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 study the catheter-related infection (CRI) in cancer patients treated with central venous catheterization. Methods A prospective study with 196 cancer patients was conducted to analyze the types of catheter-related infection and pathogen, as well as the relationship between CRI and the following factors: insert location, gender, age, remained time, or bone marrow suppression. Results Of the total 196 cases, 16 cases were diagnosed as CRI and the CRI rate was 8.2%. The types of CRI were five cases of pathogen colonization, four cases of insert location infection and seven cases of catheter-related bloodstream infection. Of the total 244 specimens, 20 were positive including 7 pathogenic bacteria in either Gram positive or Gram negative types, the dominating pathogens were staphylococcus aureus, staphylococcus epidermidis, acinetobacter baumannii and klebsiella pneumoniae. CRI was related to both insert location and age which were both the independent risk factors. Conclusion The concept of prevention should be set up, and the comprehensive measures should be taken to reduce CRI, such as choosing an appropriate insert location and complying with a strict catheter insert standard.
ObjectiveTo investigate the species and resistance phenotypes of the pathogens causing catheter-associated urinary tract infection (CAUTI) in critically ill patients in West China Hospital of Sichuan University, and to provide the basis for the prevention and treatment of this kind of infection. MethodsThe clinical data and findings of the laboratory examination of the patients, who were admitted to intensive care units and suffered from CAUTI in our hospital during January 2012 to December 2014, were retrospectively analyzed. The pathogens isolates from the urine specimens of the patients with CAUTI and their resistance phenotypes were analyzed. ResultsThree hundred and seventy patients suffering from CAUTI were included in this study. Five hundred and seventeen strains of pathogens were isolated from the urine specimens of these patients, including 222 isolates (42.9%) of fungus, 181 isolates (35.0%) of gram negative bacteria, and 114 isolates (22.0%) of gram positive bacteria. In terms of species distribution, Candida albicans (105 isolates, 20.3%), C.glabrata (78 isolates, 15.1%) and C.glabrata (30 isolates, 5.8%) were the predominant fungus. Among the gram negative bacteria, Escherichia coli (81 isolates, 15.7%), Klebsiella pneumoniae (37 isolates, 7.2%), and Acinetobacter calcoaceticus-baumannii complex (23 isolates, 4.4%) were the main species. Enterococcus faecium (79 isolates, 15.3%) and E.faecalis (13 isolates, 2.5%) were the frequently isolated gram positive bacteria. Analysis of the resistance phenotype showed that the resistance rates to itraconazole, voriconazole and fluconazole of Candida spp. were above 10%. Thirty percent of the isolates of E.coli and K.pneumoniae, and 60% of the isolates of A.calcoaceticus-Baumannii complex were resistant to many of the regular antibiotics. Imipenem resistance rate of A.calcoaceticus-Baumannii complex was 60.8%. Sixty percent of the isolates of E.faecium and E.faecalis were resistant to many of the regular antibiotics. The vancomycin-resistant isolates accounted for 16.5% of E.faecium and 31.0% of E.faecalis. ConclusionCandida species are the major pathogens for CAUTI in critically ill patients in our hospital and show the resistance to azoles. We should focus on the drug resistance of gram negative bacteria and gram positive bacteria. The rational use of antibiotics and application of effective infection control measures are important to decrease the CAUTI.
ObjectiveTo investigate the therapeutic evaluation of catheter-directed thrombolysis combined with multiple minimally invasive techniques of endovascular in the treatment of lower extremity atherosclerotic occlusive disease (ASO). MethodsClinical data of 64 ASO patients who were treated in our hospital from June 2011 to October 2014 were analyzed retrospectively. These patients were divided into two groups according to the therapies:33 patients were treated by catheter-directed thrombolysis combined with multiple minimally invasive techniques of endovascular (combination group), and the other 31 patients were only treated by multiple minimally invasive techniques of endovascular (intervention group). Comparison of the clinical indexes was performed between the 2 groups, including vascular patency rate, stent implantation rate, operation time, ankle-brachial index (ABI), saturation of blood oxygen (SO2) of toes, temperature of foot skin, amputation rate, and hospitalization expense. ResultsIn the same group (combination group and intervention group), compared with the time point of before operation, the clinical indexes of the ABI, SO2 of toes, and temperature of foot skin were higher after operation (P<0.05). At the same time point of before or after operation, there were no significant difference between the 2 groups in ABI, SO2 of toes, and temperature of foot skin (P>0.05). The vascular patency rate of combination group was higher than that of intervention group[97.0% (32/33) vs. 83.9% (26/31)], P<0.05. But the stent implantation rate[18.2% (6/33) vs. 64.5% (20/31)], amputation rate[3.0% (1/33) vs. 16.1% (5/31)], operative time[(2.0±0.5) h vs. (4.0±1.1) h], and hospitalization expense of patients who got successful limb salvage[(8 500±1 200) yuan vs. (34 000±2 100) yuan] of combination group were all lower or shorter than those of intervention group (P<0.05). After operation, there were no complication happened in the patients who got successful limb salvage of combination group (n=32) and intervention group (n=26). In postoperative 6 months, the lower extremity arteries in patients who got successful limb salvage of combination group and intervention group were all examined by color ultrasonography, and they were unblocked. The lower extremity arteries of the patients with stent implantation in the 2 groups suffered with no obvious stenosis or occlusion. ConclusionsCatheter-directed thrombolysis combined with multiple minimally invasive techniques of endovascular can become one of the surgical treatment methods for ASO, which characterized by minimally invasive operation, effective treatment, and low cost.
Objiective To observe the changes of angioarchitecture of the optic disc in cats eyes after radial optic neurotomy (RON). Methods Ten healthy adult cats underwent unilateral RON randomly in 10 eyes as an operative group in this trial, and the fellow eyes (10 eyes) were as the control group. The colophony samples of vascular angioarchitecture of 20 eyes were made 90 days after RON and scanning electron microscope was used for the observation. Results In the control group, the blood supply of optic disc mainly came from the short posterior ciliary artery, pial artery, and retinal ciliary artery; the choroidal capillary vessels around the optic disc didnprime;t take part in the blood supply in the anterior area; retinal central vascular system and complete arterial circle around the optic nerves were not seen. In the operative group, a V-shaped defect of the optic disc, retinal blood vessels and choroidal capillary vessels was found at the surgical incision; no injury of retinal blood vessels and choroidal large vascular layer was seen, and neovascularization at the incision were not found. Conclusions RON may cause the vascular defect of the optic disc and the peripheral local tissues. The depth of the incision doesnprime;t reach the Zinn-Hallers circle. (Chin J Ocul Fundus Dis, 2007, 23: 170-172)
Objective To understand the urination of orthopedic patients after the catheter is removed, and to explore the influencing factors of urodynia. Methods Convenient sampling was used to select 160 patients who underwent surgical treatment in the Department of Orthopedics of West China Hospital of Sichuan University from May to July 2020. Questionnaires were used to investigate the patients’ urination, and a logistic regression model was used to analyze the factors affecting the patients’ urodynia. Results A total of 157 patients urinated by themselves after the catheter was removed, with 102 cases of dysuria. Logistic regression results showed that male [odds ratio (OR)=0.282, 95% confidence interval (CI) (0.092, 0.863), P=0.027], Huaxi Emotional-Distress Index score [OR=1.407, 95%CI (1.124, 1.760), P=0.003] and intraoperative fluid infusion [OR=1.001, 95%CI (1.000, 1.002), P=0.014] were the influencing factors of urodynia. Conclusion Most orthopedic patients can urinate on their own after the catheter is removed, but more than half of the patients experience urodynia during the first urination. Gender, mental status and intraoperative fluid infusion are the influencing factors of urodynia after urinary catheter removal after surgery. Clinical nursing staff should identify these influencing factors as soon as possible and carry out targeted interventions to improve the success rate of patients with urination after removing the catheter, reduce the incidence of urodynia, and promote the recovery of patients.
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.
ObjectiveTo investigate the incidence and trendency of healthcare-associated infections (HAIs) in a pediatric intensive care unit (ICU) of a hospital, identify the main objectives of infection control, and formulate corresponding preventive and control measures.MethodsA prospective targeted monitoring method was adopted to investigate HAIs in the pediatric ICU of a hospital from January 2013 to December 2018.ResultsFrom January 2013 to December 2018, the number of target ICU patients was 11 898, the number of patient-days was 55 159; 226 HAIs occurred, the HAI case rate was 1.90%, the incidence of HAI per 1 000 patient-days was 4.10‰, and the adjusted incidence of HAI per 1 000 patient-days was 1.21‰. The main infection site was respiratory tract [83 cases (36.7%)], with ventilator-associated pneumonia in 73 cases (32.3%); secondly, 69 patients (30.5%) had bloodstream infection, among which 48 (21.2%) had non-catheter-related bloodstream infection.ConclusionHospital targeted monitoring is helpful to grasp the situation and trend of HAIs, define the main target of infection control, and formulate corresponding preventive and control measures, which can effectively reduce the incidence of HAIs.