This study is aiming to establish an efficient metabolite extraction method for exploration of molecular mechanisms of desiccation tolerance of the resurrection plant Boea hygrometrica using a metabolomics approach. The extracts of metabolite in B. hygrometrica using methanol solution (method A) and methanol-chloroform-water solution (method B) were analyzed by gas chromatography-mass spectrometry (GC-MS). The total numbers of chromatographic peaks, extraction efficiency, retention time and the peak stability were compared. The results showed that for fresh materials, the total chromatographic peak number of method B is more than that of method A; the extraction efficiency of nine representative metabolites by method B is higher than that by method A; the comparison of 10 random chromatographic peaks revealed that the relative standard deviation (RSD) values of the retention time are less than 1% for both methods, whereas the RSD values of the extraction efficiency is different. The percentage of peaks that owned RSD values of the extraction efficiency higher than 10% is 50% for method A and 100% for method B. In addition, method B was also efficient for dry materials from B. hygrometrica. The number of chromatographic peaks, RSD value of retention time and extraction efficiency of dry materials was similar to that of fresh materials using method B, but decreased sharply using method A. Putting together, our study provided evidence that method B is an efficient extraction method for further analysis of metabolites from this resurrection species.
Chemical Fractionation ; methods ; Chloroform ; Gas Chromatography-Mass Spectrometry ; Magnoliopsida ; chemistry ; Metabolome ; Metabolomics ; Methanol ; Plant Extracts ; chemistry ; Solvents

Objective This paper reports a modified method for the isolation of rat thoracic aortic vascular smooth muscle cells based on combined enzyme double digestion.Methods An enzyme mixture containing collagenase II, soybean trypsin inhibitor and elastase was prepared and used to remove the aortic tunica intima and tunica adventitia, and then the tunica media was subjected to second digestion using the same enzyme mixture and to isolate vascular smooth muscle cells.Results The isolated VSMCs were cultured in vitro and the growing cells had an elongated spindle-shape, reached confluence within a week, and displaying a typical hill-and-valley pattern.After 1 week, the cells were passaged.Contractile smooth muscle markers(α-SMA, myosin-II and β-tubulin) were highly expressed in the isolated cells.More than 90% of the cells significantly expressed alpha smooth muscle actin and myosin-II.Conclusions Themethod established in this study has advantages of simple and easy to operate, and good reproducibility, with a high activity and purity of the separated cells.It can ensure to obtain a large amount of contraction-type vascular smooth muscle cells within a short time.

Objective: To investigate the use of e-cigarette and analyze its influencing factors among high school students in Wuhan City, so as to provide insights into developing control measures for adolescents. Methods: From September to November 2019, high school students in Wuhan City were selected by the multi-stage stratified cluster random sampling method, and basic information, e-cigarette use and second-hand smoke exposure were collected through questionnaire surveys. Proportions of e-cigarette current use and attempt to use were analyzed, and factors affecting the current use of e-cigarettes were identified using a multivariable logistic regression model. Results: Totally 5 700 questionnaires were allocated, and 5 602 valid questionnaires were recovered, with an effective rate of 98.28%. The respondents included 2 925 males (52.21%) and 2 677 females (47.79%); 4 033 high school students (71.99%) and 1 569 vocational high school students (28.01%). The proportion of attempt to use e-cigarettes was 9.23%, and the proportion of current e-cigarette use was 2.03%. Multivariable logistic regression analysis identified male (OR=2.112, 95%CI: 1.285-3.471), vocational high school (OR=1.967, 95%CI: 1.214-3.186), private high school (OR=9.684, 95%CI: 5.648-16.605), family second-hand smoke exposure (OR=3.064, 95%CI: 1.741-5.392), second-hand smoke exposure in public places (OR=4.402, 95%CI: 1.687-11.484) and having close friends who were smokers (OR=6.432, 95%CI: 3.219-12.852) as factors affecting current e-cigarette use among high school students. Conclusions The proportion of current e-cigarette use among high school students in Wuhan City was 2.03%. Male, vocational high school, private high school, second-hand smoke exposure and having close friends who were smokers may be promoting factors for e-cigarette use.

Objective To explore the protective effect of resveratrol against oxidative damage to cultivated fibroblasts irradiated with UVB. Methods Fibroblasts from normal human skin cultured in vitro were divided into 5 groups (a normal control group, a group irradiated with UVB, a group treated with resveratrol before UVB irradiation, and a group treated after irradiation). A monolayer of fibroblasts was irradiated with UVB at 60 mJ/cm2. The vitality of the cells was measured using the methylthiazol tetrazolium (MTT) method. The activity of superoxide dis-mutase (SOD) and malondialdehyde (MDA) content were determined using enzyme biochemistry. Results Resveratrol over 100 μM inhibited the proliferation of fibroblasts. Resveratrol under 100 μM improved the proliferation of cells. The optimal concentration was 50 μM. UVB irradiation decreased the vitality of the cells and SOD activity, and it significantly enhanced MDA content. Conclusions Resveratrol treatment before or after UVB irradiation elevates the survival rate of fibroblasts, enhances the activity of SOD, and decreases MDA content. Resveratrol at low concentration could improve the proliferation of fibroblasts, and at high concentration could inhibit their proliferation. Res-veratol at 50 μM relieves the inhibited proliferation of fibroblasts damaged by UVB irradiation.

Objective To investigate the effects of rabbit carotid artery cannula on atherosclerosis formation and β-catenin expression.Methods Fifteen 2-month-old New Zealand rabbits weighing(2.0±0.2)kg were randomly divided into three groups,high-fat diet group,left common carotid artery cannula group and left common carotid artery cannula plus high-fat diet group,5 cases in each group,and taking the right blood vessels in the left common carotid artery cannula group served as the control group.The animals were sacrificed after 9-week feeding,and the total rabbit carotid artery in each group was taken;the real-time PCR and immunohistochemistry were used to detect β-catenin mRNA and protein expression targeting in rabbit common carotid artery tissue.Results The real-time PCR results showed that the β-catenin mRNA expression in the left common carotid artery cannula group was higher than that in the control group,high fat diet group and the left common carotid artery cannula + high-fat diet group.The immunohistochemistry results showed that,except for the control group,various groups had the β-catenin protein location in the cytoplasma,moreover which in the left carotid artery cannula group and left common carotid artery cannula +high-fat diet group mainly located in the area of intimal hyperplasia.Conclusion β-catenin is highly expressed in the atherosclerotic vessel wall caused by rabbit carotid artery cannula.

Objective To study the therapeutic effect of the complex mixture of luteolin and rutin ( MLR) on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin ( MPTP) induced Parkinson’ s disease ( PD) mouse model. Methods Seventy-two C57BL/6 mice were divided into six groups randomly ( n=12 in each group): the normal control , model control , madopar (50 mg·kg-1) group, MLR at low (140 mg·kg-1), middle (280 mg·kg-1) and high (560 mg·kg-1) dose groups. PD mouse models were established by intraperitoneal injection of MPTP ( 30 mg · kg-1 ) . Pole test and traction performance were recorded to access the body coordinate capability and strength. The tyrosine hydroxylase (TH), dopamine transport protein ( DAT) , and glial fibrillary acidic protein ( GFAP ) positive cells were detected by immunohistochemical method. Dopamine ( DA ) , dihydroxyphenylacetic acid ( DOPAC ) , homovanilic acid ( HVA ) , 5-hydroxytryptamine ( 5-HT ) and 5-hydroxyindoleacetic acid (5-HIAA) in striatum were quantified by HPLC-ECD. Results MLR significantly ameliorated mouse motor coordination ability (P<0. 01 or P<0. 05). MLR at 280 and 560 mg·kg-1 could increase TH-positive neurons by 69. 00%and 77. 95% compared with the normal control group (P<0. 01) and DAT-positive neurons by 68. 53% and 70. 40% compared with the normal control group(P<0. 05), and decrease GFAP-postive astrocyte reactivity. The treatment with MLR at three doses attenuated the monoamine neurotransmitter disorder. Conclusion MLR markedly improves MPTP caused movement coordinate ability injury in mice and exerts therapeutic action on PD by regulating neurotransmitters in brain, inhibiting the inflammatory reaction and alleviating the neuron injury.

Objective To evaluate the efficiency of our self-designed enriched filtrator of stem cells in screening and collecting enriched human bone marrow stem cells (BMSCs) for rapid construction of bioactive materials.Methods Bone marrow blood from 32 patients was circulated in our self-designed enriched filtratorand filtered through beta-tricalcium phosphate(β-TCP).The numbers of BMSCs were counted pre-and post-filtration to assess the enrichment efficiency.The numbers of nucleated cells,red blood cells and platelets were compared pre-and post-filtration to assess the effects of the enriched filtrator of stem cells on blood cells.Bacteriological examinations were performed to test the safety of the device.Scanning electron microscopy was carried out to observe the adhesion of BMSCs on β-TCP.Results There was a significant difference between the pre-fihration colony number of BMSCs cultured (232.8 ± 128.6/mL) and the post-filtration one (37.8 ± 34.9/mL) (t =10.743,P ＜ 0.001).The efficiency of BMSCs enrichment of the device was 85.6％ ±8.4％ (from 63.8％ to 97.1％).There were no significant differences between pre-filtration and post-filtration in the numbers of nucleated cells [(16.8 ± 5.4) × 109/mL versus (16.3 ±5.0) ×l09/mL],red blood cells [(3.9±0.6) ×1012/mL versus (3.8±0.6) ×1012/mL] and platelets [(120.4 ± 54.6) × 109/mL versus (123.0 ± 51.8) × 109/mL] (P ＞ 0.05).All the bacteriological examinations of the bone marrow samples were negative.After in vitro osteogenic induction culture,formation of numerous BMSCs colonies was observed on the β-TCP by scanning electron microscopy.Conclusion Since our self-designed enriched filtrator can efficiently collect enriched BMSCs and combine them with β-TCP simultaneously,the device is capable of rapid and safe construction of bioactive materials.

Aim To explore the therapeutical effect and mechanism of baicalein on two 6-hydroxydopamine (6-OHDA ) induced Parkinson′s disease (PD ) rat models,which received unilateral lesions of the left medial forebrain bundle (MFB ) or caudate putamen (CPu ) made by stereotaxic injection of 6-OHDA (MFB-M,CPu-M).Methods PD rat models were established by microinjection of 6-OHDA into MFB or CPu.The anti-tremor effect of baicalein on PD rat models was examined.Spontaneous activity was recor-ded. Dopamine (DA ), dihydroxyphenylacetic acid (DOPAC)and homovanilic acid (HVA)in striatum were quantified by HPLC-ECD.The tyrosine hydroxy-lase (TH)and OX-42 positive cells were detected by immunohistochemical method.The morphological vari-ation of the neurons was confirmed by analysis at an ul-trastructural level.Results Baicalein significantly in-creased the spontaneous activity in CPu-M.The elec-tromyography (EMG ) recordings revealed that com-pared with 6-OHDA group,the tremor frequency in ba-icalein group was decreased by 55% in MFB-M,and by 60% in CPu-M.6-OHDA treatment decreased DA levels in the striatum,while treatment with baicalein attenuated the DA decreases in CPu-M.Moreover,ba-icalein treatment could increase TH-positive neurons and decrease OX-42-postive microglia compared with 6-OHDA group in both MFB-M and CPu-M.Conclu-sions In the present study,it is illustrated that ①microinjection of 6-OHDA into the MFB and the CPu could cause different pathological changes of PD, which is important for efficacy evaluation;②baicalein showed the ability to alleviate the behavior symptoms in PD-rats at different stages by improving motor function and attenuating muscle tremor;③therapeutic effect of baicalein was produced by inhibiting the inflammatory medium production and release,anti-apoptosis,chan-ging dopamine catabolism, and inhibiting dopamine turnover.

Objective: To understand delay in treatment student tuberculosis (TB) patients in Wuhan from 2011 to 2018, and to provide a reference for prevention and control measures of tuberculosis in schools. Methods: The medical cases of students with tuberculosis in Wuhan recorded by the national tuberculosis management information system during 2011-2018 were analyzed for the delay and change trend of medical treatment of students with tuberculosis, and Logistic regression was used to analyze influencing factors. Results: The median number of student TB patients seeking treatment in Wuhan was 9(3,21) days, and 51.14% of student TB patients were delayed. From 2011 to 2018, the overall delay rate of TB patients in students decreased slowly and then gradually increased, decrea sing from the highest rate (56.4%) in 2011 to the lowest one (45.6%) in 2014, and then gradually increased to 53.8% in 2018, there was no significant difference in the delay rate between the first and the second four years(χ2=2.84, P=0.09); The delay rate of different gender students was the same as that of the whole. The fluctuation of female students was slightly obvious, but the difference was not statistically significant(χ2=0.07, P>0.05); Among the students of different age groups, the delay rate of 13-18 years old was the lowest(χ2=87.23, P<0.01). The delay rate of 19-22 years old and university groups of different school levels showed a slow decline, while that of other age groups and other school level groups showed an overall upward trend. The results of multivariate logistic regression analysis showed that the main factors affecting the delay rate of medical treatment were the remote urban area(OR=1.84, 95%CI=1.56-2.16), the ethnic minorities (OR=2.73, 95%CI=1.46-5.09) and the age of 13-18(OR=0.57, 95%CI=0.33-1.00). Conclusion Delay in treatment of TB patients in Wuhan is not optimistic, with more than half of the students have delayed in treatment, the main factors that affect the high rate of TB patients’ treatment delay are the students in the far urban areas and ethnic minorities. It is very important to take targeted prevention and control measures to guide the students to actively seek medical treatment according to the time, the place and the person.

China ; Humans ; Neurology ; Pediatrics