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Ovarian cancer is the most fatal malignant tumor in female reproductive system tumors.In most women, it is diagnosed in a late stage, which largely leads to the poor prognosis of ovarian cancer.Breast cancer susceptibility gene (BRCA) is an important DNA homologous repair gene, which plays a major role in the normal cellular DNA repair mechanism.Its mutation will lead to homologous recombination defects, which will affect the stability of the genome and lead to occurrence of tumors.In recent years, BRCA genetic testing has become a key step in the risk assessment, prognosis, treatment and prevention of ovarian cancer.
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The real-time reported data of treated patients from July 2017 to June 2019 Nanxiang Hospital of Jiading District were collected from chest pain center platform. The results showed that the average time of completing ECG examination from the first medical contact was 1.3 to 6.9 min with a median of 1.9 min (1.7, 2.2), meeting the quality control requirements (10 min); the time required to obtain troponin test results was 13.0 to 48.4 min with a median of 14.1 min (13.4, 18.1), meeting the requirements for quality control of 20 min; time from entry to transfer out of PCI patients was 19.0-100.0 min, with median 37.2 (29.3, 66.6) min, basically reaching quality control (30 min); the entering catheter chamber rate of STEMI patients was 50.0% to 100.0% with a median of 100.0% (73.3%, 100.0%), meeting the requirements of quality control (≥50%). Through the active construction, the main quality control indicators were well reached, the reported cases were basically stable, and the disease distribution was basically reasonable in the primary-level chest pain centers. Informed notification of transshipment and subsequent management of low-risk chest pain patients need to be further strengthened. It is suggested that the construction of chest pain centers should establish long-term normal working mechanism, strengthening the control of key quality control indicators, to play the important role of the regional treatment system.
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OBJECTIVES@#To evaluate the response to cardiac resynchronization therapy (CRT) and the correlation between CRT and pulmonary artery hemodynamic parameters.@*METHODS@#The patients with chronic heart failure indicator for CRT were enrolled. The left ventricular end-systolic volume (LVESV) was measured by echocardiography and New York Heart Association (NYHA) classification was evaluated between one week before and six months after CRT. Mean pulmonary artery pressure (mPAP), pulmonary artery systolic pressure (PASP) and pulmonary vascular resistance (PVR) were measured by right heart catheterization. Left ventricular reverse remodeling (LVRR) is defined as a decrease of 15% or more in LVESV at the 6th month after CRT; Clinical response is defined as a decrease of NYHA classification at or above grade 1 at the 6th month after CRT. Pulmonary hypertension (PH) was defined as mPAP≥25 mmHg. According to the response, patients were divided into 3 groups: group A (LVRR+clinical response), group B (no LVRR+clinical response) and group C (no LVRR+no clinical response). The changes of NYHA classification, echocardiographic and pulmonary hemodynamic parameters were observed in the 3 groups. The Kaplan-Meier survival curve was used to analyze the differences in all-cause mortality, combined end-point events of death or re-hospitalization due to heart failure among different groups.@*RESULTS@#A total of 45 patients with CRT implantation [aged (63.27±9.55) years, 36 males] were included. The average follow-up period was (33.76±11.50) months. Thirty-one patients (68.89%) were in group A, 9 of whom with PH. Eight patients (17.78%) were in group B, 7 of whom with PH. Six patients were in group C, all with PH. Cardiac function including NYHA classification, echocardiographic and pulmonary hemodynamic parameters had been significantly improved in group A after CRT implantation (0.05). There were no significant changes in NYHA classification, echocardiographic and pulmonary hemodynamic parameters in group C (>0.05). Compared with group C, group A and group B had lower all-cause mortality (=0.005) and lower incidence of composite endpoint events (=0.001).@*CONCLUSIONS@#Patients with LVRR and clinical response after CRT have a good prognosis. Patients with clinical response but without LVRR have a better prognosis than those without clinical response and LVRR, which may be related to the decrease of pulmonary hemodynamic parameters such as mPAP and TPG.
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Aged , Humans , Male , Middle Aged , Cardiac Resynchronization Therapy , Heart Failure , Therapeutics , Hemodynamics , Pulmonary Artery , Treatment Outcome , Ventricular RemodelingABSTRACT
OBJECTIVE: To establish the method for the content determination of related substance in Terazosin hydrochloride tablets. METHODS: HPLC and principal component self-control with correction factor were adopted. The determination was performed on Agilent Zorbax Eclipse XDB C18 column with mobile phase consisted of acetonitrile-perchloric acid solution (20 ∶ 80, V/V) at the flow rate of 1.0 mL/min. The detection wavelength was set at 246 nm, and sample size was 20 μL. The column temperature was 50 ℃. The linear equations of terazosin hydrochloride, impurity A, B, C were drawn. The correction factors of each impurity related to terazosin hydrochloride were calculated by slope, and relative retention time was used to determine the position of impurities. The contents of impurity A, B and C in 3 batches of Terazosin hydrochloride tablets were determined and compared with the results of impurity control method. RESULTS: The relative retention time of impurity A, B, C was 0.39, 0.74, 2.77, respectively; the linear range of them were 0.25-3.0 μg/mL, respectively. The correction factors were 0.75, 1.09, 0.84, respectively. The detection limits were 0.35, 0.51, 0.43 ng, and the limits of quantification were 0.70, 1.02, 0.86 ng, respectively. The contents of impurity A, B and C in 3 batches of Terazosin hydrochloride tablets were 0.11%-0.13%, 0.03% and 0.09%-0.12%; impurity B did not detected. The results are consistent with the determination of impurity control method. CONCLUSIONS: The method is simple, rapid and accurate for the content determination of related substances A, B, C in Terazosin hydrochloride tablets.
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Lung cancer has the highest mortality rate among all malignant tumors. The key to reducing lung cancer mortality is the accurate diagnosis of pulmonary nodules in early-stage lung cancer. Computer-aided diagnostic techniques are considered to have potential beyond human experts for accurate diagnosis of early pulmonary nodules. The detection and classification of pulmonary nodules based on deep learning technology can continuously improve the accuracy of diagnosis through self-learning, and is an important means to achieve computer-aided diagnosis. First, we systematically introduced the application of two dimension convolutional neural network (2D-CNN), three dimension convolutional neural network (3D-CNN) and faster regions convolutional neural network (Faster R-CNN) techniques in the detection of pulmonary nodules. Then we introduced the application of 2D-CNN, 3D-CNN, multi-stream multi-scale convolutional neural network (MMCNN), deep convolutional generative adversarial networks (DCGAN) and transfer learning technology in classification of pulmonary nodules. Finally, we conducted a comprehensive comparative analysis of different deep learning methods in the detection and classification of pulmonary nodules.
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Humans , Deep Learning , Multiple Pulmonary Nodules , Neural Networks, Computer , Solitary Pulmonary Nodule , Tomography, X-Ray ComputedABSTRACT
Objective To investigate the change and clinical significance of growth differentiation factor 15 (GDF15) and gastric cancer antigen 724 (CA724) in serum of patients with gastric cancer .Methods Serum levels of GDF15 in serum of 30 patients with gastric cancer were detected by ELISA .The levels of serum gas-tric CA724 were measured by electrochemiluminescence ,and 32 cases of benign gastric lesions and 30 healthy controls were compared .Results The levels of serum GDF15 and CA724 in patients with gastric cancer were (1 .58 ± 0 .53)ng/mL and (40 .80 ± 5 .20) IU/mL ,respectively .The levels of serum GDF15 and CA724 in pa-tients with benign gastric lesions were (0 .26 ± 0 .11) ng/mL ,(12 .90 ± 2 .30) IU/mL (P<0 .01) .The levels of GDF15 and CA724 in normal control group were (0 .17 ± 0 .08)ng/mL and (3 .80 ± 0 .90)IU/mL respectively . The levels of serum GDF15 and CA724 in gastric cancer group were significantly higher than those in benign gastric lesions and normal control group (P<0 .01) .The level of serum GDF15 was closely related to tumor size ,TNM stage and lymph node metastasis (P<0 .05) .The sensitivity of gastric cancer was 83 .3% ,the spe-cificity was 83 .9% and the AUC was 0 .837 .The sensitivity of CA724 was 90 .0% ,the specificity was 76 .5%and the AUC was 0 .886 .The combined detection of AUC was 0 .920 ,which was significantly higher than that of single detection .Conclusion GDF15 is associated with the development and progression of gastric cancer .Combined detection of GDF15 and CA724 in serum is helpful for early diagnosis of gastric cancer and progno-sis .
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Objective To compare the changes of tear film and corneal sensitivity between femtosecond laser-assisted cataract surgery and microincisional phacoemulsification surgery.Methods A prospective cohort study was performed.Fifty-three eyes of 53 patients with age-related cataract were included in Wuhan Aier Eye Hospital from March to July 2015.The patients were divided into femtosecond laser group and microincisional group.The 26 eyes in the femtosecond laser group received femtosecond laser-assisted cataract surgery,and 27 eyes in the microincisional group received 2.2 mm microincisional phacoemulsification surgery.The patients were followed-up for 3 months.The data of corneal sensitivity,Schirmer test values,non-invasive tear film break-up time (NIBUT) and subjective score for dryness were collected in 7 days,1 month and 3 months after surgery,and these results were compared between the two groups.This study was approved by Ethic Committee of Wuhan Aier Ophthalmology Hospital,and written informed consent was obtained from each patient prior to surgery.Results The operations were finished smoothly in all the eyes.In preoperative and 7 days,1 month,3 months after operation,the upper corneal sensitivity threshold and NIBUT between the two groups appeared a first reduction and a subsequent rise,the same phenomenon occurred in Schirmer test of the microincisional group and the subjective scores for dryness of the femtosecond laser group with statistically significant differences in the overall comparison (all at P< 0.05).No significant differences were seen in the central corneal sensitivity of the two groups and Schirmer test in the femtosecond laser group (all at P>0.05).The dry eye scores increased gradually with the time expand after operation in the microincisional group,the difference was statistically significant (P<0.05).In postoperative 3 months,these parameters were restored preoperative values.However,the dry eye scores in the microincisional group were still higher than that before operation,and the difference was statistically significant (P<0.05).Conclusions Compared with the 2.2 mm microincisional phacoemulsification surgery,femtosecond laser-assisted cataract surgery brings less disturbance to lacrimal secretion,slight symptoms of dry eye and more comfortable sensation.
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Objective To compare the anesthetic effect of atocaine adrenaline injection and lidocaine injec -tion in wisdom tooth extraction .Methods 220 patients with wisdom teeth were selected as the research subjects . According to the random table method ,the patients were randomly divided into research group 110 cases ( a total of wisdom teeth 132,maxillary wisdom teeth 47,mandibular wisdom tooth 85) and the control group 110 cases(a total of wisdom teeth 129,maxillary wisdom teeth 51,mandibular wisdom tooth 78).The research group received atocaine adrenaline injection ,the control group received lidocaine hydrochloride injection .The effects of anesthesia ,VAS score and adverse reactions in the two groups were observed .Results The total effective rate of maxillary wisdom tooth anesthesia in the research group(100.00%) was significantly higher than that in the control group (90.20%),and the difference between the two groups was statistically significant (χ2 =6.358,P<0.05).The total effective rate of mandibular wisdom tooth anesthesia in the research group (100.00%) was significantly higher than that in the control group(92.31%),and the difference between the two groups was statistically significant (χ2 =6.788,P<0.05).The VAS score of maxillary wisdom tooth anesthesia in the research group [(2.57 ±0.65)points]was significantly lower than that in the control group [(2.87 ±0.63) points],and the difference between the two groups was statistically significant(t=2.319,P<0.05).In the control group,there was 1 case of hematoma caused by blood vessels ,and no complications occurred in the research group .Conclusion The anesthetic effects of articaine adrenaline injection in wisdom tooth extraction is better than lidocaine hydrochloride injection , without any complications , it is worthy of clinical promotion .
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Objective To assess effects of stress dynamic CT myocardial perfusion imaging (CT-MPI) combined with coronary CT angiography (CCTA) on the diagnosis of myocardial perfusion defects in coronary artery disease (CAD). Methods Patients with CAD diagnosed by CCTA underwent ATP stress CT-MPI examination. Single-photon emission computed tomography (SPECT) myocardial perfusion imaging (SPECT-MPI) was performed within one week and set as the reference standard. CT-MPI results were qualitatively analyzed, and myocardial blood flow (MBF), myocardial blood volume (MBV) as well as time to peak (TTP) were quantified according to CT-MPI. Effects of CCTA, CT-MPI, and CT-MPI combined with CCTA on predicting myocardial perfusion defects were assessed in comparison with NMPI. Results Thirty patients [(54.8±8.4)years] were enrolled in our study, 20 were men (68%). MBF [(79.3±18.0) versus (135.1± 35.2) ml·100 ml-1·min-1] and MBV [(8.9±2.9) versus (13.8±8.9) ml/100 ml] were significantly decreased in hypoperfused segments compared with normal segments, while TTP was increased in hypoperfused segments [(13.9 ± 2.5)s] compared with normal segments [(9.1 ± 2.1)s] (t=0.302, 0.866 and 0.024 respectively, all P values<0.01). The sensitivity, specificity of CT-MPI for identifying segments with perfusion defects were 91.3%(147/161), 84.6%(281/332), respectively. On a per-vessel basis, the area under the receiver operating characteristic curve for predicting myocardial perfusion defects were 0.635(95%CI:0.517—0.753) for CCTA, 0.709(95%CI:0.599—0.819)for CT-MPI, and 0.837(95%CI:0.749—0.925)for CT-MPI combined with CCTA, respectively. Conclusions The performance of stress dynamic CT-MPI in the diagnosis of myocardial perfusion defects in CAD was good. One-stop examination of CT-MPI combined with CCTA improves the diagnostic accuracy for identifying flow-obstructing stenosis.
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This article summarizes the literature about studies onToll-like receptor 2and acupuncture points and explores thecorrelationand action of Toll-like receptor 2inacupuncture information start-up and conduction mechanism from the biological characteristics of Toll-like receptor 2and the relationship of Toll-like receptor 2-mediated signal pathways tomast cells, acupuncture effects and the immuno-neuro-endocrine network.
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Spastic-paralysis impacts the function of upper extremity in most apoplexy patients.Assessing spasticity condition precisely,timely and effectively plays an important role in guiding rehabilitation treatment,therapeutic evaluation and prognosis of functional outcome.Owing to the limitations of subjective scales,the issue of quantifing the spasticity objectively has become a new research direction in recent years.By reviewing the literatures,we summed up the application of objective assessment tools,which were based on rehabilitation engineering,electrophysiology or biomechanics,in assessing the function of upper extremity with spastic paralysis,as well as put forward the research directions of prospects,in order to give a reference to the systematic integration of assessment tools in the near future.
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Objective To study curative efficacy of Pramipexole in treatment of Parkinson's Disease and its effects on Serum C-reactionprotein (CRP), cystatin C, brain derived neurotrophic factor(BDNF) and oxidative stress.Methods 100 patients of Parkinson's Disease who received therapy from February 2014 to April 2016 in our hospital were selected as research objects were randomLy divided into control group and observation group, 50 patients in each group.The control group was treated with Dobasidate tablets;The observation group was given pramipexole tablets on the basis of the control group.Then the clinical effects and adverse reactions were compared.CRP, Cystatin C, BDNF and oxidative stress were measured. Results After treatment, the total effective rate of the observation group [(90.00% (45/50)] was significantly higher than the control group[74.00%(37/50)] (P<0.05);The symptoms of palpitation, depression, nausea, anorexia, diarrhea and vomiting were lower in observation group than in observation group, and the differences of depression, anorexia and vomiting were statistically significant(P<0.05);The BDNF index of the observation group (13.38 ±1.93)ng/L was higher than that of the control group (12.03 ±1.83) ng/L(P<0.05);CRP (3.53 ±0.97) mg/L and Cystatin C index (0.98 ±0.12) mg/L were lower than the control group(4.62 ±1.15)、(1.06 ±0.14) mg/L (P<0.05).After treatment, the glutathione peroxidate (GPX),superroxide dismutase(SOD),glutathione(GSH)and catalase(CAT) of oxidative stress in observation group were higher than those in control group,the difference was statistically significant (P<0.05).Conclusion Pramipexole in the treatment of Parkinson's disease is obvious, can effectively improve the serum CRP, Cystatin C and BDNF index, high security, it is worth promoting the application.
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Objective To investigate the clinical therapeutic effect of electroacupuncture on ischemic stroke by recording the therapeutic effect score and performing transcranial Doppler (TCD) detection in electroacupuncture-treated convalescent stroke patients before and after treatment.Methods Sixty convalescent patients with ischemic stroke were randomly allocated to treatment and control groups, 30 cases each. The control group received routine medication and the treatment group, electroacupuncture in addition. In the two groups, pre- and post-treatment neurological defect scores were recorded and the clinical therapeutic effects were observed after four weeks of treatment. Pre- and post-treatment middle cerebral artery (MCA) pulsation indices (RI), resistance indices (RI) and ratios of systolic to end-diastolic (S/D) blood flow velocity detected by TCD were compared between the two groups. Results The total efficacy rate was 80.0% in the treatment group and 50.0% in the control group; there was a statistically significant difference between the two groups (P<0.01). There was a statistically significant pre-/post-treatment difference in the MESSS score in the two groups (P<0.01). There was a statistically significant post-treatment difference in the MESSS score between the treatment and control groups (P<0.05). There were statistically significant pre-/post-treatment differences in TCD indices (PI, RI and S/D) on the affected side in the two groups (P<0.05). There were statistically significant post-treatment differences in TCD indices on the affected side between the treatment and control groups (P<0.05).Conclusion Electro- acupuncture can promote rehabilitation in convalescent stroke patients.
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Objective To explore the relationship of the expression level of monocyte Chemotactic protein-1 and nuclear factor κB(NF κB)with atrial fibrosis of atrial fibrillation patients with rheumatic heart disease.Methods About 200 mg right atrial tissue were taken from 26 patients with rheumatic heart disease undergoing valve replacement surgery and divided into sinus rhythm (SR) group (n=12) and atrial fibrillation (AF) group (n=14).Masson staining and atrial myocardial fibrosis markers were used to determine the level of fibrosis.The mRNA levels of cytokines in atrial tissue were measured by reverse transcription polymerase chain reaction (RT-PCR)The protein level of MCP-1 and phosphorylation of NFκB in atrial tissue were detected by Western blotting.Results Compared with SR group,the AF group showed that collagen volume fraction (AF:0.42 ± 0.03;SR:0.13 ± 0.02),the mRNA levels of myocardial fibrosis markers such as transforming growth factor (TGF)-β1,type Ⅰ collagen and type Ⅲ collagen,and cytokines such as IL-1β and TNF-α,and the protein level of MCP-1 (AF:0.170±0.003;SR:0.040±0.005) and level of phosphorylation of NF-κB (AF:0.35 ± 0.02;SR:0.12 ± 0.03) were significantly increased (all P<0.05).In addition,the expression levels of cytokines,the protein expression level of MCP1 and the phosphorylation level of NF-κB were positively correlated with collagen volume fraction of the right atrial myocardial tissue (all P<0.05).Conclusions Activation of NF-κB may induce the expression of MCP-1 in the myocardial tissue of patients with rheumatic heart disease,and sequentially stimulate the secretion of cytokines and extracellular matrix deposition,and finally participate in the occurrence and persistence of atrial fibrillation.
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<p><b>OBJECTIVE</b>To compare the features of patients with pulmonary hypertension due to left heart disease classified according to transpulmonary gradient (TGP) or diastolic pressure difference (DPD).</p><p><b>METHODS</b>Thirty-three patients with pulmonary hypertension due to left heart disease diagnosed by right heart catheterization were enrolled. Patients were divided into two groups according to TPG: 17 patients with TPG ≤ 12 mmHg (1 mmHg = 0.133 kPa) and 16 patients with TPG > 12 mmHg; or divided into two groups according to DPD: 23 patients with DPD < 7 mmHg and 10 patients with DPD ≥ 7 mmHg. McNemar's method was used to test the agreement of the two classification methods.</p><p><b>RESULTS</b>Below are the patients features according to the classification by TPG: central venous pressure ((9.0 ± 2.5) vs. (12.7 ± 5.4) mmHg), mean right atria pressure ((9.1 ± 2.4) vs. (12.8 ± 5.2) mmHg), right heart systolic pressure ((45.5 ± 9.8) vs. (66.8 ± 15.4) mmHg), right heart mean pressure ((22.6 ± 5.2) vs. (33.1 ± 7.5) mmHg), pulmonary systolic pressure ((44.2 ± 10.3) vs. (64.8 ± 14.2) mmHg), pulmonary diastolic pressure ((24.2 ± 4.5) vs. (33.1 ± 8.3) mmHg), pulmonary mean pressure ((32.3 ± 5.7) vs. (45.8 ± 8.6) mmHg), cardiac index ((2.6 ± 1.0) vs. (1.9 ± 0.9) L · min(-1) · m(-2)), right heart EF ((31.2 ± 12.6)% vs. (22.6 ± 7.1) %) and pulmonary vascular resistance ((2.3 ± 0.8) vs. (6.3 ± 2.6) Wood) were significantly different between the two groups (all P < 0.05). According to the classification of DPD, only right heart diastolic pressure ((7.4 ± 3.7) vs. (11.5 ± 5.7) mmHg), pulmonary diastolic pressure ((25.9 ± 6.4) vs. (34.7 ± 8.0) mmHg) and pulmonary vascular resistance ((3.3 ± 2.0) vs. (6.2 ± 3.4) Wood) were significantly different between the two groups (all P < 0.05). These was a weak agreement (κ = 0.386 6, 95% CI: 0.092 2-0.681 0) between the two classification methods.</p><p><b>CONCLUSION</b>TPG classification is superior to DPD classification for pulmonary hypertension patients due to left heart disease on identifying the hemodynamic differences.</p>
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Humans , Blood Pressure , Cardiac Catheterization , Diastole , Heart , Heart Failure , Hemodynamics , Hypertension, Pulmonary , Vascular ResistanceABSTRACT
<p><b>OBJECTIVE</b>To explore the technique of temperature control on the abdomen in penetrating moxibustion through observing moxibustion time on the abdomen, abdominal skin temperature and effect after moxibustion so as to provide the safe, effective and easily applicable method for penetrating moxibustion.</p><p><b>METHODS</b>Thirty-two patients were selected in an observation group, 32 healthy persons in a control group. In the observation group, the penetrating moxibustion was applied to the corresponding acupoint locations according to different symptoms. In the control group, moxibustion was used on the abdomen around the umbilicus. The skin temperature was recorded once every minute. The skin temperature of known heat sensation, the time of known heat sensation, the known reduced temperature, the time of temperature reducing, the skin temperature difference, the duration of penetrating moxibustion and the reaction of moxibustion from participants were recorded.</p><p><b>RESULTS</b>The differences in the skin temperature of known heat sensation, the time of known heat sensation and the duration of penetrating moxibustion were significant statistically in comparison between the observation group and the control group (all P<0.01). The differences in the known reduced temperature, the time of temperature reducing and the skin tem- perature difference were not significant (all P>0.05). The differences were significant statistically in skin rashes and moxibustion reaction (gastrointestinal peristalsis, chills, ant climbing feeling and hunger, etc.) between the two groups (P<0.01). The differences were not significant statistically in flushing, sweating and blisters (all P>0.05).</p><p><b>CONCLUSION</b>(1) The level of temperature sensitivity in the observation group is lower than that in the control group. During penetrating moxibustion, the sensations such as gastrointestinal peristalsis, chills, ant climbing feeling and hunger appear easily, suggesting the positive self-adjustment in the body. (2) During penetrating moxibustion, the warm feeling is penetrated not just from the epidermis to the abdominal cavity and lumbar region, but also up to thehead and down to the knee. (3) The flushing, sweating and skin rashes are the important indices for the effectiveness of penetrating moxibustion. (4) The temperature control is the core technique of penetrating moxibustion. The penetrating moxibustion in 28 min to 32 min and the temperature controlled in 43 degrees C to 45 degrees C can solve the moxibustion smoky impact to the environment, but also relieve pains of the patients.</p>
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Adult , Female , Humans , Male , Middle Aged , Young Adult , Abdomen , Physiology , Acupuncture Points , Case-Control Studies , Moxibustion , Skin Temperature , ThermosensingABSTRACT
A series of tetrahydroisoquinoline derivatives were prepared and their peroxisome proliferator-activated receptor (PPAR) alpha/gamma agonistic activities were evaluated to obtain more potent PPAR agonist. All of them were new compounds, and their structures were confirmed by 1H NMR and HR-MS. Three compounds exhibited higher agonistic activities of PPARgamma than that of the comparison, six compounds exhibited higher agonistic activities of PPARalpha than that of the comparison, and compound 8a was discovered as a highly potent PPARalpha/gamma agonist that is much more active than that of WY14643 and rosiglitazone. The development of potent PPAR agonists may offer a new choice for the treatment of diabetes.
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Three plasmids (pGenesil-P1, pGenesil-P2, pGenesil-P3) with different p27Kip1-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endothelial cells (bCEC) were investigated. Plasmid expressing irrelevant shRNA with a random combination was used as negative control (pGenesil-HK). The recombination of four plamids was confirmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kip1 was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kip1 mRNA and p27Kip1 protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group (non-transfected group). pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The proliferation rates of the pGenesil-P3 group, the pGenesil-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kip1 on bCEC cell cycle was detected by flow cytometry (FCM). Compared with the control groups, the proliferation rate of the pGenesil-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kip1 could down-regulate the expression of p27Kip1 effectively and increase the proliferation of bCEC. RNA interference (RNAi) may be an effective means to promote the proliferation of CEC.
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Cell Proliferation , Cornea/cytology , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Endothelial Cells/cytology , Endothelial Cells/metabolism , Gene Expression Regulation , Models, Biological , Plasmids/metabolism , RNA Interference , RNA, Messenger/metabolism , RNA, Small Interfering/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tetrazolium Salts/pharmacology , Thiazoles/pharmacology , TransfectionABSTRACT
The effects of sodium salicylate on the expression of heat shock protein 27 (HSP27) during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells (HLB-3) were divided into 3 groups: control group (group A), oxidation injury group (group B) and sodium salicylate group (group C). Apoptosis of human lens epithelial cells cultured in vitro was induced in the presence of 150 micromol/L H2O2. Cells viability and the expression of HSP27 were analyzed. Viability of the cells was measured by methyl thiazole tetrazolium (MTF) chromatometry. The expression of HSP27 in HLB-3 cells was detected by using immunohistochemistry and image analysis system. Sodium salicylate could induce the expression of HSP27, and the cells viability in group C was significantly higher than in group B (0.2667+/-0.01414 vs 0.2150+/-0.01080, P=0.012<0.05). The average gray value of HSP27 in group B was less than that in group C (P=0.000<0.05). The increased expression of HSP27 by sodium salicylate might play an important role in the protection of hydrogen peroxide-induced injury of human lens epithelial cells, suggesting that sodium salicylate could suppress, at least in part, the apoptosis of human lens epithelial cells.
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The effects of sodium salicylate on the expression of heat shock protein 27 (HSP27)during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells (HLB-3) were divided into 3 groups: control group (group A), oxidation injury group (group B) and sodium salicylate group (group C). Apoptosis of human lens epithelial cells cultured in vitro was induced in the presence of 150 μmol/L H2O2. Cells viability and the expression of HSP27 were analyzed. Viability of the cells was measured by methyl thiazole tetrazolium (MTT)chromatometry. The expression of HSP27 in HLB-3 cells was detected by using immunohistochemistry and image analysis system. Sodium salicylate could induce the expression of HSP27, and the cells viability in group C was significantly higher than in group B (0.2667±0.01414 vs 0.2150±0.01080, P=0.012<0.05). The average gray value of HSP27 in group B was less than that in group C (P=0.000<0.05). The increased expression of HSP27 by sodium salicylate might play an important role in the protection of hydrogen peroxide-induced injury of human lens epithelial cells,suggesting that sodium salicylate could suppress, at least in part, the apoptosis of human lens epithelial cells.