Advances in the research of plant-derived natural products against retinoblastoma.

Retinoblastoma (RB) is a highly aggressive ocular tumor, and due to socioeconomic and medical constraints, many children receive treatment only in the metaphase and advanced clinical stages, resulting in high rates of blindness and disability. Although several approaches exist in the treatment of RB, some children with the disease do not have satisfactory results because of various factors. Plant-derived natural products have shown definite therapeutic effects in the treatment of various tumors and are also widely used in the study of RB. We review plant-derived natural products used in the study of anti-RB to provide ideas for the clinical application of these drugs and the development of new therapeutic drugs.

Anti-proliferation and apoptosis-inducing effects of sodium aescinate on retinoblastoma Y79 cells.

AIM: To investigate the anti-proliferation and apoptosis-inducing effects of sodium aescinate (SA) on retinoblastoma Y79 cells and its mechanism. METHODS: Y79 cells were cultured at different drug concentrations for different periods of time (24, 48, and 72h). The inhibitory effect of SA on proliferation of Y79 cells was detected by the cell counting kit-8 (CCK-8) assay, and the morphology of Y79 cells in each group was observed under an inverted microscope. An IC50 of 48h was selected for subsequent experiments. After pretreatment with SA for 24 and 48h, cellular DNA distribution and apoptosis were detected by flow cytometry. Real-time qunatitative polymerase chain reaction (RT-qPCR) and Western blot were used to assess changes in related genes (CDK1, CyclinB1, Bax, Bcl-2, caspase-9, caspase-8, and caspase-3). RESULTS: SA inhibited proliferation and induced apoptosis of Y79 cells in a time-dependent and concentration-dependent manner. Following its intervention in the cell cycle pathway, SA can inhibit the expression of CDK1 and CyclinB1 at the mRNA and protein levels, and block cells in the G2/M phase. In caspase-related apoptotic pathways, up-regulation of Bax and down-regulation of Bcl-2 caused caspase-9 to self-cleave and further activate caspase-3. What's more, the caspase-8-mediated extrinsic apoptosis pathway was activated, and the activated caspase-8 was released into the cytoplasm to activate caspase-3, which as a member of the downstream apoptotic effect group, initiates a caspase-cascade reaction that induces cell apoptosis. CONCLUSION: SA inhibits the proliferation of Y79 cells by arresting the cell cycle at the G2/M phase, and induces apoptosis via the caspase-related apoptosis pathway, indicating that SA may have promising potential as a chemotherapeutic drug.

Study on the possible factors leading to complications of hydroxyapatite orbital implantation / 国际眼科杂志(Guoji Yanke Zazhi)

·Hydroxyapatite ( HA ) orbital implant has been widely used since it was developed due to its many advantages. It is one of the ideal materials to fill the orbital volume after enucleation or evicseration. However, it still causes complications, and some complications such as severe eye exposure may require reimplant surgery. The paper reviews the clinical application of HA orbital implant, the possible factors leading to complications of HA orbital implantation and the treatments for several common complications. The review aims to provide some help to readers.

[Effects of glutamine combined with ulinastatin on inflammatory response of patients with severe burn injury].

OBJECTIVE: To observe the effects of glutamine combined with ulinastatin on inflammatory response of patients with severe burn injury. METHODS: Sixty patients with severe burn injury admitted to our burn wards from January 2010 to December 2011 conforming to the study criteria were divided into control group (C, n = 20), glutamine group (G, n = 20), and glutamine combined with ulinastatin group (G + U, n = 20) according to the random number table. Another 10 healthy volunteers were chosen as normal control group (NC). Isonitrogenous and isocaloric nutrition supports were given to patients in groups C, G, and G + U from post burn day (PBD) 2. 0.3 g/kg protein in the form of glutamine dipeptide was given to patients in group G for 10 days. 0.3 g/kg protein was given to patients in group G + U for 10 days with the same amount of glutamine dipeptide as that in group G, followed by intravenous injection of 100 kU ulinastatin (once per 8 hours) for 7 days during 10 days. The nitrogen concentration of 24 h urine was determined with Kieldahl nitrogen determination method, and nitrogen balance was calculated one day before treatment and ten days after treatment. Meanwhile, the levels of D-lactate in serum was determined by colorimetric method, the levels of diamine oxidase (DAO), TNF-α, and IL-6 by enzyme-linked immunosorbent assay, and LPS level by kinetic turbidimetric assay with TAL. Above-mentioned indexes were also examined in group NC. The wound healing rate on PBD 30, total hospital stay days, and the incidence of burn sepsis of all burn patients were recorded. Data were processed with one-way analysis of variance, LSD test, t test, and chi-square test. RESULTS: Compared with that in group C [(-5.40 ± 1.67) g/d], nitrogen balance in group G was significantly increased ten days after treatment [(-1.35 ± 0.59) g/d, P < 0.01]. The serum levels of D-lactate, DAO, LPS, TNF-α, and IL-6 in group G ten days after treatment were significantly lower than those in group C (P < 0.05 or P < 0.01). No statistically significant difference was observed in nitrogen balance and the serum levels of D-lactate, DAO between group G + U and group G (P values all above 0.05). The serum levels of LPS, TNF-α, and IL-6 in group G + U ten days after treatment were respectively (0.167 ± 0.064) EU/mL, (43 ± 14) pg/mL, (139 ± 23) pg/mL, which were significantly lower than those in group G [(0.240 ± 0.079) EU/mL, (59 ± 8) pg/mL, (195 ± 31) pg/mL, respectively, P < 0.05 or P < 0.01]. The would healing rate on PBD 30 and total hospital stay days in group G were respectively higher and shorter than those in group C (P values all below 0.01), but no statistically significant difference in the incidence of burn sepsis was found between them (P > 0.05). The would healing rate on PBD 30 in group G+U [(96 ± 4)%] was enhanced, and total hospital stay days [(41 ± 4) d] were lowered than those in group G [(88 ± 7)%, (49 ± 5)d, P values all below 0.01]. The incidence of burn sepsis of patients in group G + U (5%) was significantly lower than that in group C (35%, χ(2) = 6.234, P < 0.05). CONCLUSIONS: Glutamine combined with ulinastatin treatment can alleviate damage to intestine after severe burn injury, lower the serum level of inflammatory cytokines, promote wound healing, and reduce the incidence of burn sepsis.

Toxicity reduction in an industrial nitro-aromatic wastewater plant: an assessment and a proposed improvement.

In this study, the performance of a typical Chinese industrial nitro-aromatic wastewater project (operational capacity: 3,000 m(3)d(-1)) was evaluated using chemical properties and toxicity data. Additionally, the relationship between the removal of organic pollutants and toxicity reduction was investigated throughout the whole-process wastewater treatment. Current advanced treatment reduced the dissolved organic carbon by 40% compared with biologically treated wastewater effluent (BTWE), but the acute toxicity and early life-stage toxicity increased significantly. For instance, the acute toxicity of the current advanced treated wastewater was 450% greater than that of the untreated BTWE. With the aim of effectively decreasing the toxicity of the effluent, several efficient adsorption technologies were assessed and compared for further treatment of BTWE. Coagulation and/or oxidation coupled with activated carbon adsorption, hypercrosslinked resin adsorption, or MIEX(®) technology was helpful for improving chemical indices and reducing toxicity. Among these adsorption treatment technologies, hypercrosslinked resin adsorption was more effective at removing most of the toxicants than MIEX(®) technology, and it also had better regeneration efficiency and mechanical properties compared with activated carbon. Therefore, hypercrosslinked resin adsorption may be a promising technology for enhancing organic pollutant removal and toxicity reduction of BTWE from nitro-aromatic factories.

Spontaneous rodent models of diabetes and diabetic retinopathy.

Diabetes is a complex and heterogeneous disorder presently affecting more than 100 million people worldwide and causing serious socio-economic problems. Spontaneous rodent models of diabetes mellitus have proved invaluable in understanding the pathogenesis, complications, and genetic or environmental influences that increase the risks of diabetes. We have reviewed here in the development and characterization of spontaneous rodent models that displayed most features commonly associated with diabetic retinopathy.

VEGF expression and cell apoptosis in NOD mouse retina.

AIM: To investigate retinal vascular endothelial growth factor (VEGF) level and retinal cells apoptosis in the early stage of diabetic NOD mouse retina. METHODS: Animals were divided into non-diabetes group, (control) (2-, 4-, 6-, 8- and 12-week sub-groups, n=30) and diabetes group (2-, 4-, 6-, 8- and 12-week sub-groups, n=30). Enzyme-linked immunosorbent assay (ELISA) was performed to detect VEGF level in both serum and retina. Transmission electron microscope method was used to examine retinal cell apoptosis. RESULTS: Compared with the control group, VEGF levels in serum and retina were increased significantly in the NOD group (12 weeks: 4.9±0.4µg/g vs 0.19±0.1µg/g in serum sample, P<0.01; 165±9µg/g vs 17±5µg/g in retinal sample, P<0.01). There exists a positive correlation between serum VEGF and retinal VEGF levels in the early diabetic NOD mice (γ=0.9902, P=0.001). The number of the cells apoptosis in the ganglion cells and endothelium can also been found increased significantly in the NOD group (P<0.01). CONCLUSION: The high VEGF expression may be contributed to increased retinal cells apoptosis. Many factors associated with retinal VEGF expression might involve in the early diabetes stage.

Effect of intravitreal bevacizumab injection before vitrectomy on proliferative diabetic retinopathy.

AIM: To evaluate the effect of intravitreal bevacizumab (IVB) injection 1 week before pars plana vitrectomy (PPV) in proliferative diabetic retinopathy (PDR) patients. METHODS: A retrospective research was done on 46 PDR patients who were divided into PPV group (n=28) and IVB group (n=18, PPV with preoperative IVB). Bevacizumab was injected 1 week before PPV. Main outcome measures were visual acuity, incidence of iatrogenic retinal breaks, intraoperative and postoperative bleeding. RESULTS: At 1 month after surgery, visual acuity in PPV (82.1%) and IVB group (88.9%) improved significantly (P<0.01) and the difference between the two groups was not significant (P>0.05). Iatrogenic retinal breaks were reported in 18 cases (64.3%) in PPV group and 4 cases (22.2%) in IVB group (P<0.05). Intraoperative bleeding was encountered in all cases in PPV group and 7 cases (39%) in IVB group (P<0.01). Postoperative bleeding was reported in 9 cases (32.1%) in PPV group and none in IVB group (P<0.01). CONCLUSION: IVB injection before PPV is helpful in reducing iatrogenic retinal breaks, intraoperative and postoperative bleeding in PDR patients.

[Expression of hypoxia-inducible factor-1alpha, vascular endothelial growth factor and sFlt-1 in preeclampsia placenta].

OBJECTIVE: To investigate the expression and correlation of hypoxia-inducible factor-1alpha (HIF-1alpha), vascular endothelial growth factor (VEGF) and sFlt-1 in the preeclampsia placenta, and discuss their significance in the pathogenesis of preeclampsia. METHODS: Placentas were collected from 20 pregnant women with preeclampsia as study group and 15 normal pregnant women as control group. The expressions of HIF-1alpha, VEGF and sFlt-1 protein were semi-quantitatively analyzed with immunohistochemical assay and mRNA level was determined using reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: (1) the expression of HIF-1alpha and sFlt-1 protein in preeclampsia group obviously increased. Strong (+++) positive expression was observed in 9 and 11 cases respectively, significantly higher than in control group (2 and 3 cases) (P < 0.05), however, VEGF expression obviously reduced in preeclampsia group (P < 0.01). (2) the level of HIF-1alpha and sFlt-1 mRNA in preeclamptic placenta was 0.604 +/- 0.013, 0.898 +/- 0.041, significantly higher than 0.208 +/- 0.007 and 0.559 +/- 0.244 in normal placenta (P < 0.05). Although the level of VEGF mRNA increased in preeclampsia placenta, it was not significantly different from that in normal placenta (P > 0.05). The ratio of VEGF mRNA/sFlt-1mRNA obviously reduced in preeclampsia group and was significantly lower than in control group (P < 0.05). (3) in preeclampsia group, HIF-1alpha mRNA expression was positively correlated with the expression of sFlt-1 mRNA (r = 0.577, P < 0.05), and negatively correlated with the ratio of VEGF mRNA/sFlt-1 mRNA (r = -0.376, P < 0.05). CONCLUSION: Abnormal high HIF-1alpha expression in preeclampsia placenta indicates that HIF-1alpha might play an important role in the pathogenesis of preeclampsia, possibly through affecting the cytotrophoblastic invasion and placental vascular reconstruction via the modulation of VEGF and sFlt-1 gene transcription.

[Mechanisms of human glutamic acid decarboxylase 65 DNA vaccine preventing diabetes in non-obese diabetic mice].

OBJECTIVE: To investigate the mechanisms of human GAD65 DNA vaccine preventing insulitis and diabetes in NOD mice. METHODS: Female NOD mice at 4 weeks of age were randomly divided into PBS (n = 21), pcDNA (n = 20), and hGAD65 (n = 21) groups. Mice in each group received two intramuscular injections of 0.05 ml PBS alone, 50 microg pcDNA3.1 and 50 microg DNA vaccine emulsified in 0.05 ml PBS 7 days apart respectively. The accumulative diabetes incidence was followed-up to 30 weeks of age in each group of NOD mice. Pancreas was removed from NOD mice at 12 weeks of age in each group (n = 10) to score insulitis severity by routine H-E staining. The apoptotic beta cells in islets were observed with double-labeling technique of TUNEL in situ combined standard sensitive avdin-biotin complex (sABC) immunohistochemical method. Their spleens were for cell culture and total RNA extraction. Spleen IL-4, IFN-gamma, NF-ATc and NF-ATp mRNA levels were tested by RT-PCR. IL-4 and IFN-gamma levels in sera and supernatants of spleen cells were measured by ELISA. RESULTS: (1) At 30 weeks of age, the diabetes incidence was 95.2%, 80.0% and 61.9% in PBS, pcDNA and hGAD65 group respectively. The diabetes incidence in the PBS group was higher than that in hGAD65 group (P = 0.008). (2) At 12 weeks of age, the insulitis scores in hGAD65 group was lower than that in PBS group (P = 0.001) and pcDNA group (P = 0.027) respectively. (3) The apoptotic beta cell rates in hGAD65 group was lower than that in PBS group (P = 0.014) and pcDNA group (P = 0.023). (4) IL-4 levels in sera, spleen IL-4 and NF-ATc mRNA level in hGAD65 group were higher than those in PBS group (all P < 0.05) and pcDNA group (all P < 0.05) respectively, NF-ATp mRNA level in hGAD65 group was lower than that in PBS group (P < 0.05). CONCLUSION: Human GAD65 DNA vaccine via downregulating NF-ATp and upregulating NF-ATc and IL-4, makes Th cells deviate to Th2, and sequently prevents insulitis, beta-cell apoptosis and diabetes onset in NOD mice.