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Objective To analyze the EGFR gene mutations and environmental exposure factors in patients with non-small cell lung cancer (NSCLC) in Bazhong City, and to provide a theoretical basis for the diagnosis and treatment of NSCLC patients. Methods A total of 356 NSCLC patients admitted to Bazhong Hospital from 2019 to 2020 were selected. All patients underwent EGFR gene detection and were divided into mutant group (n=171) and wild-type group (n=185) according to EGFR gene mutation. Environmental exposure data of patients were collected, including smoking status, smoking index, frequent frying of food, etc. Univariate analysis and logistic regression were used to analyze the environmental risk factors of EGFR gene mutations in NSCLC patients. Results A total of 171 EGFR gene mutations were detected in 356 NSCLC patients, and the mutation rate was 48.03%. The mutation rate of EGFR gene in females was significantly higher than that in males (P0.05). The mutation rate of EGFR gene in patients with adenocarcinoma was significantly higher than that in patients without adenocarcinoma (P0.05). Among the 356 NSCLC patients, there were 171 cases with EGFR gene mutations (48.03%), including 335 single mutations, 181 exon 19 mutations, 129 exon 21 L858R mutations, 12 exon 21 L861Q mutations, 8 exon 20 insertion mutations, and 5 Exon 18 mutations. There were 18 cases carrying double mutations and 3 cases carrying triple mutations. There were significant differences between the two groups in smoking status, smoking index, use of coal stove, use of smoke extraction equipment, cooking fumes, fried food intake, and family history of cancer (P<0.05). Non-smoking (OR=3.19), not using smoke exhaust equipment (OR=3.58), and using coal stove (OR=2.19) were the environmental exposure factors of EGFR mutation in NSCLC patients (P<0.05). Conclusion The EGFR gene mutation rate is high in NSCLC patients in Bazhong City, and most of them are female non-smoking patients. EGFR gene detection should be performed in NSCLS patients without smoke exhaust equipment and using coal stoves to improve the detection rate of EGFR mutation.
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Objective:To investigate the effect of epigallocatechin gallate (EGCG) on the activation of human Tenon fibroblasts (HTFs) and its mechanism.Methods:Tenon capsule tissues from nine eyes of nine advanced primary open angle glaucoma patients during trabeculectomy were obtained for primary cell culture.HTFs harvested were identified by immunofluorescence staining for vimentin and keratin.Cells at passage 4-6 were used for experiment.Viability of HTFs treated with EGCG at 0, 10, 20, 30, 40, 50, 60, 70 and 80 μmol/L was detected by cell counting kit-8 (CCK-8) assay.The cells were divided into blank control group, transforming growth factor (TGF)-β1-induced group, and EGCG-treated group, which were cultured in normal medium, medium containing 10 ng/ml TGF-β, medium containing 10 ng/ml TGF-β+ 50 μmol/L EGCG, respectively.The proliferation rate of HTFs was detected by BrdU labeling assay.Cell migration was observed by scratch wound healing assay.The expression of α-smooth muscle actin (α-SMA) was measured by immunofluorescence staining.The protein relative expression levels of Smad2/3, phosphoinositide-3-kinase (PI3K), protein kinase B (Akt) as well as the phosphorylated Smad2/3 (p-Smad2/3) and phosphorylated Akt (p-Akt) were measured by western blot.This study was approved by the Ethics Committee of Guangdong Provincial People's Hospital (NO.GDREC2019331H[R1]).Results:The HTFs harvested had spindle shape, grew regularly and were vimentin-positive.CCK-8 assay showed that there was no significant difference in the variability of HTFs treated with EGCG at 10, 20, 30, 40 and 50 μmol/L in comparison with 0 μmol/L EGCG treatment (all at P<0.05). BrdU labeling assay showed that cell proliferation in the TGF-β1-induced group was (66.37±12.65)%, which was significantly higher than (14.75±12.33)% in EGCG-treated group ( P<0.05). Three days after scratch, the relative scratch area in the TGF-β1-induced group was (47.33±12.22)%, which was significantly lower than (92.67±4.04)% in the EGCG-treated group ( P<0.05). Immunofluorescence assay showed that α-SMA fluorescence was significantly enhanced in the TGF-β1-induced group in comparison with the blank control group, which was reduced to blank control group level in EGCG-treated group.Western blot analysis showed that there were significant differences in the relative expression levels of p-Smad2/3, PI3K and p-Akt protein among the various groups ( F=58.820, 121.153, 69.289; all at P<0.001). The relative expressions of p-Smad2/3, PI3K and p-Akt in the TGF-β1-induced group were significantly higher than those in the blank control group, 10 μmol/L and 50 μmol/L EGCG-treated groups (all at P<0.05). Conclusions:EGCG can suppress TGF-β1-induced HTFs activation through Smad and PI3K/Akt signaling pathways.
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Brucellosis is a kind of animal epidemic disease that can be transmitted to human beings through skin, mucous membrane, digestive tract, respiratory tract and other ways. In recent years, the incidence of brucellosis has increased. Its pathogenesis is relatively complicated. In addition to bacteria, toxins and other factors, genetic susceptibility has gradually attracted the attention of scholars. In this paper, we summarized the previous reports and reviewed the relationship between interleukin gene polymorphism and susceptibility to brucellosis.
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Objective:To investigate the relationship between the degree of basilar artery stenosis and the short-term prognosis in patients with isolated pontine infarction.Methods:One hundred and thirty-seven patients with isolated pontine infarction within 1 month after symptom onset admitted to our hospital from April 2016 to April 2018 were consecutively included.Based on modified Rankin scale(mRS)socres, patients were divided into the good outcome group(mRS score≤2)and the poor outcome group(mRS score>2). Venous blood samples were taken for biochemical testing on admission or the next day.Baseline National Institutes of Health Stroke Scale(NIHSS)scores and demographic data were recorded and compared between the two groups.The degree of basilar artery stenosis was assessed by magnetic resonance angiography(MRA), and subjects were divided into the non-stenosis, mild stenosis, middle stenosis and severe stenosis subgroups.Results:There were 108 patients in the good outcome group and 29 in the poor outcome group.The baseline NIHSS score(2.71±0.22 vs.7.10±0.59, t=6.99, P<0.01)and total cholesterol[(4.29±0.101)mmol/L vs.(4.76±0.17)mmol/L, t=2.21, P=0.03]were lower in the good outcome group than in the poor outcome group.The proportion of patients without stenosis was higher(76 or 70.4% vs.5 or 17.2%, χ2=26.70, P<0.01)and the proportion of patients with severe stenosis were lower(4 or 3.7% vs.7 or 24.1%, P=0.002)in the good outcome group than in the poor outcome group.Binary logistics regression analysis showed that baseline NIHSS score( OR=1.658, 95% CI: 1.327-2.071, P=0.000)and degree of basilar artery stenosis( OR=2.071, 95% CI: 1.159-3.701, P=0.014)were risk factors for the short-term prognosis. Conclusions:The degree of basilar artery stenosis is a risk factor for the short-term prognosis in patients with isolated pontine infarction, and patients with severer stenosis will have a poorer prognosis.
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Objective:To investigate the influence of bundled ligation of the pancreatic stump on postoperative pancreatic fistula (POPF) of distal pancreatectomy (DP).Methods:The retrospective case-control study was conducted. The clinical data of 60 patients with diseases in pancreatic body and tail who underwent DP in the Affiliated Hospital of Inner Mongolia Medical University from January 2011 to August 2018 were collected. There were 24 males and 36 females, aged from 19 to 68 years, with a median age of 45 years. Of the 60 patients, 36 cases undergoing dissection of pancreas with Endo-GIA stapler were allocated into non-bundled group, and 24 cases undergoing bundled ligation of the pancreatic stump with No.10 or No.7 suture at the site over 1 cm of the resection site before dissection of pancreas were allocated into bundled group. Observation indicators: (1) postoperative situations; (2) analysis of risk factors for POPF of DP. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test or ANOVA test. Measurement data with skewed distribution were represented as M (range).Count data were described as absolute numbers, and comparison between groups was analyzed using the chi-square test. Univariate analysis was conducted using the chi-square test and multivariate analysis was conducted using the Logistic regression model. Results:(1) Postoperative situations: the amylase concentration, cases with biochemical fistula, cases with grade B pancreatic fistula, cases with complications, time to extubation, duration of postoperative hospital stay, total hospital expenses were (2 629±592)U/L, 14, 5, 7, (11.9±0.7)days, (13.6±0.7)days, (49 430±1 626)yuan in non-bundled group and (683±312)U/L, 3, 1, 2, (9.7±0.6)days, (11.3±0.5)days, (44 767±1 163)yuan in bundled group, respectively. There were significant differences in the amylase concentration, cases with biochemical fistula, time to extubation, duration of hospital stay, total hospital expenses between the two groups ( t=2.528, χ2=1.512, t=2.341, 2.311, 2.111, P<0.05), and there was no significant difference in the cases with grade B pancreatic fistula or cases with complications between the two groups ( χ2=1.512, 1.394, P>0.05). (2) Analysis of risk factors for POPF of DP. Results of univariate analysis showed that tumor diameter and bundled ligation of the pancreatic stump were related factors of patients undergoing pancreatic fistula after DP ( χ2=4.462, 5.061, P<0.05). Results of multivariate analysis showed that bundled ligation of the pancreatic stump was an independent influencing factor of patients undergoing pancreatic fistula after DP ( odds ratio=0.187, 95% confidence interval as 0.037-0.954, P<0.05). Conclusions:Bundled ligation of the pancreatic stump was an independent influencing factor of patients undergoing pancreatic fistula after DP. Bundled ligation of the pancreatic stump can effectively reduce the incidence of POPF, especially biochemical fistula, the time to extubation, duration of postoperative hospital stay and total hospital expenses, and promote patient recovery after DP.
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Brucellosis is an zoonosis, humans can be infected by Brucella through skin, mucous membrane, digestive tract, respiratory tract and so on. In the past, most of them recorded the infection of brucellosis from animal to human, but rarely reported the transmission of brucellosis between humans. This paper retrospectively reviews previous reports describing the spread of brucellosis between humans.
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OBJECTIVE@#To investigate the effect of β-arrestin1 overexpression on tumor progression in a NCG mouse model bearing T-cell acute lymphocytic leukemia (T-ALL) Molt-4 cell xenograft.@*METHODS@#Molt-4 cells were tagged with firefly-luciferase (F-Luc) by lentiviral infection, and fluorescence intensity of the cells was detected using a luminescence detector. Molt-4 cell lines with β-arrestin1 overexpression or knockdown were constructed by lentivirus infection and injected the tail vein in sub-lethal irradiated NCG mice. Body weight changes and survival time of the xenografted mice were observed, and the progression of T-ALL in the mice was evaluated using an fluorescence imaging system. Sixteen days after xenografting, the mice were euthanatized and tumor cell infiltration was observed in the slices of the liver and spleen.@*RESULTS@#We successfully tagged Molt-4 cells with F-Luc and overexpressed or knocked down β-arrestin1 in the tagged cells. Bioluminescent imaging showed obvious luminescence catalyzed by F-Luc in Molt-4 cells. After injection of Molt-4-Luc cells into irradiated NCG mice, a gradual enhancement of luminescence in the xenografted mice was observed over time, while the body weight of the mice decreased. Compared with the control mice, the mice xenografted with β-arrestin1-overexpressing Molt-4 cells had significantly prolonged survival time ( < 0.001), while the survival time of the mice xenografted with Molt-4 cells with β- arrestin1 knockdown was significantly shortened ( < 0.001). Histological examination revealed fewer infiltrating tumor cells in the liver and spleen of the mice xenografted with β-arrestin1-overexpressing Molt-4 cells in comparison with the mice bearing parental Molt-4 cell xenografts.@*CONCLUSIONS@#β-arrestin1 overexpression suppresses tumor progression in mice bearing Molt-4 cell xenograft.
Subject(s)
Animals , Humans , Mice , Disease Progression , Heterografts , T-Lymphocytes , Transplantation, Heterologous , beta-Arrestin 1ABSTRACT
Objective@#To monitor the WT1 mRNA level and its dynamic changes in patients with myelodysplastic syndromes (MDS) after hypomethylating agents (HMA) , as well as to assess the significance of WT1 mRNA levels and its dynamic changes in evaluating the efficacy of HMA and distinguishing the disease status of heterogeneous patients with stable disease (SD) .@*Methods@#Bone marrow or peripheral blood samples of 56 patients with MDS who underwent hypomethylating agents (≥4 cycles) from November 2009 to March 2018 were tested by real-time quantitative polymerase chain reaction (PCR) to detect the expression of WT1 mRNA, and to observe the correlation between the dynamic changes of WT1 mRNA expression and clinical efficacy and prognosis of patients.@*Results@#WT1 mRNA expression levels of MDS patients decreased significantly after 3 cycles of hypomethylating agent treatment. Besides, the WT1 mRNA expression levels of patients increased significantly after diseases progression. According to the dynamic changes of WT1 mRNA expression levels during SD, 45 cases could be further divided into increased group and non-increased group. In those SD patients with increased WT1 mRNA expression level, the ratio of suffering disease progression or transformation to AML was 95.65% (22/23) , whereas the ratio turned to be 9.09% (2/22) for the non-increased group (χ2=33.852, P<0.001) . Compared with those SD patients reporting no increase in WT1 mRNA expression level, the overall survival[17 (95%CI 11-23) months vs not reached, P<0.001] and progression-free survival [13 (95%CI 8-18) months vs not reached, P<0.001] of those SD patients reporting increase in WT1 mRNA expression level were significantly shorter.@*Conclusion@#WT1 mRNA expression level is a useful indicator to assess the efficacy of hypomethylating agents in MDS patients. Especially in patients with SD, detection of the changes in WT1 mRNA expression level is able to predict disease progression and help to make clinical decision.
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Objective To investigate the correlation between serum uric acid level and short-term outcome of acute isolated pontine infarction. Methods From April 2016 to April 2018, consecutive patients with acute isolated pontine infarction admitted to the Department of Neurology, Kaifeng Central Hospital were enrolled. The baseline clinical data were collected. Fasting venous blood was collected on the day of admission or the morning of the next day for blood biochemical tests. The National Institutes of Health Stroke Scale (NIHSS) was used to assess the severity of neurological deficit. According to the modified Rankin Scale score at discharge or 14 dafter onset, the patients were divided into good outcome group (≤2) and poor outcome group ( > 2 ). Multivariate logistic regression analysis was used to determine the independent risk factors for short-term poor outcome. Results A total of 137 patients were enrolled in the study, 108 (78.8% ) had a good outcome, and 29 (21.2% ) had a poor outcome. The baseline NIHSS score (median [ interquartile range]: 2.5 [1.0-4.0] vs. 8.5 [5.5-10.0 ]; Z= 6.092, P< 0.001 ) and total cholesterol levels (4.290 ± 0.101 mmol/L vs. 4.763 ± 0.171 mmol/L; t=2.214, P=0.028] in the good outcome group were significantly lower than those in the poor outcome group, while serum uric acid level (329.769 ± 8.122μmol/L vs. 257.103 ± 14.290μmol/L; t=4.190, P<0.001) was significantly higher than that in the poor outcome group. Multivariate logistic regression analysis showed that high serum uric acid levels were independently associated with short-term good outcomes in patients with isolated pontine infarction (odds ratio [ OR] 0.377, 95% confidence interval [ CI] 0.203-0.702; P=0.002), while high NIHSS score (OR 1.762, 95% CI 1.375-2.258; P<0.001) and hypertension (OR 5. 353, 95% CI 1.333-21.502; P= 0.018 ) were independently associated with short-term poor outcomes. Conclusion High baseline serum uric acid levels are associated with short-term good outcomes in patients with acute isolated pontine infarction.
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OBJECTIVE@#To establish a stable HEK293T cell line with c.392G>T (p.131G>V) mutation site knockout in gene using CRISPR/Cas9 technique.@*METHODS@#We designed 4 pairs of small guide RNA (sgRNA) for c.392G>T(p.131G>V) mutation site, and constructed exogenous PX458 plasmids expressing Cas9-sgRNA. The plasmids were transfected into HEK293T cells, and the cells expressing GFP fluorescent protein were separated by flow cytometry for further culture. After verification of the knockout efficiency using T7 endonuclease Ⅰ, the monoclonal cells were screened by limiting dilution and DNA sequencing to confirm the knockout. We detected the expressions of mRNA and protein and examined functional changes of the genetically modified cells.@*RESULTS@#We successfully constructed the Cas9-sgRNA exogenous PX458 plasmid based on the c.392G>T(p.131G>V) mutation site of gene. The editing efficiency of the 4 pairs of sgRNA, as detected by T7E1 enzyme digestion, was 6.74%, 12.36%, 12.54% and 2.94%. Sanger sequencing confirmed that the HEK293T cell line with stable knockout of c.392G>T(p.131G>V) was successfully constructed. The genetically modified cells expressed lower levels of mRNA and protein and showed reduced enzyme activity and proliferative capacity and increased apoptosis in response to vitamin K3 treatment.@*CONCLUSIONS@#We successfully constructed a stable HEK293T cell model with gene c.392G>T(p.131G>V) mutation site knockout to facilitate future study of gene repair.
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Humans , CRISPR-Cas Systems , HEK293 Cells , Mutation , PlasmidsABSTRACT
OBJECTIVE: To evaluate therapeutic efficacy and safety of allopurinol versus (vs.) TCM prescription for gout in the treatment of gout, and to provide evidence-based reference for clinical treatment of gout. METHODS: Retrieved from PubMed, Embase, Medline, The Cochrane library, Chinese Journal Full-text Database (CJFD), China Scientific Journal Database (CSJD) and Wanfang database, randomized controlled trials (RCTs) about therapeutic efficacy (total response rate, level of uric acid) and safety (the occurrence of ADR) of TCM prescription for gout alone or combined with allopuinol (trial group) vs. allopurinol (control group) in the treatment of gout were collected. After data extraction of clinical studies meeting inclusion criteria, Cochrane system evaluator manual 5. 1. 0 was used to evaluate the quality of included studies, and Meta-analysis was conducted by using Rev Man 5. 3 statistical software. RESULTS: A total of 20 RCTs were included, involving 1 961 patients. Results of Meta-analysis showed that total response rate of trial group was significantly higher than that of control group [OR=2. 29, 95％ CI(1. 84, 2. 84), P<0. 001], uric acid level was significantly lower control group [MD= -27. 99,95％ CI(-33. 04, -22. 93),P<0. 001],with statistical significance. The incidence of ADR in trial group was significantly lower than control group, with statistical significance (P<0. 05). CONCLUSIONS: Therapeutic efficacy and safety of TCM prescription are better than those of allopuinol for gout.
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Objective To explore the characteristics of morphology and immunology in acceleration phase and blastic phase of chronic myelogenous leukemia(CML).Methods Seventy-three cases of CML-BP bone marrow specimens were respectively conducted the morphology and related cell chemical dyeing observation for determining the FAB type.Flow cytometry was used to detect series immunological related antigens.Results Among 73 cases of FAB typing,there were 44 cases of CML-AML,21 cases of ALL and 8 cases.21 cases CML-ALL patients In the immunophenotyping by flow cytometry,among 21 cases of CML-ALL,there were 19 cases of B-ALL,2 cases of T-ALL,moreover 12 cases contained myeloid marker.Among 8 cases CML-HAL,the immunophenotypes were 6 cases of B+-My and 2 cases of T+ My.Among 44 cases CML-AML,15 cases contained T cell marker,and 2cases contained B cell marker,other cases had no cross-lineage expression.Among 73 cases of CML-BP,29 cases conducted the flow cytometry detection in the acceleration phase,in which 16 cases urgently changed to AML,and 13 cases to non-AML(9 cases of ALL and 4 cases of HAL).Among non-AML cases,2 cases had the simultaneous existence of myeloid primitive cells and precursor lymphocyte in the acceleration phase and other 9 cases were myeloid primitive cell or accompanied by lymphocyte marker.Conclusion Flow cytometry has a certain implication role for the direction and differentiation diagnosis of CML-BP.
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Objective To investigate the relationship between serum cathepsin S (Cat S) and cystatin C(Cys C) expression levels with carotid arterial plaque property in the patients with ischemic stroke(IS).Methods A total of 336 cases of atherosclerotic cerebral infarction were divided into the stable plaque and unstable plaque group according to the carotid arterial ultrasound.Contemporaneous 114 individuals undergoing healthy physical examination served as the control group.Fasting blood was collected from all subjects entering the groups for detecting serum Cat S and Cys C.Results The serum Cat S and Cys C levels and Cat S/Cys C ratio (Cat S/Cys C) had statistically significant difference between the two groups (P＜0.05),moreover the Cat S level in the unstable plaque group of the patients with cerebral infarction was increased compared with that in the stable plaque group,while serum Cys C level was decreased compared with that in the stable plaque group (P＜0.05).The Cat S levels were (75.34-±-15.45)pg/mL and (60.12±18.53)pg/mL and the Cys C levels were (0.73±0.62)mg/L and (0.93±0.53)mg/L,the Cat S/Cys C ratios were (103.68±2.52) and (64.64± 9.24) respectively,the difference between the two groups had statistical significance (P＜0.05).The Spearman correlation analysis showed that the serum Cat S level and Cat S/Cys C ratio had obviously positive correlation with carotid arterial unstable plaque (r=0.498,P＜0.05;r=0.753,P＜0.01);while the level of serum Cys C was negatively correlated with the unstable plaque (r=-0.213,P＜0.05).Conclusion Serum Cat S/Cys C level has a certain correlation with carotid arterial plaque property in ischemic stroke patients,which may become the serological indicators for predicting carotid arterial plaque.
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Objective To study the effects of PM2.5 on reproductive hormone levels and pregnancy outcome in female rats.Methods Thirty healthy 4-week old female Sprague-Dawley rats were randomly divided into the control group (normal saline), low dose of PM2.5(1.5 mg/kg) group and high dose of PM2.5 (37.5 mg/kg) group.After the blood samples were collected, the animals were exposed to PM2.5for 10 days, and then the rats were mated.On the 19th day of pregnancy, the rats were sacrificed for pregnancy outcome observation and blood samples were collected for hormone test.The blood hormone levels were detected using an ELISA kit.Results The live fetus rates in the control, low dose PM2.5 and high dose PM2.5 groups were 90.77%, 59.49% and 60.27%, respectively (P0.05), the level of FSH in the high dose group was significantly decreased (P<0.05).Conclusions PM2.5may affect pregnancy outcome through influencing the hormone levels.
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Glaucoma is an irreversible blinding disease.The aim of glaucoma therapy is to lower the intraocular pressure (IOP),and then slow down glaucoma progression.Currently,this goal can be achieved by medication,laser treatment and surgery.Selective laser trabeculoplasty (SLT) involves using a 532 nm wave length Nd:YAG laser selectively target cells of the trabecular meshwork without causing thermal or collateral damage to the surroundings structures.The exact mechanism by which SLT lowers IOP remains unknown.It has become gradually one of the most preferred methods for the treatment of open angle glaucoma.SLT seems to have a similar IOP lowering effect with argon trabeculoplasty or monotherapy of topical prostaglandin medication.Mean IOP reduction after SLT was 3.8-8.0 mmHg in 6 months to 1 year.The long-term effect of SLT in the treatment of open angle glaucoma remains to be further studied.The relationship between the range of SLT treatment and its therapeutic effect has not been recognized.SLT is efficacious and safe,also has repeatable and less adverse effects.It can be used as an efficacious primary,adjunctive and replacement therapy for treating open angle glaucoma.In this paper,we reviewed the effects of SLT as an initial and repeat treatment,also the factors that could affect therapeutic effects.
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Background Artisan phakic intraocular lens (pIOL) implantation was demonstrated to be safe and effective for the correction of high myopia in short-term.However,long-term corneal endothelium status after operation should be concerned.Objective This study was to investigate the long-term changes of corneal endothelial cell density (ECD) and morphology following Artisan pIOL implantation for the correction of high myopia.Methods Fifty-one eyes of 27 patients who received Artisan PIOL implantation for high myopia from January 2005 to December 2008 were retrospectively analyzed.The central corneal ECDs were detected by corneal endothelial cell counter before surgery and 6 months and 1 year,2,3,4,5,6 years after surgery,and the coefficient of variation (CV) and standard deviation (SD) of cellular area were evaluated.ECD loss rate with time lapse was calculated ([preoperative ECD-postoperative ECD]/preoperative ECD× 100％).Results Six years after surgery,there were 34 eyes with BCVA ≥ 1.0,14 eyes with BCVA higher than preoperative and 8 eyes with BCVA lower than preoperative.The mean intraocular pressure was (14.23±2.14)mmHg 6 years postoperative.The preoperative mean ECD was (3 184.05±233.55)/mm2,and the ECD was gradually reduced over time with the cell loss 2.34％ in 6 months,5.32％ in 1 year,6.32％ in 2 years,8.06％ in 3 years,12.59％ in 4 years,15.63％ in 5 years and 19.49％ in 6 years after operation.Before surgery,The mean CV of cellular area was 37.17±7.12,and the SD of cellular area was 118.77±21.39,and those 6 years after surgery were 32.24 ±4.62 and 125.60± 18.49,respectively.The mean CV and SD of cellular area were significantly different among different time points (P =0.000,0.036).Eight eyes with localized iris depigmentation,6 eyes with IOL dyssymmetry,3 eyes with transient high intraocular pressure and 2 eyes with Macular hemorrhage were obtained after surgery.Conclusions Corneal endothelial cells appear obvious changes in both number and morphology after Artisan pIOL implantation in high myopia.ECD is gradually reduced with time lapse,and the shape of the cells occur remodeling during 6-year fellow-up duration.Overall,endothelial specular microscopy is mandatory before and after surgery for a long term.
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Background Selective laser trabeculoplasty (SLT) can increase the outflow of aqueous humor and reduce the intraocular pressure of patients with open angle glaucoma,but its mechanism is unknown.To investigate the mechanism of SLT would improve the therapeutic effect of SLT.The aqueous outflow resistance in trabecular meshwork was affected by the extracellular matrix (ECM).Matrix metalloproteinase-3 (MMP-3) and MMP-9 were closely related to ECM degradation in trabecular meshwork.Objective This study was to observe the effects of SLT on the expressions MMP-3 and-9 in human trabecular ceils in vitro.Methods Immortallized human trabecular cells were cultured with pigment particles mixed suspension for 16 hours and incubated overnight.Then the cells were irradiated with Q switch frequency doubling 532 nm Nd:YAG laser to establish SLT-effective cells with the energy of 0.2 mJ,spot diameter of 400 μm and pulse duration of 3 ns.The expressions of MMP-3 mRNA and MMP-9 mRNA in the cells were detected by fluorescence quantitative real time PCR before and 1 hour and 4,8,12,28 hours after exposure of laser.The concentrations of MMP-3 and MMP-9 in the medium were assayed using ELISA 1 hour and 4,8,12,28 hours after exposure of laser and compared between the non-irradiation group and the irradiation group.Results The relative expressing levels of MMP-3 mRNA were 1.00,1.32±0.12,2.08±0.05,2.34±0.04,2.77± 0.05 and 2.49±0.27 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of laser SLT,showing a significant difference among the groups (F =15.966,P=0.007),and relative expressing levels of MMP-3 mRNA were significantly higher in various time points after laser irradiation than those of the non-irradiation group (all at P＜0.05).The relative expressing levels of MMP-9 mRNA were 1.00,0.91 ±0.10,1.27 ± 0.07,1.46±0.07,1.69±0.09 and 0.87±0.09 in the non-irradiation group and irradiation for 1 hour and 4,8,12,28 hours after exposure of SLT,which was considerably different among the groups (F =30.526,P =0.005),and significant increased values were seen in the 4,8 and 12 hours after irradiation compared with the non-irradiation group (all at P＜0.05),with highest expression in the irradiation for 12-hour group.The concentrations of MMP-3 and MMP-9 proteins in medium were significantly increased in various time points after laser exposure in comparison with the control group (all at P＜0.05).Conclusions The expressions of MMP-3 and MMP-9 in human trabercuolar cells upregulate and the secretion ability of human trabecular cells to MMP-3 and MMP-9 proteins improves in early stage of SLT in vitro.However,these procedures gradually diminish with the lapse of time.
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Background Selective laser trabeculoplasty (SLT) plays lowing-intraocular pressure (IOP)effect by irradiating pigmented trabecular cells selectively using 532 nm Nd:YAG laser.However,its affect pattern is not fully known.Objective This study was to establish a human trabecular cells (HTCs) model of SLT effect,and to observe the morphological changes of HTCs after they were exposed to different energies of SLT.Methods Immortalized HTCs were incubated in DMEM/F12 with pigmental granules for 16 hours (overnight) to prepare pigmented trabecular cells.The cells were irradiated with modified laser lens with the spot of 400 μm and emitting duration of 3 ns.The energy scale of 0.2 mJ was set for standardized energy and 0.1 mJ was used as the low energy.The morphology and ultrastructure of the cells were examined under the light microscope and transmission electron microscope 1 hour,4,8,12,24 hours after irradiation.Trypan blue staining and TUNEL fluorescence staining were used to evaluate the death and apoptosis of the cells after laser irradiation.Results The brown pigment particles were seen in cytoplasm around nuclei of trabecular cells after cultured for 16 hours.After laser irradiation,there was no obvious change in the shape of nonpigmented trabcular cells,but a 400 μm spot was found in the pigmented trabecular cells under the light microscope,and the pigmented particles released as the cell disruption.After 0.2 mJ laser irradiation,cell loss zone was seen at the center of the laser spot,and the positive cells for trypan blue and TUNEL staining were found;while the abnormal manifestations were slight after the 0.1 mJ irradiation.With the lapse of the time,the number of positive staining cells was gradually decreased.Twenty-four hours after laser irradiation,proliferative trabecular cells emerged and migrated to the center area of laser spot.Statistical analysis showed that the numbers of the positive cells for trypan blue and TUNEL staining were significantly reduced in the lower energy group in comparison with the standardized energy group at various time points (all at P＜0.001),and those of both energy groups were gradually reduced with lapse of time with the significant differences between any two time points (all at P＜0.01).Conclusions An in vitro laser effect model of HTCs can be established by exposing pigmented HTCs to SLT laser.After exposed to SLT,the gasification,necrosis-like death,apoptosis-like change appear at the laser center.The laser destroyed cells can be replaced by proliferative cells with the lapse of time.Low-energy laser irradiation cause a similar morphological change of the cells to high-energy irradiation,but the number of abnormal cells is much less.
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<p><b>OBJECTIVE</b>To develop an LC-MS method for identification of flavonoids in Chinese hawthorn fruits and an HPLC method for quantification of 5 flavonoids.</p><p><b>METHOD</b>Accurate molecular weights of the target flavonoids were acquired by time-of-flight mass spectrometer (TOF-MS) in both positive and negative ion modes. The flavoniods were then further confirmed by standards. An RP-HPLC method was established to quantitatively analysis five major flavonoids (chlorogenic acid, procyanidin B2, epicatechin, hyperoside and isoquercitrin) in hawthorn fruits from ten different regions.</p><p><b>RESULTS</b>The molecular formulae of 8 components were determined by TOF-MS. In the quantitative method validation, the method is linear over the studied range of 1.8-178.5, 2.2-284.8, 2.7-270. 6,1.5-150.0, 1.5-150.0 mg x L(-1) for chlorogenic acid, procyanidin B2, epicatechin, hyperoside and isoquercitrin, respectively. The correlation coefficient for each analyze was greater than 0.999. The inter-day precision of the analysis was lower than 4%, The RSDs of precision and stability were lower than 5%. The recovery rate was from 98.5% to 102.5%, and RSDs were less than 3.5%.</p><p><b>CONCLUSION</b>The method is simple, specific and reliable, and is suitable for quality control of Chinese hawthorn.</p>
Subject(s)
Chromatography, High Pressure Liquid , Methods , Crataegus , Chemistry , Flavonoids , Fruit , Chemistry , Mass Spectrometry , Quality ControlABSTRACT
Background Toric intraocular lens (Toric IOL) is a well accepted method of correct of corneal astigmatism.Axial misalignment,including the error of intraocular lens alignment during surgery and rotation of Toric IOL,is the main barrier to the precise alignment of Toric intraocular lenses.Therefore,the angle of axial misalignment is a main index to evaluate the effectiveness of Toric IOL implantation.Objective The present study is to describe a method of precisely detecting Toric IOL misalignment using postoperative digital photograph.Methods A measuring template (polar coordinates) was constructed using Adobe Photoshop software.Minimum graduate to detect the Toric axis was 1° (maximum error 0.5°).The clinical data of 24 eyes undergone Toric IOL implantation was retrospectively analyzed.Corneal astigmatism was detected by rotating Scheimpflug camera (Pentacam) or slit scan (Orbscan).Digital retro illumination picture was photographed after mydriasis.The picture was analyzed by our derived template.The steepest corneal axis and the axis of Toric IOL were then determined.The misalignment of Toric IOL was determined as the angle between the steepest corneal axis and the axis of Toric IOL.Misalignment of Toric IOL and its distribution were analyzed by statistical means.Results A method using polar coordinates and digital photo to precisely detect Toric IOL misalignment was derived.Mean Toric IOL misalignment was 10.93±6.90 degrees (0-24 degrees) in the 24 cases.The axial misalignment in all of cases were within 25 degrees,including clockwise in 14 eyes(58.33%),anticlockwise in 10 eyes (41.67%).In this study,the misalignment angle of 4 eyes were within 5 degrees;11 eyes were 5 to 10 degrees;1 eye was 10 to 15 degrees;6 eyes were 15 to 20 degrees;2 eyes were 20 to 25 degrees.Conclusion The method is feasible to the direction of Toric IOL misalignment in clinic.