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Diabetes retinopathy (DR) is a blinding ocular complication of diabetes, and its pathological mechanism is complex. The damage to the retinal neurovascular unit (NVU) and the imbalance of its coupling mechanism are important pathological foundations. Autophagy plays an important role in the progression of DR. Oxidative stress, endoplasmic reticulum stress, hypoxia, and competitive endogenous RNA regulatory networks can affect the occurrence of autophagy, and autophagy induced cell death is crucial in NVU dysfunction. Retinal neurocyte are non-renewable cells, and adaptive autophagy targeting neuronal cells may provide a new direction for early vision rescue in patients with DR. It is necessary that exploring the possible autophagy interrelationships between ganglion cells, glial cells, and vascular constituent cells, searching for targeted specific cell autophagy inhibitors or activators, and exploring the impact of autophagy on the NVU complex more comprehensively at the overall level. Adopting different autophagy intervention methods at different stages of DR may be one promising research directions for future DR.
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A novel beta-coronavirus, SARS-CoV-2, emerged in late 2019 and rapidly spread throughout the world, causing the COVID-19 pandemic. However, the origin and direct viral ancestors of SARS-CoV-2 remain elusive. Here, we discovered a new SARS-CoV-2-related virus in Yunnan province, in 2018, provisionally named PrC31, which shares 90.7% and 92.0% nucleotide identities with SARS-CoV-2 and the bat SARSr-CoV ZC45, respectively. Sequence alignment revealed that several genomic regions shared strong identity with SARS-CoV-2, phylogenetic analysis supported that PrC31 shares a common ancestor with SARS-CoV-2. The receptor binding domain of PrC31 showed only 64.2% amino acid identity with SARS-CoV-2. Recombination analysis revealed that PrC31 underwent multiple complex recombination events within the SARS-CoV and SARS-CoV-2 sub-lineages, indicating the evolution of PrC31 from yet-to-be-identified intermediate recombination strains. Combination with previous studies revealed that the beta-CoVs may possess more complicated recombination mechanism. The discovery of PrC31 supports that bats are the natural hosts of SARS-CoV-2.
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Objective:To explore the reliability of Xpert MTB/RIF (Xpert) in the diagnosis of pulmonary tuberculosis and to understand the characteristics of retreated patients, so as to provide clues for the investigation of retreated patients.Methods:905 patients sputum with suspected tuberculosis from July 2019 to December 2019 were collected from the Tuberculosis Department of Changsha Central Hospital. Xpert was used for rapid screening of tuberculosis. MGIT960 and Roche culture method were used for rapid sputum culture and drug sensitivity, respectively. Rifampin (RIF) resistance was compared between Xpert and Roche culture. The gender, age, RIF resistance, occupation and diabetes mellitus of patients with positive Xpert were compared, and the risk factors of RIF resistance were analyzed by binary logistic regression.Results:The total detection rate of Xpert was 28.7%(260/905) in 905 suspected pulmonary tuberculosis patients; RIF resistance rate accounted for 20.8%(54/260) in 260 positive patients, and the main mutation types were probe-E and probe-D. There was no significant difference between traditional drug sensitivity and Xpert method in RIF resistance detection ( P>0.05); There was no significant difference between Xpert method and traditional drug sensitive cases in predicting isoniazid (INH) resistance ( P=0.293); there were significant differences in age ( t=-3.835, P<0.05), gender, RIF resistance rate and proportion of farmers between newly treated and retreated patients (χ 2=5.091, 16.862, 4.808, P<0.05); Binary logistic regression showed that retreatment was a risk factor for RIF resistance ( OR=3.739, P=0.004). Conclusions:Xpert has high clinical value in the diagnosis of pulmonary tuberculosis and RIF resistance; The retreated patients were older, and the proportion of male farmers was larger, with higher resistance rate to RIF.
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Objective To explore the effect of short-term intensive rehabilitation training on respiratory function, motor function and the life quality of patients with obstructive sleep apnea combined with chronic obstruc-tive pulmonary disease ( OSA-COPD) . Methods Fifty-seven patients with OSA-COPD were randomly divided in-to an observation group and a control group. Both groups were treated with non-invasive positive pressure ventilation ( NPPV) , oxygen therapy and a bronchodilator, while the observation group was additionally provided with 8 weeks of intensive lung rehabilitation training, including respiratory function training and limb exercise training. Polysom-nography was used to monitor the apnea hyponea index ( AHI) , the lowest oxygen saturation level during the night ( LowSpO2 ) and the nocturnal oxygen saturation ratio for < 90% of total sleep time ( tst90) . Arterial blood gases, forced vital capacity ( FVC) and forced expiratory volume in one second ( FEV1 ) were measured. The 6-minute walk test (6MWT) and St. George's respiratory questionnaire ( SGRQ) were used to evaluate all the patients before and after the intervention. Results After 8 weeks of treatment, the average AHI, LowSpO2 , TST90 and PaO2 had improved significantly in both groups. There was no significant difference between them. After the treatment the average FVC, FEV1 and 6MWT time of the observation group were significantly better than before the treatment and the significantly better than the control group's averages. After treatment, the average SGRQ score and activity abili-ty score of the observation group were also significantly improved and significantly better than the control group's av-erages. Conclusions NPPV can effectively improve OSA-COPD patients'tolerance of short-term intensive pulmo-nary rehabilitation training. With that assistance, short-term intensive rehabilitation training can promote the recov-ery of respiratory function and motor function, and improve the life quality of patients. Therefore, such therapy is worthy of clinical promotion and application.
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Objective To explore the laboratory culture and identification of Mycobacterium tuberculosis L forms (MTB-L),isolation rate and drug resistance in smear-positive tuberculosis patients,and to improve clinical attention to MTB-L.Methods 222 smear-positive pulmonary tuberculosis patients treated in our hospital from September 2017 to December 2017 were randomly selected for MGIT 960 and 92-3TB-L liquid culture.After MGIT 960 was reported positive,acid-fast staining was performed on the precipitated smears of 92-3TB-L liquid medium for preliminary screening.The suspected L-positive strain culture was transformed into improved TSA-L solid medium to observe the colony characteristics and microscopic characteristics.The properties of the strain were confirmed by acid-fast staining and tuberculosis DNA amplification.Drug susceptibility and mutation sites of drug resistance genes were analyzed in MTB-L.Results Ⅰ-dentification of MTB-L:after the positive strain has been cultured,the colonies have the characteristics of "fried egg sample","particle-like" and " filament-like".MTB confirmed by tubercle DNA amplification experiments.Isolation rate:after cultured by MGIT 960 and Modified 92-3TB-L medium,the positive rate of single bacterial type was 50.90%,(113/222) the positive rate of both bacterial type and L type was 15.32% (34/222),and the positive rate of MTB-L type was 2.25% (5/222).Drug resistance:MTB-L was resistant to Streptomycin,Isoniazid,Rifampin,and Ethanol butylamine.No mutation was found in the drug resistance gene loci.Conclusions Clinical laboratory should routinely develop the culture of L-form of Mycobacterium tuberculosis bacteria,and increase the clinical attention to MTB-L.
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Objective To explore the effect of pulmonary rehabilitation training on the respiratory function,motor function,life quality,survival and complications of patients with non-operative lung cancer.Methods A group of 88 patients with non-operative lung cancer was randomly divided into a training group (n=45) and a control group (n=43).Both groups were given anti-tumor therapy,while the training group was additionally provided with systematic respiratory training,including breathing pattern training,cough and expectoration training,respiratory gymnastics and walking training.Forced vital capacity (FVC) and forced expiratory volume in the first second (FEV1) were measured at the outset and after 8,16 and 24 weeks of the training.The 6-minute walk test (6MWT) was administered along with the QLQ-C30 assessment of the European Organization for Research and Treatment of Cancer.Complications in both groups were also recorded and analyzed.The progression-free survival (PFS) and overall survival (OS) were followed up after the treatment.Results After 8,16 and 24 weeks of the treatment,the average FVC and FEV1 volumes and the 6MWT times of the training group were significantly better than those before treatment and significantly better than the control group averages.Indeed,no significant improvement was observed in the control group's average FVC,FEV1 or 6MWT results.After 24 weeks the treatment group's average scores on the physical function,social function,emotional function,fatigue,nausea and vomiting,pain,dyspnea,insomnia,appetite,constipation,and overall quality of life sub-scales of the QLQ-C30 had all improved significantly more than in the control group.The incidence of pulmonary complications in the control group (26%) was significantly higher than that in the training group.(11%).The median PFS and OS of the training group (14.3 and 27.3 months) were not significantly better than those of the control group,however.Conclusion Respiratory exercise training and aerobic exercise training combined with the anti-tumor therapy,while not prolonging survival,can effectively improve the life quality of patients with non-operative lung cancer,reducing the incidence of complications and promoting the recovery of respiratory function.The combination is worthy of popularization in clinical practice.
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Objective To explore the effects of ApoC3 gene on the severity of hypertriglyceridemiainduced acute pancreatitis (AP).Methods ApoC3 transgenetic mice and C57BL/6J mice AP model was induced by cerulein intraperitoneal injection,and ApoC3 transgenetic mice and C57BL/6J mice injected by normal saline solution in equal volume served as control group.Serum triglyceride and cholesterol were detected,and the pathological changes of the pancreas were observed.RT PCR method was used to examine the changes of the inflammatory factor including IL-1β,IL-6,α-SMA and TNF-α mRNA levels,which reflected the severity of the inflammation.Results Serum triglyceride and cholesterol were higher in ApoC3 transgenetic mice than in C57BL/6J mice [(3.434 ± 0.931) mmol/L vs (0.766 ± 0.120) mmol/L,(2.553 ±0.178) mmol/L vs (1.996 ± 0.080) mmol/L],and the differences were statistically different (P < 0.05).The pathological changes of the pancreas were more severe in ApoC3 transgenetic AP mice than in C57BL/6J AP mice,and the IL-1β,IL-6 and α-SMA mRNA levels in the pancreatic tissue were obviously higher in ApoC3 transgenetic AP mice than in C57BL/6J mice (1.72 ± 0.07vs 0.78 ± 0.09,1.58 ± 0.09vs 0.87 ±0.04,0.83 ± 0.05vs 0.44 ± 0.04),and the differences were statistically significant (P < 0.05),while there was no statistical difference on TNF-αmRNA level (0.70 ± 0.09vs 0.65 ± 0.08,P > 0.05).Conclusions ApoC3 gene could aggravate the severity of the inflammation in hypertriglyceridemia-induced AP.
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Objective To investigate the effect of artesunate (ASN) on the expression of Heme oxygenase-1 (HO-1) in THP-1 cells induced by the early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) antigens of Mycobacterium tuberculosis and to investigate its possible mechanism.Methods THP-1 ceils were cultured in vitro.The effects of ESAT-6 and CFP-10 on cell viability were detected by methyl thiazolyl tetrazolium (MTT) assay.THP-1 cells were pre-treated with or without ASN prior to incubation with or without ESAT-6 and CFP-10,the mRNA expression of HO-1 was detected by real time quantitative polymerase chain reaction (RT-qPCR) and Toll-like receptor 2 (TLR2) level was measured by Western blot.Results MTF assay showed that ESAT-6 and CFP-10 were non-toxic to cells in the range of 0-5 μg/ml.Compared with the control group,5 μg/ml ESAT-6 and 5 μg/ml CFP-10 could significantly increased the mRNA expression of HO-1 (P < 0.05).In addition,20 μg/ml ASN could significantly enhance the mRNA expression of HO-1 induced by ESAT-6 and CFP-10,and inhibit the expression of TLR2 induced by ESAT-6.Conclusions ASN in combination with ESAT-6 or CFP-10,may have potential value in treatment of pathogen-associated inflammatory diseases.
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Objective The establishment of the model of myocardial ischemia-reperfusion injury (MIRI) in SD rats involves lengthy time, severe damage, and death of the rats.We aimed at an efficient and rapid method for establishing an MIRI model in rats to ensure a high success rate of modeling and survival of the animals.Methods Using the trachea intubation-assisted breathing machine, we fully exposed the hearts of 80 adult SD rats (8-10 weeks old and weighing 250-300 mg) by mutilating the 2-3 intercostal muscles.Then, we rapidly positioned and ligated the left anterior descending (LAD) coronary artery for an hour, and established the model of MIRI at 48 hours after releasing the slipknot.We performed electrocardiography (ECG) before, during, and at 1 and 48 hours after operation, ligated the same part again at 48 hours postoperatively, and measured the size of the myocardial ischemia and infarction areas using Evans-TTC double staining.Results The survival rate of the 80 rats was 90% and the success rate of MIRI modeling was 86.25%.After ligation of the LAD coronary artery, ECG showed the ST segment and T wave elevation, followed by gradual decrease of the ST segment and R wave voltage, the myocardium and cardiac apex muscle grey or dark grey below the ligation slipknot.The heart rate myocardial motion amplitude were obviously reduced after ligation.Evans-TTC double staining revealed an evident myocardial infarction area at 48 hours after modeling and the ratio of the ischemia area + infarction area to the total myocardial area of the left ventricle was 0.43 to 0.55 (P>0.05) and that of the infarction area to the ischemia area + infarction area was 0.35 to 0.45 (P>0.05).Conclusion The modified rat model of MIRI can be fast and efficiently established, with a high success rate of modeling and a low mortality of the animals.
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AIM:To explore the effect of Pycnogenol on transforming growth factor-β1 ( TGF-β1)-induced he-patic stellate cell activation .METHODS:Cultured LX-2 cells were treated with 5μg/L TGF-β1 and different concentra-tions (0, 10, 25 and 50 mg/L) of Pycnogenol.The viability of the LX-2 cells under the conditions with or without autoph-agy inhibitor 3-MA and ERK inhibitor PD98059 was determined by MTT assay .The protein levels of α-SMA, ColⅠ, TIMP-1, LC3-Ⅱ/Ⅰ, beclin 1, p-ERK1/2 and ERK1/2 were detected by Western blot .RESULTS:Compared with con-trol group, 5μg/L TGF-β1 treatment elevated the cell viability , and increased the protein levels of α-SMA, ColⅠ, TIMP-1, LC3-Ⅱ/Ⅰ, beclin 1, p-ERK1/2, and ERK1/2 in the LX-2 cells (P<0.05).However, these effects were reversed by Pycnogenol pretreatment in a dose-dependent manner and the inhibitory effect of 50 mg/L Pycnogenol was the most sig-nificant in the LX-2 cells (P<0.05).Furthermore, compared with TGF-β1 group, pretreatment with 50 mg/L Pycnog-enol, 5 mmol/L 3-MA or 20 μmol/L PD98059 downregulated TGF-β1-induced cell viability and the protein levels of α-SMA and LC3-Ⅱ/Ⅰ in the LX-2 cells ( P<0.05 ) .CONCLUSION: Pycnogenol suppresses TGF-β1-induced hepatic stellate cell activation via p-ERK and autophagy inhibition .
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Objective:To investigate the value of the Holter monitoring in silent myocardial ischemia (SMI).Methods: Sixty-eight patients with silent myocardial ischemia had the Holter monitoring.Results:Sixty-eight cases were detected 263 times of the SMI, Among them 05:00-12:00 highest incidence occurs 156 times total, 52.88% of the total number; second highest incidence of the time period of 18:00-22:00, total occurred 97 times, the total number of of 32.88%; 22:00-05:00 totaling occurred 46 times, the total number of 15.59%. There are a total of 187 predisposing factors array times, 63.34% of the total number. Heart rate greater than 100 bpm, SMI total array occurred 175 times, 59.32% of the total number; heart rate>65 bpm and
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Objective To investigate the drug resistance of 1 031 Mycobacterium tuberculosis to rifampicin and isoniazide in the Center Hospital of Changsha from January 1 ,2013 to September 30 ,2014 .Methods A total of 1 031 strains with positive culture result and identified as strains of Mycobacterium tuberculosis were used absolute concentration method to do the conventional drug susceptibility ,and detected rifampicin and isoniazide resistance gene including rpoB ,katG and inhA gene locus mutation by chip technology ,the results of two methods were compared using card square test statistics .Results By gene chip method ,the sensitive strain of rifampicin was 896 ,the drug-resistant strains was 135 ,the sensitive strains of isoniazide was 901 strains ,130 drug-resistant strains .Compared with the absolute concentration method ,resistance chip detection results were consistent with rifampicin resistant strains 1 011 strains(including 894 drug-resistant strains ,and 117 sensitive strains) ,the coincidence rate was 98 .00% ,consistent with isoniazideresistant strains 1 005 strains(including 890 drug-resistant strains ,115 sensitive strains) ,the coincidence rate was 97 .48% .The most common spot of rifampin resistance related mutations of rpoB gene was 531TCG to TTG ,accounted for 51 .11% ,followed by 526CAC→TAC ,accounted for 10 .37% ,11 strains with 526TCG to TTG ,accounted for 8 .15% .Isoniazid re-sistance was caused by mutations in katG315AGC→ACC resistant strains ,accounted for 83 .85% ,inhA-15C→T mutations accoun-ted for only 12 .30% .Conclusion The results of gene chip method is highly consistent with that of absolute concentration method , could be a fast and effective method for screening rifampicin and isoniazide ,the resistant gene of Mycobacterium tuberculosis to rif-ampicin and isoniazide almost mutate in rpoB531 ,526 and katG315 in Changsha .
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Background Laser assisted in situ keratomileusis (LASIK) for myopia will change corneal asphericity and further affect the quality of vision.However,how operating parameters such as ablation depth and optic zone diameter affect corneal asphericity is still rarely reported.Objective Aim of this study was to investigate the influence of corneal ablation depth and optic zone diameter on corneal asphericity after LASIK for myopia.Methods This prospective study comprised 175 eyes of 89 patients with the spherical equivalent of (-5.93± 1.98)D and the best corrected visual acuity (BCVA) ≥ 1.0.The flap creation with the femtosecond laser combine iris recognize guided LASIK was performed on the patients who were available for the evaluation at postoperative 6 months.Corneal aspherical index (Q value) at the central corneal 6.0 mm were measured with Orbscan IIz-corneal topography before and 6 months after operation.Corneal ablation depth and optic zone diameter were recorded in operation.The changes of visual acuity and spherical equivalent before and after surgery were compared by self-control method,and the correlations between corneal ablation depth or optic zone diameter with alteration of Q value (AQ) after LASIK were assessed using multiple regression analysis.Results The mean spherical power,cylinder diopter and spherical equivalent were (-5.57 ± 1.89) D,(-0.71 ±0.55) D and (-5.93 ± 1.98) D before surgery,and those after surgery were (-0.25 ±0.30),(-0.14±0.22)D and (-0.32±0.37)D,showing significant differences between before and 6 months after LASIK (t=-32.39,-23.91,-35.18,all at P<0.01).The Q values at the central corneal 6.0 mm were -0.13 ± 0.09 (-0.47-0.08) in preoperation and 1.09 ± 0.54 (0.22-2.51) in postoperation,with a significant increase in postoperation (t=29.37,P<0.01).Corneal ablation depth was (95±28) μm and optic zone diameter was (6.32±0.26)mm.Ablation depth appeared to be positive correlation with AQ (β =0.803),and optic zone diameter showed a negative correlation with AQ (β =-0.149),with a multiple regression formula AQ =1.517+0.015×ablation depth-0.3 ×optic zone diameter.Conclusions LASIK for myopia increase corneal Q value.The increase of corneal ablation depth and decrease of optic zone diameter contribute to enlargement of corneal Q value.
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Objective To investigate whether macrophage-activating lipopeptide-2 ( MALP-2) in-duces the expression of heme oxygenase-1 ( HO-1 ) in THP-1 cells and its possible mechanism .Methods Human monocyte cells THP-1 were cultured in vitro and then were incubated with various concentrations (0, 0.01, 0.1, 1.0 or 5.0 ng/ml) of MALP-2 for 16 h, or were stimulated by 5.0 ng/ml MALP-2 for different length of time (0 h, 4 h, 8 h, 12 h, 16 h or 24 h).The expression of HO-1 at mRNA and protein levels were detected by real-time PCR analysis and Western blot assay .The enzyme activity of HO-1 was detected by colorimetric analysis.THP-1 cells were pre-incubated with 30 μmol/L of SB203580, PD98059 and SP600125 for 30 min and then were cultured with 5.0 ng/ml MALP-2 for 16 h to investigate the role of mito-gen-activated protein kinases (MAPKs) signaling pathway in HO-1 production.After incubating THP-1 cells with 5.0 ng/ml MALP-2 for different periods of time, NF-E2-related factor 2 (Nrf2) protein was detected by Western blot assay to study the effects of Nrf2 pathway on MALP-2-induced HO-1 expression.Nrf2 and HO-1 proteins were measured by Western blot assay after transfecting THP-1 cells (1×106/well) with Nrf2 siRNA at a final concentration of 100 nmol/L.Results MALP-2 enhanced the expression of HO-1 at mRNA and protein levels as well as the enzyme activity of HO-1 in THP-1 cells in a concentration-dependent manner.The expression of HO-1 protein induced by MALP-2 was significantly inhibited by 30 μmol/L MAPKs specific inhibitors ( SB203580 , PD98059 and SP600125 ) .MALP-2 induced Nrf2 translocation at a concentration of 5.0 ng/ml.The expression of Nrf2 and HO-1 proteins were significantly decreased in Nrf 2 siRNA-transfected THP-1 cells.Conclusion MAPKs and Nrf2 signaling pathways were involved in the MALP-2 induced HO-1 expression .
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Objective:To observe the molecular mechanism involved in expression of hemeoxygenase -1 (HO-1) induced by a macrophage-activating lipopeptide-2 (MALP-2).Methods:THP-1 cells were cultured in vitro and stimulated by MALP-2 for 12 h, expression of HO-1 was detected by Western blot .TLR2 and TLR6 neutralizing antibodies incubation , dominant negative plasmids transfection were used to assess the functional of TLR 2,6 in mediating HO-1 expression.Phosphorylation of c-Src and Akt were detec-ted by Western blot, and c-Src siRNA and PI3K inhibitor LY294002 were used to investigate the role of c-Src and PI3K in HO-1 ex-pression.Results:MALP-2 induced c-Src phosphorylation , and TLR2 and TLR6 neutralizing antibodies , or their dominant negatively plasmids could abrogate this effect .In addition, siRNA of c-Src could decrease the phosphorylation level of Akt , and the PI3K inhibi-tor could inhibit HO-1 expression.Conclusion: MALP-2 can induce THP-1 cells expression of HO-1 through TLR2,6/c-Src/PI3K pathways .
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Objective To investigate whether macrophage-activating lipopeptide-2 ( MALP-2) in-duces the expression of hemoxygenase-1 ( HO-1 ) in THP-1 cells and to further elucidate its possible regulatory mechanism for a better understanding of protective response upon mycoplasma infection .Methods THP-1 cells were cultured in vitro and stimulated by MALP-2 at different concentrations for 12 h.THP-1 cells were incubated with TLR 2 or TLR6 neutralizing antibodies , or transfected with their dominant negative plasmids to evaluate the effects of TLR 2 and TLR6 on HO-1 expression .Phosphorylation of Akt was detected by Western blot.PI3K inhibitor LY294002 was used to investigate the role of PI3K in HO-1 expression.Im-munofluorescence and electrophoretic mobility shift assay ( EMSA ) were performed to observe the nuclear translocation and DNA-binding activity of nuclear factor Nrf 2.Small interfering RNA ( siRNA) was used to silence the genes encoding Nrf2 and HO-1.Cobalt protoporphyrin (CoPP), an inducer of HO-1, was used to treat THP-1 cells.The expression of HO-1 was detected by Western blot .The secretion of TNF-αand IL-1βby THP-1 cells were measured by ELISA .Results MALP-2 induced the expression of HO-1 in THP-1 cells.However, the expression of HO-1 was inhibited by TLR2 and TLR6 neutralizing antibodies and expres-sion of their dominant negative plasmids .Moreover, PI3K pathway was activated by MALP-2, and with the use of PI3K inhibitor, the expression of HO-1 was decreased.The translocation of Nrf2 to the nucleus and itsDNA-binding activity were enhanced by MALP-2, but were inhibited by the treatment of PI3K inhibitor.Theexpression of HO-1 was significantly down-regulated upon the interference of Nrf2 gene expression withsiRNA.Silenced expression of HO-1 increased the level of TNF-αand IL-1β, while CoPP treatment decreasedthe secretion of MALP-2-induced cytokines.Conclusion MALP-2 might induce the expression ofHO-1 in THP-1 cells through TLR2,6/PI3K/Nrf2 pathways.The expression of HO-1 could negatively regulatethe hyper-secretion of cytokines.
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Background The excessive growth of human Tenon fibroblasts (HTFs) is a primary cause of failure of anti-glaucomatous filtering surgery.To seek a drug of anti-fibrosis is of an important significance for improving the successful rate of anti-glaucomatous filtration surgery.Objective The goal of this study was to investigate the effect of paclitaxel on proliferation of HTFs in vitro.Methods Human Tenon tissue was obtained during the anti-glaucomatous filtering surgery.HTFs were cultivated using explant method and 3-6 generations of cells were used in the experiment.The cells were identified by immunochemistry using keratin,vimentin,fibronectin and S-100.Different concentrations (0,1 ×10-s,1 × 10-7,1 × 10-6 mol/L) of paclitaxel were added into the medium,and then the cell indexes (CI) in the various groups were detected by real-time cell electronic sensing (RT-CES) 24 hours after affection of paclitaxel.Apoptosis of the cells was examined using DAPI staining,and the proportion of the cells in different cycles were assayed by flow cytorneter 12 hours after addition of paclitaxel.The expressions of matrix metalloproteinase-1 (MMP-1) protein and mRNA were detected by ELISA and real-time PCR,respectively.Results The cells migrated from the cultivated tissue within 7 days with the fibrocyte-like shape.The cells showed the positive response for vimentin and absent response for keratin,fibronectin and S-100.The CI values were 1.093 ±0.191,0.665 ± 0.093 and 0.473 ± 0.117 in the 1 × 10-8,1 × 10-7 and 1 × 10-6 mol/L paclitaxel groups,showing significant rise in comparison with the 1.514 ±0.283 of the 0 mol/L paclitaxel group (all at P =0.000).The cell nuclei were normal in the 0 mol/L paclitaxel group,however,blue-fluorescent particles and apoptotic bodied were found in the cell nuclei after affection of paclitaxel.The proportion of G2/M phase of cells were (9.20±0.80) %,(12.37±0.45)% and (13.80±0.35)% in the 1×10-8 mol/L,1×10-7 mol/L and 1×10-6 mol/L paclitaxel groups,which were higher than the (7.17±0.50) % in the 0 mol/L paclitaxel group (P=0.005,0.000,0.000).In addition,the relative expressing level of M MP-1 mRNA (MMP-1 mRNA/GAPDH mRNA) and the expression level of MMP-1 protein in the HTFs were significantly lower in the 1 ×10-8 mol/L,1 × 10-7 mol/L and 1 × 10-6 mol/L paclitaxel groups than those in the 0 mol/L group (all at P<0.05).Conclusions Paclitaxel at the concentrations of 1 × 10-8 mol/L-1 × 10-6 mol/L inhibits the proliferation of HTFs in vitro by arresting the cellular mitosis and inducing cell apoptosis.These effects probably associated with down-regulation of MMP-1 expression in the HTFs.
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The progress of research of the physical and chemical modification methods to improve the antithrombogenic property of biomedical polyurethane (PU) in the past five years is reviewed in this paper. The physical modification method includes physical blending, physical vapor deposition (PVD) and replication molding technique. Meanwhile, chemical modification method is focused on the covalent bonding to immobilized special molecular. Moreover, the covalent bonding method covered functionalizing the PU surface with tailor-made groups in the bulk and the activation of the surface to form unstable active sites for further reactions.
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Animais , Humanos , Materiais Biocompatíveis , Química , Farmacologia , Fenômenos Químicos , Fibrinolíticos , Química , Farmacologia , Poliuretanos , Química , FarmacologiaRESUMO
Objective To settle the "bottleneck" disputes existed in the key aspect of emergency treatment care of emergency department, long delayed time of medical workers or patients, perplexity of first-aid personnel, less standard in medical records. Methods Comparison experiment was carried out with the traditional model of emergency care in contrast with the new model, besides the service attitude was improved,the awareness of responsibility was strengthened and the first-aid technique was increased. Number of nursing errors and disputes, hospitalization time, satisfaction degree of patients and record trace-ability were observed. Results The nursing errors and disputes reduced by 46%, hospitalization time re-duced by an average of (7.2±0.2) minutes, satisfaction degree of patients increased by 10.99%, leaving 1673 record traceability of care. Conclusions The application of new management processes of nursing interface in emergency department can settle the "bottleneck" disputes of key interface, make the green passage of emergency patients more convenient and is conducive to safe care, less mortality, high work effi-ciency, which proves to be a reference for fellows.
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Cataract surgery is not only a surgery for vision resuming, but also a kind of refractive surgery. Besides improving the design of intraocular lens, perfecting the technique of surgery and examining far and near visual acuity, we should stress the objective evaluation including contrast sensitivity, glare sensitivity, accommodate ability, binocular function and wavefront aberation for better visual results.