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Objective:To investigate the diagnostic and diagnostic values of plasma microRNA-21-3p (miR-21-3p) and miR-551-5p expression in patients with acute pancreatitis (AP).Methods:A prospective observational study was conducted. AP patients admitted to the Third People's Hospital of Hainan Province from January 1st 2017 to December 31st 2019 were enrolled. The patients were divided into mild acute pancreatitis (MAP) group, moderate severe acute pancreatitis (MSAP) group and severe acute pancreatitis (SAP) group according to their severity. Fasting venous blood was collected from all subjects the day after admission, and real-time quantitative polymerase chain reaction (PCR) as used to detect the expression levels of plasma miR-21-3p and miR-551-5p. Rehabilitation, discharge or death were end points of study. In addition, 50 healthy people in the same period were selected as the control group. The receiver operating characteristic (ROC) curve was used to analyze the value of the expression levels of plasma miR-21-3p and miR-551-5p for the diagnosis and prognosis of SAP. Pearson correlation was used to analyze the relationship between the expressions of miR-21-3p and miR-551-5p in SAP patients.Results:A total of 164 AP patients were enrolled, including 72 MAP patients, 47 MSAP patients and 45 SAP patients. Among the SAP patients, 27 cases survived and 18 cases died. There were no deaths in MAP group and MSAP group. The levels of plasma miR-21-3p and miR-551-5p in AP group were significantly higher than those in control group [miR-21-3p (2 -ΔΔCt): 2.17±0.90 vs. 0.65±0.12, miR-551-5p (2 -ΔΔCt): 1.80±0.73 vs. 0.42±0.08, both P < 0.01]. The expression levels of plasma miR-21-3p and miR-551-5p in AP patients increased gradually with the aggravation of the disease ( F values were 11.635, 10.204 respectively, both P < 0.01), and the expression levels of plasma miR-21-3p and miR-551-5p in SAP group were significantly higher than those in MSAP group and MAP group [miR-21-3p (2 -ΔΔCt): 3.16±1.08 vs. 1.85±0.71, 1.70±0.64; miR-551-5p (2 -ΔΔCt): 2.63±0.95 vs. 1.52±0.46, 1.36±0.40; all P < 0.01]. ROC curve analysis showed that the area under the ROC curve (AUC) and 95% confidence interval (95% CI) of the joint diagnosis of SAP with miR-21-3p and miR-551-5p were significantly higher than that of miR-21-3p or miR-551-5p alone [0.898 (0.841-0.960) vs. 0.820 (0.763-0.882), 0.806 (0.748-0.867), Z1 = 4.480, Z2 = 4.916, both P < 0.05], and the sensitivity was 90.7% and the specificity was 85.0%. The expression levels of plasma miR-21-3p and miR-551-5p in the death group of SAP patients were significantly higher than those in the survival group [miR-21-3p (2 -ΔΔCt): 3.75±1.17 vs. 2.66±0.87, miR-551-5p (2 -ΔΔCt): 3.17±1.04 vs. 2.24±0.83, both P < 0.01]. ROC curve analysis showed that the AUC and 95% CI of the combined prediction of death in SAP patients with miR-21-3p and miR-551-5p were significantly higher than that of miR-21-3p or miR-551-5p alone [0.933 (0.875-0.996) vs. 0.856 (0.794-0.917), 0.816 (0.759-0.874), Z1 = 4.395, Z2 = 5.520, both P < 0.05], and the sensitivity was 95.2% and the specificity was 87.5%. Correlation analysis showed a positive correlation between the expression level of plasma miR-21-3p and miR-551-5p in SAP patients ( r = 0.827, P < 0.001). Conclusions:The increased expression of plasma miR-21-3p and miR-551-5p are positively correlated with the severity of AP patients. The combination of the two items has a better value in the diagnosis and prognosis evaluation of SAP.
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Objective:To evaluate the influence of benzalkonium chloride (BAK) in anti-glaucoma medication on intraocular pressure and ocular surface.Methods:A Meta-analysis was performed.The literature about anti-glaucoma medications published in PubMed, EMbase, Cochrane Library from January 1, 2000 to December 31, 2016 were comprehensively searched to identify eligible studies.Randomized controlled trials (RCT) for intraocular pressure and ocular surface disease in anti-glaucoma eye drops with or without BAK in various types of glaucoma or ocular hypertension patients aged >18 years were collected.EXCEL tables were used to count the relevant data and Cochrane collaboration's tool was used to assess the risk of bias system and to evaluate the quality of studies.RevMan 5.3 and Comprehensive Meta Analysis (CMA)V3 were used to perform the Meta-analysis of all RCT.Results:Thirteen RCTs were incorporated.With BAK or without BAK, anti-glaucoma eye drops could efficiently decrease intraocular pressure, meanwhile this comparison had no significant difference ( SMD: -0.00, 95% CI: -0.063-0.063, P=0.99, I2=0%). Adverse events including conjunctival hyperemia, dry eye and keratitis between the two groups were analyzed.During the follow-up period, conjunctival hyperemia, dry eye and keratitis were the main adverse reactions.A random effect model was used for conjunctival hyperemia Meta-analysis, and the fixed effect model was used for dry eye and keratitis Meta-analysis.The combined effect values of conjunctival hyperemia, keratitis and dry eye were OR=1.163, 95% CI: 0.896-1.509, P=0.257, I2=51.6%; OR=1.15, 95% CI: 0.76-1.76, P=0.50, I2=0%; OR=1.13, 95% CI: 0.79-1.61, P=0.51, I2=40%.Systematic comparison result showed that there were no statistically significant differences in the three adverse reactions between the two groups (all at P>0.05). Conclusions:This Meta-analysis indicates that the reduction in intraocular pressure achieved by BAK-free eye drops is equivalent to the effect of eye drops with BAK, and the short term use of eye drops with BAK doesn't change the incidence of conjunctival hyperemia, keratitis and dry eye.
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Allergic conjunctivitis is one of the most common eye diseases,and its incidence is ever increasing every year.The disease has unbearable ocular symptoms and repeated attacks,which seriously affect the daily life and work of patients.Ophthalmologist has not paid enough attention for the diagnosis and treatment of allergic conjunctivitis.Ophthalmologists can give appropriate laboratory examinations in combination with the medical history and signs of patients with allergic conjunctivitis,so that patients can be fully and clearly diagnosed.While giving appropriate drug treatment,the patients can get appropriate desensitization treatment by defining allergens.Ophthalmologists should take full care of the allergic conjunctivitis,especially for allergen testing and allergen-specific immunotherapy,in their practices.
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Objective To investigate the effects of transient receptor potential melastatin (TRPM) 7 on dexamethasone (Dex)-mediated cytoskeleton remodeling in human trabecular meshwork.Methods Human trabecular meshwork cells (HTMs) were primarily cultured and the cells of generation 3 to 6 were used in this study.The expression of TRPM7 protein in the cells was located using immunofluorescence technology.Dex at the dose of 0.2 mg was added into culture medium for 4 days with the final concentration of 1×10-5,1×10-6 and 1×10-7 mol/L,respectively.Western blot assay was employed to detect the relative expression level of TRPM7 protein.Cultured cells were divided into non-transfected group,siRNA transfected group,TRPM7-siRNA1 transfected group and TRPM7-siRNA2 transfected group,and the expressions of TRPM7 protein and p-cofilin protein in the cells were assayed by Western blot method.Cultured cells were divided into normal control group,Dex-treated group,siRNA transfected group and TRPM7-siRNA transfected group,and the expression of phalloidin (a cytoskeletal protein) and Vinculin (focal adhesion protein) was detected by immunofluorescence staining.In addition,cultured cells were divided into normal control group,Dex-treated group,2-APB (a Ca2+ inhibitor) treated group,ethylene glycol tetraacetic acid (EGTA) (a calcium chelator)-treated group,TRPM7-siRNA transfected group and TRPM7-siRNA+Dex group,and the [Ca2+] i in the cells was observed by Fluo-3AM immunofluorescence staining.Western blot assay was used to detect the expression of p-cofilin in the cells.Results TRPM7 was positively expressed on the cell membrane.The relative expression of TRPM7 was gradually reduced with an increase of Dex dose (F=4.210,P＜0.05),and the expression of TRPM7 was significantly decreased in 1 × 10-5 mol/L Dex group compared with the normal control group (P＜ 0.05).Western blot assay revealed that the relative expression levels of TRPM7 in the TRPM7-siRNA1 and TRPM7-siRNA2 group were significantly lower than those of non-siRNA transfected group and siRNA transfected group (all at P＜0.05).In the Dex-treated group and TRPM7-siRNA transfected group,the cells were enlarged in size with the lessened processes in comparison with the normal control group.Immunofluorescence staining showed that the actin fiber and vinculin increased in the Dex-treated group,and more spread but depolymerized actin fiber was seen in the TRPM7-siRNA transfected group.Compared with the normal control group,the fluorescence intensity of [Ca2×] i was weak in the Dex-treated group and TRPM7-siRNA transfected group.The relative expression levels of p-confilin protein was lower in the TRPM7-siRNA transfected group than that in the siRNA transfected group (0.317 ±0.031 vs.0.092±0.071) (t =5.030,P =0.007).Conclusions Dex induces the downregulation of TRPM7 expression in HTMs.The downregulation of TRPM7 probably participates in Dex-induced cytoskeletal remodeling by causing the depolymerization of actin cytoskeleton and reduction of [Ca2+] i in HTMs.
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Skeleton is one of the most common metastasis sites of malignant tumor.Bone metastases can cause bone pain,pathologic fractures,spinal cord compression and other bone related events,which seriously affect the quality of life of patients.The treatment modalities of bone metastases are diverse.External beam radiotherapy and 89 Sr are commonly used in the treatment of bone metastases,however,a single treatment cannot achieve satisfactory results.This review summarizes the advances of 89Sr combined with external beam radiotherapy for bone metastases.
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Background Blepharitis caused by Demodex infestation is very common in clinical practice.There are various methods mentioned in the study of Demodex infestation in China,but a unified introduction and evaluation of the operating procedures is lacked.A quick and accurate clinical diagnostic method for Demodex infestation needs to be further studied.Objective This study aimed to establish operation procedures for the clinical examination of eyelid Demodex infestation,which were applied to evaluate the conditions of eyelid Demodex infestation in ocular patients with discomfort.Methods One thousand and fifty-two patients with eye dryness,eye itchiness or other symptoms were selected for slit lamp examination and photographing of the eyelid margin area.Three eyelashes with associated scurf from each superior eyelid were plucked out for examination of Demodex under the microscope.Positive findings included observation of Demodex mites or eggs.Their amounts were recorded individually for all eyelash samples.Results A procedure for observing,recording and reporting eyelid Demodex infestations in patients was successfully established.By using this procedure,1 052 patients were investigated for the examination of Demodex infestations.Demodex mites or eggs were found in 582 cases (55.3％).The positive rate of Demodex infestation increased with age,and the population over 60 years group had the highest positive rate,showing a significant difference among the different age groups (x2=10.547,P=0.001).There was no significant difference in positive rate between male patients and female patients (P =0.352).The test turnaround time (TAT) for one examination was (11.4±5.2) seconds.Conclusions The operational procedure for examining the palpebral margin Demodex infestation by the slit-lamp,optical microscope,photographing and laboratory reports is established.It is simple and quick in the appliation for the clinical diagnosis of eyelid Demodex infestation.
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Background Visual adaptive mechanism of mammalian is close responsible for the development of visual cortex.The various genes with different biological functions in different developing stages of visual cortex participate in regulation of visual development.To investigate the differential expression profiles of various genes in different ages of rat cortex can offer basis and evidence for the study of visual development.Objective Present study aimed to investigate the genes that changed continuously in the postnatal developmental process of rat visual cortex by microarray analysis of visual cortex RNA.Methods Sixty clean SD rats were grouped numbered and randomized into the postnatal day 0 group (P0,n =20),before eye opening group (postnatal day 10,P10,n =15),before the critical period of visual cortex growth group (postnatal day 20,P20,n =15) and the end of development of visual cortex group(postnatal day 45,P45,n=15).The rats were sacrificed at corresponding time point respectively,and the fresh visual cortex were obtained for the extraction of total RNA and microarray analysis.Genes exhibiting changes in expression by≥2.0 folds were further confirmed using real-time PCR(RT-PCR).In order to evaluate the association of differential gene expression with growth,the postnatal stages were paired as 36 groups with the 3 pairs for each target gene (P45/P0,P20/P0,P10/P0).Results Microarray analysis showed that the gene with differential ratio ≥ 2.0 folds in rat visual cortex included Akap7,Asam,Casp3,Cxcr4,Egr1,Ennp2,Fabp7,Gpr88,Inpp5d,Rpsa,Stk32c and Vamp1.Real-time PCR verified that 24 genes form 26 probe sets had the same-phase regulating tendency,including 20 up-regulating probe sets and 6 down-regulating probe sets.The homodromous expressing tendency was seen in Akap7,Asam,Casp3,Cxcr4,Egr1,Ennp2,Fabp7,Inpp5d,Rpsa and Vamp1 genes between microarray analysis and RT-PCR.However,reverse expressions were found in the P45/P0 of Gpr88 and Stk32c genes,showing the up-regulation in the microarray analysis and down-regulation in RT-PCR.The concordant rate of gene expression between microarray analysis and RT-PCR was 94.44％.The expressing genes mainly functioned nervous system development,(metal) ion binding/transport,metabolism,regulation of neuronal synaptic plasticity.Conclusions New relevant candidate genes of age-associated rat visual cortex can be identified by microarray analysis,which provide a clue for the research of visual plasticity.
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Objective To investigate the effects of spontaneous agonal respiration on coronary perfusion pressure (CPP) during untreated cardiac arrest (ventricular fibrillation) in swine model.Methods Ten male healthy domestic swines (25.0 ± 1.5) kg were anaesthetised,intubated and mechanically ventilated.The catheterizations were separately inserted into the right atrium and thoracic aorta to monitor aortic pressure (AOP) and right atrial pressure (RAP).A pacing electrode was inserted into the right ventricle to induce ventricular fibrillation (VF).VF was induced by intra-ventricular stimulation withalternating electric current and untreated for 8 minutes.AOP and RAP were recorded until respiratory activity ceased.The CPP before and after agonal respiration was calculated and analyzed by paired-sample T test.Results All animals presented with agonal respiration from 1 to 6 minutes after VF during the first attempt.The CPP was (7.18 ±4.22) mmHg at 1 sec before agonal respiration,(11.78 ±5.16) mmHg at 0 sec after agonal respiration,(8.75 t:4.38) mmHg at 5 sec after agonal respiration and (8.23 ± 4.55)mmHg at 6 sec after agonal respiration.The CPP at 0 sec after agonal respiration was higher than that before agonal respiration (t =-3.140,P =0.012).The CPP at 5 sec after agonal respiration was higher than that at 1 sec before agonal respiration (t =-2.828,P =0.020).There was no difference in CPP between at 6 sec after agonal respiration and at 1 sec before agonal respiration (t =-1.778,P =0.109).Conclusions Agonal respiration accompanies ventricular fibrillation.After agonal respiration,the coronary perfusion pressure is increased for 5 seconds being in favor of cardiaopulmonary resuscitation.
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<p><b>OBJECTIVE</b>To investigate the effect of tetrandrine (Tet) on the expression of bax, bcl-2, and transforming growth factor-β2 (TGF-β2) mRNA in cultured human fibroblasts of Tenon's capsules (TCFS) and explore its possible mechanism.</p><p><b>METHODS</b>The third passage of TCFS cultured in vitro were exposed to 1×10(-5) mol/L Tet for 24 h, and real-time fluorescence quantitative PCR was used to detect the changes in the expressions of bax, bcl-2, and TGF-β2 mRNA.</p><p><b>RESULTS</b>The expression level of bax mRNA was obviously higher, while bcl-2 and TGF-β2 mRNA levels were significantly lower in Tet-treated TCFS than those in the control cells (P<0.05).</p><p><b>CONCLUSION</b>Tet can inhibit the proliferation of TCFS possibly by reducing the expressions of bcl-2 and TGF-β2 mRNA, enhancing the expression of bax mRNA and inducing cell apoptosis, suggesting its potential in preventing fibrous scar formation after glaucoma filtration surgery.</p>
Subject(s)
Humans , Apoptosis , Benzylisoquinolines , Pharmacology , Cell Proliferation , Cells, Cultured , Cicatrix , Metabolism , Fibroblasts , Cell Biology , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Tenon Capsule , Cell Biology , Transforming Growth Factor beta2 , Genetics , Metabolism , bcl-2-Associated X Protein , Genetics , MetabolismABSTRACT
Background Researches have demonstrated that dexamethasone (Dex) can induce the changes of the function and structure of trabecular meshwork cells,and latrunculin A (Lat A) can enhance the outflow of aqueous humour and therefore low the intraocular pressure.Objective The aim of the present study is to investigate the effects of Dex and Lat A on the expression of protein in human trabecular meshwork cells.Methods Human trabecular meshwork cells were primarily cultured in DMEM using expand culture method and the fifth generation of cells were used to this experiment.Dex and/or Lat A were added in medium as 10~(-6)mol/L Dex group(Dex treating for 24 hours),Dex+Lat A group(10~(-6)mol/L Dex+2mmol/L Lat A for 24 hours),Lat A group(2mmol/L Lat A for 24 hours) and DMEM culture group.Two dimensional gel electrophoresis(2 DE) was used to compare the protein expressions among these four groups.Subsequently protein spots with different intensity were selected for mass spectrometry analysis.Results Four gel patterns of two dimensional gel electrophoresis of human trabecular meshwork cells from Dex,Dex+Lat A,Lat A and control groups were obtained.A good isolated result for majority of proteins in human trabecular meshwork cells was found in all of the four groups.An obvious expression difference of proteins in human trabecular meshwork cells was seen among the different culture conditions.Twenty four kinds proteins were identified by GDPiMALDI TOF MS,including cytoskeleton related proteins,heat shock proteins,redox related proteins,and proteins participating in carbohydrate metabolism.The expressions of aldehyde dehydrogenase(ADLH)and Rab were increased in Lat A group and decreased in Dex group,but HSP27 and hCRMP2 showed the contrary outcome.Conclusion This study construct the pattern of protein expression in human trabecular meshwork cells by using 2 DE.Dex and Lat A impact the protein expressions in human trabecular meshwork cells.
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Background Elevated intraocular pressure leads to the loss of retinal ganglion cells and vigorous reaction of retinal glial cells.The expression of nestin in retinal glial cells secondary to hypertention and its significance are unclear.ObjectiveThis study aim to investigate the expression of nestin in retinal glial cells (RGCs) in ocular hypertention rats.Methods The ocular hypertention models were established by cauterizing the limbus-draining veins in the right eyes of 42 SD rats,and a conjunctival incision in the left eyes of the rats served as the sham group.The intraocular pressure (IOP) was measured with the Tono-Pen XL tonometer.The number of RGCs in the rats with ocular hypertention was counted.The expression of the nestin protein in RGCs was semi-quantitatively analyzed using Western by immunochemistry.Double immunofluorescence was carried out to evaluate the the confocal laser scaning microscope.Results Significant differences were found in the IOP between the model group and the sham group at various time points (P＜0.05).In 1 week to 3 weeks after operation,the number of RGCs significantly declined in the model group compared with the sham group (P＜0.05).Immunochemistry showed that from 2 hours through 1 week after operation,the expression of nestin was gradually enhanced in the model group in comparison with the sham group.Western blot revealed that the expression of the nestin protein reflected a similar tendency to that of immunofluorescence.The increased introcular pressure as manifested by the induced expression of nestin.Immunoelectron microscopy also confirmed the induced expression of nestin especially at their end-feet suggests a potential neuroprotective mechanism in neuronal degeneration.Nestin may be a useful biomarker for retinal injury study.
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BackgroundExcessive conjunctival scar formation is a main cause of filtering surgical failure in glaucoma.It has been reported that the failure rate of filtering surgery is a tough problem in the research of glaucoma.Research showed that tetrandrine (Tet) suppress the proliferation of cultured human fibroblasts of Tenons capsule (TCFs) in vitro,but its possible mechanism is still unclear up to now.ObjectiveThe aim of this study is to investigate the inducement effects of Tet on the apoptosis of fibroblasts of Tenons capsule and mechanism.MethodsThe Tenon's capsule was obtained from donor eyes of Zhongshan Eye Center Eye Bank for further use.Human Tenon's capsule fibroblasts were cultured by explant culture method in mixed medium in vitro and the third to seventh generations of cells were collected for the experiment.The subcultured cells were identified by morphology observation and Vimentin staining.The apoptosis of cultured fibroblasts in Tet-treated group and control group was studied by using TUNEL (TdT- mediated dUTP nick end labling,TUNEL),flow cytometry (FCM) and transmission electron microscope.The cell number in different cellular cycle was calculated in Ted-treated group and control group.Results The cultured cells reached confluence in two weeks after cultured and presented the spindle or triangle shape with the radial-like or vortexin-like arrangement .The cells showed the positive staining for Vimentin.Apoptosis changes of cultured cell and apoptosis bodies were seen under the transmission electron microscope.Apoptotic cell nuclei were observed in Tet group by TUNEL.FCM result showed that the cells at G_0/G_1 phase decreased by 22.2%,and the cells at S and G_2/M phases increased by 20.53% and 1.6% in Ted-treated group.Significant differences in the numbers of cells in different cellular cycles and cell numbers of apoptosis were found between Tet-treated group and control group(P=0.000).ConclusionTet can inhibit the proliferation of human TCFs by inducing apoptosis in vitro.
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Objective To observe the protection of human lens epithelial cells(HLEC) by different polar alkaloids extracted from Herba Dendrobii(HD).Methods We extacted the Herba Dendrobii powder with ethanol,and then treated the extract with falling-film concentration,acidification,salting out,chloroform extraction,and washing with water.Different polar alkaloids were extracted from HD after the above treatment.The protective effect of HD alkaloids was observed on HLEC,which were cultured with DMEM medium containing 10% fetal calf serum.Ten groups were set up for the experiment: normal control group,model group,high-and low-dose water-soluble alkaloids groups,high-and low-dose fat-soluble alkaloids groups,high-and low-dose low-polar alkaloids group,and high-and low-dose weak-polar alkaloids groups.The high-dose dosage of the alkaloids was 25.0 ?g/L and low-dose dosage was 12.5 ?g/L.Methyl thiazolyl tetrazolium(MTT) assay was used to evaluate the proliferation of HLEC under the different conditions of interventions.Results The single-factor experiments showed that the highest extracting rate of HD alkaloids was obtained under the conditions of extracting the powder with 80% ethanol for 3 times and for 3 hours.The results of protective experiment showed that the proliferation of HLEC in the model group was inhibited by hydrogen peroxide(H2O2),and the inhibitive rate was lower in low-dose fat-soluble alkaloids group than that in the model group(P
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AIM: To identify the differences of gene expression between human age-related cataract and clear lenses. METHODS: The RNA were extracted from human age related cataract and clear lens epithelial cells, labeled with cy3/cy5 as probes, then were hybridized to cDNA chip containing 8 064 genes. The differential expressions of the genes were screened. Furthermore, a primary classification of these genes function was given. The expression levels of the identified genes were further evaluated by real time polymerase chain reaction. RESULTS: 286 genes expression were observed to increase and 438 genes expression were observed to decrease in cataractous lens epithelial cells as compared with normal lens. According to functional analysis, the changed genes in cataract lens are associated with lens structural components, cytoskeleton, cell cycle, apoptosis and stress responses. CONCLUSION: These data suggest that there are differences in gene expression between cataract and clear human lens epithelial cells. The majority of genes changed in cataract exhibited decreased expression. Processes associated with the down-regulated genes may reflect the inability of the lens to maintain its homeostasis and transparency.