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Objective:To evaluate the application value of patient-based real-time quality control (PBRTQC) algorithms in intralaboratory comparison between various hematology analyzers.Method:From April 1 st 2020 to March 31 th 2021, data of white blood cell (WBC) counts and daily comparison results of fresh venous blood, measured by five hematology analyzers, were collected at the Department of Laboratory Medicine in Hebei Children′s Hospital. First, the professional intelligent PBRTQC software system was applied to conduct the parameter setting, program establishment, and performance verification. Three concentration ranges of WBC were selected, low concentration (2.5-4.5)×10 9/L, medium concentration (6.0-8.0)×10 9/L and high concentration (12.0-14.0)×10 9/L for the comparison. Next, WBC counts were calculated with both of the EWMA and median methods, the results were then analyzed by PBRTQC using the module of"intralaboratory comparison of hematology analyzers". Finally, bias of intralaboratory comparison among various hematology analyzers analyzed by means of EWMA and daily comparison results of fresh venous blood were compared. Based on the standard of WS/T 406-2012,allowable error ±7.50% in WBC counts was set as the relative bias standard among different instruments. Results:(1) A total of 38 313 sample results were included, there were 70 warning results out of these samples based on the EWMA quality control method established on the data of patients with white blood cell count in our laboratory, with an early warning rate of 0.183‰, a probability of error detection of 100%, and a probability of false loss of control of 0. EWMA quality control efficiency met the quality objectives. (2) In the comparison monitoring of the results of 5 blood cell analyzers at high concentrations, the coincidence rate between EWMA and median method were both 100% (46/46) in weekly and monthly comparison, and EWMA could maintain a relatively stable monitoring efficiency in daily comparison. (3) In the selected natural month, the consistency rate between EWMA method and fresh blood comparison method was 95.24% (20/21).Conclusion:PBRTQC can be used as a valuable supplementary tool of IQC to continuously and effectively monitor the consistency of data derived from intralaboratory hematology analyzers with different bands and types, which can not only reduce the risk of quality and operating costs, but also improve the efficiency of laboratory management.
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Objective:To investigate the effect of interleukin-33 (IL-33) on lipopolysaccharide (LPS)-induced permeability of rat cardiac microvascular endothelial cells (RCMECs).Methods:RCMECs were cultured in vitro to be divided into control group, LPS group, IL-33 group and LPS+IL-33 group. The effect of IL-33 on the proliferation of RCMECs was detected by cell counting reagent (CCK8). Fluorescein isothiocyanate (FITC)-dextran assay was used to evaluate the permeability of RCMECs. The expression of vascular endothelial calmodulin, ras homologous gene family (Rho) member A (RhoA) and phosphorylated Rho-associated coiled-coil-containing protein kinase (p-ROCK2) proteins were tested by western blot. High-throughput sequencing and gene ontology (GO) were performed for gene expression in LPS and LPS+IL-33 groups.Results:No significant effect of IL-33 at 10-50 ng/ml on the proliferation of RCMECs was observed ( P>0.05). Compared with the control group, the permeability of RCMECs (permeability coefficient ratio 1.404±0.029 vs. 1.000±0.200, P<0.05) was significantly increased in LPS group and the expression of vascular endothelial calmodulin (relative gray value 0.429 5±0.012 9 vs. 0.594 9±0.014 2, P<0.05) was down-regulated, while the permeability of monolayers (permeability coefficient ratio, 0.948±0.013, P<0.01) was decreased in LPS+IL-33 group and the expression of vascular endothelial calmodulin (relative grayscale value 0.549 1±0.012 0, P<0.005) was up-regulated compared with the LPS group. High-throughput sequencing data revealed that the differential genes downregulated in the LPS and LPS+IL-33 groups were associated with cytoskeleton and Rho signaling pathway. Compared with the control group, RhoA (relative gray value 0.211 4±0.009 9 vs. 0.135 0±0.007 6, P<0.000 1) and p-ROCK (relative gray value 0.656 3±0.013 2 vs. 0.503 6±0.036 2, P<0.000 1) protein expression was upregulated in the LPS group. When compared with LPS group, RhoA (relative gray value 0.157 7±0.010 7, P=0.000 2), p-ROCK (relative gray value 0.427 7±0.003 8, P<0.000 1) protein expression was decreased in LPS+IL-33 group. Conclusion:IL-33 may improve LPS-induced hyperpermeability of RCMECs by inhibiting RhoA and p-ROCK protein expression in Rho/Rho-associated coiled-coil-containing protein kinase signaling pathway.
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Vanin-1 is an amidohydrolase that catalyses the conversion of pantetheine into the amino-thiol cysteamine and pantothenic acid (coenzyme A precursor), which plays a vital role in multiple physiological and pathological processes. In this study, an enzyme-activated near-infrared (NIR) fluorescent probe (DDAV) has been constructed for sensitively detecting Vanin-1 activity in complicated biosamples on the basis of its catalytic characteristics. DDAV exhibited a high selectivity and sensitivity toward Vanin-1 and was successfully applied to the early diagnosis of kidney injury in cisplatin-induced kidney injury model. In addition, DDAV could serve as a visual tool for in situ imaging endogenous Vanin-1 in vivo. More importantly, Enterococcus faecalis 20247 which possessed high expression of Vanin-1 was screened out from intestinal bacteria using DDAV, provided useful guidance for the rational use of NSAIDs in clinic. Finally, oleuropein as a potent natural inhibitor for Vanin-1 was discovered from herbal medicines library using a high-throughput screening method using DDAV, which held great promise for clinical therapy of inflammatory bowel disease.
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Objective:To study the characteristics and management of peripancreatic effusion in chronic pancreatitis.Methods:Data of 32 patients with chronic pancreatitis and 141 acute pancreatitis admitted to the First Affiliated Hospital of Guangxi Medical University from January 2018 to December 2019 were collected. According to the Atlanta classification, the peripancreatic effusion was divided into four categories: acute peripancreatic fluid collection (APFC), acute necrotic collection(ANC), pancreatic pseudocyst (PPC) and walled-off necrosis (WON). The general information, clinical manifestations, medical history, laboratory examination indicators and treatment of the four types of patients were recorded and analyzed.Results:Among the 32 patients with chronic pancreatitis complicated with peripancreatic effusion, 27 patients (84.4%) were diagnosed as having PPC, 3 patients (9.4%) WON and 2 (6.2%) APFC. No chronic pancreatitis with ANC was found. The incidence of PPC was higher in patients with chronic pancreatitis than those with acute pancreatitis [84.4% (27/32) VS 31.2% (44/141), P<0.01], and the APFC was lower [6.2% (2/32) VS 24.8% (35/141), P=0.021]. The incidence of ANC was also lower [0.0% (0/32) VS 36.9% (52/141), P<0.01], and there was no significant difference in the incidence of WON [9.4% (3/32) VS 7.1% (10/141), P=0.944]. Compared with patients with peripancreatic effusion of chronic pancreatitis, acute pancreatitis showed a higher proportion of clinical manifestations: fever [19.1% (27/141) VS 3.1% (1/32)], nausea [59.6% (84/141) VS 21.9% (7/32)], vomit [56.7% (80/141) VS 21.9% (7/32)], tenderness [79.4% (112/141) VS 34.4% (11/32)], rebounding pain [42.6% (60/141) VS 0.0% (0/32)], increase of C reactive protein [95.7% (135/141) VS 40.6% (13/32)] ( P< 0.05), and the mean hospital stay was longer (13 days VS 11 days, P=0.048). Imaging examination showed that the proportion of lesions >5 cm in diameter in PPC patients with acute pancreatitis was higher than those with chronic pancreatitis [70.5% (31/44) VS 29.6% (8/27), P=0.001]. WON in chronic pancreatitis patients was limited to the pancreas [3/3 VS 1/10, P =0.014]. In terms of treatment strategies, 25 patients (78.1%) received conservative treatment in 32 chronic pancreatitis. There was no significant difference in treatment strategy between patients with acute pancreatitis and those with chronic pancreatitis. Conclusion:In the peripancreatic effusion of chronic pancreatitis, PPC is the most common. Peripancreatic effusion is mainly treated conservatively. There is no difference in treatment among different types of peripancreatic effusion in chronic pancreatitis. However, compared with chronic pancreatitis, peripancreatic effusion in acute pancreatitis may need more active intervention.
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Objective:To construct a mutant strain of hypervirulent Klebsiella pneumoniae NTHU-K2044 with hfq gene deletion and to analyze its biological characteristics. Methods:The hfq gene of NTUH-K2044 was knocked out by homologous recombination technology to construct △ hfq mutant strain. Its biological characteristics including growth rate, environmental stress tolerance, biofilm formation, capsular polysaccharide synthesis, resistance to neutrophil phagocytosis and lethality to Galleria mellonella larvae were analyzed by comparing with the wild-type strain using phenotypic experiments. Results:The △ hfq mutant strain of hypervirulent Klebsiella pneumoniae NTHU-K2044 was successfully constructed. Phenotypic experiments showed that the △ hfq mutant strain had significantly slower growth rate, smaller colonies and decreased hypermucoviscosity. Its growth was significantly inhibited under different environmental stress conditions such as pH9, pH5.5, 0.7 mmol/L SDS, 5% NaCl, 0.1% H 2O 2 and high temperature of 50℃. In terms of virulence and pathogenicity, the △ hfq mutant strain showed decreased ability to form biofilm and capsule, significantly down-regulated expression of magA and rmpA genes required for capsule synthesis, lower survival rate in the neutrophil bactericidal test and obviously reduced lethality to Galleria mellonella larvae. Conclusions:As a RNA chaperone, Hfq protein could participate in post-transcriptional regulation and play an important role in regulating the physiology, environmental adaptability and virulence of hypervirulent Klebsiella pneumoniae. This study provided reference for further study on hypervirulent Klebsiella pneumoniae.
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Objective:To evaluate the status and influencing factors of psychiatric comorbidities of patients with epilepsy (PWEs) in Hubei province during the outbreak of COVID-19.Methods:From February 23, 2020 to March 5, 2020, a network questionnaire survey (including demographic characteristics, seizures, Generalized Anxiety Disorder Scale-7 score, Patient Health Questionnaire-9 score, Insomnia Severity Index score) was conducted among 570 PWEs who visited the Epilepsy Center of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology during April 1, 2019 and January 20, 2020. SPSS 22.0 software was used for correlation analysis of sociodemographic characteristics, epilepsy related factors, perceived threat to the COVID-19 and psychiatric comorbidity (depression, anxiety and insomnia) of PWEs during the COVID-19 epidemic.Results:A total of 362 valid questionnaires were included for analysis (the response rate was 63.51%,362/570). Thirty-four (9.4%), forty-seven (13.0%) and seventy-one (19.6%) patients suffered from anxiety, depression and insomnia, respectively. Patients with seizure frequency ≥2 times/month before the epidemic ( OR=3.395,95% CI 1.561-7.384, P=0.002), poor subjective quality of life during the epidemic ( OR=10.753,95% CI 1.938-59.654, P=0.024), and moderate to severe worry about bad impact of the epidemic on epilepsy ( OR=3.077, 95% CI 1.382-6.853, P=0.006) were more likely to be anxious. Patients with poor subjective quality of life during the epidemic ( OR=6.188, 95% CI 1.317-29.079, P=0.021) were more likely to be depressed. Patients with COVID-19 related symptoms ( OR=3.609, 95% CI 1.674-7.778, P=0.001), children ( OR=3.090, 95% CI 1.759-5.431, P<0.001), seizure frequency ≥2 times/month before the epidemic ( OR=1.907, 95% CI 1.017-3.575, P=0.044), and moderate to severe worry about unanticipated seizures ( OR=2.555, 95% CI 1.370-4.764, P=0.003) were more likely to suffer from insomnia. Conclusions:During the COVID-19 epidemic, parts of PWEs suffered from anxiety, depression and insomnia. PWEs with poor subjective quality of life, high frequency of epileptic seizures before the epidemic, excessive worry about bad impact of the epidemic on epilepsy and excessive worry about unanticipated seizures were prone to anxiety, depression and insomnia.
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Objective:To evaluate the efficacy and safety of mycophenolate mofetil(MMF) in patients with active moderate to severe thyroid associated ophthalmopathy(TAO) refractory to multiple intravenous glucocorticoid(GC).Methods:Fifty-two patients with active moderate to severe TAO that was refractory to multiple intravenous GC were treated with MMF 0.5g orally, 2/d. To evaluate the overall response rate of TAO patients, the improvement of more than 3 items including clinical disease activity score(CAS), soft tissue involvement, proptosis, diplopia, decrease of eye movements, visual acuity and other improvements were defined as response.Results:After 12 weeks of MMF treatment, the overall response rate of TAO patients was 75.0%, and then increased to 88.5% significantly at the 24th weeks. At the 12th weeks, CAS decreased from(5.06±1.21) to(2.52±1.13), and then continued to decrease to(2.02±0.92) at the 24th week( P<0.05), the response rates were 82.7% and 90.4%, respectively. In addition, after 12 weeks of treatment, 58.1% of patients with diplopia improved significantly, and the response rate was 74.2% at the 24th weeks. Similarly, the degree of proptosis decreased significantly at the 12th and 24th weeks, and the response rates were 53.8% and 69.2%, respectively. No serious adverse events occurred during the treatment. Conclusion:The MMF therapy is efficient and safe for patients with active moderate to severe corticosteroid-resistant TAO.
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Hunan University of Chinese Medicine has improved the implementation of animal ethics education from the aspects of carrying out elective courses, emphasizing the implementation of pre-class learning, integrating ethics education into experimental teaching, cultivating students' good medical literacy in a subtle way, and further deepening their learning by offering elective courses of animal ethics. The results show that the opening of elective courses and the enforcement of experimental animal ethics education in experimental classes have greatly strengthened the students' experimental animal ethics, which is conducive to the formation of students' medical literacy.
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Objective:To investigate the hierarchical management scheme of cervical adenocarcinoma in situ (AIS) based on cervical conization margin state.Methods:All medical records of 249 patients diagnosed as AIS by loop electrosurgical excision procedure (LEEP) conization from Jan. 2010 to Dec. 2015 in Obstetrics and Gynecology Hospital of Fudan University were retrospectively reviewed, to explore the relationship between the status of the resection margin and the residual lesion after LEEP, and the multivariate logistic regression method was used to analyze the related factors that affect the residual lesion after LEEP in cervical AIS patients.Results:(1) The age of 249 cervical AIS patients was (40±8) years old (range: 23-71 years old). Of the 249 patients, 19 (7.6%, 19/249) had residual lesions; 69 cases were pathologically diagnosed as AIS after LEEP, and the residual lesion rate was 13.0% (9/69), which was significantly higher than that of AIS + high-grade squamous intraepithelial lesion [5.6% (10/180); χ2=3.968, P=0.046]; 33 cases were multifocal lesions, the residual rate of lesions was 21.2% (7/33), which was significantly higher than that of single focal lesions patients [5.6% (12/216); χ2=7.858, P=0.005]; 181 patients underwent endocervical curettage (ECC) before surgery, the residual rate of lesions in ECC-positive patients was 14.0% (14/100) , significantly higher than that of ECC-negative patients [4.9% (4/81); χ2=4.103, P=0.043]. (2) Among 249 cases of AIS patients, the positive rate of resection margins after LEEP was 35.3% (88/249); the residual rate of lesions in patients with positive resection margins (14.8%, 13/88) was significantly higher than those with negative margins [3.8%(6/156); χ2=9.355, P=0.002]. The age of patients underwent total hysterectomy after LEEP was (43±7) years old, which was significantly higher than that of patients who did not undergo total hysterectomy [(37±8) years old; t=6.518, P<0.01].Among the patients underwent total hysterectomy after LEEP, 3 cases (2.0%, 3/152) had fertility requirements, while 38 cases (39.2%, 38/97) did not underwent total hysterectomy, the difference between the two groups was statistically significant ( χ2=59.579, P<0.01). Among the 152 patients who underwent total hysterectomy after LEEP, the residual rate of lesions was 11.8% (18/152); the residual rate of lesions in patients with positive resection margins was significantly higher than that of patients with negative resection margins [18.8% (12/64) vs 7.0% (6/86); χ2=4.861, P=0.028]. The median follow-up time of 97 patients who did not undergo total hysterectomy after LEEP was 32 months (range: 4-70 months). During the follow-up period, 3 cases of cervical AIS recurrence (3.1%, 3/97) and were followed by hysterectomy,no invasive adenocarcinoma were seen. (3) Multivariate logistic regression analysis showed that the positive resection margin ( OR=4.098, 95% CI: 1.235-13.595, P=0.021), multifocal lesions ( OR=5.464, 95% CI: 1.494-19.981, P=0.010) were independent risk factors that affected the residual lesions in patients with cervical AIS after LEEP. Conclusions:The cervical AIS patients after LEEP conization suggested be stratified by cone margin state as the first-line stratified index, age and fertility needs as the second-line stratified management index. The individualized management plan should be developed based on comprehensive assessment of high-risk factors of residual lesions.
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By analyzing the shift of microbial communities under different iron/sulfur ratios, the response of metallurgical microorganisms to energy substrates was investigated based on molecular ecological networks. High-throughput sequencing of microbial samples from different domesticated batches was conducted to analyze the changes in community composition, alpha and beta diversity. Based on the molecular ecological network, the interactions between microorganisms under different iron/sulfur ratios were explored. Keystones were identified to analyze the community response to energy substrates. In the process of domestication based on different energy substrates, the dominant species in the in iron-rich and sulfur-less community were Acidithiobacillus ferrooxidans and A. ferriphilus. A. thiooxidans accounted for up to 90% in the sulfur-rich and iron-less community after 3 domesticating batches. The results of alpha and beta diversity analysis show that the domestication process of sulfur-rich and iron-less substrates reduced the diversity of microbial communities. Molecular ecological network analysis shows that the keystones were all rare species with low abundance. During the domestication by sulfur-rich and iron-less energy substrates, the bacterial species had a closer symbiotic relationship and the community was more stable. Through this domestication experiment, the impact of different energy substrates on microbial aggregation was clarified. Domesticating metallurgical microorganisms by using sulfur-rich and iron-less energy substrates made the microbial colonies to be more stable, which was conducive to the oxidation of iron and sulfur, promoting the dissolution of sulfide minerals. Our findings provide a reference for the directional domestication of metallurgical microorganisms.
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Acidithiobacillus/genetics , Iron , Minerals , Oxidation-Reduction , SulfurABSTRACT
Objective@#To develop a method for rapid detection of influenza virus and its subtypes based on flow fluorescence technology, and then detect the clinical specimens by our established method .@*Methods@#We designed degenerate primers and specific probes, synthesized plasmids, used Luminex platform to establish detection method and later detected 430 clinical specimens, and compared the result with those of Fujian Center for Disease Control and prevention.@*Results@#A method for the simultaneous determination of influenza viruses A, B, C and its subtypes (H116, N19) was established. The time consumption was 3.5 hours, with good specificity, high sensitivity and feasible stability. The detection result of 430 clinical specimens showed high consistency with the result of Fujian Center for Disease Control and Prevention.@*Conclusions@#We established a method for simultaneous determination of influenza viruses and its subtypes, high sensitivity, specificity and stability.
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This study was aimed to investigate the association between dyslipidemia and thyroid associated ophthalmopathy (TAO). We evaluated the relationship between dyslipidemia and TAO in 218 patients with Graves′ disease (GD) and found that the serum total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) in the GD subjects with TAO (n=110) were significantly increased [(5.32±1.39) mmol/L vs. (3.18±2.12) mmol/L, (2.98±0.75) mmol/L vs. (1.25±0.98) mmol/L] than those in the GD subjects without TAO (n=108). TC and LDL-C were positively correlated with the Clinical disease activity score (CAS) [TC (r=0.7, P=0.03),LDL-C (r=0.82, P=0.03)], and the levels of TC (OR=2.56, P=0.02) and LDL-C(OR=2.01, P=0.015) were positively associated with TAO. These suggested that high serum cholesterol level is a novel risk factor for TAO, and management of blood lipids should be included in the treatment of TAO.
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Objective@#To investigate the ultrasonic features of fetal isolated coronary artery fistula (CAF).@*Methods@#A total of 13 cases of fetal isolated CAF from December 2011 to March 2018 were retrospectively analyzed. Data of echocardiography and follow-up were collected.All cases were divided into retrograde group and no retrograde group based on the presence of diastolic retrograde flow in aortic arch. The inner diameters of affected coronary arteries (CA) and aortic annulus (AO) were measured, and the CA to AO ratios (CA/AO) were then calculated.Correlations between CA, CA/AO and the presence of retrograde flow were analyzed. Adverse birth outcomes including abortion, enlarged cardiac cavity, pulmonary hypertension were recorded at follow-up. Correlation between diastolic retrograde flow in aortic arch and the rate of adverse birth outcomes was analyzed.@*Results@#①Thirteen cases of fetal CAF were characterized by the varying degrees of dilation of affected coronary arteries. Thirteen cases were presented with abnormal blood flow in the large arteries or cardiac cavities: biphasic continuous flow pattern in atrio-coronary fistula and biphasic bidirectional continuous flow pattern with a diastolic dominant flow in coronary artery to ventricle fistula. Seven cases were presented with retrograde holodiastolic flow in aortic arch. ②The CA/AO ratios of retrograde group was higher than in no retrograde group [(0.63±0.24)mm vs (0.39±0.09)mm, P=0.047], there was no significant difference of CA between the two groups [(3.00±1.25)mm vs (2.03±0.62)mm, P=0.115]. ③The rate of adverse birth outcomes in retrograde group was higher than in no retrograde group (80% vs 20%, χ2=4.80, P=0.028).@*Conclusions@#Fetal isolated CAF has distinct ultrasonic features. Abnormal CA/AO ratio and diastolic retrograde flow in aortic arch are significant ultrasonic features and has prognostic values.
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Objective To investigate the ultrasonic features of fetal isolated coronary artery fistula ( CAF) . Methods A total of 13 cases of fetal isolated CAF from December 2011 to M arch 2018 were retrospectively analyzed . Data of echocardiography and follow‐up were collected .All cases were divided into retrograde group and no retrograde group based on the presence of diastolic retrograde flow in aortic arch . T he inner diameters of affected coronary arteries ( CA ) and aortic annulus ( AO ) were measured ,and the CA to AO ratios ( CA/AO ) were then calculated .Correlations between CA ,CA/AO and the presence of retrograde flow were analyzed . Adverse birth outcomes including abortion , enlarged cardiac cavity , pulmonary hypertension were recorded at follow‐up . Correlation between diastolic retrograde flow in aortic arch and the rate of adverse birth outcomes was analyzed . Results ① T hirteen cases of fetal CAF were characterized by the varying degrees of dilation of affected coronary arteries . T hirteen cases were presented with abnormal blood flow in the large arteries or cardiac cavities :biphasic continuous flow pattern in atrio‐ coronary fistula and biphasic bidirectional continuous flow pattern with a diastolic dominant flow in coronary artery to ventricle fistula . Seven cases were presented with retrograde holodiastolic flow in aortic arch . ②T he CA/AO ratios of retrograde group was higher than in no retrograde group [ ( 0 .63 ± 0 .24 ) mm vs ( 0 .39 ± 0 .09) mm , P ＝0 .047] ,there was no significant difference of CA between the two groups [ ( 3 .00 ± 1 .25) mm vs ( 2 .03 ± 0 .62) mm , P ＝0 .115] . ③T he rate of adverse birth outcomes in retrograde group was higher than in no retrograde group ( 80% vs 20% ,χ2 ＝4 .80 , P ＝0 .028) . Conclusions Fetal isolated CAF has distinct ultrasonic features . Abnormal CA/AO ratio and diastolic retrograde flow in aortic arch are significant ultrasonic features and has prognostic values .
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OBJECTIVE:To establish a method for the determination of drug loading and encapsulation efficiency of Epirubi-cin hydrochloride-sorafenib-loaded Polylactic Acid-glycolic Acid Polymer(PLGA)embolic microspheres. METHODS:HPLC meth-od was adopted to determine the contents of epirubicin hydrochloride and sorafenib in the preparation,and then drug loading and encapsulation efficiency were calculated by formula. The determination was performed on Phenomenex Luna 5u C8(2) 100A col-umn with mobile phase consisted of methanol-water(containing 0.05% trifluoroacetic acid and 0.14% dium dodecyl sulfate)(75:25,V/V)at the flow rate of 1.0 mL/min. The detection wavelength was set at 252 nm,and the column temperature was maintained at 25℃. The injection volume was 10μL. RESULTS:The linear ranges were 2.020-101.00μg/mL for epirubicin hydrochloride(r=0.9998)and 2.048-102.40 μg/mL for sorafenib(r=0.9997),respectively. The limits of quantification were 3.2970,2.5468 μg/mL, respectively. The detection limits were 0.9891,0.7641 μg/mL,respectively. RSDs of precision,stability and repeatability tests were all less than 2.0%. The recoveries were 96.41%-101.80%(RSD=1.64%,n=9),99.46%-101.45%(RSD=0.70%,n=9),re-spectively. Drug loading of two components in 3 batches of samples were no lower than 1.17%,encapsulation efficiency no lower than 58%. CONCLUSIONS:The method is simple,accurate,can be used to determine drug loading and encapsulation efficiency of Epirubicin hydrochloride-sorafenib PLGA embolic microspheres.
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With citric acid as carbon source and different amino acids as nitrogen source,the nitrogen-doped carbon was synthesized by one-step hydrothermal method under the condition of no catalyst.The pre-experiments show that arginine-doped carbon dots(CDs-Arg) with relatively high fluorescence quantum yield (33.25％) were picked out for further study.Furthermore,we studied the physical and chemical properties of CDs-Arg through a series of spectral,electric potential,particle size,X ray and elemental analysis experiments.At the same time,the stabilities of the nanoparticles towards different pH,temperatures,excitation lasers or redox conditions were studied.And the MTr and in vivo distribution experiments were also conducted for the toxicity and metabolism studies.The experimental results showed that the arginine doped carbon dots has high fluorescence efficiency,good stability,and extremely low toxicity.And the water-soluble small particles of CDs-Arg nanoparticles showed that the nanoparticles can be excreted through the glomerulus.These results show that the CDs-Arg Nanoparticles are biocompatible nanoparticles and have potential applications in biological imaging and can serve as a monitor of drug metabolism.
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Objective To investigate the significance of variation of concentrations of serum IL-1,IL-6, TNF-α and cyclooxygenase-2 in patients with post-traumatic stress disorder (PTSD). Methods Thirty-two PTSD patients were chosen as the observation group and another 26 normal patients were taken as the control group. The concentrations of serum IL-1,IL-6,TNF-α and COX-2 in patients of both groups were detected by Enzyme-linked immunosorbent assay (ELISA) and Western blot and were compared in these 2 groups. Results The concentrations of serum IL-1 , IL-6 and TNF-α were significantly higher in the observation group than those in the control group (P < 0.01);and the level of COX-2 in the observation group was significantly higher than that of the control group(P < 0.01). Conclusions Abnormal immune states are awared in PTSD patients and the IL-1,IL-6,TNF-α and COX-2 might play an important role in the pathogenesis of PTSD.
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Objective To study the effects of electromagnetic field (EMF) on the activation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) pathways in mesenchymal stem cells (MSCs) and their interaction, and to explore the cellular signal transduction mechanism of the biological effects induced by EMF.Methods The 3rd-passage rat bone marrow MSCs were randomly divided into a control group, an EMF group, an EMF + PD98059 group and an EMF + SB202190 group.Cells in the EMF group were cultured in the electromagnetic field, those in the EMF + PD98059 and EMF + SB202190 groups cultured in the electromagnetic field after PD98059 or SB202190 was added, and those in the control group were cultured normally.Different groups of cells were exposed to electromagnetic fields (15 Hz, 1 mT, sine wave form) for different exposure duration.The activated phosphorylated and non-phosphorylated p38MAPK were measured using Western blotting analysis with their specific corresponding antibodies.The alkaline phosphatase (ALP) activity in cells in different groups was detected according to the instructions of ALP kit.MTT assay was applied to investigate the proliferation of cells.Results Electromagnetic fields could rapidly induce the activation of p38 MAPK (P ＜ 0.05) and the phosphorylation of p38 MAPK elevated after 15 min exposure to EMF.The phosphorylation of p38 MAPK was significantly lower in the EMF + SB202190 group than that of the EMF group.After 5 days of EMF exposure, the ALP activity of cells was significantly improved, and the effect could be inhibited by SB202190.The bone marrow mesenchymal stem cells proliferation increased significantly after being exposed to EMF for 3 days, and it could not be blocked by SB202190.Phosphorylation of ERK and MAPK increased significantly when the p38 MAPK pathway was blocked by SB202190 and exposed to EMF for 5 minutes, and it also increased significantly when the ERK MAPK pathway was blocked by PD98059 and received 30 minutes of EMF exposure.Conclusion EMF can quickly activate ERK and p38 MAPK pathways to induce cell proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.Moreover, in EMF there is a mutual interference between ERK and p38 MAPK pathways.
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Objective To investigate the role of scavenger receptor class B type 1 (SR-B1) signaling pathway in serum amyloid A (SAA)-induced angiogenesis in rheumatoid arthritis (RA).Methods The expression and location of SR-B1 in RA and osteoarthritis (OA) synovial tissues were observed by immunohistochemistry.And SR-B1 expression in the resting human umbilical vein endothelial cells (HUVECs) was detected by immunoflourescence.Wound repair assessement and tube formation assessement were employed to evaluate the effect on cell migration and tube formation stimulated by SAA and/or anti-SR-B1 antibody.The t-test and one-way analysis of variance (ANOVA) were used for statistical analysis.Results ① SR-B1 was significantly highly expressed in RA tissue samples (A=6 788±819) when compared to the minimal expression in OA (A =31 849±6 977,t=3.567,P＜0.01).Positive staining of SR-B1 was observed in RA synovial vascular endothelial cells and perivascular areas.② Strong staining for SR-B1 was observed in all HUVECs tested.③ Significant wound healing induced by SAA (MI=2.50±0.17) was found compared with the untreated controls (MI=1.00±0.09,q=14.38,P＜0.01),and the effects were inhibited in the presence of anti-SR-B1 antibody (MI=1.16±0.14,q=13.02,P＜0.01).④ Compared to the untreated group (branch point number:6.6±0.8),there was an enhanced formation of branched and capillary-hke tube structure followed by SAA stimulation (branch point number:19.0±1.1,q=25.04,P＜0.01) after culturing for 72 h,whereas,tube formation decreased markedly upon pre-treated with anti-SR-B1 antibody (branch point number:7.6±1.3,vs SAA,q =23.32,P＜0.01).Conclusion Our present study suggests that serum amyloid A may induce angiogenesis via SR-B1 signaling pathway in RA.
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TCM experimental teaching has a very important position in university training process, in which the laboratory standardized management and the strengthening of the teaching quality monitoring play important roles. According to the the experimental teaching reforms and requirements enacted in recent years by the Ministry of Education of the People’s Republic of China, Hunan TCM University has conducted a series of standardized management of exploration and practice in the laboratory.