ObjectiveTo explore the light response, retinal inflammation and apoptosis of the retinal ganglion cells (RGCs) 1 year after the new type of channelrhodopsin PsCatCh2.0 was transfected into the retina of rd1 mice. MethodsTwenty-four male rd1 mice were randomly divided into rd1 experimental group and rd1 control group, 12 mice in each group. 1.5 μl of recombinant adeno-associated virus (rAAV)2/2-cytomegalovirus (CMV)-PsCatCh2.0-enhanced green fluorescent protein (EGFP) was injected into the vitreous cavity 1 mm below the corneoscleral limbus of mice in the rd1 experimental group, and the same dose of recombinant virus was injected 2 weeks later at temporal side 1 mm below the corneoscleral limbus. One year after virus injection, the light response of RGCs expressing PsCatCh2.0 was recorded by patch clamp technique; the expression of PsCatCh2.0 in the retina was evaluated by immunofluorescence staining; the transfection efficiency of recombinant virus was evaluated by the transfection efficiency of virus and the number of RGCs. Hematoxylin-eosin staining was performed to measure the inner retinal thickness. Western blotting was used to detect the protein expression of nuclear factor (NF)-κB p65 in retina; real-time quantitative polymerase chain reaction was used to detect the relative expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and Bax mRNA. Terminal deoxynucleotidyl transferase kit was used to observe the apoptosis of retinal cells in each group of mice. ResultsOne year after the intravitreal injection of recombinant virus, PsCatCh2.0-expressing RGCs can still generate 30 pA photocurrent. The virus PsCatCh2.0-EGFP was mainly transfected into RGCs, and partly transfected into amacrine cells, almost no transfection was seen in bipolar and horizontal cells. There were no significant differences in the number of RGCs and thickness of the inner retina between the rd1 experimental group and the rd1 control group (F=14.35, 0.05; P＞0.05), while the rd1 experimental group NF-κB p65 protein expression, TNF-α and IL-6 mRNA quantification were significantly lower than those of rd1 control group (F=4.61, 5.91, 5.78; P＜0.05). The number of red fluorescent apoptotic cells in the retina of mice in the rd1 experimental group was less than that in the rd1 control group, and the Bax mRNA expression was lower than that in the rd1 control group, and the difference was statistically significant (F=7.52, P＜0.01). ConclusionOne year after intravitreal injection of recombinant virus, the PsCatCh2.0 expressing RGCs can still generate photocurrent. Long term transfection and expression of PsCatCh2.0 has no obvious cytotoxic effect on RGCs, nor it increases the inflammatory effect of the retina of rd1 mice with retinal degeneration.
ObjectiveTo explore the light sensitivity and kinetic of the new optogenetics tools Channelrhodopsin-XXM2.0 (XXM2.0) and Channelrhodopsin-PsCatCh2.0 (PsCatCh2.0), and analyze whether they could be used to restore the visual function by optogenetics.MethodsMolecular biology techniques were used to link the gene fragments of XXM2.0 and PsCatCh2.0 to the vector pCIG(c)-msFoxn3 containing ampicillin resistant screening gene and reporter gene to form new plasmid pCIG(c)-msFoxn3-XXM2.0 and pCIG(c)-msFoxn3-PsCatCh2.0. The constructed plasmids were transfected into HEK 293T cells, and light responses were recorded in the whole cell mode with the HEKA patch clamp system. The photocurrent was recorded under three light intensity included 2.7×1016, 4.7×1015, and 6.4×1014 photons/(cm2·s). And then, XXM2.0 and PsCatCh2.0 were stimulated with 2.7×1016 photons/(cm2·s) and fully recovered. The opening and closing time constants were analyzed with Clampfit 10.6 software. At the same light intensity, photocurrents of XXM2.0 and PsCatCh2.0 were recorded by the light pulse stimulating of 2-32 Hz. The current attenuation was analyzed at long intervals of 4000 ms and short intervals of 200 ms after repeated stimulation. Comparisons between groups were performed by independent samples t test.ResultsRestriction endonuclease sites of EcoRⅠ and EcoRⅤ were successfully introduced at XXM2.0 and PsCatCh2.0 sequences. When the digestion was completed, they were ligated by T4 DNA ligase to construct new plasmids pCIG(c)-msFoxn3-XXM2.0 and pCIG (c)-msFoxn3-PsCatCh2.0, and then transfected on HEK 293T cells. The light intensity dependence was showed in XXM2.0 and PsCatCh2.0. The greater light intensity was accompanied by the greater photocurrent. Under the light intensity 6.4×1014 photons/(cm2·s) below the retinal safety threshold, large photocurrent was still generated in XXM2.0 and PsCatCh2.0 with 92.8±142.0 and 13.9±5.6 pA (t=1.24, 1.24; P=0.28, 0.29). The opening time constants of XXM2.0 and PsCatCh2.0 were 23.9±6.7 and 2.4±0.8 ms, and the closing time constants were 5803.0±568.2 and 219.9±25.6 ms. Compared with PsCatCh2.0, the opening and closing time constant of XXM2.0 were both larger than PsCatCh2.0. The differences were statistically significant (t=7.10, 31.60; P=0.00, 0.00). In terms of response frequency, XXM2.0 and PsCatCh2.0 could follow to 32 Hz high-frequency pulsed light stimulation, and all could respond to repeated light stimulation at a long (4000 ms) and a short time (200 ms) interval with the small current decay rate.ConclusionXXM2.0 and PsCatCh2.0 could be activated under light intensity with safety for the retina, and could respond to high frequency (at least 32 Hz) pulsed light stimuli with low current attenuation, which could meet the characteristics of opsins required to restore the visual function by optogenetics.
PURPOSE: To explore the pathogenesis of anisometropic and amblyopias. METHODS:To carry out on monocular and binocular atropinized cat models during the developmental period for anisometropia and ametropia ,and measure the cytosomal sectional area and some parameters of the dendric field from the dorsal lateral geniculate nuclei (dLGN)of adult cats by using Golgi-Cox staining. RESULIS:The changes of cytosomal sectional areas and parameters about dendric fields in the dLGN of experimental cats were as following:significant differences between cells of dLGN's A1 lamina by the monocular atropinized eyes and normal ones, binocular atropinized eyea and normal ones;no significant difference between tbat driven by the monoular and binocular atropinized eyes. CONCLUSIONS:There might be resemble pathogenesis between anisomelropic and ametropic amblyopias. （Chin J Ocul Fundus Dis,1996,12:153-156）
Objective To explore and evaluate the protective effects of soluble fms-like tyrosine kinase recptor-1(sFlt-1) gene 2-3 and 2-4 transcellular region on retinal vascular leakage and phosphatidyl inositol-3 kinase (PI3K)/Akt pathway under hypoxia and/or hyperglycemia. Methods The plasmids pcDNA3.1-EGFP/sFlt-1(2-3) and pcDNA3.1-EGFP/sFlt-1(2-4) were constructed.. Two of these plasmids with carboxy methylation glucan (CMD) magnetic particles were transfected into human umbilical vein endothelial cells (HUVEC), which were cultured under hypoxia and/or hyperglycemia. The blood retinal barrier was evaluated by Evans blue permeation (EBP). Then the expression of p-Akt mRNA and protein were detected by reverse transcriptionpolymerase chain reaction (RT-PCR) and Western blot separately. Results In the diabetic rabbits, The blood retinal barrier breakdown was detected by the retinal vascular leakage of EBP. The expression of p-Akt mRNA and protein in hypoxia and hyperglycemia groups were also obviously increased. These changes were largely prevented by transfection the plasmids pcDNA3.1-EGFP/sFlt-1(2-3) and pcDNA3.1-EGFP/sFlt-1(2-4) (P＜0.01 in both groups). Conclusion Both sFlt-1(2-3) and sFlt-1(2-4) can make the retinal blood vessels less leaky and may be beneficial in preventing vision loss from diabetic retinopathy.
Objective To evaluate the effect of optimizing the management measures of cataract ambulatory surgery. Methods The patients who underwent cataract phacoemulsification combined with intraocular lens implantation in the Ambulatory Surgery Center of East District of Beijing Tongren Hospital affiliated to Capital Medical University were selected. Patients between January and December 2021 (after the optimization of ambulatory surgery process) were included, and patients between January and December 2020 (before the optimization of ambulatory surgery process) were included as control. The three factors of age, gender and surgical eye type were used as predictive variables for propensity score matching. The proportion of patients who completed the surgery according to the scheduled time, the proportion of eye drops used according to the doctor’s instructions and the number of hospital visits before and after the optimization of the ambulatory surgery process were compared with the patients who successfully matched. Results A total of 28306 patients were included, including 13284 before and 15022 after process optimization. There were 13467 males and 14839 females, with a median age of 70 (60, 78) years. There was no statistically significant difference in the age of patients before the process optimization (P>0.05), but there was statistically significant difference in gender and surgical eye (P<0.05). After the propensity score matching, a total of 12932 pairs of patients were matched successfully. After the propensity score matching, there was no statistically significant difference between the two groups in age, gender and surgical eye (P>0.05). After the process optimization, the proportion of patients who completed surgery on schedule (98.8% vs. 93.3%) and used eyedrops according to the doctor’s instructions (97.4% vs. 93.0%) were higher than that before the process optimization, and the proportion of patients who came to hospital more than 3 times (0.7% vs. 1.9%) was lower than that before the process optimization (P<0.05). Conclusion The optimized ambulatory surgery process can help patients complete the surgery according to the scheduled time and use eye medication according to the doctor’s instructions, and can reduce the number of patients coming to the hospital.
ObjectiveTo evaluate tip location of peripherally inserted central catheter (PICC) by using the carina as evaluation mark, so as to provide clinical references for blinding manipulation of PICC. MethodsA retrospective analysis about the effects of tip location of PICC by using the carina as evaluation mark on inpatients with malignant tumour was conducted in tumour centre of the West China Hospital from March 2013 to April 2014. Chest X radiography was applied to evaluate the tip location of PICC. Statistical analysis was performed using SPSS software (version 19.0) and the outcomes were presented by frequency and percentage. ResultsA total of 612 cases of blinding manipulation of PICC were evaluated, of which, 502 (82%) were located in the central venous and 110 (18%) were located in the noncentral venous. The numbers of those with the catheter tips located in the non-central venous were 38 in the right atrium, 33 in the internal jugular vein, 24 in the innominate vein, 8 in the axillary vein, and 7 in the subclavian vein, respectively. ConclusionThe effects of blinding manipulation of peripherally-inserted central catheter by using the carina as evaluation mark are relatively good. But it still needs to be improved.
Objective Healthcare risk has become a popular topic in medical research field all over the world. With the increasing of healthcare safety adverse events and medical conflict cases every year, publications about this topic have been rapidly increased in China, but research on the essentials and categories of healthcare risk are still lacked. This paper aims to summarize the essentials and categories of healthcare risk research systematically through analyzing domestic studies in healthcare risk over the past decade, so as to provide decision-making evidences and policy suggenstions on healthcare risk management in China. Methods We searched the China Academic Journal Network Publishing Database of CNKI (1999-2010), in which those papers’ title containing the phrase of healthcare risk had been brought into our research, then we summerized the domestic concepts and contents in healthcare risk by essential analysis and frequency statistics. Results 255 papers were included, including 26 Class B papers (accounting for 10.20%), six Class C papers (accounting for 2.35%) and 223 Class D papers (accounting for 87.45%); but there were only 79 papers (accounting for 35.11% ) which had a clear definition of healthcare risk. From these papers, we found the essentials of healthcare risk included object, consequences, process and causes, and the research category of healthcare risk included prevention and treatment of risk, causes and effects of factors, risk sharing mechanisms, healthcare risk of special diseases, concept and characteristics, education and training, management method, risks of medical appliances, the species and object of healthcare risk, the category of healthcare risk, the research methods of healthcare risk etc. Conclusion At present, the study of healthcare risk focuses on the risk control and how to release risks from the results, besides, systematic studies are lacked, and the unified definition of healthcare risk has not been formed, Therefore, we recommend that systematic study of the healthcare risks should be strengthened, the definition of healthcare risk should be unified, and the early warning, monitoring, evaluating, and identifying methods should be positively developed, providing decision-making evidences to establish healthcare risk management system in China.
Objective To prepare cationic Vancomycin hydrochloride multivesicular liposome (MVL) and to inspect its quality. Methods Cationic Vancomycin hydrochloride MVLs were prepared by double emulsion method, and the storing solution of Vancomycin was prepared. The analysis method of Vancomycin in vitro was established; the specificity, precision, and resorption rate were estimated. Reverse phase high performance liquid chromatography (RP-HPLC) was used to determine the concentration of Vancomycin, encapsulation efficiency, and release characteristics in vitro. The formulation and pharmaceutical process were optimized by single factor experiments and orthogonal experimental design with the factor of encapsulation efficiency as the criteria. The liposome morphology was observed by optical microscopy and transmission electron microscopy. The particle size and Zeta potential were determined by Malvern instrument. The stability was analyzed by dynamic analysis. Results An RP-HPLC method was established for the assay of Vancomycin. The analysis method was precise, simple, and reliable for the quality control of Vancomycin. Vancomycin hydrochloride MVLs were round and well-distributed. The average particle size and the encapsulation efficiency were 3.3 μm and 24.9%, respectively. Zeta potential was 24.53 mV, and 90.5% of Vancomycin hydrochloride was released after 264 hours in normal saline under 37℃. Cationic Vancomycin MVLs were stored for 1 month at 4 ℃, which mantained good stability. Conclusion Cationic Vancomycin hydrochloride MVLs have good appearance, high encapsulation efficiency, good stability, and significant sustained release properties.
Objective To compare the effectiveness of aspheric intraocular lenses(IOL) versus spherical IOL in the treatment of cataract. Method Randomized controlled trials comparing aspheric IOL with spherical IOL were identified by searching PubMed (2000 to October, 2008), EMbase (2000 to October, 2008), MEDLINE (2000 to October, 2008), and The Cochrane Library (Issue 3, 2008). Two reviewers independently assessed trials for eligibility and quality, as well as the extracted data. Data were synthesized using RevMan software (release 5) provided by the Cochrane Collaboration. Results A total of 14 trials (1383 eyes) were included for systematic review, and 11 out of 14 studies were included in the meta-analysis. Subgroup analyses were used according to different aberrations of aspheric IOL. The results showed a significant difference in the mean best corrected visual acuity at 3 months after cataract surgery in the AcrySof IQ IOL group with WMD -0.02, 95% CI -0.03 to -0.01(Plt;0.0001). It showed a significantly worse difference in the mean of the best corrected visual acuity at 3 months after cataract surgery in the AcrySof IQ IOL group with WMD –0.02, 95%CI –0.03 to –0.01 (Plt;0.000 1). It showed a significant worse in the mean the best corrected visual acuity in the Tecnis Z9000 IOL group with WMD 0.02, 95%CI 0.01 to 0.03 (P=0.002); and there was no significant difference between the two groups with WMD 0.00, 95%CI –0.02 to –0.03 (P=0.71). The results did display markedly less spherical aberration and total higher order aberrations than eyes implanted with the traditional spheric IOL in all subgroups (WMD –0.06, 95%CI –0.07 to –0.06, Plt;0.000 01, WMD –0.06, 95%CI –0.07 to –0.02, Plt;0.000 1, respectively). The majority of studies suggested significant improvement in the aspheric IOL group in contrast sensitivity, especially at mesopic low spatial frequencies, although some trials showed no significant difference. Conclusion The effectiveness of aspheric IOL is superior to spherical IOL during cataract surgery. No differences in aspheric IOL with different aberration are found in this series, and further study is required.
【Abstract】Objective To investigate the diagnostic and therapeutic value of selective angiography and embolization treatment in massive biliary hemorrhage in emergency. Methods Eleven cases of massive biliary bleeding were selectively angiographied and embolized with thread segment and gelatin sponge grain by Seldinger technique in emergency. Results Massive biliary bleeding in this series included bleeding after biliary surgery (7), post-laparoscopic cholecystectomy bleeding (1), bleeding from traumatic injury to liver (1), and rupture of huge hepatic cancer (2). There were 9 cases with abnormal angiographic findings including contrast medium leakage (9), tumor vessels and tumor tissue staining (2) and signs of pseudo-aneurysm formation (4). All the bleedings were stopped after emergency intervention but 2 cases with recurrent massive bleeding after 48 hours. Conclusion Emergency trans-catheter embolization treatment is an effective and safe method for biliary massive hemorrhage. It is also important to keep the biliary tract unobstructed.