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Arrhythmogenic cardiomyopathy (ACM), a fatal heart disease characterized by fibroadipocytic replacement of cardiac myocytes, accounts for 20% of sudden cardiac death and lacks effective treatment. It is often caused by mutations in desmosome proteins, with Desmoglein-2 (DSG2) mutations as a common etiology. However, the mechanism underlying the accumulation of fibrofatty in ACM remains unknown, which impedes the development of curative treatment. Here we investigated the fat accumulation and the underlying mechanism in a mouse model of ACM induced by cardiac-specific knockout of Dsg2 (CS-Dsg2 -/-). Heart failure and cardiac lipid accumulation were observed in CS-Dsg2 -/- mice. We demonstrated that these phenotypes were caused by decline of fatty acid (FA) β-oxidation resulted from impaired mammalian target of rapamycin (mTOR) signaling. Rapamycin worsened while overexpression of mTOR and 4EBP1 rescued the FA β-oxidation pathway in CS-Dsg2 -/- mice. Reactivation of PPARα by fenofibrate or AAV9-Pparα significantly alleviated the lipid accumulation and restored cardiac function. Our results suggest that impaired mTOR-4EBP1-PPARα-dependent FA β-oxidation contributes to myocardial lipid accumulation in ACM and PPARα may be a potential target for curative treatment of ACM.
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Objective:To investigate the prevalence of metabolic syndrome(MS) among adult residents with different characteristics and the relationship between serum uric acid(SUA) level and MS using the data of Chinese Adult Chronic Diseases and Nutrition Surveillance(2018) program in Anhui.Methods:Multi-stage stratified cluster random sampling was used to select participants aged 18 and over for questionnaires, physical measurements, and laboratory tests. The complex weighted method was used to estimate the prevalence of MS among residents with different characteristics. Logistic regression model based on complex sampling data was used to analyze the relationship between SUA and MS. Receiver operating characteristic(ROC) curve was used to evaluate the reliability of SUA in diagnosing MS and determine the optimal cutoff point.Results:A total of 7 182 participants were included and the prevalence of MS among adult residents was 29.46%. The prevalence of MS was higher in females(33.76%) than that in males(25.28%), and the difference was statistically significant( P<0.05). After adjusting for other factors, for every 10 μmol/L increase in SUA, the risk of MS increased by 4% in males( OR=1.040, 95% CI 1.019-1.061) and 7% in females( OR=1.070, 95% CI 1.059-1.082). The area under the curve(AUC) for SUA in diagnosing MS was 0.816(95% CI 0.806-0.826), with a sensitivity of 0.761 and specificity of 0.727. The optimal cutoff point for SUA was 450 μmol/L. Conclusion:The prevalence of MS among adult residents in Anhui Province is 29.46%. SUA is a risk factor for MS, and increasing SUA level indicated a higher risk of MS. The optimal cutoff value of SUA may be helpful in diagnosing MS.
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Objective:To explore the safety and efficacy of continuous pair triangular flap plasty in the treatment of webbed penis.Methods:The clinical data of 5 patients with webbed penis treated by continuous pair triangular flap plasty in Tianjin Baodi Hospital from November 2020 to January 2021 were retrospectively analyzed. The average age was 18.2 (15-24) years old. Physical examination showed a webbed fusion between the ventral skin of the penis and the scrotal skin, the penoscrotal angle disappeared and penile erection was limited. The web extended to or exceeded the middle of the penis in the 5 cases. None of the 5 patients had other penile malformations. Taking the ventral midline of the penis and the junction of the penis and scrotum as the origin, a " Z-shaped" incision was made above and below it respectively, and two triangular flaps with the same size and shape but opposite direction were formed respectively. Then the triangular flaps were switched and sutured.Results:The average operation time of five patients was 13(10-20) minutes. The operative average blood loss was 1.6(1-3)ml. All patients had good wound healing and no surgical complications. Three months after operation, the appearance of the penis was satisfactory and erection was unrestricted.Conclusions:Continuous pair triangular flap plasty overcomes the disadvantages of traditional operation, and has the advantages of simple operation, good operation results and few complications. It is a good choice for the treatment of webbed penis.
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Objective:By exploring the source of children's total fluoride intake and the relationship between children's total fluoride intake and dental fluorosis prevalence, to calculate the benchmark dose (BMD) of children's total fluoride intake and its 95% confidence lower limit (BMDL), and to provide the basis for revision of "Total Fluoride Intake for Inhabitants" (WS/T 87-2016).Methods:The villages that had water improvement for 5 years and more in 6 provinces of Jiangsu, Shandong, Hebei, Anhui, Henan and Shaanxi were selected as survey sites in April 2014. The water fluoride content of these villages was 0.3 - 3.0 mg/L, tap water samples from the centralized water supply were collected, and fluoride content was detected by ion selective electrode method. Children aged 8 to 12 years were selected, children's dental fluorosis was checked by Dean's method. Children's dietary and drinking water volume were surveyed by duplicate portion study (measurement for 3 d), and dietary fluoride content was detected according to the "Method for Determination of Fluorine in Foods". The mean and standard deviation of drinking water fluoride intake, dietary fluoride intake, and total fluoride intake were measured. According to the dose-response relationship between children's total fluoride intake and the detection rate of dental fluorosis, BMD and BMDL were calculated, and the reference dose (RfD) was calculated based on BMDL.Results:The mean of water fluoride of all 29 villages was 1.26 mg/L (from 0.41 to 2.85 mg/L). Totally 3 043 children aged 8 to 12 years were checked, and the detection rate of dental fluorosis was 30.2% (919/3 034). The lowest detection rate of dental fluorosis was 2.0% (2/100) and the highest was 71.4% (30/42) in the 29 villages. The children's dietary and drinking water volume of 769 person-time aged 8 to 12 years were surveyed, the children's daily drinking water fluoride intake, dietary fluoride intake, and total fluoride intake were (0.83 ± 0.66), (1.13 ± 0.61) and (1.96 ± 0.89) mg/d, respectively. The BMD of children's daily total fluoride intake was 2.43 mg, the BMDL was 2.21 mg, and the RfD was 2.21 mg.Conclusion:The BMD of 8 to 12 years old children's daily total fluoride intake is the same as the allowable limit (2.4 mg) of the national standard "Total Fluoride Intake for Inhabitants" (WS/T 87-2016).
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Objective:To compare the effects of Dynesys dynamic fixation and fusion fixation on the sagittal parameters of lumbar degenerative diseases of 2-4 segments.Methods:A total of 45 patients with 2-4 segments lumbar degenerative diseases, who underwent decompression and internal fixation in our hospital from March 2014 to July 2019, were retrospectively analyzed. There were 23 males and 22 females with an average age of 55±18 years (range 15-86 years). Dynesys dynamic fixation was performed in 22 cases and fusion fixation in 23 cases. Lumbar anteroposterior, flexion and extension lateral and standing full-length spine radiographs were taken before surgery, 2 weeks postoperatively, 3 months postoperatively and at the last follow-up. Sagittal alignment parameters were measured on those radiographs and compared between two groups. Lumbar parameters included lumbar lordosis (LL), lordosis of the fixed segments (LFx), lordosis of the adjacent level to fixed segment (LAdj) and lumbar range of motion (ROM). Pelvic parameters included pelvic incidence (PI), pelvic tilt (PT) and sacral slope (SS).Results:All cases were followed up for 12-50 months with an average of 18.5 months. There was no significant intra- and inter-group difference of LL value between Dynesys dynamic fixation and fusion fixation group before and after surgery ( P>0.05). The overall difference of LFx between groups was not statistically significant ( F=0.700, P=0.406). There was statistically significant variance between time points ( F=7.960, P<0.001) and an interaction effect between group and time ( F=3.940, P=0.006). The LFx of the fusion fixation group was more than that of the dynamic fixation group at 3 months postoperatively and at the last follow-up ( P<0.05). The difference of LAdj value between the two groups was not statistically significant ( F=0.520, P=0.476), while the difference between time points was significant ( F=4.810, P=0.002) with interaction effect between group and time ( F=3.560, P=0.010). Postoperative ROM values of the two groups showed statistically significant differences in group effect, time effect and interaction effect, respectively ( F=4.770, P=0.034; F=18.510, P<0.001; F=5.940, P=0.002). Dynamic fixation group's ROM was more than that of the fusion fixation group at 3 months postoperatively and at the last follow-up ( P<0.05). There was no significant difference of PI between groups and between different time points ( F=0.580, P=0.451; F=0.750; P=0.477). There was no interaction effect in PI between group and time ( F=0.120, P=0.886). There was no significant difference of PT and SS between two groups ( F=0.320, P=0.576; F=0.020, P=0.901). Both time effect and interaction effect were statistically significant ( P<0.05). One complication of unilateral S 1 screw loosening happened in one patient with dynamic fixed at 2 years after surgery. There was no complication in the fusion fixation group during the follow-up. Conclusion:Both Dynesys dynamic fixation and fusion fixation can effectively reconstruct the lumbar lordosis. The fixed segments' lordosis of Dynesys was less than that of fusion fixation from 3 months after operation. Theoretically, the similar increasing trends of LAdj after fixation implied that the degeneration of adjacent segments may occur in both fixation systems. The dynamic fixation can retain more ROM than the fusion fixation. Both two fixation systems have similar influence on the pelvic parameters.
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Based on the statistics of 350 technical evaluations of changes in licensing items of class Ⅱ passive and active medical devices completed in Henan province from July 2017 to November 2018, this paper summarized and analysed the common problems and requirements listed in the correction notifications of the technical evaluation, and put forward relevant countermeasures or suggestions, with a view to further speeding up the evaluation and approval of medical devices.
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Equipment and Supplies , Reference Standards , Government Regulation , LicensureABSTRACT
To analyze the changes of early procalcitonin (PCT) and hypersensitive C-reactive protein (hs-CRP) in patients with acute cerebral infarction, and to explore the predictive value of both for acute cerebral infarction with infection. Methods 206 acute cerebral infarction patients admitted to the department of neurology of Feicheng Mining Center Hospital from May 2014 to May 2019 were enrolled. Clinical data of patients and serum PCT and hs-CRP levels at 24, 48 and 72 hours after onset were collected. Patients were divided into infected group (n = 69) and non-infected group (n = 137) according to whether infection occurred within 5 days after onset. And 60 healthy people in the same period were selected as the healthy control group. The trends of serum PCT and hs-CRP levels in each group were analyzed. The receiver operating characteristic (ROC) curve was used to analyze the values of serum PCT and hs-CRP levels in identifying acute cerebral infarction with infection. Results The serum level of PCT at 24, 48 and 72 hours in the infected group and the non-infected group were significantly higher than those in the healthy control group, and the serum level of PCT at 48 hours and 72 hours in the infected group were significantly higher than those in the non-infected group (μg/L: 0.28±0.08 vs. 0.19±0.03, 0.31±0.07 vs. 0.15±0.06, both P < 0.05). Compared with 24 hours, the serum PCT level in the infected group at 48 hours and 72 hours were significantly increased, but decreased in the non-infected group. The serum hs-CRP level in the infected group at 24, 48 and 72 hours were significantly higher than those in the non-infected group and the healthy control group (mg/L: 5.86±1.73 vs. 5.45±1.08, 5.25±1.33; 8.01±2.41 vs. 5.67±2.13, 5.25±1.33; 14.25±2.19 vs. 12.30±1.87, 5.25±1.33; all P < 0.05). And the serum hs-CRP level in the non-infected group at 72 hours was significantly higher than that in the healthy control group. Compared with 24 hours, the serum hs-CRP level in the infected group and non-infected group at 48 hours and 72 hours were significantly increased. It was shown by ROC curve analysis that serum PCT and hs-CRP levels at 24 hours had no predictive value for infection in patients with acute cerebral infarction [area under ROC curve (AUC) was 0.440, 0.576 respectively, both P > 0.05]. At 48 hours, the AUC of serum PCT in diagnosis of acute cerebral infarction with infection was 0.850 [95% confidence interval (95%CI) = 0.784-0.916], the sensitivity and specificity were 66.7% and 97.8% when the cut-off of PCT was 0.25 μg/L; the AUC of serum hs-CRP was 0.759 (95%CI = 0.689-0.830), the sensitivity and specificity were 66.7% and 76.6% when the cut-off of hs-CRP was 6.80 mg/L; the AUC of PCT combined with hs-CRP was 0.911 (95%CI = 0.859-0.964), the sensitivity was 90.5%, the specificity was 86.9%. At 72 hours, the AUC of serum PCT in diagnosis of acute cerebral infarction with infection was 0.952 (95%CI = 0.916-0.989), the sensitivity and specificity were 89.9% and 93.4% when the cut-off of PCT was 0.23 μg/L; the AUC of serum hs-CRP was 0.753 (95%CI = 0.678-0.828), the sensitivity and specificity were 60.9% and 83.2% when the cut-off of hs-CRP was 14.01 mg/L; the AUC of PCT combined with hs-CRP was 0.954 (95%CI = 0.918-0.991), the sensitivity was 97.1%, and the specificity was 89.8%. The results showed that the diagnostic value of serum PCT at 48 hours and 72 hours were higher than those of hs-CRP, and the predictive value of PCT combined with hs-CRP was higher than those of single index. Conclusion Acute cerebral infarction itself has an effect on serum PCT level; serum PCT level above 0.23 μg/L at 72 hours after onset and reference to serum hs-CRP level have a high predictive value for the diagnosis of infection in patients with acute cerebral infarction.
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Objective@#To evaluate the accuracy of transarticular screw fixation using intraoperative three-dimensional fluoroscopy-based navigation (ITFN) and to evaluate the clinical outcomes of this treatment method.@*Methods@#Data of 56 patients(26 males and 30 females) with atlantoaxial instability who were treated by C1, 2 transarticular screw fixation using ITFN from November 2005 to October 2015 were retrospectively analyzed. The mean age of the patients was 44.5 years (range, 9-68 years). There were 44 cases with congenital malformation, 4 with old odontoid fracture, 7 with spontaneous dislocation, and 1 with rheumatoid arthritis. C2 isthmus width and height were measured on preoperatively obtained CT scans, and screw positioning was evaluated on postoperatively obtained CT scans, and classified into three types: ideal position (type I), acceptable position (type II) and unacceptable position (type III). A novel grading system is proposed based on previous study and grading system, and the difficulty of placing C1, 2 transarticular screw using ITFN was classified into three types: easy (total score 0), median (total score 1) and hard (total score 2, 3). Pain scores were assessed using the visual analogue scale. Myelopathy was assessed using the Nurick scale and Odom’s criteria.@*Results@#The isthmus width was 5.46±1.86 mm on the right side and 5.38±1.36 mm on the left side. The isthmus height was 4.89±1.33 mm on the right side and 4.97±1.17 mm on the left side. According to the grading system, 78, 11, and 23 of the sides were classified into easy, median and hard groups respectively. One hundred and seven transarticular screws were placed in 56 patients, and 71.03% of which were ideal screws, and 28.97% were acceptable screws. Five patients had unilateral screws placed. There was no significant difference in screw positioning among the three groups (χ2=0.46, 0.54, 1.18; P=0.50, 0.46,0.28). The mean follow-up period was 44.7 months (range, 6-120 months). At the latest follow-up, according to Nurick score, there are 30 patients scoring 0, 25 patients scoring 1, and 1 patient scoring 2. According to Odom’s criteria, outcomes were as follows: excellent, 66.1%; good, 26.8%; fair, 7.1%; and poor, 0%. All patients with preoperative neck pain had symptom relief or improvement, with more than 89.33% improvement in visual analogue scale scores. No dural laceration, injury to the vertebral artery, spinal cord, or hypoglossal nerve were noted.@*Conclusion@#ITFN is a safe, accurate, and effective tool for transarticular screw placement in patients with atlantoaxial instability.
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Objective To explore the iodine nutritional status of people after adjusting iodine content in iodized salt in Anhui Province. Methods In 2014, 30 counties (cities, districts) were selected in Anhui Province according to the probability ratio sampling method (PPS), and one school was selected in each county (city, district), and 50 children aged 8 - 10 years were selected in each school. Urine and household salt samples were collected, urine and salt iodine levels were tested, and thyroid was examined using B-ultrasound. In the township where the school was located, 20 pregnant women's urine samples and household salt samples were collected for urine and salt iodine detection. The salt iodine level was measured by direct titration, Chuan salt and other fortified salt iodine levels were determined by arbitration method (GB/T 13025.7-2012). Urinary iodine was determined by arsenic-cerium catalytic spectrophotometry (WS/T 107-2006). Results A total of 1575 children's household salt samples were collected, the median salt iodine was 23.77 mg/kg, the qualified iodized salt consumption rate was 93.65%(1475/1575); 600 household salt samples were collected from pregnant women, the salt iodine median was 23.50 mg/kg, the consumption rate of qualified iodine salt was 96.33%( 578/600 ) . A total of 1575 urine samples were collected from children, the median urinary iodine was 242.20 μg/L; six hundred urine samples were collected from pregnant women, and the median urinary iodine was 158.15μg/L, 43.33%(13/30) of the counties (cities, districts) pregnant wowen median urinary iodine < 150 μg/L. A total of 1575 thyroid glands were examined in children aged 8 to 10 years, and the thyroid enlargement rate was 3.75% (59/1575). Conclusion The iodine nutrition of children aged 8 - 10 years in Anhui Province is higher than the appropriate level, but the iodine is not in the excessive state, and the iodine nutrition of pregnant women in some counties (cities, districts) is insufficient.
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AIM:To screen and verify the effective ingredient of traditional Chines medicine (TCM) Q0409 in improving learning and memory ability of mice.METHODS:The mouse learning and memory impairment model was induced by intraperitoneal injection of scopolamine.The mice in each group were given the corresponding drug by gavage at the same time for 14 d in succession.Morris water maze test was used to measure the learning and memory ability of the mice, and then the hippocampal tissue homogenate was taken to determine the activity of acetylcholinesterase (AChE).The animals were divided into 8 groups according to L8(27) orthogonal table.The variance analysis and factorial analysis were used to analyze the pharmacological effects of seven kinds of single herbal in TCM Q0409 and determine the screening ingredients.The animals were divided into 6 groups according to the results of the preliminary screening results, and further testing and validation of TCM Q0409 screening ingredients were performed to get the final simplified ingredients.RESULTS:Three medicinal herbs of Polygalae, Panax ginseng and Acori graminei rhizome were screened by the orthogonal results of Morris water maze test and the activity of AChE in mouse hippocampal tissues.The simplified ingredients of TCM Q0409 were obtained through the variance results of Morris water maze test and the activity of AChE in mouse hippocampal tissues.CONCLUSION:Polygala and ginseng were eventually determined as simplified ingredients of TCM Q0409 and it was verified that they improve the learning and memory ability of the mice with learning and memory impairment.
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Protein arginine methyltransferases ( PRMTs) play crucial roles in the methylation of a series pro-tein substrates.PRMT5 is a type Ⅱ methyltransferase that symmetrically methylates arginine residues of histone and non-histone substrates, thereby regulating a variety of cellular processes through epigenetic control of target gene expression or post-translational modification of signaling molecules.Recently, accumulated evidence has suggested that PRMT5 may function as an oncogene.This review is aimed to summarize the oncogenic role of PRMT5 and its regulatory mechanisms in tumors.
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AIM: To observe the effects of berberine (Ber) on enterocyte apoptosis in septic mice and its pos-sible mechanism.METHODS: Male C57BL/6 mice (8 ~10 weeks old) were randomly divided into sham group, cecal ligation and puncture (CLP) group, CLP +Ber group and sham +Ber group.The mice in CLP group underwent CLP ope-ration, and the mice in sham groups suffered a similar operation except the ligation and puncture.After the sham or CLP operation, the mice were administered intragastrically with distilled water or berberine (50 mg/kg) within 2 h.After 20 h, the mice were killed with excess pentobarbital sodium and the ileum tissues were removed.The histological changes of the intestine were observed and the enterocyte apoptosis was examined by determining the protein level of cleaved caspase-3. Furthermore, mitochondrial Bax, cytoplasm cytochrome C (Cyt C) and the total proteins of Bcl-2, Fas, FasL and Fas-as-sociated protein with death domain (FADD) were examined by Western blot.The mRNA expression of tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) was measured by real-time PCR.RESULTS: The extensive ileum injuries, including remarkably increased leukocytes and necrosis of intestinal villus were observed 20 h after CLP.In CLP group, the protein levels of cleaved caspase-3, cytoplasm Cyt C, as well as Fas, FasL were significantly increased, but the Bcl-2 level was decreased.Bax translocation into mitochondria was promoted.However, FADD was not changed significantly.The mRNA expression of TH and DBH was also increased sharply in CLP group.On the contrary, treatment with berberine made a considerable alleviating alteration in the ileum of the septic mice.CONCLUSION: Treatment with berberine pro-vides protective effects on intestinal injury in septic mice by reducing enterocyte apoptosis, and its possible mechanism may be involved in the inhibition of the endogenous and exogenous apoptosis pathways.
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Objective To investigate the effect of decompression with long-segment (L) or short-segment (S) fusion on the outcomes of the surgical treatment for degenerative adult scoliosis (ADS) patiens.Methods A retrospective study on 32 patients treated in our department for ADS from April 2013 to May 2015 was carried out, including 12 male and 20 female (1 : 1.7).Their average age was 66.4 (range: 51-77 years).All patients underwent decompression and fusion surgeries through posterior approach.They were divided into long-segment fusion group (L) and short-segment fusion group (S) according to fusion range.During follow-ups (FU), clinical outcomes were assessed by means of visual analog scale (VAS) and Oswestry disability index (ODI).Radiographic evaluation on full-length standing film included coronal Cobb's angle, distant between C7plumb line and center sacral vertical line (C7PL-CSVL), thoracic kyphosis (TK) angle, lumbar lordosis (LL) angle, pelvic incidence (PI), and pelvic tilt (PT), PI-LL, sagittal vertical axis(SVA).Postoperative complications were also recorded.Results All patients were followed up for average 2.2 years (range:1.5-3.5 years).No significant difference of age or gender was found between two groups (L: 22, S:10) of patients (P=0.066, 0.182).As for the fusion segments, group L (6.3±1.5) was more than group S (2.9±0.3) (P=0.001).Operation time and blood loss of group L were statistically more than group S (P<0.05).Postoperative VAS sores of back pain and leg pain as well as ODI were all improved significantly in two groups (P<0.05).At the final FU, back pain VAS was more in group L than in group S (P<0.05) , but no significant difference was found in leg pain VAS between two groups (P>0.05);at the final FU, group L's ODI showed better functional recovery than group S's[(12.8±9.3)% vs.(25.4±11.4)%, P<0.05].With no obvious difference in the two groups (P>0.05), coronal Cobb's angle corrected more satisfactional in group L than group S (P<0.05).The same situation was found in C7-CSVL correction in two goup in FU (P<0.05).Sagital balance was restored to normal alignment better in group L than group S, with increase of lumbar lordosis after surgery.The overall incidence of postoperative complications was 31.3%, including wound infection, cerebrospinal fluid leakage, transient neurological symptoms and internal fixed rod breakage, more common in group L than group S.Conclusions Decompression and fusion with internal fixation showed good clinical outcomes in the treatment of ADS.Long-segment fusion yielded better coronal and sagittal correction outcomes with higher peri-operation risks;however, short-segment fusion showed higher safety with relatively inferior correction effect.Appropriate fusion mode should be chose according to the patient's deformity features.
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AIM:To observe the effect of B-HT933, a selective α2-adrenoceptor agonist, on lipopolysaccha-ride ( LPS )-induced TNF-αproduction in neonatal rat cardiomyocytes and to explore the underlying mechanisms . METHODS:The neonatal rat cardiomyocytes were cultured .The localization of α2A-adrenoceptor in the cardiomyocytes was examined by immunofluorescence staining .The cardiomyocytes were exposed to LPS or/and B-HT933 for different time.The level of TNF-αin the supernatants and the mRNA expression of TNF-αwere detected by ELISA and real-time PCR, respectively.In addition, LPS-associated signal molecules in the cardiomyocytes were also examined by Western blotting.RESULTS: Immunofluorescence staining showed that α2A-adrenoceptors were localized in the cardiomyocytes . LPS stimulated TNF-αproduction in the cardiomyocytes in a dose and time-dependent manner .B-HT933 pretreatment sig-nificantly inhibited the expression of TNF-αat mRNA and protein levels in LPS-treated cardiomyocytes .Furthermore, LPS exposure induced IκBαand p38 phosphorylation in cardiomyocytes and only IκBαphosphorylation was prevented by B-HT933 treatment.CONCLUSION:α2A-adrenoceptors are present in neonatal rat cardiomyocytes and its agonist B -HT933 inhibits LPS-induced TNF-αproduction in cardiomyocytes via suppressing IκBαphosphorylation .
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AIM:To study the expression of glycine receptorα1 subunit in neonatal rat myocardial cells and to investigate the effect of lipopolysaccharide (LPS), hypoxia/reoxygenation, isoproterenol (ISO) and high concentration of glucose (HG) on the expression of glycine receptorα1 subunit in the neonatal rat myocardial cells.METHODS:Neonatal rat myocardial cells were cultured in vitro.The expression of glycine receptorα1 subunit was detected by Western blotting. The neonatal rat myocardial cells were treated with LPS (20 mg/L), ISO (100 μmol/L) or high concentration of glucose (25 mmol/L) for 24 h, or were exposed to hypoxia for 3 h followed by reoxygenation for 3 h.Subsequently, the cell viabil-ity was measured by CCK-8 assay, and the expression of glycine receptorα1 subunit was determined by Western blotting. RESULTS:The expression of glycine receptor α1 subunit in the neonatal rat myocardial cells was positively detectable by Western blotting.Compared with control group, no significant difference of the cell viability ( P>0.05) in LPS group, ISO group, hypoxia/reoxygenation group and HG group was observed.The expression of glycine receptor α1 subunit was in-creased (P<0.01) in LPS group, ISO group and hypoxia/reoxygenatio group, but decreased (P<0.01) in HG group. CONCLUSION:Glycine receptorα1 subunit exists in the neonatal rat myocardial cells.A certain concentration of LPS or ISO, or hypoxia/reoxygenation for a certain period upregulate the expression of glycine receptorα1 subunit, but HG down-regulates the expression of glycine receptor α1 subunit in cultured neonatal rat myocardial cells.
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[ ABSTRACT] AIM:To observe the effects of berberine and yohimbine on splenocyte apoptosis in septic mice and underlying mechanisms.METHODS:The mice were subjected to cecal ligature and puncture ( CLP) .The drugs or vehi-cle were given intragastrically 2 h after the surgery according to the following 5 groups:sham, CLP, CLP+berberine, CLP+yohimbine, and CLP+berberine+yohimbine.The apoptosis of splenocytes stained by TUNEL was observed under laser scanning confocal microscope 20 h after CLP.The splenic lymphocytes were isolated and observed using flow cytometry. The activities of caspase-3, caspase-8 and caspase-9 in splenic lymphocytes were detected, and the expression of Fas, Bim, Bcl-2 and Bax in the splenocytes was also determined by Western blotting.RESULTS:The TUNEL staining showed that the apoptotic rate of the splenocytes in septic mice 20 h after CLP was significantly higher than that in sham and CLP+yohimbine groups (P<0.05).Compared with CLP group, the proportion of apoptotic cells was decreased in septic mice in CLP+berberine+yohimbine and CLP+yohimbine groups ( P<0.05) .Flow cytometry analysis demonstrated the similar results in the apoptosis of splenocytes and T lymphocytes.However, only yohimbine treatment reduced the apoptosis of B lymphocytes in the spleen of sepsis-challenged mice.Compared with CLP group, caspase-9 activity was significantly re-duced in CLP+berberine group (P<0.05), the activities of caspase-3, caspase-8 and caspase-9 were all statistically re-duced (P<0.05) in CLP+yohimbine group and CLP+yohimbine+berberine group.CLP significantly increased the ex-pression of cytosolic Fas, Bim and mitochondrial Bax in the splenocytes, and decreased Bcl-2 expression compared with sham group.Compared with CLP group, the expression of cytosolic Bim and mitochondrial Bax in CLP+berberine group were reduced (P<0.05).Fas expression decreased only in CLP+yohimbine group (P<0.05).Berberine combined with yohimbine reduced the expression of cytosolic Fas, Bim and mitochondrial Bax in the septic mouse splenocytes ( P <0.05).CONCLUSION:Yohimbine reduces sepsis-induced splenic lymphocyte apoptosis in mice by inhibiting Fas expres-sion and in turn blocking both extrinsic and intrinsic apoptosis pathways.Berberine reduces Bim expression and inhibits caspase-9 activation, but not caspase-3 activation and apoptosis in the septic mouse splenocytes.Berberine combined with yohimbine reduces splenocyte apoptosis in the septic mice by inhibiting both extrinsic and intrinsic apoptotic pathways.
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AIM:To investigate the effects of thalidomide ( THD) on the activation of connective tissue growth factor ( CTGF) gene promoter induced by transforming growth factor β1 ( TGF-β1 ) in human embryonic lung fibroblasts ( HELF) .METHODS:DNA sequence of CTGF gene promoter was cloned into luciferase reporter gene vector to construct the recombinant eukaryotic expression vector pGL 3-CTGFP, and the recombinant vector was transfected into HELF cell line.The effects of TGF-β1 and THD on the activation of CTGF gene promoter were detected by dual-luciferase analysis . RESULTS:TGF-β1 increased the reporter gene activity dose-dependently (P<0.05), with a plateau at 5 μg/L being 2.16 folds as high as the control .TGF-β1-induced increase in the reporter gene activity was also time-dependent ( P<0.05).After exposure to TGF-β1(5 μg/L), the level of luciferase activity reached its peak at 12 h and was 2.52 folds as high as the control .THD significantly inhibited TGF-β1-induced increase in the reporter gene activity in a dose-dependent manner , but its basal activity was not changed .CONCLUSION: TGF-β1 stimulates the transcriptional activity of CTGF gene promoter in HELF cells in a dose-and time-dependent manner , while THD may inhibit the effects dose-dependently .
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A real-time polymerase chain reaction ( PCR ) micro total analysis system (μ-TAS ) integrated nucleic acid extraction, PCR amplification and real-time-fluorescent PCR detection on a same microfluidic chip was prepared for the fully automated and on-chip analysis of nucleic acid. The proposed method had the advantage such as low sample consumption, fast analysis and simple operation and so on. Micromachining technology was used to fabricate the anodic molds of integrated nucleic acid extraction microfluidic chip. A polydimethylsiloxane (PDMS) substrate with 3D channels was manufactured by a combination of molds and an injection molding method. The glass substrate and the chip were bonded together using a plasma treatment. The μ-TAS included a microfluidic control device whose micro fluidic velocity ( 0-10 mL/min ) could be adjusted, a TEC platform which the precision of temperature control was 0. 1℃ and a CCD detection module. The DNA of human blood was extracted by using a silica gel membrane method on the microfluidic chip. The processes of DNA extraction and detection were preset in the μ-TAS. Human blood lysate ( 20 μL ) were driven to the extraction chamber and was then washed. The fluidic drive speed was 2 mL/min. DNA and PCR reagents were mixed and then were driven into the PCR chamber. The fluidic drive speed was 1 mL/min. The GAPDH gene in extracted genome DNA was amplified by PCR and detected. The amplified product was verified by melting analysis. The results of nucleic acid extraction method on the chip were compared to those obtained using a standard manual centrifuge extraction method. The amplification curves were obvious. Ct values of the chip method were 25 . 3 and 26 . 9 . The denaturation temperature of all the melting was 89 . 9 ℃. The results validated that the chip-based method and device realized the extraction of nucleic acid, amplification and detection automatically.
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AIM:To explore the preliminary mechanism of senegenin ( Sen) on inhibiting hypoxia/reoxygenation ( H/R)-induced apoptosis of primary cortical neurons .METHODS:The cultured cortical neurons were randomly divided in-to normal group (control group), model group (H/R group), Sen+H/R group and Sen group.Flow cytometry was used to evaluate the effect of Sen on H/R-induced cell apoptosis .The protein levels of JNK , p-JNK, c-Jun, p-c-Jun, Bcl-2 and Bax were assessed by Western blotting .RESULTS:The apoptotic rate in H/R group was obviously higher than that in control group (P<0.05), while the apoptotic rate in Sen +H/R group was obviously lower than that in H/R group (P<0.05), suggesting that the model of apoptosis was established successfully .The results of Western blotting showed that Sen increased the expression of JNK and c-Jun, inhibited the phosphorylation of JNK and c-Jun (P<0.05), increased the protein level of Bcl-2 and inhibited the protein level of Bax in H/R treated primary cortical neurons (P<0.05).CONCLUSION:Sen has a protective effect against H/R-induced neuronal apoptosis by increasing the expression of JNK and c-Jun, inhibiting the phosphorylation of JNK and c-Jun, increasing the protein level of Bcl-2 and decreasing the protein level of Bax .
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Objective To evaluate the protective effect of perfluorocarbon emulsions (FCE) on donor lung of rats during storage.Methods Twenty-four healthy male Sprague-Dawley rats,weighing 350-400 g,were equally and randomly divided into 2 groups using a random number table:University of Wisconsin (UW) solution group (UW group) and FCE group (FCE group).After the model of lung perfusion was established according to the method described by Fischer et al,the lung and heart were removed and perfused with 4 ℃ UW or FCE preservation solutions.The lung was taken out when stored for 6 h for determination of SOD activity (by WST assay),malondialdehyde (MDA) content (by TBA assay),and activity of myeloperoxidase (MPO) and content of interleukin1 β (IL-1β),IL-6,tumor necrosis factor-apha (TNF-α) (using ELISA) in lung tissues and for microscopic examination of pathologic changes.Results MPO activity was significantly lower in UW group than in FCE group (P <0.05).There were no significant differences in the SOD activity and content of MDA,IL-1β,IL-6,TNF-α between the two groups (P > 0.05).Conclusion FCE can reduce the neutrophil infiltration in lung tissues,indicating that FCE is more superior to UW solution in reduction of injury to the donor lung of rats.