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.
Objective To investigate the effects of phacoemulsification on macula in diabetics． Methods Thirty eyes of cataract in diabetics were chosen randomly for measurement of the thickness of fovea of retina using OCT before phacoemulsification and 1 month after surgery ． The other eyes in these patients and 30 eyes of cataract in nondiabetic pati ents with phacoemulsification were as control． Results In 30 eyes of diabetics with phacoemulsification, the mean fovea thickness were (148．5plusmn;27．7) mu;m preoperatively and (219．4plusmn;68．23) mu;m postoperatively, and the difference was significant (Plt;0．05)． In 30 eyes of diabetics without surgery, the mean foveal thickness were (147.4plusmn;27.5) mu;m preoperatively and (148.2plusmn;27.3) mu;m postoperatively and the difference was not significant (Pgt;0．05)． In 30 eyes of cataract in nondiabetic patients, the mean fovea thickness were (142．37plusmn;12．7) mu;m preoperatively and (151．9plusmn;23．7) mu;m postoperatively and the difference was not significant (Pgt;0．05)． In 30 eyes of diabetic s with phacoemulsification, 11 eyes had new macula edema after surgery and 3 eye s had significant retinal thickening． In 6 eyes with macular edema before surgery, the macular edema were aggravated in 3 eyes after surgery． The macular stru ctural changes were not found in two control groups． Conclusion The thickness of retina is inreased after phacoemulsification in deabetics,and morbidity and its severity of postopevative macular edema are increas ed as well． (Chin J Ocul Fundus Dis, 2001,17:175-177)
Objective To explore the risk factors of catheter-associated bloodstream infections ( CRBSI) in intensive care unit ( ICU) of primary hospital. Methods A total of 623 patients with central venous catheters were recruited in the study. 60 of themsuffered fromCRBSI served as an observation group and other 563 cases without CRBSI served as control. Univariate analysis was used to scan possible risk factors. Then logistic regression analysis was used to exclude the confounding factors. Results The overall incidence rate of CRBSI was 9. 63% ( 60 /623) . There were significant differences in APACHE score, type of catheter, location of catheter, duration of central venous catheter, intravenous nutrition, use of steroid, times of intubation, urgent intubations, nutritional status, diabetes, and MODS between the two groups. Logistic regression analysis revealed that higher APACHE score, double-lumen catheter, femoral vein catheter, catheter indwelling more than two weeks, intravenous nutrition, intubation more than 2 times, and emergency intubation were risk factors of CRBSI. Conclusions Higher APACHE score, double-lumen catheter, femoral vein catheter, catheter indwelling more than two weeks, intravenous nutrition, intubation more than 2 times, and emergency intubation were major risk factors of CRBSI in ICU of primary hospital.
ObjectivesTo systematically review the efficacy and safety of catheter-directed thrombolysis (CDT) versus anti-coagulation (AC) for deep vein thrombosis (DVT). MethodsWe searched PubMed, EMbase, The Cochrane Library, Web of Science, WanFang Data and CNKI databases to collect randomized clinical trials (RCTs) about CDT versus AC for DVT from inception to March 2018. Two reviewers independently screened literature, extracted data and evaluated the risk of bias of included studies. Then, meta-analysis was performed using RevMan 5.3 software. ResultsA total of 5 RCTs and 989 patients were included. Meta-analysis showed that there was no significant difference between the two group in incidence of post-thrombotic syndrome (RR=0.73, 95%CI 0.49 to 1.09, P=0.13), iliofemoral venous patency rate (RR=2.57, 95%CI 0.59 to 11.24, P=0.21), bleeding (RR=2.03, 95%CI 0.50 to 8.28, P=0.32), severe bleeding (RR=1.77, 95%CI 0.91 to 3.42, P=0.09) and recurrence rate of venous thromboembolism (RR=1.00, 95%CI 0.42 to 2.36, P=0.99). However, the incidence of moderate-severe PTS decreased in CDT group was lower than that in the control group (RR=0.70, 95%CI 0.53 to 0.92, P=0.01). ConclusionsCompared with the control group, catheter-directed thrombolysis does not reduce the incidence of PTS and VTE recurrence rate, cannot improve the long-term patency of the iliofemoral vein, yet can prevent the occurrence of moderate to severe PTS. Due to limited quality and quantity of the included studies, more high quality studies are required to verify above conclusions.
Objective To investigate the species distribution and antibiotic resistance of pathogens fromcatheter-related bloodstream infections ( CRBSI) in intensive care unit( ICU) , to provide evidence for the guidance of clinical rational administration.Methods A retrospective analysis was performed to review the microbiological and susceptibility test data of all CRBSI patients in ICU from January 2009 to December 2011. The patterns of antibiotic resistance among the top seven bacteria were compared. Results 67 cases of CRBSI were detected with 81 strains, including 40 Gram-positive ( G+ ) bacteria( 49.4% ) , 38 Gram-negative( G- ) bacteria ( 46.9% ) , and 3 fungi ( 3.7% ) . The main pathogens causing CRBSI were coagulase negative Staphylococci ( 27 strains, 33.3%) , Acinetobacter baumannii ( 12 strains, 14.8% ) , Klebsiella pneumoniae( 9 strains, 11. 1% ) , Staphylococcus aureus ( 8 strains, 9. 9% ) , Pseudomonas aeruginosa ( 7 strains, 8. 6% ) , Escherichia coli ( 6 strains, 7.4% ) , suggesting that Staphylococcus epidermidis was predominant pathogenic G+ bacteria, and Acinetobacter baumannii was predominant G- bacteria. The antibiotic resistance tests demonstrated that isolated G- bacillus was highly sensitive to carbopenem, while vancomycin-resistant G+ bacteria were not found. Conclusions Within the latest 3 years, the predominant pathogens of CRBSI in ICU are Staphylococcus epidermidis and Acinetobacter baumannii. Acinetobacter baumannii exhibited high drug resistance to all antibiotics.
Objective It is difficult to treat chronic osteomyel itis due to the formation of the Staphylococcus aureus biofilms. Liposomal gentamicin-impregnated allogeneic cortical bone can inhibit the formation of the Staphylococcus aureusbiofilms. To explore the treatment of chronic osteomyel itis of rabbit by l iposomal gentamicin-impregnated allogeneic cortical bone. Methods The l iposomal gentamicin, l iposomal gentamicin-impregnated allogeneic cortical bone and gentamicinimpregnated allogeneic cortical bone were produced. Then the chronic Staphylococcus aureus osteomyel itis models of rabbit were made in left lower l imbs of 40 6-month-old rabbits and the right lower l imbs were used as controls. After 2 weeks, the observations of gross and X-ray were done. Four rabbits died within 10 days after the models were made and other 36 rabbits were devided into 6 groups: group A (no antibiotics), group B (intravenous injection of gentamicin), group C (intravenous injection of l i posomal gentamicin), group D (implantation of gentamicin-impregnated allogeneic cortical bone), group E (implantation of l i posomal gentamicin-impregnated allogeneic cortical bone), and group F (implantation of allogeneic cortical bone). After 2 weeks of treatment, the bacterial culture, X-ray and HE staining were done. Results The chronic Staphylococcus aureus osteomyel itis model of rabbit was made successfully. The X-ray showed dissolution of bone and periosteal reaction in groups A, B, C, and F, and no obvious dissolution of bone and periosteal reaction in groups D and E. The Norden scores were (2.5 ± 0.3), (2.1 ± 0.2), (1.5 ± 0.3), (1.5 ± 0.2), (0.9 ± 0.3), and (2.7 ± 0.3) points in groups A-F, respectively; showing significant differences between group A and groups B-E (P lt; 0.05), between groups B, E, F and other groups (P lt; 0.05). The results of blood and marrow cultures for Staphylococcus aureus were positive in groups A and F, and negative in other 4 groups; the results of bone marrow culture for Staphylococcus aureus were positive in 6 rabbits of group B, 4 rabbits of group C and 3 rabitts of group D; and the results were negative in group E. HE staining showed: in groups A and F, abscess and dead bone formed, and no new bone formation were observed; in groups B and C, different degrees of neutrophil accumulation was seen; in group D, some neutrophil accumulation occurred, and osteoprogenitor cells and osteoclasts were seen around implanted bone; and in group E, no neutrophil accumulation was observed, a lot of granulation tissues formed, and osteoprogenitor cells and osteoclasts were seen around implanted bone. Conclusion Implantation of l iposomal gentamicin-impregnated allogeneic cortical bone has remarkly better effect in treating chronic osteomyel itis than intravenous injection of l iposomal gentamicin and implantation of gentamicin-impregnated allogeneic cortical bone.
ObjectiveTo assess the therapeutic effect of catheter thrombolysis combined with inferior vena cava (IVC) filter placement in treatment of deep venous thrombosis (DVT) complicated with floating IVC thrombus. MethodsThe clinical data of 16 patients with DVT complicated with floating IVC thrombus from July 2013 to November 2014 in this hospital were collected. These patients were treated with the catheter thrombolysis combined with IVC filter placement, the IVC filter was placed via jugular vein, the catheter thrombolysis was performed by the side of the small saphenous vein, the amount of urokinase was (60-80) ×104 U/d. Results①The floating IVC thrombi of 13 patients were disappeared following catheter thrombolysis combined with IVC filter placement therapy, a small amount of visible thrombi were adhered on the recycled IVC filter, the lower limb swelling was relieved, the IVC could effectively open.②The IVC filters of 2 patients could not be recycled due to the adhesion of floating IVC thrombus and lumen of IVC resulting in luminal stenosis.③The floating IVC thrombus of 1 patient was disappeared, the IVC filter could not be recy-cled due to a large of thrombi adhered on the IVC filter. The lower limb swelling was slowly relieved. The complications such as severe pulmonary embolism didn't happen in all the patients during treatment and following-up. ConclusionThe limited data preliminarily shows that it is an effective and safe method by catheter thrombolysis combined with IVC filter placement in treatment of DVT complicated with floating IVC thrombus.
ObjectiveTo observe the clinical manifestations of a Wagner syndrome (WS) family. MethodsA retrospective clinical study. Four patients (the proband, his father, sister, and brother) and one family member (the proband's mother) from a WS family diagnosed by clinical examination in Chengdu Aidi Eye Hospital in June 2023 were included in the study. The proband's medical history was examined in detail, followed by best corrected visual acuity (BCVA), fundus color photography, optical coherence tomography (OCT), and OCT angiography (OCTA). The proband underwent full field electroretinogram (ERG) examination. The proband and his sister and brother underwent blood glucose, blood pressure, hearing, face, joint, exercise and general physical examination at the same time. Peripheral venous blood was collected from the proband and 4 other family members. The proband extracts genomic DNA samples, conducts target region capture, library construction and high-throughput sequencing after qualified quality control. The suspected pathogenic mutation sites were verified by Sanger. According to the selected mutation sites, other family members in this family were co-isolated and verified. The pathogenicity of the mutation site was analyzed using the guidelines of the American College of Medical Genetics and Genomics (ACMG). ResultsProband (Ⅱ-1) was 23 years old female. Both eyes BCVA were 0.1. The waveforms of ERG in both eyes were basically normal, and some amplitudes were reduced. Sister of the proband (Ⅱ-2) was 20 years old. Both eyes BCVA 1.0. Fundus examination showed no obvious abnormality. Brother of the proband (Ⅱ-3) was 19 years old. The left eye underwent pars plana vitrectomy combined with silicone oil filling 2 years ago due to retinal detachment and severe vitreous hyperplasia. BCVA light sensitivity, complicated cataract, and fundus opacity were observed. Right eye BCVA was 0.1. The lenses of the proband and his younger sister and brother were pointed and wedged, and the younger brother was heavier. Vitreous cavity of lens. The retina color of both eyes and the right eye of the younger brother of the protor was dark, with flaky dark areas on the side of the nose and the posterior pole, and the symmetrical retinal veil membrane hyperplasia and pulling on the periphery, showing small retinal splits. The choroidal retina showed focal and segmental symmetrically large atrophy. The optic disc was tilted. By OCT examination, the ellipsoid band was partially missing and broken, and the thickness of the choroid layer was reduced. Retinal cortical atrophy in 1 eye (younger brother of proband). By OCTA examination, the mesovascular layer of choroid was atrophied seriously and the blood density decreased. The results of laboratory and general examination of the three siblings showed no obvious abnormalities. The results of genetic testing showed that the proband, his father (Ⅱ-1), his sister and his brother carried a heterozygous mutation of the VCAN gene c.9264A>G (p.Pro3088=). According to ACMG guidelines, the pathogenicity of this variant was unknown. The mother of proband (Ⅰ-2) was wild type. ConclusionsThe abnormal manifestations of WS eyes are diverse, and both anterior and posterior segments could be involved. The pathogenicity of the heterozygous variation of VCAN gene c.9264A>G (P.RO3088 =) in this family is unknown.
Objective To analysis the effect of lens opacity on the measurement of retinal vessel oxygen saturation. Methods This was a cross sectional study. Forty four eyes of 44 patients with different degrees of lens opacity were enrolled. There were 23 males and 21 females. The patients aged from 48 to 84 years, with the mean age of (71.8±10.3) years. The mean best corrected visual acuity was 0.65±0.22. The mean intraocular pressure was (14.2±4.3) mmHg (1 mmHg=0.133 kPa). The mean equivalent spherical degree was (−0.05±2.10) D. The opitical quality analysis system was applied to measure intraocular objective scattering index (OSI) caused by lens opacity. According to the OSI, the opacity of lens was divided into four groups. Patients with OSI value ＜1.0 was grouped to level 1, which indicated that the lens were basically transparent; patients with OSI value between 1.0 and 3.0 was grouped to level 2, which indicated early cataract; patients with OSI value between 3.0 and 7.0 was grouped to level 3, which indicated progressive cataract; patients with OSI value ＞7.0 was grouped to level 4, which indicated the mature stage of cataract. The retinal oximeter Oxymap T1 was used to capture the fundus images under different wavelengths. Pearson correlation analysis was used to analyze the correlation between retinal oxygen saturation and age, intraocular pressure, equivalent spherical degree and OSI. One way ANOVA was used to analyze the difference of retinal oxygen saturation among groups. Results The mean retinal arterial oxygen saturation, venous oxygen saturation and arteriovenous difference was (90.70±6.46)%, (47.34±13.51)%, (43.36±10.09)%, respectively. The correlations of retinal arterial oxygen saturation, venous oxygen saturation and arteriovenous difference with age, intraocular pressure, equivalent spherical degree was not statistically significant (all P＞0.05). The retinal arterial oxygen saturation and venous oxygen saturation was negatively correlated with OSI (r=−0.462,−0.500; P=0.002, 0.001), the arteriovenous difference and OSI was positively correlated (r=0.373, P=0.013). According to lens opacity, there were 11 eyes in level 1, 9 eyes in level 2, 14 eyes in level 3, 10 eyes in level 4. There were significant differences of retinal artery and venous oxygen saturation among different lens opacity levels (F=5.340, 4.710; P=0.003, 0.007); meanwhile, the arteriovenous difference was not significantly different (F=2.048, P=0.123). The retinal arterial oxygen saturation and venous oxygen saturation was significantly lower in the level 4 lens opacity group than any other three groups (all P＜0.05), but there was no statistically significant difference among level 1 to level 3 lens opacity group. Conclusion The effect of lens opacity of level 1 to level 3 is limited on the measurement of retinal oxygen saturation, but level 4 lens opacity will cause decrease of retinal artery and venous oxygen saturation.