNA damage in E. fetida were more significant than those of S-metolachlor at the same exposure concentration and time. Rac- and S-metolachlor did not result in severe lipid peroxidation. The toxic effects of both herbicides on E. fetida gradually decreased after 7 days as the exposure was prolonged. At the same concentration, S-metolachlor degrades faster than Rac-metolachlor. These results suggest that Rac-metolachlor has a greater effect on E. fetida than S-metolachlor, providing a significant reference for the rational use of metolachlor.

Therapeutic effect on Alveolar echinococcosis by targeting EM-Leucine aminopeptidase.

Alveolar echinococcosis (AE) is a parasitic disease caused by E. multilocularis metacestodes and it is highly prevalent in the northern hemisphere. We have previously found that vaccination with E. multilocularis Leucine aminopeptidase (EM-LAP) induced specific immune response and had an inhibiting effect on the parasites. In this study, the therapeutic effect of recombinant EM-LAP (rEM-LAP) on AE was evaluated and verified using Ubenimex, a broad-spectrum inhibitor of LAP. The results reveal that rEM-LAP could inhibit cyst growth and invasion and induce specific immunity response in BALB/c mice infected with E. multilocularis protoscoleces. The ultrasonic, MRI, and morphological results show that treatment with rEM-LAP inhibits E. multilocularis infection and reduces cyst weight, number, fibrosis and invasion. The same effect is observed for the treatment with Ubenimex by inhibiting LAP activity. The indirect ELISA shows that rEM-LAP could induce specific immunity response and produce high levels of IgG, IgG1, IgG2a, IgM, and IgA, and the serum levels of IFN-γ and IL-4 are significantly increased compared to the control groups, indicating that treatment with rEM-LAP leads to a Th1 and Th2 mixed-type immune response. This study suggests that EM-LAP could be a potential therapeutic target of E. multilocularis infection.

miR-21 regulates the proliferation and migration of non-small cell lung cancer A549 cells by targeting PDCD4 gene / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the effects of miR-21 targeting PDCD4 (programmed cell death factor 4) on proliferation and migration of non-small cell lung cancer (NSCLC) A549 cells and the possible mechanism. Methods: The miR-21 mimics, miR-21 inhibitors and miR-NC plasmids were transfected into A549 cells in logarithmic growth phase by liposome transfection technology. Forty-eight hours after transfection, the transfection efficiency was observed under a fluorescence microscope, and the mRNA expression levels of miR-21 and PDCD4 in A549 cells were detected by qPCR. Dual luciferase reporter gene experiment was used to detect the targeting relationship between miR-21 and PDCD4, MTT method was used to detect cell proliferation, Transwell chamber method was used to detect cell migration ability, and ELISA was used to detect the content of TNF-α in each group of cell culture fluids. WB was used to detect the protein expression levels of PDCD4, NF-κB p65 and p-NF-κB p65 in cells. Results: The A549 cell line with miR-21 over-expression or knockdown was successfully constructed. Dual luciferase reporter gene assay confirmed that miR-21 targetedly inhibited PDCD4 expression. Over-expression of miR-21 could significantly inhibit the mRNA expression of PDCD4 in A549 cells (P<0.01), promote cell proliferation and migration (P<0.05 or P<0.01), increase the secretion level of TNF-α (P<0.01), down-regulate the expression of PDCD4 protein (P<0.01), and up-regulate p-NF-κB p65 protein level (P<0.05). The effect of silencing miR-21 on cells was opposite to the effect of miR-21 over-expression. Conclusion: Over-expression of miR-21 can promote the proliferation and migration ability of A549 cells, which may be related to its targeted inhibition of PDCD4 and activating the NF-κB/TNF-α pathway.

Bioinspired Artificial Nanodecoys for Hepatitis†B Virus.

A facile route is presented for fabricating a new class of nanomimics that overexpress hepatitis B virus (HBV) receptor by a natural biosynthetic procedure against HBV infection. A nine-transmembrane HBV-specific receptor, human sodium taurocholate co-transporting polypeptide (hNTCP), was engineered to naturally immobilize it onto the cellular surface and subsequently trigger the budding of hNTCP-anchoring membrane vesicles (hNTCP-MVs) that favor the HBV virion. hNTCP-MVs could rapidly block HBV infection in cell models. Furthermore, hNTCP-MVs treatment could effectively prevent viral infection, spreading, and replication in a human-liver-chimeric mouse model of HBV infection. Our findings demonstrate the receptor-mediated antiviral effect of hNTCP-MVs to trick HBV and offer novel opportunities for further development of antiviral strategies in nanomedicine.

Papaverine inhibits lipopolysaccharide-induced microglial activation by suppressing NF-κB signaling pathway.

OBJECTIVE: To investigate the effects of papaverine (PAP) on lipopolysaccharide (LPS)-induced microglial activation and its possible mechanisms. MATERIALS AND METHODS: BV2 microglial cells were first pretreated with PAP (0, 0.4, 2, 10, and 50 µg/mL) and then received LPS stimulation. Transcription and production of proinflammatory factors (IL1ß, TNFα, iNOS, and COX-2) were used to evaluate microglial activation. The transcriptional changes undergone by M1/M2a/M2b markers were used to evaluate phenotype transformation of BV2 cells. Immunofluorescent staining and Western blot were used to detect the location and expression of P65 and p-IKK in the presence or absence of PAP pretreatment. RESULTS: Pretreatment with PAP significantly inhibited the expression of IL1ß and TNFα, and suppressed the transcription of M1/M2b markers Il1rn, Socs3, Nos2 and Ptgs2, but upregulated the transcription of M2a markers (Arg1 and Mrc1) in a dose-dependent manner. In addition, PAP pretreatment significantly decreased the expression of p-IKK and inhibited the nuclear translocation of P65 after LPS stimulation. CONCLUSION: PAP not only suppressed the LPS-induced microglial activity by inhibiting transcription/production of proinflammatory factors, but also promoted the transformation of activated BV2 cells from cytotoxic phenotypes (M1/M2b) to a neuroprotective phenotype (M2a). These effects were probably mediated by NF-κB signaling pathway. Thus, it would be a promising candidate for the treatment of neurodegenerative diseases.

Effects of low-level laser irradiation on proliferation and functional protein expression in human RPE cells.

Low-level laser irradiation (LLLI) modulates a set of biological effects in many cell types such as fibroblasts, keratinocytes, and stem cells. However, no study to date has reported the effects of LLLI on retinal pigment epithelia (RPE) cells. The aim of this study was to investigate whether LLLI could enhance the proliferation of RPE cells and increase the expression of RPE functional genes/proteins. Human ARPE-19 cells were seeded overnight and treated with 8 J/cm(2) of LLLI. Cell proliferation was measured by CCK8 assay and cell cycle distribution was evaluated by FACS. The transcription of cell cycle-specific genes and RPE functional genes was quantified by RT-PCR. Moreover, the expression of ZO-1 and CRALBP were evaluated by immunostaining. A dose of 8 J/cm(2) of LLLI significantly increased proliferation and promoted cell cycle progression while upregulating the transcription of CDK4 and CCND1 and decreasing the transcription of CDKN2A, CDKN2C, and CDKN1B in human ARPE-19 cells. Additionally, LLLI enhanced the expression of ZO-1 and CRALBP in human ARPE-19 cells. In conclusion, LLLI could enhance the proliferative ability of human ARPE-19 cells by modulating cyclin D1, CDK4, and a group of cyclin-dependent kinase inhibitors. It also could increase the expression of RPE-specific proteins. Thus, LLLI may be a potential approach for the treatment of RPE degenerative diseases.

Comparison of dexamethasone intravitreal implant and intravitreal triamcinolone acetonide for the treatment of pseudophakic cystoid macular edema in diabetic patients.

BACKGROUND AND OBJECTIVE: Our objective was to investigate the efficacy and safety of dexa methasone (DEX) implant for the treatment of pseudophakic cystoid macular edema (PCME) in diabetic patients. STUDY DESIGN: This was a prospective, non-randomized, interventional case series of 43 participants. Eighteen patients were enrolled in the DEX implant group and 25 were enrolled in an intravitreal triamcinolone acetonide (IVTA) group. MAIN OUTCOME MEASURES: The primary efficacy measurement was the percentage of patients who gained improvements of more than ten letters in best corrected visual acuity (BCVA) during 6 months of follow-up. Other efficacy measurements included change in BCVA, change in central macular thickness (CMT), and number of retreatments. The primary safety evaluation was the percentage of patients with intraocular hypertension and variation in intraocular pressure (IOP) during 6 months of follow-up. Other adverse events, such as conjunctival hemorrhage, eye pain, secondary infection, endophthalmitis, noninfectious inflammation, retinal detachment, and implant migration, were also recorded during follow-up. RESULTS: At month 1, we observed that the percentage of patients gaining improvement of more than ten letters was similar in both groups (P = 0.625). As patients in the IVTA group were retreated several times, this effect persisted throughout the study (P = 0.941 at month 2, P = 0.553 at month 3, P = 0.856 at month 6). Variations in CMT were noticed at week 1 and reached their maximum at month 1. No significant difference was found between the two groups (P = 0.831 at week 1, P = 0.783 at month 1). At month 1, the variation in IOP reached its maximum in the DEX implant group and then decreased slightly. However, in the IVTA group, it increased continuously throughout the study. Conjunctival hemorrhage and eye pain were found in both groups, but both were rated as mild in severity, and no significant difference was found (P = 0.184, P = 0.766, respectively). CONCLUSION: Both IVTA and DEX implants could effectively restore visual function and recover morphological change in diabetic patients with PCME for at least 6 months, but repeated intravitreal injection was required in the IVTA group. DEX implant is well tolerated. We suggest that intravitreal injection of DEX implant is a promising new therapeutic option for diabetic patients with PCME.

Subfoveal choroidal thickness after photodynamic therapy in patients with acute idiopathic central serous chorioretinopathy.

BACKGROUND: The purpose of this study was to evaluate changes in subfoveal choroidal thickness after photodynamic therapy in patients with acute idiopathic central serous chorioretinopathy (ICSCR). METHODS: This was a retrospective observational study conducted in 63 participants. The primary outcome measure was subfoveal choroidal thickness at baseline and 3 days, one week, 4 weeks, and 12 weeks after photodynamic therapy. The secondary outcome measure was indocyanine green angiography at baseline and 4 weeks and 12 weeks after photodynamic therapy. RESULTS: Four weeks after photodynamic therapy, 20 (64.51%) symptomatic eyes showed hypofluorescence corresponding to the area of photodynamic therapy irradiation at the posterior pole. The mean subfoveal choroidal thickness increased significantly from 422±132 µm at baseline to 478±163 µm at day 3 after treatment (P=0.022) and then decreased to 362±113 µm at week 4 (P<0.001) and 339±135 µm at week 12 (P<0.001). CONCLUSION: The subfoveal choroid in patients with acute ICSCR is thicker than in the normal population, and in symptomatic eyes is significantly thicker than in fellow eyes. Photodynamic therapy using a one third dose of verteporfin may decrease choroidal vascular hyperpermeability and choroidal thickness in patients with acute ICSCR.

Secondary glaucoma as initial manifestation of ring melanoma: a case report and review of literature.

Melanomas account for over 70% of adult malignancies in the eye and occur primarily in the choroid. Melanomas rarely originate in the ciliary body, with an annual incidence of approximately 1.6 cases per million. While the incidence rate of these tumors is low, malignant melanomas metastasize at early stages of disease development and show poor prognoses. Malignant melanomas of the ciliary body are often deeply hidden and have complex clinical manifestations, which are easily misdiagnosed and affect the prognosis. Here, we report a case of monocular ciliary body melanoma in an elderly Asian woman. Using this case as an example, we perform a systematic review of the disease's clinical symptoms, signs, diagnoses, differential diagnoses, treatment and prognosis.

Stem cell-based therapies for age-related macular degeneration: current status and prospects.

Age-related macular degeneration (AMD) is one of the major causes of irreversible blindness both in developed and developing countries. During the past decades, the managements of neovascular AMD (wet AMD) have dramatically progressed. However, still no effective treatment for non-neovascular AMD (dry AMD) which was characterized by geographic macular atrophy. Recent advances in stem cell sciences have demonstrated that retinal pigment epithelium (RPE) cells can be generated from several types of stem cells (including embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells, et al) by cell co-culturing or defined factors. Additionally, studies also showed that visual function could be recovered by transplantation of these cells into subretinal space in vivo. Moreover, the United States Food and Drug Administration already approved several clinical trials to evaluate the efficiencies of stem cell based cell transplantation for dry AMD patients. Till now, a few patients enrolled in these studies achieved promising outcomes. This review will summarize recent advances in stem cell based RPE differentiation, transplantation, and the preliminary results of clinical trials. The obstacles and prospects in this field will also be discussed.

The effect of eradicating Helicobacter pylori on idiopathic central serous chorioretinopathy patients.

PURPOSE: To evaluate the effect of Helicobacter pylori (H. pylori) eradication on the remission of acute idiopathic central serous chorioretinopathy (ICSCR). STUDY DESIGN: A prospective, randomized, placebo-controlled study of 53 participants. MAIN OUTCOME MEASURE: Twenty-seven acute ICSCR patients tested positive for H. pylori were given an eradication H. pylori therapy, and another 26 patients with the same diagnosis received matching placebo medication. All participants were tested for the following items: (1) disappearance rate of subretinal fluid (SRF); (2) best-corrected visual acuity (BCVA); and (3) central retinal sensitivity at baseline, 2 weeks, 4 weeks, 8 weeks, and 12 weeks after treatment. The difference between the two groups was analyzed by PASW statistics version 18.0. RESULTS: At each follow-up, the disappearance rate of SRF in the active treatment group seemed slightly better than in the control group, but no statistically significant differences were observed (P > 0.05 at each follow-up). The BCVA between the two groups also did not demonstrate statistically significant differences (P > 0.05 at each follow-up). Unlike the BCVA and the disappearance rate of SRF, we compared the change in central retinal sensitivity at 12 weeks after treatment; a statistical difference was observed (P = 0.042). CONCLUSION: Our findings suggested that H. pylori eradication does not improve BCVA and the disappearance rate of SRF, but it could improve the central retinal sensitivity in acute ICSCR patients. We recommend that chronic ICSCR patients and more sensitive methods for H. pylori diagnosis should be involved in evaluating the effect of H. pylori eradication.

[Expression of mGluR1 at primary visual cortex of monocular deprivation amblyopia rat and the observing of ultrastructure].

OBJECTIVE: To explore the regulation of expression of mGluR1 and the changes of neuron ultrastructure at primary visual cortex of monocular deprivation amblyopia rat within cortical period. METHODS: Taking randomized concurrent controlled trail. Establishing the model of monocular deprivation amblyopia rat. After proving the model successful by PVEP, all of the rats were randomly divided into three groups: normal visual cortex, experimental visual cortex and experimental opposite visual cortex. Immunocytochemical technology, electron microscope, photography microscope, computer image analysis, SPSS 11.5 and ANOV were used to get the results. RESULTS: Compared with the layer IV of normal visual cortex and experimental opposite visual cortex, the area of immunopositive neurons in layer IV of experimental visual cortex are deficiency, there is significant difference between them (P < 0.01). There are no significant differences between the other four corresponding layers (P > 0.05). Morphological abnormals were found in layer IV of experimental visual cortex by observing of ultrastructure. CONCLUSION: The expression of mGluR1 in layer IV of primary visual cortex of monocular deprivation amblyopia is reduced. There are morphological abnormals happened in layer IV of primary visual cortex of monocular derivation amblyopia. Reduced afference of nerve pulse because of monocular deprivation leads to the expression deficiency of mGluR1 in layer IV of the primary visual cortex, then synaptic plasticity happened, then neurons atrophy occurred may be one of the etiopathogenesis of amblyopia.