AIM: To analyze the effects of two kinds of anti-vascular endothelial growth factor (VEGF) drugs, conbercept and ranibizumab, on proliferative diabetic retinopathy (PDR) patients as pre-treatment for pars plana vitrectomy (PPV).METHODS: From June 2016 to December 2016, 62 patients (64 eyes) aged 41-59 years old diagnosed with PDR with nonclearing vitreous hemorrhage (VH) and/or tractional retinal detachment (TRD) requiring PPV were enrolled in our study.Patients were treated with intravitreal injection of anti-VEGF drugs 0.50mg (0.05mL) 3d before PPV.Then the standard 23G minimally invasive sclera three-channel vitrectomy was performed where there were no significant complications after the injection of anti-VEGF drugs.The operation time, intraoperative bleeding, iatrogenic retinal breaks, the use of endodiathermy and silicone oil, and postoperative complications were recorded and analyzed.We compared and analyzed the visual acuity and macular thickness before and 1mo after the surgery with the preoperative data.RESULTS: Both conbercept and ranibizumab could improve the postoperative visual acuity and reduce the postoperative macular thickness of PPV.There was no significant difference between the impacts of two kinds of anti-VEGF drug pre-treatment on operation time, intraoperative bleeding, iatrogenic retinal breaks, the use of endodiathermy, silicone oil filling and postoperative vitreous secondary hemorrhage.CONCLUSION: The effects of conbercept and ranibizumab pre-treatment were similar.PPV combined with anti-VEGF pre-treatment could improve postoperative visual acuity and macular edema.The choice of conbercept or ranibizumab should be made flexibly according to the actual situation of patients.

Objective To discuss the intraoperative and postoperative effects of conbercept combined with pars plana vitrectomy (PPV) for proliferative diabetic retinopathy (PDR) patients.Methods From January 2016 to December 2016,123 PDR patients (149 eyes) aged 41-65 years old with nonclearing vitreous hemorrhage (VH) and/or tractional retinal detachment (TRD) were enrolled in this study.According to whether preoperative intravitreal injection conbercept,the patients were divided into two groups:Conbercept group (64 cases,78 eyes),Control group (59 cases,71 eyes).Conbercept group was treated with intravitreal injection of Conbercept 0.50 mg (0.05 mL) at 3 days before PPV.Then the standard 23G minimally invasive sclera three-channel vitrectomy was performed if no significant complications after the injection of conbercept.The operation time,intraoperative bleeding,iatrogenic retinal hole,use of endodiathermy and silicone oil,and postoperative complications were recorded and analyzed.The visual acuity and macuiar thickness were compared before and 1 month after the surgery.Results Two groups had no difference on age,sex,HbA1 C,duration,VH ≥ three degree(56/78,45/71),TRD.Conbercept pretreatment could significantly reduce the bleeding during PPV (43/78,49/71),the probability of iatrogeuic retinal holes (11/78,21/71),reduce intraocular electrocoagulation using (57/78,62/71) and silicone oil (43/78,51/71),and then shorten the operation time (58.63 ± 21.66)s and (72.69 ± 22.48)s,and it could significantly improve the postoperative visual acuity (0.23 ± 0.15,0.16 ± 0.11) and macular edema thickness (260.95 ± 27.44) μm and (330.81 ± 36.62)μm,while reduce the incidence of second bleeding (3/78,10/71).Conclusion Conbercept pre-treatment combined with PPV for PDR is a positive and effective treatment,which has good clinical application significance.

ObjectiveTo establish a central nervous system axonal mechanical transection model in vitro and observe the axonal regeneration style and speed following the transection. MethodsThe cortical explants the mice were cultured in vitro,of which the axon and dendrite parts were marked using immunofluorescence.The model was built by mechanically transecting the axons and removing the severed axons.The axonal regeneration style was observed by tracing the undeveloped cellular bodies adhering to the residual axonal surface.The growth speed of the regenerated axons and normal axons were measured as well.Results ( 1 ) After mechauically transecting the axons of explants,the axonal regeneration was largely founded at the transected line.The undeveloped cellular bodies went over the transected line and spread to the distal area with axonai regeneration. (2) The axonal growth speed was (118 ± 32) μm/d and (72 ±41) μm/d in the transection group,but was (41 ± 17) μm/d and (32 ± 19) μm/d in the control group at 24 and 48 hours respectively. ConclusionThe regeneration style of the transected axons is the extension of the axonal stumps,rather than sprouting growth,and the growth speed is faster than that of the normal axons.

Objective To establish a model fit for axonal regeneration research after its mechanical injury.Methods Cortical explants from mice were planted on culture dishes by microglass pipettes or routine glass pipettes.The cell body and dendrites in axonal area were detected by immunofluorescence and RT-PCR.Besides,purity of regenerated axons was also tested by immunofluorescence and RT-PCR after mechanical transection of axons under microscopy.Results Compared with explants planting by routine glass pipettes,in the outside 1/2 axons of explants planted by micro-glass pipettes,the immunofluorescence and RT-PCR showed negative nucleus and dendrites.In the regenerated axons following mechanical transection of explants planted by micro-glass pipette,the immunofluorescene and RT-PCR showed no regenerated axons nucleus mixed into the dendrites and nucleus.Conclusions Explants planted by micro-glass pipette obtains enough pure axons and regenerated axons.The establishment of models of axonal mechanical transection lays foundation for its molecular study after trauma.

Objective To investigate effect of polymorphism of apolipoprotein E (APOE) gene on early apoptosis of astrocytes after hypoxic injury.Methods Astrocytes separated from APOE wild mice and APOE transgenic mice (ε3,ε4) were primarily cultured,and then purified and identified.Models of astrocyte hypoxic injury were set up by hypoxia.Morphological changes of astrocytes and mitochondria were observed by transmission electron microscope.Early apoptosis rate and changes of mitochondrial membrane potential were detected by flow cytometry.Results Cell foot process tumidness and mitochondria with irregular outline,vacuoles and irregular cristae were observed in each group by electron microscopy at six hours after hypoxia.There were no significant differences of cellular form changes among groups.Early apoptosis rate and decreasing degree of mitochondrial membrane potential in APOFε4 group were significantly higher than those in APOEε3 group and APOE wild group (P ＜ 0.05).Conclusion Compared with astrocytes from APOEε3 group and APOE wild group,mitochondrial membrane potential in astrocytes from APOEε4 group at early period after hypoxia declines more significantly,as may be one of causes for more astrocyte apoptosis.

Objective To investigate the effect of lentivirus-mediated siRNA interference of USP39 on proliferation and migration of mice vascular smooth muscle cell in vitro.Methods Five siRNAs of siControl, siRNAUSP39-70, siRNAUSP39-71, siRNAUSP39-72 and siRNAUSP39-73 were designed and sythezied,mice VSMCs were infected with the lentivirus for delivering siRNAUSP39-73, and the stably transfected cells were selected by puromycin.The interference efficiency of siRNAUSP39-73 was assessed with Western blot.The effect of USP39 interference on the proliferation of VSMCs was determined by cells counting and MTT assay.Transwell assay was used to detect the migration of VSMCs.Results Recombinant lentiviral vector siRNAUSP39-73 was successfully transfected into mice VSMCs.Comparing with siControl group and Normal group, USP39 protein level of siRNAUSP39-73 VSMCs were decreased(P<0.05), and the proliferation and migration ability were all inhibited(P<0.05).Conclusion Targeted down-regulation of USP39 expression can inhibit the proliferation and migration of mice VSMCs in vitro.

Objective To establish a RP-HPLC method investigate the processing technique and mechanism of Eucommiae Cortex.Methods The RP-HPLC method was applied to simultaneously determining six ingredients,geniposidic acid,geniposide,genipin,chlorogenic acid,(+)-pinoresinol-di-β-D-glucopyranoside,and(+)-syringaresinol-di-β-D-glucopyranoside,in the different processed barks of Eucommia ulmoides.Results The valid method with good accuracy could be well used to study the processing technique of E.ulmoides;Besides,target ingredients in E.ulmoide were decreased within 6 h when they were processed.Conclusion Established RP-HPLC is a reliable method which could be used to research the processing technique of the barks ofE.ulmoides.Moreover,the result of this study could be provided with significant evidence of processed barks of E.ulmoides.

Objective:To investigate the correlation between fluid attenuated inversion recovery vascular hyperintensities (FVH) -diffusion weighted imaging (DWI) mismatch and the outcomes after endovascular mechanical thrombectomy (EMT) in patients with acute middle cerebral artery M1 segment occlusive stroke.Methods:Patients with middle cerebral artery M1 segment occlusive stroke who received EMT treatment and whose FLAIR images showed FVH in the Affiliated Hospital of Yangzhou University from January 2016 to June 2020 were enrolled retrospectively. The demographics and basic clinical information of the patients were collected. The modified Rankin Scale was used to evaluate the outcomes at 3 months after the onset of symptoms. 0-2 was defined as a good outcome, and >2 was defined as a poor outcome. Multivariate logistic regression analysis was used to determine the independent influencing factors of clinical outcome. Results:A total of 77 patients were enrolled in the study. Their age was 67.16±9.63 years, 51 were males (66.23%). The baseline National Institutes of Health Stroke Scale (NIHSS) score was 14.16±7.49. Forty patients (51.95%) had a good outcome, and 37 (48.05%) had a poor outcome. Univariate analysis showed that the proportion of patients with FVH-DWI mismatch in the good outcome group was significantly higher than that in the poor outcome group (60.00% vs. 29.73%; χ2=7.103, P=0.008), and baseline NIHSS score (11.60±4.44 vs. 16.92±9.05; t=-3.312, P=0.001) and the proportion of patients with hypertension (65.00% vs. 86.49%; χ2=4.774, P=0.029) were significantly lower than those in the poor outcome group. Multivariate logistic regression analysis showed that FVH-DWI mismatch was independently associated with the good outcomes (odds ratio [ OR] 0.345, 95% confidence interval [ CI] 0.121-0.984; P=0.047), baseline NIHSS score was independently associated with the poor outcomes ( OR 1.133, 95% CI 1.036-1.239; P=0.006). Conclusion:FVH-DWI mismatch was independently associated with the good outcomes after EMT treatment in patients with acute middle cerebral artery M1 segment occlusive stroke.

Objective To study if 5-Aza-2’-deoxycytidine along or together with 4-phenylbutyric acid could affect miR-196b expression levels in chronic myeloid leukemia cells .Methods K562 cells were treated with DNA methylation inhibitor 5-Aza-2’-deoxycytidine, histone deacetylase inhibitors 4-phenylbutyric acid separately and the combined treatment with both of them, then expression levels of miR-196b were detected using Real-time PCR.Results The half inhibition concentration of 4-phenylbutyric acid was 1.58mmol/L.Comparing with the expression level of miR-196b in normal human bone marrow cells, the expression levels of miR-196b were significantly lower in Aza group , PBA group and negative control cells and nearly consistent among three groups , and as high as normal cells in combined treatment group . Conclusion The expression level of miR-196b in K562 cells could not return to normal treated with 5-Aza-2 ’-deoxycytidine or 4-phenylbutyric acid separately , while could restore normal when treated with both agents , indicating that miR-196b expression level in K562 cells is related with both DNA methylation and histone acetylation .

Objective To study the application of Cell Counting Kit-8(CCK-8) in detecting the growth inhibiting effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cell .Methods The proliferation of K562 cells was detected by CCK-8 with different concentrations of 5-Aza-2 ’-deoxycytidine and the cell cycle and apoptosis of K 562 cells were detected after K562 treated by 50% inhibitory concentration of 5-Aza-2 ’-deoxycytidine .Results The 50% inhibitory concentration of 5-Aza-2’-deoxycytidine was 15.55nmol/L, after treated with this concentration , K562 cells showed that G2 phase arrest occurred , proliferation inhibited and apoptosis peaks appeared .Conclusion Inhibition of proliferation of K562 cells with different concentrations of 5-Aza-2’-deoxycytidine varied in a dose-dependent relationship , and 5-Aza-2’-deoxycytidine could promote apoptosis of K 562 cells.CCK-8 can be used in detecting the effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cells .