Objective Targeted adenoviral gene delivery from peripheral nerves was used to integrally analyse the characterization and time course of LacZ gene (AdLacZ) retrograde transfer to spinal cord and transgene product anterograde labeling ofperipheral nerve. Methods Recombinant replication-defective adenovirus containing AdLacZ was administrated to the cut proximal stumps of median and tibial nerves in Wister rats. Then the transected nerve was repaired with 10-0 nylon sutures. At different time point postinfection the spinal cords of C5 to T1 attached with DRGs and brachial plexuses, or L2 to L6 attached with DRGs and lumbosacralplexuses were removed. The removed spinal cord and DRGs were cut into 50 μm serialcoronal sections and processed for X-gal staining and immunohistochemical staining. The whole specimens of brachial or lumbosacral plexuses attaching with theirperipheral nerves were processed for X-gal staining. The number of X-gal stained neurons was counted and the initial detected time of retrograde labeling, peaktime and persisting period of gene expression in DRG sensory neurons, spinal cord motor neurons and peripheral nerves were studied. Results The gene transfer was specifically targeted to the particular segments of spinal cord andDRGs, and transgene expression was strictly unilaterally corresponding to the infected nerves. Within the same nerve models, the initial detected time of gene expression was earliest in DRG neurons, then in the motor neurons and latest in peripheral nerves. The persisting duration of β-gal staining was shortest in motor neurons, then in sensory neurons and longest in peripheral nerves. The initial detected time of β-gal staining in median nerve models was earlier in mediannerve models compared with that in the tibial nerve models. Although the initial detected time and the beginning of peak duration of β-gal staining were not same, the decreasing time of β-gal staining in motor and sensory neurons of thetwo nerve models were started at about the same day 8 post-infection. The labeled neurons were more in tibial nerve-models than that in median nerve models. Within the same models, the labeled sensory neurons of DRGs were morethan labeled motor neurons of ventral horn. The β-gal staining was tenser in median nerves than that in tibial nerves. However the persisting time of β-gal staining was longer in tibial nerve models. Conclusion The b gene expression in neurons and PNS renders this system particularly attractive for neuroanatomical tracing studies. Furthermore this gene delivery method allowing specific targeting of motor and sensory neurons without damaging the spinal cord might offer potentialities for the gene therapy of peripheral nerve injury.
Objective To construct the expression vector of HLA-G-shRNA and investigate the effect of HLA-GshRNA from NK cell lysis. Methods Four HLA-G shRNA plasmids were constructed and transiently transfected to Bel-7402 cell lines， the levels of mRNA and protein of HLA-G were detected by Real-Time PCR and Western blot. The cytotoxicity of NK-92MI cells against the transfected cells was analyzed by LDH releasing assay. Results The gel electrophoresis and sequencing showed that the inserted sequence was identical to the one which we designed, and no aberrations such as mutation，deletion or insertion occurred. The expressions of HLA-G confirmed by Real Time-PCR and Western blot were significantly down-regulated. Bel-7402 cell lines transfected HLA-G shRNA showed higher lytic activity （P＜0.01）. After KIR2DL4 receptor blocked，lytic activity of NK-92 MI cell were decreased （P＜0.01）. Conclusions HLA-G shRNA plasmids are successfully constructed and HLA-G down-regulated can increase NK cytolysis against Bel-7402 cell. After HLA-G combines with KIR2DL4 receptor at the surface of NK cells， the inhibition effect is transferred.
Objective To observe the rate of iris vessels exposure and analyze its relevant factors in normal full-term neonates. Methods A retrospective study. 1855 normal full term neonates, including 947 boys and 908 girls, were enrolled. The mean gestational age (GA) was (38.84±1.10) weeks and mean birth weight (BW) was (3 396.52±402.08) g. There were 1235 neonates from normal term vaginal delivery, 402 cases of cesarean delivery and 218 cases of forceps delivery. All neonates were examined with hand-held portable slit lamp biomicroscopy within 1 to 3 days after birth by two trained ophthalmologist respectively. Iris vessels exposure was defined as radial red blood vessels along iris fibers. Infants were divided into iris vessels exposure group and iris vessels unexposed group according to the findings of slit lamp biomicroscopy. 78 infants with iris vessels exposure were followed up for 42 days after birth till the iris vessels can’t be seen under microscope. The differences between the two groups were compared for gender, mode of delivery (MOD), GA, BW and body length (BL). Multiple logistic regressions were used to determine the factors related to iris vessels exposure. Results There were 298 neonates with iris vessels exposure among 1855 neonates and the rate was 16.1%. 1557 neonates (83.9%) had unexposed iris vessels. There were no different in gender (χ2=0.551) and MOD (χ2=3.036) between iris vessels exposure group and unexposed group (P>0.05), while the differences in GA (χ2=47.216), BW (t=4.603) and BL (t=3.936) between the two groups were statistically significant (P=0.000). Multiple logistic regression analysis revealed that only GA (β=−0.291, odds ratio=0.747, 95% confidence interval: 0.656 - 0.851, P=0.000) was correlated to iris vessels exposure significantly. The iris vessels couldn’t be seen in 77 of 78 infants with iris vessels exposure when followed up to 42 days. Conclusions The iris vessels exposure in normal full-term neonates is frequently observed. There is a significant inverse correlation between GA and iris vessels exposure.
ObjectiveTo analyze the influencing factors on clinical response to conbercept for diabetic macular edema (DME).MethodsA total of 51 patients (51 eyes) with DME who underwent intravitreal injection of conbercept were included in this retrospective study. The general information (age, sex, body mass index, smoking history, drinking history), blood glucose indicators (duration of diabetes, fasting blood glucose, HbA1c), blood pressure indicators (history of hypertension, systolic blood pressure, diastolic blood pressure), lipid indicators [total cholesterol (TC), high-density lipoprotein (HDL), apolipoprotein A (APOA)], biochemical indicators [neutrophil concentration, hemoglobin (HB), serum creatinine (Scr)] were collected. The best corrected visual acuity (BCVA) and macular central macular thickness (CMT) before and after treatment were comparatively analyzed. CMT reduced not less than 20% and BCVA increased by 2 lines as effective standards. Univariate analysis and multivariate logistic regression analysis were used to determine the factors affecting the efficacy of intravitreal injection of conbercept in patients with DME.ResultsUnivariate analysis showed that diastolic blood pressure, HDL, serum neutrophil concentration, baseline CMT and baseline BCVA were associated with edema regression (P＜0.05); HbA1c was associated with vision improvement (P＜0.05). Multivariate logistic regression analysis showed that there was a history of smoking (OR=0.122, 95% CI 0.017 − 0.887), low diastolic blood pressure (OR=0.850, 95%CI0.748 − 0.966), low HDL (OR=0.007, 95%CI 0.000 1 − 0.440), thin baseline CMT (OR=0.986, 95%CI0.977 − 0.995) were independent risk factors for failure outcome of edema regression (P＜0.05); long duration of diabetes (OR=1.191, 95%CI 1.011 − 1.404), high APOA (OR=1.007, 95% CI 1.000 − 1.013) were independent risk factors for failure outcome of vision improvement. Age, fasting blood glucose, systolic blood pressure, TC, HB, Scr and other indicators had no effect on the efficacy of edema regression and vision improvement after treatment (P＞0.05).ConclusionsSmoking history, long duration of diabetes, low diastolic blood pressure, low HDL level, high APOA level and thin baseline CMT are independent risk factors for the treatment of DME with intravitreal injection of conbercept.
ObjectiveTo observe the application value with color Doppler flow imaging (CDFI) in the detection of macular diseases in senile cataract.MethodsA cross-sectional descriptive study. From December 1 to December 30, 2017, 720 senile cataract patients (1380 eyes) diagnosed in Beijing Tongren Eye Center were included in this study. There were 330 males (629 eyes) and 390 females (751 eyes), with the mean age of 70.35±19.05 years. There were 716 right eyes and 664 left eyes, with the visual acuity 0.05-0.8. The fundus of all the patients can be observed. All patients were examined by OCT using Cirrus HD-Model 5000 produced by German Carl Zeiss company and CDFI using MyLab90 color Doppler ultrasound system produced by ESAOTE S.P.A. Comparing the detection rate of macular lesions between the two methods by using χ2 test. Using statistical methods to evaluate diagnostic tests, a crosstabulation was made to compare the consistency of frequency-domain OCT and CDFI in the diagnosis of macular lesions. Logistic regression analysis of correlation between detection of CDFI macular lesions and central foveal thickness (CFT), average thickness (AT), with or without macular anterior membrane, retinal folds, retinal cystoid edema, hard exudation, macular hole or lamellar macular hole, superficial detachment of neuroepithelium, detachment of pigment epithelium, vitreous macular traction.ResultsTwo hundreds and thirty-nine eyes (17.3%) showed macular disease by OCT, and 161 eyes (11.7%) showed the abnormal echo in the macular region by CDFI. The detection rate of the macular disease between two methods were statistically significant (χ2=851.661, P＜0.001), however, the consistency was well (κ=0.766, P=0.000). Logistic analysis showed that the detection rate of macular lesions was related to retinal folds, cysts in retina and CFT (r=1.396, 1.041, 0.12; P＜0.01).ConclusionCDFI can effectively detect macular lesions that affect the visual prognosis of senile cataract patients.
Objective To summarize the characteristics of color doppler flow imaging (CDFI) of ocular toxocariasis (OT) in children. MethodsA retrospective clinical study. From July 2014 to June 2020, 61 OT patients with 61 eyes diagnosed through clinical and laboratory testing in the Department of Ophthalmology of Beijing Tongren Hospital of Capital Medical University were included in the study. There were 45 males with 45 eyes and 16 females with 16 eye (male: female=2.81:1). Age were (6.93±2.50) years. The right eye and left eye were 29 and 32 eyes, respectively. Both eyes of the patient underwent two-dimensional ultrasound and CDFI examination. Two dimensional ultrasound was used to estimate the axial length (AL) of the affected eyes and healthy eyes on the opposite side. Among them, 52 cases were measured for AL using optical biometry and/or A-mode ultrasound. Vitreoretinal surgery was performed within one week after ultrasound examination. Two-dimensional ultrasound was used to observe the morphology of vitreous opacity, its connection to the eyeball wall, and whether posterior vitreous detachment and retinal detachment have occurred. CDFI examination was used to observe the presence of blood flow signals on the pathological membrane. The detection rates of different forms of vitreous opacity and traction retinal detachment were calculated. The location of proliferative lesions in the eye was analyzed. Paired t-test was performed to compare the AL of the affected eye and the healthy eye on the opposite side. Perform Kappa consistency test on the location of proliferative lesions was used during CDFI examination and vitreoretinal surgery. ResultsAll affected eyes have varying degrees of vitreous opacity. Among them, 23 eyes (37.7%, 23/61) showed typical "Christmas tree" like turbidity; 27 eyes (44.3%, 27/61) had clustered and striped echoes; 9 eyes (14.8%, 9/61) had weak punctate and strip echoes. Two eyes (3.3%, 2/61) showed a large amount of dense punctate and strip-shaped echoes. There were 50 eyes (82.0%, 50/61) with traction retinal detachment, of which 46 eyes (92.0%, 46/50) had visible blood flow signals on the detached retina, and the remaining 4 eyes (8.0%, 4/50) had no blood flow signals. During CDFI and surgery, there were 5 (8.2%, 5/61) and 4 (6.6%, 4/61) eyes with visible proliferative lesions in the periphery, respectively; 18 (29.5%, 18/61) and 14 (23.0%, 14/61) eyes were distributed in the posterior pole, respectively; there were 38 (62.3%, 38/61) and 43 (70.5%, 43/61) eyes with both peripheral and posterior polar regions, respectively. The consistency between CDFI and surgery in detecting the location of proliferative lesions was good (κ=0.832, 95% confidence interval 0.691-0.973, P＜0.001). The two-dimensional ultrasound measurement results showed that the AL of the affected eye was shorter than that of the contralateral healthy eye in 46 cases (75.4%, 46/61). Among the 52 patients who underwent AL biometry, the AL of the affected eye was shorter than that of the contralateral healthy eye by (0.63±0.68) mm, and the difference was statistically significant (t=－6.738, P＜0.05). ConclusionsCDFI can clearly display various intraocular lesions (vitreous opacity and traction retinal detachment) and eyeball sizes in children with OT. Vitreous opacity is often manifested as "Christmas tree" like, clustered, strip-shaped.
ObjectiveTo analyze the ultrasonographic features of adenoma of the nonpigmented ciliary epithelium (ANPCE). MethodsA retrospective series of case studies. From January 2014 to October 2021, 31 patients (31 eyes) with ANPCE (ANPCE group) were diagnosed in the eye center of Beijing Tongren Eye Center of Beijing Tongren Hospital, Capital Medical University, and 17 patients (17 eyes) with ciliary body melanoma (control group) diagnosed at the same time were selected as the control group. There was no significant difference in age (t=－0.564) and sex composition ratio (χ2=0.182) between the two groups (P=0.576, 0.670). All patients underwent ultrasound biomicroscopy to obtain the measurement parameters: tumor height, maximum basal diameter, maximum diameter, ratio of maximum diameter to basal diameter and ratio of maximum diameter to height; tumor location, shape, internal echogenicity intensity, echogenicity uniformity, degree of sound attenuation, invasion of iris, anterior displacement of the iris, lens subluxation were observed. The measurement parameters and observation indexes of the two groups were compared by independent sample t-test and χ2 test. Receiver operating characteristic (ROC) curve was drawn, area under the ROC curve (AUC) was determined, and parameter indicators with differential diagnosis value were screened. ResultsThe maximum diameter, height, maximum basal diameter, ratio of the maximum diameter to the maximum basal diameter, and the ratio of the maximum diameter to the height of the tumors in the ANPCE group and the control group were 5.64±0.98 mm, 4.24±0.59 mm, 3.66±0.71 mm, 1.58±0.34, 1.34±0.19 and 7.82±2.03 mm, 4.47±2.44 mm, 7.02±1.96 mm, 1.13±0.16, 2.09±1.06. The maximum diameter, the maximum basal diameter, and the ratio of the maximum diameter to the height of the tumor in the ANPCE group were all smaller than those of the control group, and the ratio of the maximum diameter to the maximum basal diameter was greater than that of the control group, and the differences were statistically significant (t=－4.159, －6.808,－2.924, 6.257; P<0.05). The tumors in the ANPCE group were mainly spherical (87.1%, 27/31), with no significant acoustic attenuation (77.4%, 24/31), less invading the root iris (77.4%, 24/31), and the tumors were mostly located in the ciliary body coronal (74.2%, 23/31); tumors in the control group were mainly hemispherical (47.1%, 8/17) or spherical (47.1%, 8/17), with significant sound attenuation (76.5%, 13/17), most of the tumors invaded the iris (70.6%, 12/17), and the tumors were mostly located from the pars plana to the coronal (76.5%, 13/17). There were statistically significant differences in the position, shape, sound attenuation degree, and whether it invaded the iris between the two groups of eyes (χ2=15.132, 19.767, 13.118, 10.581; P<0.05). The results of ROC curve analysis showed that the ratio of the largest diameter to the largest base diameter, the degree of sound attenuation and the AUC of whether the iris was violated were higher, which were 0.881, 0.769, and 0.740, respectively. ConclusionsUltrasound biomicroscopy is helpful in the diagnosis and differential diagnosis of ANPCE and ciliary body melanoma. The ratio of maximum diameter to maximum basal diameter, the degree of sound attenuation and whether it invades the root iris are important parameters to distinguish the two tumors.