Objective To observe the different changes of macular microstructure in patients with large idiopathic macular hole (IMH) treated with vitrectomy combined with internal limiting membrane (ILM) transplantation or not.Methods Forty eyes in 40 consecutive patients with giant IMH (≥500 μm) were included in the study.Twenty eyes received vitrectomy with ILM transplantation (ILM transplantation group) and others with ILM peel off (ILM removal group).During the operation,a proper size of the ILM was removed and filled in the bottom of the macular hole.The age,duration of disease and the ocular laterality of the two groups of patients were not statistically significant (P＞0.05).Minimum resolution angle in logarithmic (logMAR) best corrected visual acuity (BCVA) and frequency domain optical coherence tomography (SD-OCT) scan were examined.There was no statistically significant difference in logMAR BCVA,average defect diameter of photoreceptor ellipsoid (IS/OS) and average defect diameter of external limiting membrane (ELM) between two groups (t=0.128,1.452,1.321;P＞0.05).The logMAR BCVA and SD-OCT were examined on 1,3,6,12 months postoperatively.Results On 1 month after the surgery,there was no statistically significant difference in logMAR BCVA,average defect diameter of IS/OS and average defect diameter of ELM between two groups (t=1.226,1.435,1.018;P＞0.05).On 3,6,12 months after the surgery,compared with ILM removal group,the logMAR BCVA (t=2.059,2.871,2.415) increased and the average defect diameter of IS/OS (t =2.070,2.110,2.121) and ELM (t =2.034,3.647,3.556) significantly reduced in ILM transplantation group (P＜0.05).On 1 month after the surgery,there was statistically significant difference in CRT between two groups (t=2.113,P＜0.05).On 3,6,12 months after the surgery,there was no statistically significant difference in CRT between two groups (t=0.428,0.847,0.849;P＞0.05).Conclusion Compared with vitrectomy combined with ILM peeling surgery,the diameter of IS/OS and ELM defect were significantly decreased after vitrectomy combined with ILM transplantation in the patients with large IMH.

Objective To establish a tandem affinity purification(TAP) system of innate immune-regulatory protein PKR and analyze PKR function, for the future screen and identification of novel PKR-interaction proteins. Methods PKR gene was amplified by PCR, and then cloned into a mammalian expression vector pcTAP-A. Recombinant pcTAP-PKR was transfected into PKR knock-down(PKRkd) HeLa cells by LipofectAMINE 2000,and the PKR overexpressed HeLa cells were harvested for mitogen-activated protein kinases(MAPK) activation analysis. Cell extracts of PKR overexpressed cells were purified using TAP kit and examined by Western blot. Results Cal modulin resin(CBP) and streptavidin resin(SBP) tagged PKR was detected in PKRkd HeLa cells as early as 24 h upon transfection with pcTAP-PKR, and its expression decreased at later time points. The overexpression of PKR was autophosphorylated, and thus involved in the regulation of MAPK actviation. After small-scale TAP kit purification, PKR protein was detectable by Western blot. Conclusion We have successfully established a TAP system that over-expresses functional PKR, providing a useful tool for the future study on the identification of PKR interacting proteins.

Objective To investigate the effect of CTLA4-Ig chimera protein on mice mortality, histopathological changes, viral fiters, expression of CTLA4 protein on infiltrated T lymphocyte and the balance of Thl/Th2 in mice myocarditis caused by coxsackie virus B3 (CVB3). Methods A total of 106 four to six week-old male BALB/c mice were used in the experiments, which were divided into CTLA4-Ig group (n = 16), CVB3 group (n=40), IgG group (n =40) and normal control group(n = 10) randomly. The mice in CVB3 group, IgG group and CTLA4-Ig group were inoculated intraperitoneally with 0. 15 ml CVB3 and the mice in norreal control group with 0. 15 ml Eagle. The mice in IgG group and CTLA4-1g group were inoculated with IgG (0. I mg/kg) and CTLA4-Ig(0. 1 mg/kg) at 6 h and 72 h post inoculation(p, i. ), respectively, The surplus mice in each group were sacrificed at day 7 p.i. Light microscope was used to quantify the inflammation. The expression of CVB3 mRNA in mycardium were semi-quantified by real-time quantitative polymerase chain reaction (RQ-PCR). The expression of CTLA4 protein were analyzed by immunohistochemistry. The levels of IL-2, IL-4 and 1FN-γ in serum were measured by ELISA. Results The mice mortality, histopathological score and CVB3 mRNA in CTLA4-Ig group were lower than that in CVB3 group ( P < 0.05, P < 0.01, P < 0. 05, respectively). The expression of CTLA4 was significantly increased in CTLA4-Ig therapy group (P < 0.05 ). The serum level of IFN-γ of mice in CVB3 group were significantly higher than that in normal control group( P < 0.01 ). The serum level of IL-4 of mice in CVB3 group were much lower than that in normal control group( P < 0.01 ). The serum level of IL-2 in CVB3 group had no statistical significance with that in normal control group ( P > 0.05 ). The serum level of IFN-γ in mice of CTLA4-Ig group were much lower than that in CVB3 group ( P <0.01 ) and lgG group (P < 0. 01 ). The serum level of IL-4 of mice in CTLA4-Ig group were significantly higher than that in CVB3 group (P<0.01) and IgG group (P<0.01). The serum level of IL-2 in CTLA4-Ig group had no statistical significance with that in CVB 3 control group and lgG group ( P > 0. 0 5 ) . Conclusion CTLA4-Ig may relieve inflammation and reduce mice mortality by blocking the costimulation signals for T lymphocyte activation and reinforcing Th2 response.

Objective:To investigate the role of neutrophil CD 11b (nCD 11b), soluble CD 14 subtype (sCD 14-St) and mitochondrial coupling factor-6 (CF-6) in the risk stratification of disease outcome in neonatal sepsis and its clinical significance. Methods:The clinical data of 121 septic neonates from July 2019 to March 2020 in Shanxi Children′s Hospital were retrospectively analyzed. According to the neonatal critical illness score (NCIS), the neonates were divided into non-critical group (NCIS>90 scores) with 35 cases, critical group (NCIS 70 to 90 scores) with 49 cases, very critical group (NCIS<70 scores) with 37 cases. There were 25 cases with poor prognosis (death), and 96 cases with good prognosis (survival). The C-reactive protein (CRP), procalcitonin (PCT), nCD 11b, sCD 14-St and CF-6 before treatment were detected. The correlation between nCD 11b, sCD 14-St, CF-6 and disease severity was analyzed by Spearman method; the value of nCD 11b, sCD 14-St and CF-6 in predicting poor disease outcome in sepsis neonates was analyzed by the receiver operating characteristic (ROC) curve. Results:The nCD 11b, sCD 14-St, CF-6, PCT and CRP in critical group and very critical group were significantly higher than those in non-critical group: (414.68 ± 93.29) and (532.74 ± 101.85) MFI vs. (325.45 ± 71.90) MFI, (892.40 ± 113.72) and (1 249.53 ± 95.41) ng/L vs. (784.66 ± 103.72) ng/L, (84.79 ± 28.35) and (121.66 ± 34.27) ng/L vs. (42.59 ± 13.51) ng/L, (19.24 ± 6.30) and (34.96 ± 11.95) μg/L vs. (8.89 ± 2.24) μg/L, (109.49 ± 36.77) and (247.13 ± 82.06) mg/L vs. (56.84 ± 17.25) mg/L; the indexes in very critical group were significantly higher than those in critical group, and there were statistical differences ( P<0.05). Spearman correlation analysis result showed that nCD 11b, sCD 14-St and CF-6 were positively correlated with disease severity in sepsis neonates ( r = 0.719, 0.813 and 0.823; P<0.01). The nCD 11b, sCD 14-St, CF-6, PCT and CRP in poor prognosis neonates were significantly higher than those in good prognosis neonates: (618.58 ± 146.92) MFI vs. (374.55 ± 120.03) MFI, (1 516.91 ± 194.38) ng/L vs. (828.13 ± 175.67) ng/L, (165.84 ± 25.63) ng/L vs. (62.51 ± 16.75) ng/L, (43.46 ± 10.14) μg/L vs. (20.19 ± 6.30) μg/L and (321.09 ± 94.56) mg/L vs. (88.24 ± 29.19) mg/L, and there were statistical differences ( P<0.01). ROC curve analysis result showed that the area under the curve (AUC) of nCD 11b, sCD 14-St and CF-6 for predicting poor disease outcome in sepsis neonates were 0.763, 0.796 and 0.838 (95% CI 0.678 to 0.836, 0.713 to 0.864 and 0.760 to 0.899), and the AUC of combination the 2 indexes was 0.921 (95% CI 0.858 to 0.962). Conclusions:The nCD 11b, sCD 14-St and CF-6 are associated with the disease severity and prognosis in sepsis neonates, and can be used as markers for risk stratification of disease outcome and assessment prognosis.

Background Giant idiopathic macular hole (IMH) severely affects visual acuity and increases operative difficulty during the surgery,so modifying or optimizing the operation method is helpful for improving the prognosis.Objective This study aimed to evaluate the efficacy and safty of vitrectomy combined with free internal limiting membrane transplantation for large IMH.Methods A prospective serial cases-observational study was carried out under the informed consent of each patient.Forty-two eyes of consecutive 42 patients with IMH of mean diameter (814.31±112.95)μm were included in the Second Hospital of Hebei Medical University from January 2013 to November 2015.All the eyes received vitrectomy combined with free internal limiting membrane transplantation and 12％ C3F8 filling.The best corrected visual acuity (BCVA) (LogMAR),inner segment/outer segment (IS/OS) defect range,external limiting membrane defect range,retinal thickness at macular fovea were measured with slit lamp microscope,indirect ophthalmoscope and spectral-domain optical coherence tomography (SD-OCT) before surgery and 1,3,6 and 12 months after surgery.Results IMH complete closure in 97.6％ eyes (41/44) at 12 months after surgery.BCVA was improved after operation and showed a significant difference among various time points (F =28.032,P＜0.001).The IS/OS defect range was (1 112.00±45.44),(859.00±84.55),(649.00±52.47),(486.00±46.88) and (320.00±45.13) μm before surgery and 1,3,6 and 12 months after surgery,showing a significant difference among different time points (F=38.761,P＜0.001),and the IS/OS defect range was gradually shrinked after operation compared with that before operation (all at P＜0.05).The mean defect range of external limiting membrane was (1 038.00 ±39.63),(748.00±64.12),(585.00±48.88),(438.00±42.84) and (265.00±28.97)μm before surgery and 1,3,6 and 12 months after surgery,with a significant difference among various time points (F=36.459,P＜0.001),and the mean defect range of external limiting membrane was evidently reduced (all at P＜0.05).The foveal retinal thickness value was increased at 3,6 and 12 months after surgery,which was significantly higher than that 1 month after surgery (all at P＜0.05).The high reflect signal was faded away at 3 months after operation,indicating that implanted internal limiting membrane was decomposed and metabolized.Conclusions Vitrectomy combined with autologous internal limiting membrane transplantation seems to be safe and effective for large IMH.