Cloning and prokaryotic expression analysis of AlCMK from Atractylodes lancea / 药学学报

4-(Cytidine 5′-diphospho)-2-C-methyl-D-erythritol kinase (CMK) was one of the key enzymes in the methylerythritol-4-phosphate (MEP) pathway to generate terpenoids. In this study, based on the transcriptome data of Atractylodes lancea, the sequence of the CMK gene was cloned, named AlCMK (GenBank accession number OM283293). The results showed that AlCMK contains a 1 230 bp open reading frame (ORF) encoding 409 amino acids. The deduced protein had a theoretical molecular weight of 44 752.53 and an isoelectric point of 6.67. Transmembrane structure analysis showed that there was no transmembrane structure, and the secondary structure of AlCMK was predicted to be mainly composed of random coil. Homologous alignment revealed that AlCMK shared high sequence identity with the CMK proteins of Tanacetum cinerariifolium, Osmanthus fragrans, Eucommia ulmoides, Lonicera japonica and Salvia miltiorrhiza. Phylogenetic analysis indicated that AlCMK protein had the higher homology with CMK protein of Compositae. The pET-32a-AlCMK prokaryotic expression vector was constructed and a fusion protein with molecular mass of about 65 kDa was expressed in the E. coli BL21 (DE3). The qRT-PCR was used to analyze the expression pattern of AlCMK gene in different tissues and after MeJA treatment. Meanwhile, the enzyme activity was determined by ELISA kit. The results showed that AlCMK gene was tissue-expressed in different origins and its expression was induced by MeJA, and the results of the enzyme activity assay showed that the AlCMK enzyme activity in different regions was higher in the leaves. The subcellular localization showed that AlCMK was located in the chloroplast. This study provides a reference for further elucidating the biological function of AlCMK gene in terpenoid synthesis pathway in Atractylodes lancea.

Dihydromyricetin reverses Herceptin resistance by up-regulating miR-98-5p and inhibiting IGF1R/HER2 dimer formation in SKBR3 cells / 南方医科大学学报

OBJECTIVE@#To explore the effect of dihydromyricetin on the expression of miR-98-5p and its mechanism in the development of Herceptin resistance in SKBR3 cells.@*METHODS@#The expression of IGF2 and miR-98-5p and their interaction relationship were analyzed by bioinformatics analysis through TargetScan online databases. SKBR3 cells and drug-resistant SKBR3-R cells were cultured in cell experiments. Xenograft tumor mice were constructed by SKBR3 and SKBR3-R cells. Proteins were detected by western blotting and immunohistochemistry. Transfected cells were constructed by shRNA lentivirus vectors. RT-QPCR was used to detect RNA. Cell proliferation was detected by MTS method. Cell jnvasion was detected by Transwell assay. Luciferase reporting assays were used to verify RNA interactions. IGF-1R/HER2 heterodimer was determined by immunocoprecipitation.@*RESULTS@#The expression of IGF2, p-IGF1R, p-Akt and p-S6K in SKBR3-R cells were significantly higher than those in SKBR3 cells, while the expression of PTEN protein was lower in SKBR3-R cells (P < 0.05). IGF1R/HER2 heterodimer in SKBR3-R cells was significantly increased (P < 0.01).The expression of IGF2 and invasion ability were significantly reduced while transfected with miR-98-5p in SKBR3-R cells (P < 0.05), but the IGF2 mRNA were no difference in both cells (P > 0.05). The expression of miR-98-5p was up-regulated and IGF2 was decreased in drug-resistant xenograft tumor mice after feeding with dihydromyricetin, and the tumor became more sensitivity to Herceptin (P < 0.05).@*CONCLUSION@#Dihydromyricetin could induce the expression of miR-98-5p, which binds to IGF2 mRNA to reduce IGF2 expression, inhibit the IGF-1R/HER2 formation, thereby reversing cell resistance to Herceptin in SKBR3-R cells.

The value of myocardial work in detecting the reduction of left ventricular global systolic function in acute myocardial infarction patients with preserved ejection fraction / 中华心血管病杂志

Objective: To investigate the diagnosis value of myocardial work (MW) in evaluating left ventricular global systolic function among acute myocardial infarction (AMI) patients with preserved ejection fraction (LVEF). Methods: This study was a diagnostic trial in a prospective case-control design. AMI patients with preserved LVEF were enrolled as AMI(LVEF>50%) group and age and sex-matched healthy individuals undergoing healthy checkup in our hospital were collected as control group. Two-dimensional dynamic images, including standard apical two-chamber, long-axis and four-chamber views in 3 consecutive cardiac cycles were acquired. General clinical data, routine echocardiography and myocardial work parameters were obtained from all subjects. The indices were compared between the two groups. Intra-observer and inter-observer repeatability of myocardial work parameters were evaluated by intra-group correlation coefficient (ICC). Receiver operator characteristic (ROC) curve was used to determine the diagnostic value of global constructive work (GCW), global wasted work (GWW), global work efficiency (GWE) and global work index (GWI) on the reduction of left ventricular global systolic function in AMI(LVEF>50%) group. Results: There were 30 patients in AMI(LVEF>50%) group, the age was (67.3±9.7)years, and 14 cases were female(46.7%). Thirty participants were included in the control group, the age was (68.1±8.6)years, and 12 cases were female (40.0%). Compared with the control group, left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), interventricular septum thickness (IVSD), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), peak early diastolic velocity of mitral orifice/tissue Doppler velocity of posterior mitral annulus (E/e), left ventricular mass (LVM), left ventricular mass index (LVMI) were significantly higher, while E and e values were significantly lower in AMI (LVEF>50%) group (all P<0.05). Compared with the control group, GCW ((1 145.9±440.1)mmHg% (1 mmHg=0.133 kPa) vs. (1 425.7±355.4)mmHg%), GWE ((80.9±9.5)% vs. (87.3±5.5)%), GWI ((1 001.3±416.2)mmHg% vs. (1 247.6±341.7)mmHg%) and the absolute value of long axis integral strain (GLS) ((8.5±3.4)% vs. (11.4±3.7)%) were significantly lower, while peak strain dispersion(PSD)((101.3±66.4)ms vs. (74.7±31.9)ms) was significantly higher in AMI(LVEF>50%) group (P all<0 05). There was no significant difference in GWW((177.2±71.1)mmHg% vs. (155.7±64.6)mmHg%) between the two groups (P>0.05). The reproducibility of GCW, GWW, GWE and GWI within and between observers were satisfactory (all ICC>0.75). ROC curve analysis showed that all four parameters, GCW, GWW, GWE, and GWI, could be used for the diagnosis of reduced left ventricular systolic function in patients with AMI (LVEF>50%), and their areas under the ROC curve were 0.896,0.929,0.808,0.862. Conclusion: Myocardial work assessment is valuable on diagnosing left ventricular global systolic function reduction in AMI patients with preserved LVEF.

The predictive value of 18F-FDG PET-CT in the conversion from thoracoscopic lobectomy to thoracotomy for non-small cell lung cancer / 肿瘤研究与临床

Objective:To investigate the predictive value of 18F-FDG PET-CT in the conversion from thoracoscopic lobectomy to thoracotomy for non-small cell lung cancer (NSCLC). Methods:The clinical data, CT and PET-CT images of 193 patients with primary NSCLC undergoing thoracoscopic surgery from January 2014 to June 2021 at China-Japan Friendship Hospital were retrospectively analyzed. All patients were divided into 45 cases who were converted to thoracotomy (the conversional group) and 148 cases who were not converted to thoracotomy (the non-conversional group). Univariate analysis was performed on clinicopathological characteristics and image parameters. Multivariate logistic regression was used to analyze the factors affecting the conversion to thoracotomy. Taking the final conversion to thoracotomy or not as the gold standard, the predictive effect of variables in the conversion to thoracotomy was analyzed through the receiver operating characteristic (ROC) curve. Delong test was used to compare the area under the curve (AUC) predicted by all variables.Results:In the conversional group, the proportion of peribronchial lymph node (PLN), peribronchial cuffs of soft (PCS), pleural calcification, pulmonary nodule calcification, PLN or PCS calcification or increased density on chest CT was higher than that in the non-conversional group (all P<0.05); and the maximum standardized uptake value (SUV max) of PET in the conversional group was higher compared with that in the non-conversional group ( P<0.001). Univariate logistic regression analysis suggested age ( OR = 4.663,95% CI 2.191- 9.923, P < 0.001) and PLN or PCS density of chest CT scan ( OR = 2.824, 95% CI 1.791-7.303, P < 0.001) were independent influencing factors of the conversion from thoracoscopic lobectomy to thoracotomy. ROC analysis showed that the effect of the conversion to thoracotomy predicted by the combination of 18F-FDG PET and chest CT [AUC = 0.891 (95% CI 0.831-0.951); the optimal cut-off value of SUV max and CT was 3.45, 70 Hu: the sensitivity was 84.4%, the specificity was 83.8%] was better than that by chest CT alone [AUC = 0.678 (95% CI 0.591-0.766); the optimal cut-off value of CT was 70 Hu: the sensitivity was 62.2%, the specificity was 62.8%; P < 0.001] and by age [AUC = 0.625 (95% CI 0.532-0.719); the optimal cut-off value was 65.5 years: the sensitivity was 75.6%, the specificity was 60.1%; P < 0.001]. Conclusions:PLN or PCS density on chest scan and age are valuable in predicting the conversion from thoracoscopic lobectomy to thoracotomy for NSCLC patients. The combination of PET and CT has an additional role in predicting the conversion to thoracotomy during thoracoscopic lobectomy.

High-intensity inspiratory muscle resistance training can improve the exercise capacity and life quality of persons with bronchiectasis / 中华物理医学与康复杂志

Objective:To explore any effect of high-intensity inspiratory muscle resistance training on exercise capacity and life quality for persons with bronchiectasis.Methods:Sixty patients with bronchiectasis were randomly divided into an observation group and a control group, each of 30. The observation group received two 30-minute sessions of inspiratory muscle resistance training daily using the PowerBreak inspiratory muscle trainer 3 days/week for 8 weeks. The intensity was 70% of the maximum inspiratory pressure (MIP). The control group underwent the same training with the intensity at 10% of the MIP. The severity of illness, pulmonary function, respiratory muscle strength and endurance, exercise capacity and life quality of the two groups were evaluated before and after the intervention.Results:Compared with before the intervention, the average MIP in the observation group and the average distance they walked in the 6min walk test (6MWT) improved significantly. Their average social factor score on the Leicester cough questionnaire had increased significantly, while their average heart rate and self-perceived exertion during the 6MWT had decreased significantly. There were no significant differences in any of these indicators for the control group.Conclusions:High-intensity inspiratory muscle resistance training can significantly improve the exercise capacity and life quality of patients with bronchiectasis. The treatment is worthy of further research and application in the clinic.

Clinical characteristics of lung cancer associated acute ischemic stroke and atrial fibrillation associated acute ischemic stroke / 中国医师杂志

Objective:To analyze the clinical characteristics of lung cancer associated acute ischemic stroke (LCA-AIS) and atrial fibrillation associated acute ischemic stroke (AFA-AIS).Methods:From January 1, 2014 to December 31, 2018, 46 patients diagnosed with LCA-AIS (LCA-AIS group)in Hainan Hospital of Chinese PLA General Hospital were selected, and 46 patients diagnosed with AFA-AIS (AFA-AIS group) were matched according to age and sex.The general situation, laboratory test results and imaging results of the two groups were analyzed.Results:(1) The neurological deficit symptoms in AFA-AIS group were more serious than those in LCA-AIS group; there was significant difference in National Institutes of Health Stroke Scale (NIHSS) score and the Modified Rankin Scale (mRs) score between the two groups ( P=0.001, P=0.003). (2)The D-D polymer concentration in LCA-AIS group was significantly higher than that in AFA-AIS group ( P<0.001), but the hemoglobin, erythrocyte count and hematocrit were significantly lower than those in AFA-AIS group (all P<0.001). (3)There was no significant difference in imaging classification and the number of infarct basins between LCA-AIS group and AFA-AIS group ( P>0.05). LCA-AIS patients was more likely to have poly-period acute ischemic lesions ( P=0.015), while AFA-AIS had significantly larger infarct diameter and more likely to be complicated with acute hemorrhagic stroke or bleeding ( P<0.001). Conclusions:The clinical characteristics of LCA-AIS and AFA-AIS are similar, so it is necessary to distinguish LCA-AIS from AFA-AIS in combination with neurological impairment, laboratory tests and imaging findings to avoid misdiagnosis.

Serum mitochondrial tsRNA serves as a novel biomarker for hepatocarcinoma diagnosis / 医学前沿

Hepatocellular carcinoma (HCC), which makes up the majority of liver cancer, is induced by the infection of hepatitis B/C virus. Biomarkers are needed to facilitate the early detection of HCC, which is often diagnosed too late for effective therapy. The tRNA-derived small RNAs (tsRNAs) play vital roles in tumorigenesis and are stable in circulation. However, the diagnostic values and biological functions of circulating tsRNAs, especially for HCC, are still unknown. In this study, we first utilized RNA sequencing followed by quantitative reverse-transcription PCR to analyze tsRNA signatures in HCC serum. We identified tRF-Gln-TTG-006, which was remarkably upregulated in HCC serum (training cohort: 24 HCC patients vs. 24 healthy controls). In the validation stage, we found that tRF-Gln-TTG-006 signature could distinguish HCC cases from healthy subjects with high sensitivity (80.4%) and specificity (79.4%) even in the early stage (Stage I: sensitivity, 79.0%; specificity, 74.8%; 155 healthy controls vs. 153 HCC patients from two cohorts). Moreover, in vitro studies indicated that circulating tRF-Gln-TTG-006 was released from tumor cells, and its biological function was predicted by bioinformatics assay and validated by colony formation and apoptosis assays. In summary, our study demonstrated that serum tsRNA signature may serve as a novel biomarker of HCC.

Preparation of tanshinone Ⅱ_A-glycyrrhetinic acid solid lipid nanoparticles and its inhibitory effect on acne / 中国中药杂志

The optimal prescription of tanshinone Ⅱ_A(TSN)-glycyrrhetinic acid(GA) solid lipid nanoparticles(GT-SLNs) was explored and evaluated in vivo and in vitro, and its effect on acne after oral administration was investigated. The preparation processing and prescription were optimized and verified by single factor and response surface methodology. The in vitro release of GA and TSN in GT-SLNs was determined by ultra-performance liquid chromatography(UPLC). The effect of GT-SLNs on acne was investigated by the levels of sex hormones in mice, ear swelling model, and tissue changes in sebaceous glands, and the pharmacokinetics was evaluated. The 24-hour cumulative release rates of GA and TSN in SLNs were 65.87%±5.63% and 36.13%±2.31% respectively. After oral administration of GT-SLNs and the mixture of GA and TSN(GT-Mix), the AUC_(0-t) and AUC_(0-∞) of TSN in GT-SLNs were 1.98 times and 4.77 times those in the GT-Mix group, respectively, and the peak concentration of TSN in the GT-SLNs group was 17.2 times that in the GT-Mix group. After intragastric administration of GT-SLNs, the serum levels of testosterone(T) and the ratio of testosterone to estradiol(T/E2) in the GT-SLNs group significantly declined, and the sebaceous glands of mice were atrophied to a certain extent. The results demonstrated that obtained GT-SLNs with good encapsulation efficiency and uniform particle size could promote the release of GA and TSN. GT-SLNs displayed therapeutic efficacy on acne manifested by androgen increase, abnormal sebaceous gland secretion, and inflammatory damage.

A new method for isolating gas vesicles from Microcystis for ultrasound contrast / 生物工程学报

Gas vesicles are a unique class of gas-filled protein nanostructures which are commonly found in cyanobacteria and Halobacterium. The gas vesicles may scatter sound waves and generate harmonic signals, which enabled them to have the potential to become a novel ultrasound contrast agent. However, the current hypertonic cracking method for isolating gas vesicles contains tedious operational procedures and is of low yield, thus not suitable for large-scale application. To overcome these technical challenges, we developed a rapid and efficient method for isolating gas vesicles from Microcystis. The new H2O2-based method increased the yield by three times and shortened the operation time from 24 hours to 7 hours. The H2O2 method is not only suitable for isolation of gas vesicles from laboratory-cultured Microcystis, but also suitable for colonial Microcystis covered with gelatinous sheath. The gas vesicles isolated by H2O2 method showed good performance in ultrasound contrast imaging. In conclusion, this new method shows great potential for large-scale application due to its high efficiency and wide adaptability, and provides technical support for developing gas vesicles into a biosynthetic ultrasonic contrast agent.

Impact of dispatcher-assisted cardiopulmonary resuscitation on survival after out-of-hospital cardiac arrest: A Meta-analysis / 中华急诊医学杂志

Objective:To evaluate the outcome of the patients receiving dispatcher-assisted cardiopulmonary resuscitation (DA-CPR) delivered by first-responders who witnessed the out-of-hospital cardiac arrest (OHCA) before the Emergency Medical Service (EMS) arrived.Methods:We performed a search of the relevant literature exploring major scientific databases. We assessed the quality of the included cohort study according to the Cochrane Handbook for Systematic Reviews of Interventions Version 5.1.0. Meta-analysis was performed on three outcome indicators (recovery of spontaneous circulation survival to hospital discharge and survival with favourable neurologic outcome) using the Revman5.3 software.Results:A total of 21 studies with 349 822 patients were selected for the meta-analysis, including 182 125 patients in the DA-CPR group and 167 697 in the CPR-only group. The meta-analysis showed no significant difference between the DA-CPR and CPR-only groups in ROSC [ RR=1.10, 95% confidence interval ( CI): 0.94-1.29, P=0.24], survival to hospital discharge ( RR=1.10, 95% CI: 0.90-1.34, P=0.34) and survival with favourable neurologic outcome ( RR=1.01, 95% CI: 0.79-1.28, P=0.97) of the patients in America, Japan and Korea. However, there was a significant difference between the DA-CPR and the CPR-only groups in ROSC ( RR=2.61, 95% CI:1.53-4.46, P=0.0005), survival to hospital discharge( RR=6.08, 95% CI: 1.84-20.04, P=0.003), and survival with favourable neurologic outcome( RR=9.76, 95% CI: 1.87-51.02, P=0.007) of the patients in China. Conclusions:The overall effect of DA-CPR is significantly different for each country. In detail, DA-CPR offers a survival advantage (Return of spontaneous circulation, survival to hospital discharge and survival with favourable neurologic outcome) over CPR alone in China but no advantage in developed countries.

Research progress of microbe gut brain axis / 中国综合临床

The emergence of gut microbes has become one of the key regulatory factors of gut brain function, and has made people realize the importance of microbe gut brain axis.The gut microbiota and brain are interconnected through immune system, tryptophan metabolism, vagus nerve and endocrine system.Many factors can affect the composition of gut microbiota, including infection, mode of delivery, use of antibiotics, stress, and host genetics.Recent studies have shown that gut microbiota may be associated with many diseases, including autism, irritable bowel syndrome and so on.Future research will focus on understanding the underlying mechanisms of the microbial gut brain axis and attempt to elucidate microbial based intervention and treatment strategies for neuropsychiatric diseases.

Different development phase of transcriptome analysis from Crataegus pinnatifida Bge. and cloning, structure and function prediction of squalene epoxidase involved in triterpenic acid biosynthesis / 药学学报

italic>Crataegus pinnatifida is a traditional Chinese medicine, which contains organic acids, triterpenoid acids and other active components, has important medicinal and edible value. In order to study the difference of gene expression level in different developmental stages of hawthorn and explore the genes of active ingredient biosynthesis in Crataegus pinnatifida, high-throughput Illumina HiSeq 2000 technology were used to conduct transcriptome sequencing and bioinformatics analysis on Crataegus pinnatifida fruits from the same origin at different developmental stages. 78 496 Unigenes with an average length of 941 nt were obtained by Trinity software. Among them, 58 395 Unigenes can be annotated by NR, NT, Swiss prot, KEGG, COG, GO and other public databases. KEGG pathway analysis showed that 52 Unigenes encoding 15 key enzymes involved in the citric acid cycle. There are 62 Unigenes were involved in the triterpene biosynthesis pathway of Crataegus pinnatifida. Two key enzymes SQE of triterpenoid metabolism pathway in Crataegus pinnatifida were cloned and performed bioinformatic analysis. The results showed that ORF of CpSQE1 and CpSQE2 were 1 594 bp and 1 597 bp, respectively, encoding 530 and 531 amino acids. The molecular weight of proteins was 57.6 kDa and 57.5 kDa. Bioinformatics analysis showed that both CpSQE1 and CpSQE2 proteins have a PLN02985 superfamily conserved domain, belonging to the squalene monooxygenase superfamily. The phylogenetic tree shows that CpSQE1 and CpSQE2 are clustered together with SQE with squalene epoxidase function in other plants. This study provides a research basis for further exploring the key genes in the biosynthesis process of hawthorn active ingredients and analyzing the regulation pathway of its active ingredient biosynthesis.

Analysis of triterpenoic acids in different medicinal parts of Poria cocos (Schw.) Wolf using supercritical fluid chromatography / 药学学报

Qualitative and quantitative methods were used to establish the quality of different medicinal parts of Poria cocos (Poriae Cutis, rubra Poria, white Poria, Poria cum Radix Pini) by using ultra-performance convergence chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry (UPC2-PDA-Q-TOF/MSE). A total of 18 chromatographic peaks were detected from Poria cocos by UPC2-PDA. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used to compare the four medicinal parts. The results showed that there were significant differences in the components of different medicinal parts, and the main triterpenoic acids were poricoic acid A, poricoic acid B, dehydroeburicoic acid, and dehydrotrametenolic acid. When combined with the common active component polyporenic acid C, a method for determination of five triterpenoic acids in different parts of Poria cocos was established. These components could be separated within 15 min, and the amount of methanol was 3.63% of that of HPLC method. Taking the five triterpenoid acids as an index, the content of triterpenoid acids in different parts of Poria cocos from high to low were Poriae Cutis, rubra Poria, white Poria and Poria cum Radix Pini. The method is simple, rapid, and uses minimal solvent. The mobile phase of environment-friendly gas carbon dioxide has unique advantages in reducing environmental pollution, which can provide a basis for the development and standard formulation of Poria cocos and its related products.

Availability of school health staff and health equipment in primary and secondary schools in Jinan / 中国学校卫生

Objective@#To understand the situation of school health staff and the allocation of health equipment in primary and secondary schools in Jinan.@*Methods@#A questionnaire survey was conducted in CDC and public primary and secondary schools to describe the situation of the school health staff and health equipment in primary and secondary schools.@*Results@#There were 23 school health staffs in CDC, among which 26.1% were full time. The total staffing ratio of school doctors was 13.9%, with the compliance rate of 19.8%. The total allocation rate of health care teachers was 74.7%, 92.2% of them were part time. The total staffing ratio of school doctors in boarding schools was 20.0%, with a staffing compliance rate of 12.5%. The total allocation rate of school doctors and health care teachers in boarding schools was 66.7%. 16.8% of public primary and secondary schools and 23.3% of public boarding primary and secondary schools did not have any health staffs. 27.6% of public primary and secondary schools are equipped with infirmary, 70.8% with health rooms, and 10.3% without any. The configuration of necessary equipment in infirmary and health care rooms were less than 100%.@*Conclusion@#There is a serious shortage of school health staffs and health equipment in primary and secondary schools in Jinan. Attention and investment in school health should be strengthened, and resources should be rationally allocated to ensure the health of primary and secondary school students.

Effect of paclitaxel on mast cell-CCL2-macrophage axis in rats with pulmonary hypertension / 中华麻醉学杂志

Objective:To evaluate the effect of paclitaxel on the mast cell-CCL2-macrophage axis in rats with pulmonary hypertension.Methods:Thirty SPF-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 180-220 g, were divided into 3 groups ( n=10 each) using a random number table method: control group (group C), pulmonary hypertension group (group PH), and paclitaxel group (group PTX). The model of pulmonary hypertension was established by subcutaneous injection of monocrotaline 60 mg/kg in rats.At 25 days after establishing the models, paclitaxel 2 mg/kg was injected via the tail vein once every four days, for 4 times in total in group PTX.The equal volume of normal saline was injected in the remaining 2 groups.The mean pulmonary artery pressure (mPAP) was performed at 40 days after establishing the model.The heart was removed and dried, the right ventricle (RV) and left ventricle plus ventricular septum (LV+ S) was weighed, and the Fulton index [RV/(LV+ S)] was calculated.The inferior lobe of left lung was taken, the ratio of media wall thickness of pulmonary vessels was calculated by HE staining, the number of Tryptase + , CD68 + , CD163 + , and Ki67 + cells was recorded by immunohistochemistry, the mean value was calculated, the percentage of Ki67-positive cells in blood vessels was recorded, and the proportion of muscularized blood vessels was calculated.The content of CCL2 was measured by enzyme-linked immunosorbent assay, and the expression of cleaved caspase-3 and Cyclin D1 was detected by Western blot. Results:Compared with group C, the mPAP, Fulton index, ratio of media wall thickness, proportion of muscularized blood vessels, the number of Tryptase + , CD68 + and CD163 + cells and percentage of Ki67 + cells were significantly increased, and the expression of cleaved caspase-3 was down-regulated in PH and PTX groups ( P<0.05), the expression of Cyclin D1 was significantly up-regulated in group PH ( P<0.05), and no significant change was found in group PTX ( P>0.05). Compared with group PH, the mPAP, Fulton index, ratio of media wall thickness, percentage of muscularized blood vessels, the number of Tryptase + , CD68 + and CD163 + cells and percentage of Ki67 + cells were significantly decreased, the expression of CCL2 and Cyclin D1 was down-regulated, and the expression of cleaved caspase-3 was up-regulated in group PTX ( P<0.05). Conclusion:The mechanism by which paclitaxel alleviates pulmonary hypertension is related to inhibiting the mast cell-CCL2-macrophage axis in rats.

A comparative study of real-time three-dimensional ultrasound automatic quantification and magnetic resonance imaging on left atrial volume and function in patients with acute myocardial infarction / 中华超声影像学杂志

Objective:To study the changes of left atrial volume and function in patients with acute myocardial infarction (AMI) by three-dimensional echocardiography (3DE) and magnetic resonance imaging (MRI).Methods:Thirty-one patients with AMI in Henan People′s Hospital from March to October 2020 were selected as AMI group and 30 healthy subjects were selected as control group.The left atrial maximum volume (LAVmax), minimum volume (LAVmin), presystolic volume (LAVpre), volume index(LAVI), emptying volume (LAEV), ejection fraction (LAEF), long axis and circumferential strain (LASr, LASr-c; LAScd, LAScd-c; LASct, LASct-c) were measured by two-dimensional echocardiography (2DE), 3DE left atrial automatic quantitative technique and CMR.The differences of left atrial indices between two groups and among 2DE, 3DE left atrial automatic quantitative and CMR techniques were compared. Pearson correlation coefficient and Bland-Altman analysis were used to compare 2DE, 3DE left atrial automatic quantitative and CMR. Intra-observer and inter-observer repeatability of 2DE, 3DE left atrial automatic quantitative technique and CMR were evaluated by intra-group correlation coefficient (ICC).Results:①Compared with the control group: LAVmax, LAVmin, LAVpre, LAVI, LASct and LASct-c by 3DE left atrial automatic quantitative increased, while LAEV, LAEF, LASr, LASr-c, LAScd and LAScd-c by 3DE left atrial automatic quantitative technique decreased in AMI group (all P<0.05). ②LAVmax, LAVmin, LAVpre, LASr, LAScd, LASct measured by 3DE left atrial automatic quantitative were more strongly related to CMR than that measured by 2DE(all P<0.05). Compared with CMR, 2DE underestimated LAVmax, LAVmin, LAVpre, LASr, LAScd, LASct by (8.01±4.01)ml, (9.03±4.15)ml, (7.26±2.09 )ml, (7.26±1.23)%, (5.02±1.08)%, (6.24±0.43)%(all P<0.05); 3DE left atrial automatic quantitative technique underestimated LAVmax, LAVmin, LAVpre, LASr, LAScd, LASct by (1.67±0.62)ml, (1.95±0.90)ml, (2.52±0.76)ml, (1.97±0.59)%, (2.03±0.39)%, (1.02±0.30)% (all P>0.05); The time-consuming of 2DE and 3DE left atrial automatic quantitative technique was reduced, and the time-consuming of 3DE left atrial automatic quantitative technique was less than 2DE [(12.18±3.24)s vs (73.34±10.37)s]. ③The reproducibility of 2DE, 3DE left atrial automatic quantitative technique and CMR measurement within and between observers were good. Conclusions:3DE left atrial automatic quantitative technique can effectively evaluate the changes of left atrial volume and function in patients with AMI. Compared with 2DE and CMR, it is simple, rapid, accurate and repeatable, which provides a new and effective method for clinical study of cardiovascular diseases.

Clinical laboratory features of Meigs' syndrome: a retrospective study from 2009 to 2018 / 医学前沿

Meigs' syndrome (MS), a rare complication of benign ovarian tumors, is easily misdiagnosed as ovarian cancer (OC). We retrospectively reviewed the clinical laboratory data of patients diagnosed with MS from 2009 to 2018. Serum carbohydrate antigen 125 and HE4 levels were higher in the MS group than in the ovarian thecoma-fibroma (OTF) and healthy control groups (all P < 0.05). However, the serum HE4 levels were lower in the MS group than in the OC group (P < 0.001). A routine blood test showed that the absolute counts and percentages of lymphocytes were significantly lower in the MS group than in the OTF and control groups (all P < 0.05). However, these variables were higher in the MS group than in the OC group (both P < 0.05). The neutrophil-to-lymphocyte ratio (NLR) was also significantly lower, whereas the lymphocyte-to-monocyte ratio was higher in the MS group than in the OC group (both P < 0.05). The NLR, platelet-to-lymphocyte ratio, and systemic immune index were significantly higher in the MS group than in the OTF and control groups (all P < 0.05). The hypoxia-inducible factor-1 mRNA levels were also significantly higher, whereas the glucose transporter 1, lactate dehydrogenase, and enolase 1 mRNA levels were lower in peripheral CD4

Simvastatin Blocks Reinstatement of Cocaine-induced Conditioned Place Preference in Male Mice with Brain Lipidome Remodeling / 神经科学通报·英文版

Drug-associated reward memories are conducive to intense craving and often trigger relapse. Simvastatin has been shown to regulate lipids that are involved in memory formation but its influence on other cognitive processes is elusive. Here, we used a mass spectrometry-based lipidomic method to evaluate the impact of simvastatin on the mouse brain in a cocaine-induced reinstatement paradigm. We found that simvastatin blocked the reinstatement of cocaine-induced conditioned place preference (CPP) without affecting CPP acquisition. Specifically, only simvastatin administered during extinction prevented cocaine-primed reinstatement. Global lipidome analysis showed that the nucleus accumbens was the region with the greatest degree of change caused by simvastatin. The metabolism of fatty-acids, phospholipids, and triacylglycerol was profoundly affected. Simvastatin reversed most of the effects on phospholipids induced by cocaine. The correlation matrix showed that cocaine and simvastatin significantly reshaped the lipid metabolic pathways in specific brain regions. Furthermore, simvastatin almost reversed all changes in the fatty acyl profile and unsaturation caused by cocaine. In summary, pre-extinction treatment with simvastatin facilitates cocaine extinction and prevents cocaine relapse with brain lipidome remodeling.

Cloning and prokaryotic expression of 3-ketoacyl-CoA thiolase gene AIKAT from Atractylodes lancea / 中国中药杂志

In this study, the gene encoding the key enzyme 3-ketoacyl-CoA thiolase(KAT) in the fatty acid β-oxidation pathway of Atractylodes lancea was cloned. Meanwhile, bioinformatics analysis, prokaryotic expression and gene expression analysis were carried out, which laid a foundation for the study of fatty acid β-oxidation mechanism of A. lancea. The full-length sequence of the gene was cloned by RT-PCR with the specific primers designed according to the sequence information of KAT gene in the transcriptomic data of A. lancea and designated as AIKAT(GenBank accession number MW665111). The results showed that the open reading frame(ORF) of AIKAT was 1 323 bp, encoding 440 amino acid. The deduced protein had a theoretical molecular weight of 46 344.36 and an isoelectric point of 8.92. AIKAT was predicted to be a stable alkaline protein without transmembrane segment. The secondary structure of AIKAT was predicted to be mainly composed of α-helix. The tertiary structure of AIKAT protein was predicted by homology modeling method. Homologous alignment revealed that AIKAT shared high sequence identity with the KAT proteins(AaKAT2, CcKAT2, RgKAT and AtKAT, respectively) of Artemisia annua, Cynara cardunculus var. scolymus, Rehmannia glutinosa and Arabidopsis thaliana. The phylogenetic analysis showed that AIKAT clustered with CcKAT2, confirming the homology of 3-ketoacyl-CoA thiolase genes in Compositae. The prokaryotic expression vector pET-32 a-AIKAT was constructed and transformed into Escherichia coli BL21(DE3) for protein expression. The target protein was successfully expressed as a soluble protein of about 64 kDa. A real-time quantitative PCR analysis was performed to profile the AIKAT expression in different tissues of A. lancea. The results demonstrated that the expression level of AIKAT was the highest in rhizome, followed by that in leaves and stems. In this study, the full-length cDNA of AIKAT was cloned and expressed in E. coli BL21(DE3), and qRT-PCR showed the differential expression of this gene in different tissues, which laid a foundation for further research on the molecular mechanism of fatty acid β-oxidation in A. lancea.

Expression and Significance of Low-Density Lipoprotein-Related Receptors 5 and 6 in the Wnt/β-Catenin Signaling Pathway in Childhood Acute Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the significance of low-density lipoprotein receptor-related protein 5 and 6 (LRP5/6) in the Wnt/β-catenin signaling pathway in the pathogenesis and prognosis of childhood acute lymphoblastic leukemia (ALL).@*METHODS@#A total of 43 children who were newly diagnosed and achieved complete remission after remission induction therapy were enrolled. The children before treatment were included in incipient group, and after treatment when achieved complete remission included in remission group. A total of 39 children with immune thrombocytopenia were enrolled in control group. Three milliliter bone marrow samples were collected from above-mentioned each group. QRT-PCR was used to determine the mRNA expression of LRP5 and LRP6 in blood mononuclear cells of bone marrow. Western blot was used to detect the protein expression of LRP5 and LRP6. According to the protein expression levels of LRP5 and LRP6, the children were divided into low-expression group and high-expression group, and the clinical biological characteristics were compared between these two groups. Survival analysis was performed by Kaplan-Meier method.@*RESULTS@#Both mRNA and protein expression levels of LRP5 and 6 were upregulated in the incipient group compared with the control and remission group (P<0.05). The mRNA and protein expressions of LRP5 and LRP6 in the high-risk group were higher than those in the medium-risk group (P<0.05), it is the same as in the medium-risk group than the low-risk group (P<0.05). The mRNA and protein expressions of LRP5 and 6 positively correlated with risk degree in the incipient group (r@*CONCLUSION@#The high expression of LRP5/6 may be one of the pathogenesis of childhood ALL, and the degree of LRP5/6 increase may be related to the risk level.