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.
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.
Objective To observe the effect of cationic liposomal ceftazidime (CLC) combined with nano-hydroxyapatite/β-tricalcium phosphate (n-HA/β-TCP) in the treatment of chronic osteomyelitis of rabbits. Methods Thirty healthy New Zealand white rabbits (4-6 months old; weighing, 2-3 kg) were selected to prepare the chronic osteomyelitis models. After 4 weeks, the gross observation, X-ray examination, and bacteriological and histopathological examinations were done; the models were made successfully in 27 rabbits. Of 27 rabbits, 24 were randomly divided into 4 groups (n=6): only debridement was performed in group A; ceftazidime was given (90 mg/kg), twice a day for 8 weeks after debridement in group B; ceftazidime and n-HA/β-TC were implanted after debridement in group C; and CLC and n-HA/β-TCP were implanted after debridement in group D. Before and after treatments, X-ray examination was done, and Norden score was recorded. At 8 weeks after treatment, the specimens were harvested for gross observation and for gross bone pathological score (GBPS) using Rissing standard; half of the specimens was used for histological observation and Smeltzer scoring, the other half for bacteriological examination and calculation of the positive rate of bacteria culture. Results At 8 weeks after treatment, Norden score of group D was significantly lower than that of groups A, B, and C (P lt; 0.05), but no significant difference was found among groups A, B, and C (P gt; 0.05). At 8 weeks after treatment, sinus healed in groups C and D, but sinus was observed in groups A and B; the GBPS scores of groups C and D were significantly lower than those of groups A and B (P lt; 0.05). The Smeltzer scores of groups C and D were significantly lower than those of groups A and B (P lt; 0.05). The positive rates of bacteria culture of groups C (0) and D (0) were significantly lower than those of group A (25.0%) and group B (16.7%) (P lt; 0.05). Conclusion CLC combined with n-HA/β-TCP has good effect in treating chronic osteomyelitis of rabbits, and it has better effect in treating chronic osteomyelitis of rabbits than ceftazidime with n-HA/β-TCP.
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.
Objective To investigate the expression of cerb B2 and CathepsinD in gastric carcinoma and its correlation with the biological behavior of gastric carcinoma (GC). MethodsThe expression was studied by immunohistochemical technique. The expression of cerb B2 and CathepsinD were analyzed with their relation to histologic types, depth of invasion, growth pattern, lymph node metastasis and prognosis of gastric carcinoma. ResultsThirtynine of the 102 gastric carcinoma specimens (38.24%) were positive for cerb B2 and correlated with depth of invasion (P＜0.05) and lymph node metastasis (P＜0.05); eightythree of the 102 gastric carcinoma specimens (81.37%) were positive for CathepsinD and correlated with depth of invasion (P＜0.05), growth pattern (P＜0.05), lymph node metastasis (P＜0.05) and blood vessels cancer embolus (P＜0.05). Prognosis of patients with gastric carcinoma with positive expression of cerb B2 or CathepsinD was poor. The 5year survival rate was significantly lower in gastric carcinoma patients with positive expression of cerb B2 or CathepsinD. Conclusion cerb B2 and CathepsinD are highly related to growth, invasion, metastasis and prognosis of gastric carcinoma.
ObjectiveTo investigate the changes of amount of interstitial cells of Cajal (ICC) and expression of stem cell factor (SCF)/c-Kit in the process of cathartic colon induced by emodin in mice. MethodsA modified cathartic colon mouse model was established. Seventy-two healthy male Kunming (KM) mice were randomly divided into the blank control group and sustained drug delivery group.Morphological changes of colon in mice were observed; frozen section immunofluorescence was used to observed changes of amount of ICC; serum concentrations of SCF were examined by ELISA; Western blot was employed for observation of expression of SCF/c-Kit in colon. ResultsAfter the mice model were completed, the weight of mouse, length and diameter of entire colon were all reduced compared with the blank control group. The amount of ICC appeared to decline in the beginning of the first 6 weeks with emodin used, and significant decreased in 10-12 weeks. The serum concentrations of SCF first began to decline in 4 weeks with emodin used, and significantly decreased in 6 weeks, and continued at a low level after 8 weeks. The expression of c-Kit in colon began to decline in 4 weeks with emodin used and significantly reduced after 8 weeks. Conciusions The amount of ICC appear to slowly decline in the beginning of the first 12 weeks with emodin used, and significant decrease after 12 weeks.The serum concentrations of SCF and expression of c-Kit in colon have the dynamic changes in the meanwhile, and the changes of SCF are earlier than that of c-Kit. The trend of amount ofICC may have a certain relationship with changes of SCF and c-Kit.
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.
Objectives To systematic review the safety of routine preoperative testings versus selectivepreoperative testings for cataract surgery. Methods We searched the Cochrane Central Register of Controlled Trials-Central (which contains the Cochrane Eyes and Vision Group Trials Register) in the Cochrane Library (Issue 3, 2008), MEDLINE (1966 to Oct. 2008), EMBASE (1980 to Oct. 2008), CMB-disk (1979 to Oct. 2008). We also hand-searched related journals. All the search was restricted in English and Chinese. Methodological quality of randomized controlled trials (RCTs) was evaluated by simple evaluate method that recommended by the Cochrane Collaboration. Data extracted by two reviewers with designed extraction form. RevMan 5.0 software was used for data management and analysis. Obtained evidence the safety of routine preoperative testings versus selective preoperative testings.Result Four RCTs involving 20 490 participants required cataract surgery were included by total retrieve and riddling. The result of metaanalysis showed that there was no significant difference between the two groups in the rates of intraoperative systemic events [RR=1.05, 95%CI (0.89, 1.24), P=0.59], postoperative systemic events [RR=0.97, 95%CI (0.80, 1.18), P=0.77], intraoperative ocular events [RR=0.99, 95%CI (0.74, 1.33), P=0.97] and postoperative ocular events [RR=1.11, 95%CI (0.76, 1.60), P=0.59]. Conclusion To compare routine preoperative testings group with selective preoperative testings group, there was no significant difference in the rates of intraoperative systemic events, postoperative systemic events, intraoperative ocular events and postoperative ocular events. However the application of the results to the patients with incapacitating systemic diseases needs further study.
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.
ObjectivesTo systematically review the influence for catheter mechanical dysfunction of different peritoneal dialysis catheterization methods.MethodsPubMed, EMbase, The Cochrane Library, CNKI, WanFang Data and VIP databases were searched to collect randomized controlled trials (RCTs) and cohort studies on comparisons of different peritoneal dialysis catheterization from inception to March 31st, 2018. Two reviewers independently screened literature, extracted data and assessed risk of bias of included studies. Network meta-analysis was then performed by using ADDIS 1.16.6 software.ResultsA total of 33 studies (9 RCTs and 24 cohort studies) involving 3 301 patients were included. Network meta-analysis showed that the incidence of catheter mechanical dysfunctionwas the least and had statistically significant difference compared with that in percutaneous catheterization (OR=3.60; 95%CI, 1.64 and 15.38) and open surgery catheterization (OR=5.86; 95%CI, 2.68 and 14.53). Percutaneous catheterization was superior to open surgery catheterization, but there was no significant difference.ConclusionsLaparoscopic catheterization may be the best technique for catheter insertion in peritoneal dialysis considering catheter dysfunction. Each technology has its own advantages. Choice of insertion method should be based on the characteristics of both the patient and the insertion techniques.