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In recent years, with the vigorous marketing of e-cigarette manufacturers, e-cigarettes have become a popular tobacco product for adolescents. The problem of e-cigarette environmental exposure among adolescents is also getting worse and its associated adverse impacts cannot be ignored. However, domestic research on the environmental exposure to e-cigarettes among youth is insufficient, and experience on e-cigarette regulation is also limited. This review first briefly introduced the definition and sources of e-cigarette environment exposure, then focused on the differences of e-cigarette environmental exposure among adolescents with different characteristics to identify possible influencing factors, as well as the impacts of e-cigarette environmental exposure on adolescents, and finally summarized international countermeasures to prevent e-cigarette environmental exposure in adolescents, aiming to provide directional guidance for China to conduct e-cigarette control activities among adolescents.
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Ischemic stroke is one of the primary causes of death and disability worldwide. Neutrophils can release depolymerized chromatin and proteins to form neutrophil extracellular traps (NETs) and participate in intravascular thrombus formation. In recent years, NETs have received increasing attention in the study of acute ischemic stroke. The results indicate that NETs play an important role in the pathogenesis of acute ischemic stroke. This review presented recent advances in the study of NETs in acute ischemic stroke.
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Objective:To investigate the preparation methods and quality control of 177Lu-labeled radiopharmaceuticals, and conduct preliminary clinical application research. Methods:177Lu-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA)- D-Phe1-Tyr3-octreotide (TOC) and 177Lu-prostate specific membrane antigen (PSMA)-I&T were labeled by manual labeling and automatic labeling, respectively. Factors such as the amount of precursor and nuclide, reaction temperature, pH value, reaction time, labeling yield and specific activity were investigated. Quality control of the products were carried out, such as clarity, pH value, sterility, bacterial endotoxin and stability in vitro. 177Lu-PSMA-I&T was applied to the treatment of prostate cancer patients, and the efficacy was evaluated by SPECT/CT imaging. Paired t test was used to analyze the data. Results:The amount of precursor and nuclide, reaction temperature, pH value and reaction time of the two methods were basically the same, both with high yield and specific activity. The yield of 177Lu-DOTA-TOC automatic labeling was significantly higher than that of manual labeling (99.2±0.4)% vs (95.3±1.5)% ( t=7.17, P<0.001), and the specific activity were (91.6±13.7) vs (89.1±13.2) GBq/μmol. The yield of 177Lu-PSMA-I&T automatic labeling was also significantly higher than that of manual labeling (99.6±0.3)% vs (95.7±1.3)% ( t=8.24, P<0.001), and the specific activity were (96.1±14.3) vs (93.2±13.8) GBq/μmol. The labeled products were colorless clear solution with pH value of 6.5-7.0. The sterility and bacterial endotoxin met the requirements. The radiochemical purity of the labeled products was more than 95% after 48 h, which showed good stability. The clinical application of 177Lu-PSMA-I&T in patients with prostate cancer showed that both primary and metastatic lesions had good uptake. Conclusions:The labeling of 177Lu radiopharmaceuticals is simple and has high yield and stability. The application of automatic labeling can simplify the process, improve the yield and reduce irradiation.
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Peptides that are composed of dextrorotary (d)-amino acids have gained increasing attention as a potential therapeutic class. However, our understanding of the in vivo fate of d-peptides is limited. This highlights the need for whole-body, quantitative tracking of d-peptides to better understand how they interact with the living body. Here, we used mouse models to track the movement of a programmed death-ligand 1 (PD-L1)-targeting d-dodecapeptide antagonist (DPA) using positron emission tomography (PET). More specifically, we profiled the metabolic routes of [64Cu]DPA and investigated the tumor engagement of [64Cu/68Ga]DPA in mouse models. Our results revealed that intact [64Cu/68Ga]DPA was primarily eliminated by the kidneys and had a notable accumulation in tumors. Moreover, a single dose of [64Cu]DPA effectively delayed tumor growth and improved the survival of mice. Collectively, these results not only deepen our knowledge of the in vivo fate of d-peptides, but also underscore the utility of d-peptides as radiopharmaceuticals.
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Polymerase chain reaction (PCR) is the gold standard for nucleic acid amplification in molecular diagnostics. The PCR includes multiple reaction stages (denaturation, annealing, and extension), and a complicated thermalcycler is required to repetitively provide different temperatures for different stages for 30-40 cycles within at least 1-2 hours. Due to the complicated devices and the long amplification time, it is difficult to adopt conventional PCR in point-of-care testing (POCT). Comparing to conventional PCR, isothermal amplification is able to provide a much faster and more convenient nucleic acid detection because of highly efficient amplification at a constant reaction temperature provided by a simple heating device. When isothermal amplification is combined with microfluidics, a more competent platform for POCT can be established. For example, various diagnosis devices based on isothermal amplification have been used to rapidly and conveniently detect SARS-CoV-2 viruses. This review summarized the recent development and applications of the microfluidics-based isothermal amplification. First, different typical isothermal amplification methods and related detection methods have been introduced. Subsequently, different types of microfluidic systems with isothermal amplification were discussed based on their characteristics, for example, functionality, system structure, flow control, and operation principles. Furthermore, detection of pathogens (e.g. SARS-CoV-2 viruses) based on isothermal amplification was introduced. Finally, the combination of isothermal amplification with other new technologies, e.g. CRISPR, has been introduced as well.
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COVID-19/diagnosis , Humans , Microfluidics , Nucleic Acid Amplification Techniques , Polymerase Chain Reaction , SARS-CoV-2/geneticsABSTRACT
Objective:To evaluate the therapeutic effect of 177Lu-prostate specific membrane antigen (PSMA)-I&T on prostate cancer. Methods:The culture medium of 1.85, 18.50, 185.00, 555.00 and 925.00 MBq/L 177Lu-PSMA-I&T was added into LNCaP cells (200 μl/well, 5 experimental groups and 1 control group, 3 replicates in each group) for 24 h, and the cell viability in each group was detected. The culture medium of 3.7 MBq 177Lu-PSMA-I&T was added into LNCaP cells (1 experimental group, 1 control group, 3 replicates in each group) for 48 h to detect the changes of cell cycle. LNCaP cells (3 experimental groups and 1 control group, 3 replicates in each group) were added into the culture medium of 3.7, 19.5 and 37.0 MBq 177Lu-PSMA-I&T for 48 h to detect cell apoptosis. Tumor-bearing mice models were established (BALB/c-nu/nu nude mice, n=32). The changes of tumor volume and body mass of tumor-bearing mice were observed within 20 d after treatment. On the 7th day after treatment, tumor tissues of tumor-bearing mice were stained with HE staining and fluorescently stained with Ki-67 protein, and apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL) assay. On the 20th day after treatment, pathological analysis was performed on the main viscera of the tumor-bearing mice. One-way analysis of variance, the least significant difference t test and paired t test were used to analyze the data. Results:Compared with the control group ((100.00±12.35)%), the cell survival rates were significantly decreased after 177Lu-PSMA-I&T intervention in 185.00, 555.00, 925.00 MBq/L groups ((57.56±6.35)%, (38.65±3.39)%, (27.95±4.48)%; F=78.91, t values: 8.312-14.106, all P<0.01). Cell survival rates were significantly reduced in 185.00 MBq/L group at different time points (24, 48 and 72 h; F=78.28, t values: 6.628-14.384, all P<0.01). The proportion of LNCaP cells in G2/M phase was increased from (12.36±0.28)% to (19.92±0.48)% ( t=17.180, P<0.01). The apoptosis rates of cells were significantly increased in 18.5 and 37.0 MBq groups ( F=71.86, t values: -6.138, -13.050, both P<0.05). The difference of relative tumor volume (RTV%) was statistically significant among 3.7, 14.8 and 29.6 MBq groups and control group (136.7±7.4, 59.2±23.8, 47.3±13.8 vs 240.3±3.7; F=78.20, t values: 7.549-13.345, all P<0.01). But there was no significant difference in body mass of tumor-bearing mice among groups. Compared with the control group, the positive rates of Ki-67 staining cells ((37.23±3.04)% vs (14.89±3.80)%, (5.60±1.83)%, (3.46±0.71)%) and TUNEL-fluorescein isothiocyanate (TUNEL-FITC) staining ((0.74±0.18)% vs (1.61±0.30)%, (3.19±0.44)%, (3.54±0.47)%) in tumor tissues of 3.7, 14.8 and 29.6 MBq groups were statistically significant ( F=103.91, t values: 10.429-15.762; F=38.66, t values: from -9.312 to -2.881, all P<0.01). Conclusions:177Lu-PSMA-I&T has a good therapeutic effect on prostate cancer, with no obvious therapeutic side effects. Therefore, 177Lu-PSMA-I&T is expected to be an ideal drug for treating prostate cancer.
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Objective:To investigate the regulatory effects of endogenous nitric oxide (NO) on the activity of superoxide dismutase-1 (SOD1) and apoptosis of human umbilical vein endothelial cells (HUVECs).Methods:HUVECs were taken as the research object.The endothelial NO synthase (eNOS) short hairpin RNA(shRNA) lentivirus was employed to transfect HUVECs to knock down eNOS.HUVECs were divided in 4 groups: the scramble group, the eNOS shRNA group, the eNOS shRNA + sodium nitroprusside(SNP) group and the eNOS shRNA+ SNP+ tris (2-carboxyethyl) phosphine hydrochloride (TCEP) group.The protein expressions of eNOS and SOD1 dimer/monomer in cells were detected by western blot.The activity of SOD was detected by the enzyme-linked immunosorbent assay.The NO content in cells was detected with NO fluorescence probe.The level of superoxide anion in HUVECs was detected with dihydropyridine (DHE). The terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) assay was adopted to detect the apoptosis of HUVECs in situ.Results:Compared with the scramble group, the endogenous NO content (2.690±0.420 vs.15.029±2.193, P<0.01), eNOS protein expression (1.000±0.778 vs.3.141±0.199, P<0.01), SOD1 dimer/monomer ratio (4.6±1.0 vs.7.6±2.0, P<0.05) and SOD activity [(0.432±0.254) Carmen′s unit/10 4 cell vs.(1.000±0.116) Carmen′s unit/10 4 cell, P<0.01] were significantly decreased, while the level of intracellular superoxide anion (11.180±1.560 vs.6.146±1.007, P<0.01) and HUVECs apoptosis [75.0 (55.0, 100.0)% vs.0 (0, 0)%, P<0.01] were significantly increased in the eNOS shRNA group.Compared with the eNOS shRNA group, the content of endogenous NO (16.705±0.116 vs.2.690±0.420, P<0.01), the ratio of SOD1 dimer/monomer (7.3±2.0 vs.4.6±1.0, P<0.05) and the activity of SOD [(0.737±0.060) Carmen′s unit/10 4 cell vs.(0.432±0.254) Carmen′s unit/10 4 cell, P<0.05] were significantly increased, while the level of superoxide anion (6.897±1.648 vs.11.180±1.560, P<0.01) and the HUVECs apoptosis [0 (0, 0)% vs.75.0 (55.0, 100.0)%, P<0.01] were significantly decreased in the eNOS shRNA+ SNP group.Compared with the eNOS shRNA + SNP group, the ratio of SOD1 dimer/monomer (4.4±0.9 vs.7.3±2.0, P<0.05) and the activity of SOD [(0.214±0.084) Carmen′s unit/10 4 cell vs.(0.737±0.060) Carmen′s unit/10 4 cell, P<0.01] were significantly decreased, while the level of superoxide anion (10.917±1.552 vs.6.897±1.640, P<0.01) and the apoptosis level of HUVECs[63.6 (55.0, 90.0)% vs.0 (0, 0)%, P<0.01] were significantly increased in the eNOS shRNA+ SNP+ TCEP group.However, there was no significant difference in the NO content (16.112±0.926 vs.16.705±0.116, P>0.05). Conclusions:Endogenous NO could effectively antagonize the apoptosis of endothelial cells by increasing the cysteine-dependent SOD1 dimer/monomer ratio, enhancing SOD activity and inhibiting the accumulation of reactive oxygen species.
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Objective:To improve the awareness of primary myeloid sarcoma.Methods:The clinical data of one primary myeloid sarcoma patient with first symptom involving nasopharynx who was admitted to the Second Affiliated Hospital of Anhui Medical University in June 2017 were retrospectively analyzed, and the related literature was reviewed.Results:The patient presented with nasal congestion with tinnitus; it took half a year for the myeloid sarcoma diagnosis to be clear with biopsy and immunohistochemical examinations for many times. The patient was treated with chemotherapy regimens for acute myeloid leukemia (AML). Isolated myeloid sarcoma relapsed with skin masses after remission for more than 2 years. Further improvement of the bone marrow-related examinations showed that the bone marrow was not involved, and it was still isolated myeloid sarcoma. The skin mass disappeared completely after the induction chemotherapy of AML. Six months after the recurrence, the patient was in stable condition and was still under follow-up treatment.Conclusions:The primary nasopharyngeal myeloid sarcoma lacks specificity and is easy to be misdiagnosed. The prognosis of isolated myeloid sarcoma is better than that of AML. Isolated recurrence after chemotherapy remission is rare, and usually progresses to the leukemia stage quickly. Early and correct diagnosis is very important for the prognosis, and clinicians should improve the awareness of primary myeloid sarcoma.
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Objective@#To conduct a comparative study on the specific effects of systematic sex education on adolescent students in terms of sexual cognition, sexual values, and sexual adaptation, and to provide the support for conducting a comprehensive education in middle schools.@*Methods@#A whole group sampling method was used to select 3 369 middle and high school students from six general and vocational middle schools in Sichuan Province, which were divided into systematic sex education schools and non systematic sex education schools, and a comparative study was conducted using the Adolescent Mental Health Scale.@*Results@#The results showed that the adolescent students who received systematic sex education were significantly different from those who did not receive systematic sex education in terms of sex related cognition (7.18±6.24, 5.65±7.40), sexual values(7.60±1.17，7.30±1.24), and sexual adjustment (11.49±1.29，11.10±1.41). All differed significantly ( t =5.95, 6.80,7.57, P <0.01). The students who received systematic education in junior middle school were higher than those who received non systematic education in sex related cognition, sexual values and sexual adaptation ( P <0.01). However, in senior high school, the differences in systematic education are only shown in sexual values control and self adaptation in sexual adjustment ( P <0.01). There were significant differences in sex related cognition, sexual values and sexual adjustment between male and female students who received systematic education and non systematic education ( P <0.01).@*Conclusion@#Systematic sex education is more beneficial to the psychosexual health of adolescent students than non systematic sex education in schools.
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Objective:To investigate the predictive value of regional leptomeningeal collateral (rLMC) score on CT angiography (CTA) for the outcome after endovascular therapy in patients with late window anterior circulation stroke.Methods:Patients with acute anterior circulation large vessel occlusive stroke received endovascular treatment 6 to 24 h after onset in the Department of Neurology, the First Affiliated Hospital of Soochow University from January 2018 to July 2021 were enrolled retrospectively. At 3 months after onset, the outcome was evaluated according to the modified Rankin Scale. A score of ≤ 2 was defined as good outcome, and a score of >2 was defined as poor outcome. The clinical data, non-enhanced CT and CTA parameters of the patients were collected. Multivariate logistic analysis was used to determine the independent influencing factors of poor outcomes after endovascular treatment. Results:A total of 74 patients with acute anterior circulation large vessel occlusive stroke treated with endovascular treatment in the late window were enrolled. Their age was 64.41±12.98 years, 43 were males (58.1%). The baseline National Institutes of Health Stroke Scale (NIHSS) score was 13.18±5.22, and the median time from onset to puncture was 527.5 min. Fifty-three patients (71.6%) chose direct thrombectomy and 21 (28.4%) chose intravenous thrombolysis and bridging thrombectomy. Thirty-six patients (48.6%) had a good outcome and 38 (51.4%) had a poor outcome, including 4 (5.4%) died. Univariate analysis showed that there were significant differences in age, atrial fibrillation, fasting blood glucose, NIHSS score, Alberta Stroke Program Early CT Score (ASPECTS), rLMC score and clot burden score between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that higher ASPECTS (odds ratio 0.352, 95% confidence interval 0.157-0.791; P=0.012) and rLMC score (odds ratio 0.550, 95% confidence interval 0.329-0.919; P=0.022) were the independent predictors of good outcomes after endovascular therapy. Conclusion:ASPECTS and rLMC scores were the independent predictors of clinical outcomes after endovascular therapy in patients with late window anterior circulation large vessel occlusive stroke. It had certain guiding value for the decision-making of endovascular treatment in such patients.
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Objective: The objective of this study was to investigate the molecular mechanisms involved in rapamycin-induced inhibition of tumor growth. Materials and Methods: Murine S180 sarcoma cells were subcutaneously injected into mice, and the tumor-bearing mice were randomly divided into three groups (vehicle control, 2 mg/kg rapamycin, and 4 mg/kg rapamycin). The effect of rapamycin on tumor growth was determined by measuring tumor volume. Mammalian target of rapamycin (mTOR), Beclin1, ULK1, LC3, Notch1, CD133, and CD90 expressions was confirmed using confocal microscopy and Western blotting. Results: The tumor growth inhibition rates induced by high-dose and low-dose rapamycin were 48.8% and 30.1%, respectively. Beclin1 and ULK1 expressions and the LC3-II/LC3-I ratio in tumor tissues were altered by rapamycin, whereas mTOR, Notch1, CD133, and CD90 expressions were significantly inhibited by rapamycin in immunofluorescence assays. Western blotting also showed similar results. Conclusion: Tumor growth delay induced by rapamycin may be associated with the suppression of the cancer stem cell phenotype (Notch1, CD133, and CD90) and promotion of autophagy (mTOR, Beclin1, ULK1, and LC3-II/LC3-I ratio) in the murine S180 sarcoma model
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Objective@#Symptomatic intracranial hemorrhage (sICH) is one of the severe complications of ischemic stroke thrombolysis. Several prognostic scales have been developed to predict the risk of sICH. The performance of seven scales was compared in a single center cohort.@*Methods@#Data of patients with consecutive ischemic stroke who received 0.9 mg/kg intravenous recombinant tissue plasminogen activator (rt-PA) thrombolysis within 4.5 h time window from stroke onset were collected. Seven scales that can provide an estimate of risk of sICH were identified and evaluated: Hemorrhage After Thrombolysis (HAT), blood Sugar, Early infarct signs, (hyper) Dense cerebral artery sign, Age, National Institutes of Health (NIH) Stroke Scale (SEDAN), Stroke Prognostication using Age and NIH Stroke Scale (SPAN)-100, Safe Implementation of Thrombolysis in Stroke (SITS), Total Health Risks In Vascular Events (THRIVE), Glucose at presentation, Race (Asia), Age, Sex (male), systolic blood Pressure at presentation, and Severity of stroke at presentation (NIH Stroke Scale; GRASPS) and Multicenter Stroke Survey (MSS). The area under the receiver operating characteristic curve (AUROC) was calculated and Logistic regression and the Hosmer-Lemeshow test were also performed.@*Results@#The current study included 293 patients, of whom 7.85% (23/293) had sICH by National Institute of Neurological Disorders and Stroke (SICHNINDS), 5.46% (16/293) by Europe Cooperative Acute Stroke Study Ⅱ (SICHECASSⅡ) and 4.44% (13/293) by Safe Implementation of Thrombolysis in Stroke (SICHSITS) criteria. SEDAN had the highest AUROC for predicting sICH: sICHNINDS: AUROC=0.843, OR=3.167, 95%CI 2.106-4.762, P<0.01; sICHECASSⅡ: AUROC=0.797, OR=2.509, 95%CI 1.652-3.812, P<0.01; sICHSITS: AUROC=0.784, OR=2.172, 95%CI 1.405-3.357, P<0.01. And SPAN-100 had the lowest AUROC among all the seven scales and was only associated with risk of SICHNINDS in regression analysis. Furthermore, when sub-grouped the cohort into anterior circulation infarction and posterior circulation infarction, regression analysis suggested that all the seven scales were however not associated with sICH risk in patients with posterior circulation infarction.@*Conclusions@#SEDAN constantly had the highest predictive power, SPAN-100 had the worst. The seven scales studied could not predict sICH in posterior circulation infarction.
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Objective@#To synthesis 177Lu-prostate specific membrane antigen (PSMA)-I&T with automated module, evaluate the biodistribution and pharmacokinetics in mice and study the targeting property in human prostate cancer cell line LNCaP Clone FGC.@*Methods@#The iQS-TS automated module was applied in labeling 177Lu-PSMA-I&T. Radiochemical purity and stability were determined with high performance liquid chromatography (HPLC). The biodistribution was observed in normal ICR mice and U-SPECT/CT imaging was performed in LNCaP Clone FGC tumor-bearing mice. Independent-sample t test was used to analyze the data.@*Results@#177Lu-PSMA-I&T was stable in vitro and in vivo, with the radiolabeled yield of (91.5±4.9)% and radiochemical purity >99%. The half maximal inhibitory concentration (IC50) of 177Lu-PSMA-I&T binding to LNCaP Clone FGC cells was (26.74±3.53) nmol/L. The uptake of 177Lu-PSMA-I&T by LNCaP Clone FGC cells increased with time and significantly decreased after the inhibitor addition (t values: 4.301-27.483, all P<0.05). 177Lu-PSMA-I&T was cleared from blood rapidly and predominantly excreted by kidneys. Significant radioactive uptake was observed in tumors with a long retention time.@*Conclusion@#177Lu-PSMA-I&T can be produced in a convenient and efficient procedure using iQS-TS automated module, with good biological properties and excellent affinity and targeting property towards prostate cancer cells, which making it a potential radiopharmaceutical for prostate cancer therapy.
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Objective To investigate the safety and efficacy of 177Lu-prostate specific membrane antigen (PSMA)-617 in the treatment of metastatic castration-resistant prostate cancer (mCRPC).Methods From August 2017 to September 2018,11 patients(average age 70.6 years) with mCRPC who underwent 177Lu-PSMA-617 therapy in Nanjing First Hospital were studied.All patients underwent 68Ga-PSMA-11 PET/CT before therapy to assess the tumor radioactive uptake.Blood routine examination and renal function test results were documented before and after therapy to assess the safety.The efficacy was reflected by the changes of prostate specific antigen (PSA) levels and maximum standardized uptake value (SUVmax) on 68Ga-PSMA-11 PET/CT imaging.Paired t test and Wilcoxon's sign rank test were used to analyze the data.Results No acute side effects were observed after therapy of 177Lu-PSMA-617.There were no statistically significant differences after therapy in WBC counts,RBC counts,and PLT,as well as Hb levels (t values:-0.28-1.11,all P＞ 0.05).No kidney toxicity was found.The PSA level after 177Lu-PSMA-617 therapy was significantly lower than that before therapy (80.70 (14.29,1 538.00) μg/L vs 604.60 (88.41,3 980.00) μg/L;u =59,P =0.023).Of the 11 patients,only 2 had elevated PSA levels and disease progression,while the other 9 patients had varying decreases,of which 2/11 decreased by ＞30％ and 7/11 decreased by ＞50％.After therapy,SUVmax of metastatic lesions and metastatic lymph nodes were decreased in 9 and 2 patients respectively.Conclusions 177Lu-PSMA-617 has a good therapeutic value for mCRPC.It is safe and has no obvious side effects.
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Objective To investigate the effect of silencing troponin Ⅰ3 (Tnni3) gene expression on biological property of rat embryonic H9C2 cardiomyocytes.Methods The rat embryonic H9C2 cardiomyocytes were cultured and divided into 2 groups:control group transfected with negative control small interfering RNA (NC-siRNA group) and experimental group transfected with Tnni3 small interfering RNA (Tnni3-siRNA group).At 48 h,72 h after transfection,the cells were collected,and real time quantitative polymerase chain reaction (qPCR)was used to detect the mRNA expressions of Tnni3 and Caspase-3,and Western blot was used to detect the protein expressions of Tnni3,Cyclin A1 and Cyclin B1.Annexin V-fluorescein isothiocyanate(FITC) apoptosis detection kit was used to analyze cell apoptosis.Cell proliferation was measured by Cell Counting Kit-8 (CCK-8) solution and cell cycle was detected by flow cytometry.Results At 48 h post-transfection with Tnni3-siRNA,H9C2 cells exhibited a significant decrease in Tnni3 mRNA (0.27 ± 0.05 vs.1.00 ± 0.00) and protein (0.18 ± 0.03 vs.1.00 ± 0.00) compared with those transfected with NC-siRNA,and the differences were statistically significant (t =25.26,47.40,all P ＜ 0.01).Apoptotic cells were observed in the NC-siRNA group and the Tnai3-siRNA group.At 72 h post-transfection,the percentage of apoptotic cells significantly increased in H9C2 cells transfected with Tnni3-siRNA [(11.30 ± 1.85) ％ vs.(0.33 ± 0.15) ％] compared with those transfected with NC-siRNA,an increased expression of Caspase-3 mRNA was also observed in Tnni3-siRNA-transfected H9C2 cells (1.39 ±0.13 vs.1.00 ±0.00),and the differences were statistically significant (t =10.24,5.19,all P ＜ 0.01).Compared with NC-siRNA-transfected H9C2 cells,a time-dependent reduction in cell proliferation was observed in Tnni3-siRNA-transfected H9C2 cells (48 h:0.32 ± 0.06 vs.0.46 ± 0.03;72 h:0.31 ± 0.01 vs.0.63 ±0.04;96 h:0.36 ± 0.01 vs 0.75 ± 0.04),and the differences were statistically significant (t =3.62,13.45,16.39,all P ＜ 0.01).At 72 h post-transfection with Tnni3-siRNA,the percentage of G1 phase,S phase and G2 phase cells was (71.25 ± 3.82) ％,(18.28 ± 2.78) ％ and (9.94 ± 1.09) ％,respectively.There was a significant increase in the proportion of G2 phase cells [(9.94 ± 1.09) ％ vs.(4.54 ±0.99) ％] in H9C2 cells transfected with Tnni3-siRNA compared with those transfected with NC-siRNA,an increased expression of Cyclin A1 protein (1.89 ±0.09 vs.1.00 ±0.00) and a decreased expression of Cyclin B1 protein (0.47 ± 0.06 vs.1.00 ± 0.00) were observed in Tnni3-siRNA-transfected H9C2 cells,respectively,and the differences were statistically significant (t =6.35,17.12,15.32,all P＜0.01).Conclusions Silencing Tnni3 gene expression in rat embryonic H9C2 cardiomyocytes can induce cell apoptosis,suppress cell proliferation,and led to G2 cell cycle arrest.
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Objective To investigate the relationship between total cerebral small vessel disease (CSVD) burden and intracranial hemorrhage transformation (HT) after intravenous thrombolysis in patients with acute ischemic stroke (AIS).Methods One hundred and fifty-four patients who suffered from ischemic stroke within 4.5 hours of onset and received recombinant tissue plasminogen activator thrombolytic therapy in the emergency green channel of the First Affiliated Hospital of Soochow University from August 2016 to January 2018 were enrolled.HT examined by computed tomography scan within 24 hours after thrombolysis was included.The magnetic resonance imaging examination was performed within 48 hours.The patients were divided into two groups:HT group and control group according to the presence or absence of HT.Periventricular white-matter hyperintensities (WMH) with Fazekas score of 3 or deep WMH with Fasekas score of 2 or 3 was recorded as 1 point,MRI of cerebral microbleeds (CMBs) or lacunar infarction (LI) was recorded as 1 point respectively,and peripheral vascular space (PVS) in basal ganglia graded 2-4 (≥11)was counted 1 point.Single-factor analysis was used to compare total CSVD burden score,baseline data and clinical data between the two groups.Multivariate Logistic regression analysis was performed to explore the relationship between total CSVD burden score and HT.Results The age of the 154 patients was 66.00(59.00,74.25) years,males accounted for 66.9％ (103/154),onset to treatment time (OTT) was 174.50 (131.50,200.00) minutes and the NIHSS score before thrombolytic therapy was 6.00 (3.00,10.25).There were 43 cases (27.9％) with moderate to severe WMH,35 cases (22.7％) with CMBs,52 cases (33.8％) with PVS graded 2-4,and 96 cases (62.3％) with LI.There were 21 enrolled patients (13.6％) who suffered from HT.Symptomatic intracranial hemorrhage occurred in nine cases (5.8％).In the multivariate Logistic regression model,the results demonstrated that baseline diastolic pressure (OR=1.072,95％CI 1.027-1.118,P=0.001)and atrial fibrillation (OR=28.564,95％CI 6.217-131.241,P=0.000) were independently associated with HT.After using the mild CSVD burden score as a reference,moderate CSVD burden (OR=0.810,95％ CI 0.154-4.257,P=0.804) was not associated with HT after thrombolysis,and severe CSVD burden (OR=8.429,95％ CI 1.643-43.227,P=0.011) was independently associated with HT.Conclusions The severity of total CSVD burden in patients with AIS was closely related to HT after thrombolysis.Severe CSVD was an independent risk factor for HT after thrombolysis.
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Objective@#To investigate the effect of silencing troponin I3 (Tnni3) gene expression on biological property of rat embryonic H9C2 cardiomyocytes.@*Methods@#The rat embryonic H9C2 cardiomyocytes were cultured and divided into 2 groups: control group transfected with negative control small interfering RNA (NC-siRNA group) and experimental group transfected with Tnni3 small interfering RNA (Tnni3-siRNA group). At 48 h, 72 h after transfection, the cells were collected, and real time quantitative polymerase chain reaction (qPCR)was used to detect the mRNA expressions of Tnni3 and Caspase-3, and Western blot was used to detect the protein expressions of Tnni3, Cyclin A1 and Cyclin B1.Annexin V-fluorescein isothiocyanate(FITC) apoptosis detection kit was used to analyze cell apoptosis.Cell proliferation was measured by Cell Counting Kit-8 (CCK-8) solution and cell cycle was detected by flow cytometry.@*Results@#At 48 h post-transfection with Tnni3-siRNA, H9C2 cells exhibited a significant decrease in Tnni3 mRNA (0.27±0.05 vs. 1.00±0.00) and protein (0.18±0.03 vs. 1.00±0.00) compared with those transfected with NC-siRNA, and the differences were statistically significant (t=25.26, 47.40, all P<0.01). Apoptotic cells were observed in the NC-siRNA group and the Tnni3-siRNA group.At 72 h post-transfection, the percentage of apoptotic cells significantly increased in H9C2 cells transfected with Tnni3-siRNA [(11.30±1.85)% vs. (0.33±0.15)%] compared with those transfected with NC-siRNA, an increased expression of Caspase-3 mRNA was also observed in Tnni3-siRNA-transfected H9C2 cells (1.39±0.13 vs. 1.00±0.00), and the differences were statistically significant (t=10.24, 5.19, all P<0.01). Compared with NC-siRNA-transfected H9C2 cells, a time-dependent reduction in cell proliferation was observed in Tnni3-siRNA-transfected H9C2 cells (48 h: 0.32±0.06 vs. 0.46±0.03; 72 h: 0.31±0.01 vs. 0.63±0.04; 96 h: 0.36±0.01 vs 0.75±0.04), and the differences were statistically significant (t=3.62, 13.45, 16.39, all P<0.01). At 72 h post-transfection with Tnni3-siRNA, the percentage of G1 phase, S phase and G2 phase cells was (71.25±3.82)%, (18.28±2.78)% and (9.94±1.09)%, respectively.There was a significant increase in the proportion of G2 phase cells [(9.94±1.09)% vs. (4.54±0.99)%] in H9C2 cells transfected with Tnni3-siRNA compared with those transfected with NC-siRNA, an increased expression of Cyclin A1 protein (1.89±0.09 vs.1.00±0.00) and a decreased expression of Cyclin B1 protein (0.47±0.06 vs.1.00±0.00) were observed in Tnni3-siRNA-transfected H9C2 cells, respectively, and the differences were statistically significant (t=6.35, 17.12, 15.32, all P<0.01).@*Conclusions@#Silencing Tnni3 gene expression in rat embryonic H9C2 cardiomyocytes can induce cell apoptosis, suppress cell proliferation, and led to G2 cell cycle arrest.
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In the development of Shanghai′s health services, tertiary public hospitals play an important role, and their positioning deserves in-depth research.The authors sorted out the relevant laws and policies of the state and Shanghai, clarified the relevant policy basis of tertiary public hospitals in the development of health service industry on the premise of public welfare, and proposed the functional orientation of these hospitals in the industry. Their roles include namely, proper provision of special medical services, focused efforts on the transformation of medical scientific and technological achievements, active cooperation with private medical institutions on contract basis, and promotion of the cluster development of health service industry chain.At the same time, the authors put forward policy suggestions to support the development of health industry in Shanghai′s tertiary public hospitals.
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Homocysteine is a risk factor for ischemic stroke.The association between homocysteine and ischemic stroke has been becoming a hot spot of research all over the world,but has not yet got a consistent result up to now.In this article,we reviewed the pathogenic mechanism,intervention,new treatment strategies,and summarized the current progress of the association of homocysteine with ischemic stroke.
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Objective This study was to observe the meibomian gland scores,the miss rate of meibomian,break-up time of tear film (BUT) changes after long-term wearing orthokeratolog,and to evaluate orthokeratolog impact on meibomian gland function and the stability of the tear film.Methods A prospective case-controlled study was performed.Fifty patients 98 eyes with myopia were enrolled.According to the myopia correction mode,the patients were divided into control group (24 cases) and experimental group (overnight orthokeratolog treatment) group (26 cases).All subjects were underwent the examinations sequentially as follows:evaluation of ocular surface disease symptoms using the Ocular Surface Disease Index (OSDI),lid margin and ocular surface examination by slit lamp microscrope,infrared meibomian photography and BUT.All measurements were repeated at 3 months,6 months,24-months follow-up visit.All patients signed informed consent before examination.Results After wearing orthok-contact lens for 24 months,the meibomian gland score was 4.54±1.10 and 2.29±0.75 in the experimental group and control group,respectively,the difference was statistically significant (P＜0.05).The BUT was (6.35 ± 1.52) s 24 months after wearing orthokeratolog,which was less than (9.38 ± 1.88) s in the control group,the difference was statistically significant (P＜ 0.05).The miss rate of meibomian gland was 24.39 ± 5.54 and 22.81 ± 5.23 in the experimental group and control group,respectively.There was no statistically significant difference among the the miss rate of meibomian gland between groups and within groups (all at P＞0.05).Conclusions Long-term wearing orthokeratolog has negetive effects on meibomian gland function and the stability of the tear film.