Objective To construct a cDNA expression library from unfed female tick Haemaphysalis longicornis for screening and cloning potential antigenic genes.Methods Total RNA was isolated from unfed female ticks,mRNA was purified and a library of oligo(dT)-primed cDNA with added directional EcoR Ⅰ/Hind Ⅲ linkers was constructed from the purified mRNA.The constructed cDNA was ligated to the EcoRⅠ/HindⅢ arms of the ?SCREEN vector.Pure phage stocks were harvested by plaque purification and converted to plasmid subclones by plating phage on host strain BM25.8.Recombinant plasmids that were subcloned to E.coli BM25.8 were isolated and transformed into E.coli JM109.Recombinant plasmids abstracted from JM109 were identified by PCR and sequencing.Rusults The recombinant phage DNA was packaged by using phage-marker packaging extracts,resulting in a primary cDNA library with a size of 1.8?106 pfu.Data showed 100% of the library were recombinant and the titer of the amplified library was 2.4?109 pfu/ml.Forty-two clones of encoding immunodominant antigens were obtained from the cDNA library.Sequence analysis revealed 12 unique cDNA sequences and the encoded putative proteins showed similarities to H.longicornis tropomyosin mRNA,Rhipicephalus annulatus unknown larval protein mRNA,chromosome 2R of Drosophila melanogaster,mitochondrial DNA of H.flava,clones HqL09 unkown mRNA and Hq05 mRNA of H.qinghaiensis,and myosin alkali light chain protein mRNA.Conclusion The cDNA expression library from unfed female H.longicornis was successfully constructed and screening of protective genes may provide candidate antigens of the tick.

Objective To investigate the quality of life of army recruits in their basic military training period,and to analyze the influence of job burnout and work satisfaction on their quality of life.Methods A total of 625 recruits enrolled in 2014 of Xinjiang Army troop were chosen as subjects in this study by stratified cluster random sampling.Quality of life was assessed by Chinese version SF-36(Short-Form Health Survey Scale),job burnout was assessed by Chinese version Maslach Burnout inventory general survey (MBI-GS) and work satisfaction was assessed by Chinese version Minnesota Satisfaction Questionnaire (MSQ).Results The scores of PCS and MCS about 597 recruits were (85.93±12.62) and (81.10±14.12) respectively.According to demographic characteristics,the score of physiological function was lower than that of non-smoking group (t=2.009,P＜0.05),the score of role physical was lower than that of non-smoking group (t=2.617,P＜0.05),and the score of PCS was lower than that of non-smoking group (t=2.141,P＜0.05).As to except reported health transition,there were negative correlations between emotional exhaustion,depersonalization and other SF-36 scales (r=-0.344～-0.661,P＜0.01) respectively.There were negative correlations between personal accomplishment and vitality (r=-0.204,P＜0.05),role emotional (r=-0.239,P＜0.05),mental health (r=-0.289,P＜0.05) and MCS (r=-0.276,P＜0.05) respectively.Work satisfaction and quality of life was positively correlated (r=0.243～0.635,P＜0.01).As to the independent variable,regression analysis showed that emotional exhaustion (β=-4.732,P＜0.01),Minnesota external satisfaction (β=0.783,P＜0.01) influenced PCS,the difference was statistically significant.Emotional exhaustion(β=-6.534,P＜0.01),pre-enlistment place of residence (β=-5.319,P＜ 0.05),Minnesota external satisfaction (β=0.813,P＜0.01)influenced MCS,the difference was statistically significant.Conclusion Job burnout and work satisfaction influence the quality of life about the army recruits,the more job burnout signify the lower quality of life,and its influence on mental health is more significant.The higher work satisfaction signify the higher quality of life.

Pregnane X receptor (PXR), a member of nuclear receptor superfamily, plays an important role in xenobiotic and endogenous metabolism, endocrine balance, and cell proliferation, etc. Previous study has shown that pregnenolone 16α-carbonitrile (PCN), a mouse PXR agonist, could induce liver enlargement. And we found that the change in hepatocytes exhibits regional distribution characteristics: hepatocyte enlargement occurs around the central vein (CV) area, while hepatocyte proliferation occurs around the portal vein (PV) area. In this study, the dynamic changes of hepatocytes during PXR-induced liver enlargement were determined. Serum and liver samples from male C57BL/6 mice were collected for biochemical and pathological analysis after PCN treatment for 1, 2, 3, 5 days, respectively. The animal experiment was approved by the Animal Ethics Committee of Sun Yat-Sen University. The results showed that with the increase in the PCN treatment days, the feature of this regional change of hepatocyte around the CV and PV areas became more and more obvious. At the same time, the factors related to hepatocyte enlargement, such as the expression of PXR downstream genes and the hepatic content of triglyceride (TG), has gradually increased. The upregulation of proliferation-related proteins and downregulation of cyclin-dependent kinases inhibitor proteins were observed in the early stage of PCN treatment, suggesting that hepatocyte proliferation occurs earlier than hepatocyte enlargement during PXR-induced liver enlargement. This study reveals the dynamic change of hepatocytes during PXR-induced liver enlargement and provides a new insight in liver enlargement promoted via PXR activation.

navirus disease 2019 (COVID-19) in the patients younger than 18 years old infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant, and to provide a basis for determining the chest CT changes and efficacy of COVID-19 caused by Omicron virus variant in patients younger than 18 years old. Methods　The clinical and imaging data of 30 cases of patients younger than 18 years old infected with COVID-19 Omicron variant, who admitted to the Third People's Hospital of Shenzhen from February 11 to March 26, 2022 were collected and retrospectively analyzed. The clinical manifestations, imaging features and dynamic changes of lesions were summarized. Results A total of 41 intrapulmonary lesions in 30 patients with COVID-19 caused by SARS-CoV-2 Omicron variant. The main manifestations were patchy or nodular ground-glass opacities and/or consolidation, with focal subpleural distribution, lesions mainly occur in the right lung (70.73%, 29/41). There were 42 lesion morphologies, with 22 (52.38%) striped shadows and 16 (38.10%) nodular shadows, with small lamellar and patchy shadows predominating. There were 36 lesion density variations, with ground glass shadows being the most common, with a total of 24 ground glass shadows (66.66%) in each lobe of the lung, and also 6 consolidation lesions (16.67%) and 6 mixed ground glass opacity and consolidation lesions (16.67%). With the progression of the disease, lesions gradually enlarged, appeared on the 2nd day (312.93 mm3), peaked on the 9th day (1 837.18 mm3). The average absorption time of the lesions was (16±3) days, and there was no significant difference between the absorption time of patchy and nodular lesions (ground glass and/or consolidation) (t=0.853, P>0.05). The lesions showed focal ground-glass opacity in the early stage, 77.78% lesions were absorbed after treatment in the late stage. Inflammatory nodules were absorbed slowly (9-19 days), without residual fibrotic changes. Conclusions　The imaging manifestations of COVID-19 in patients younger than 18 years old infected with SARS-CoV-2 Omicron variant have certain characteristics, showed patchy or nodular ground glass opacities and/or consolidation, mainly distributed in the subpleural area, with small and few lesions and slow change, didn't remain fibrosis. Being familiar with its clinical and imaging manifestations can assist in early diagnosis, but confirming the diagnosis requires a combination of epidemiological history, clinical symptoms, SARS-CoV-2 nucleic acid and radiological manifestations.

Objective To investigate the discrepancy of aldosterone synthesis process and potential regulation abnormality between aldosterone-producing adenoma(APA) and normal adrenal(NA) with microarray. Methods cRNA probes labelled with biotin were prepared from mRNA of APAs(APA group,n=10) or NAs(control group,n=7).The probes were hybridized with oligonucleotide microarray of target gene expression profile.Expression levels were read from the fluorescent intensity scanned.The difference of gene expression profile was analyzed by computer software.Differentially expressed genes were verified by real-time RT-PCR. Results Compared with control group,97 genes were up-regulated and 168 genes were down-regulated in APA group.In the genes related to steroid hormone synthesis,only CYP11B2 was significantly up-regulated.In the physiologic regulators of aldosterone synthesis,CYB5A,CYP17A1,DUSP1 and HMGCR were down-regulated,while RENBP and NR1H2 were up-regulated.As a key enzyme in the biosynthesis of cortisol,the expression of CYP17A1 gene was inhibited. Conclusion Among the aldosterone synthesis related enzyme and corresponding regulatory genes in APA,CYP11B2 may be a key synthetase,and the suppressed physiologic regulators of aldosterone synthesis may indicate the existence of neoplastic modulation.

OBJECTIVETo observe the effect of orexin-A on the recovery and cognitive function of aged rats after ketamine anesthesia.

METHODSFifty-five aged rats were divided randomly into control group, model control group, 1 nmol/L Orexin-A group, and 4 nmol/L Orexin-A group. In the latter 3 groups, the rats received an intraperitoneal injection of ketamine at 100 mg/kg, and normal saline was injected in the control group. Ten minutes after the injections, the rats received intraventricular injections of artificial cerebrospinal fluid (control and model control group) or of 10 microl 1 or 4 nmol/L Orexin-A as indicated. The behavioral changes of the rats were assessed by the duration of loss of righting reflex (LORR). Electroencephalogram (EEG) recordings were used to evaluate the changes in rat brain activity by comparison of the percent of sigma wave in EEG before and after the intraventricular injections. Morris water maze was used to test the learning and spatial localization abilities of the rats.

RESULTSKetamine resulted in obvious impairment of learning and memory abilities of the aged rats. Orexin-A at 4 nmol/L induced significant decrease in the duration of LORR and marked reduction of sigma activities in anesthetic rats (P<0.05), and obviously improved the learning and spatial localization abilities of the rats after anesthesia (P<0.05).

CONCLUSIONOrexin-A can promote the recovery and improve the cognitive function of aged rats after ketamine anesthesia.

Aging ; Anesthesia Recovery Period ; Anesthetics, Dissociative ; Animals ; Cognition ; drug effects ; Delayed Emergence from Anesthesia ; prevention & control ; Intracellular Signaling Peptides and Proteins ; pharmacology ; Ketamine ; Male ; Neuropeptides ; pharmacology ; Orexins ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore biological exposure markers, we investigated the chromium content in peripheral erythrocytes from occupational population with broad ranges of soluble chromate exposure, as the candidate biomarker may provide the scientific evidence for health risk assessment in occupational chromate-exposed population.

METHODSA cross-sectional study was conducted in chromate exposed workers employed at a chromate factory in a district of Jinan city, Shandong Province. The studied population contained 114 workers from different processes of the chromate plants, which included 74 males and 40 females, with an age range from 25 to 52 years old, averaging at (35.83 +/- 6.14) years old; the length of service was ranging from 1 year to 37 years, an average of (14.20 +/- 6.77) years. In addition, 30 farmers in the countryside one hundred kilometers away from the factory, without exposure to chromate matched with exposed subjects by age, gender and smoking status were identified as a control group, which included 22 men and 8 women, with age ranging from 25 years old to 47 years old, having an average age of (36.13 +/- 6.17) years old. Personal information on age, chromate exposure, medical history, smoking habit and alcohol consumption was obtained at an interview. The air concentration of personal exposure was determined by individual sampling for 8 hours per day as shift work, and chromium was assayed by atomic absorption spectrometry. The chromium content in the erythrocytes from peripheral blood was determined by graphite furnace atomic absorption spectrometry. The potential plasma reduction capacity was determined by dibenzene anthracoamid dihydrazide spectrophotometry. The content of total vitamin C and reductive ascorbic acid were determined by 2, 4-dinitrobenzene hydrazine. The data were analyzed by SPSS10.0 software for statistical significance.

RESULTS(1) The results showed that the chromium levels in erythrocytes in the exposed group [(15.79 +/- 31.01) microg/L] were significantly higher than those in the control group [(3.21 +/- 2.20) microg/L] (P < 0.01). (2) There existed a dose-response relationship between the personal airborne chromate concentration and the chromium content in erythrocytes. As airborne chromate concentration lowered to 106.00 microg/m(3), the chromium content in erythrocytes increased, depending on the air concentration of chromate. (3) Correlation analysis showed that there was a significant positive correlation between airborne chromate concentration and the chromium content in erythrocytes (P < 0.01). (4) In multiple regression analysis, it was found that the potential plasma reduction capacity and reductive ascorbic acid may be a good indicator for oxidative stress produced by chromate exposure and be used to evaluate the effects on intracellular uptake of chromium (VI).

CONCLUSIONOur findings suggested that the chromium content in erythrocytes should be used as an effective exposed biomarker in the risk assessment for occupational chromate-exposure.

Adult ; Air Pollutants, Occupational ; analysis ; Biomarkers ; blood ; Chromates ; analysis ; Chromium ; blood ; Erythrocytes ; chemistry ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; analysis

BACKGROUNDTo repair late median nerve injury, many methods have been used in the past years. The aim of this study was to review a thirteen-year experience in restoration of thumb opposition by transposing flexor pollicis brevis muscle.

METHODSFrom July 1992 to August 2005, 63 patients without thumb opposition because of late median never injury were treated by transposing the flexor pollicis brevis muscle. All the patients had received primary nerve repair after the jnjury. The interval between the injury and the second operation was (1.87 +/- 2.31) years (6 months to 4.2 years). The patients were followed up for 3 to 48 [months mean (22.93 +/- 2.31) months]. A functional evaluation system designed in 1992 were used to estimate the outcomes of the patients.

RESULTSAll the patients gained excellent functional results without complications and disabilities during follow-up.

CONCLUSIONSRestoration of thumb opposition by transposing flexsor pollicis brevis muscle has the following advantages: 1. Operative trauma is minimal; 2. It is not necessary to transpose other tendons; 3. Except for the thumb in opposition, movements of other fingers and the wrist are not restricted postoperatively.

Adolescent ; Adult ; Biomechanical Phenomena ; Female ; Forearm Injuries ; physiopathology ; surgery ; Humans ; Male ; Median Nerve ; injuries ; Middle Aged ; Muscle, Skeletal ; surgery ; Tendon Transfer ; Thumb ; physiopathology ; surgery ; Wrist Injuries ; physiopathology ; surgery

OBJECTIVETo investigate the biological effects of anti-beta(3) adrenoceptor (beta(3)-AR) autoantibody in the serum of patients with heart failure, which may contribute to a new therapeutic clue for heart failure.

METHODSThe synthetic peptide of the second extracellular loop of the beta(3)-AR was used as the antigen to screen sera of patients with heart failure and of healthy controls by using enzyme-linked immunosorbent assay. IgG in the patients group of positive autoantibody sera was prepared by using a MabTrap Kit (Amersham) following the manufacturer's instructions. The effects of IgG per each group both on contractile response of adult isolated cardiomyocytes and on beating frequency of cultured neonatal rat cardiomyocytes were observed.

RESULTSThe positive rate of anti-beta(3)-AR autoantibody was 26.7% (mean antibody titer: 1:43.27 +/- 2.71) or 11.0% (mean antibody titer: 1:14.59 +/- 1.61) in patients or healthy subjects, respectively P < 0.05. Compared with the control group, the autoantibody against beta(3)-AR from the patients group decreased cell shortening amplitude/cell shortening 3.84% +/- 0.33%, the velocity of shortening -0.47 microm/s +/- 0.07 microm/s and relengthening 0.17 microm/s +/- 0.02 microm/s in adult isolated cardiac myocytes, respectively. The autoantibody in the patients group decreased the beating rate in cultured neonatal rat cardiac myocytes by 47.1 beats/min +/- 8.11 beats/min, which could have a 6-hour continuance. This decreasing was not modified by Nadolol (beta(1)-AR and beta(2)-AR antagonist) in pretreating myocytes, but was nearly prevented by Bupranolol (nonselective beta-AR antagonist) or beta(3)-AR specific antigen.

CONCLUSIONIt seems reasonable to state that a high titer of the autoantibody against beta(3)-AR in the serum in patients with heart failure, which could have a negative inotropic and chronotropic effect, may be a part of pathophysiological mechanisms of heart failure.

Adult ; Animals ; Autoantibodies ; immunology ; metabolism ; Case-Control Studies ; Cells, Cultured ; Female ; Heart Failure ; immunology ; metabolism ; Humans ; Immunoglobulin G ; immunology ; Male ; Middle Aged ; Myocytes, Cardiac ; immunology ; Rats ; Rats, Wistar ; Receptors, Adrenergic, beta-3 ; immunology

Objective To study the proliferative inhibition and apoptosis promoting activity of oxymatrine on human lung cancer cells (A549) and human hepatocellular carcinoma cells (HepG-2) in vitro,and make a comparison.Methods Effect of oxymatrine on the proliferation activity of two kinds of tumor cells were compared by MTT method and growth curve method.HE staining,Hoechst 33258 fluorescent staining and transmission electron microscopy were used to compare morphological changes.Results MTr results showed that,compared with control group,the survival rate of oxymatrine 200,400,and 800 μg/mL concentration group in A549 and HepG-2 cells decreased significantly (P ＜ 0.05,0.01),half inhibitory concentration (IC50) were 1055.45 and 774.11 g/mL respectively;The growth curve showed that,compared with control group,the number of cells in oxymatrine group of HepG-2 cells cultured for fourth and fifth day and A549 cells cultured for fifth day decreased significantly (P ＜ 0.05,0.01).Oxymatrine affected morphological changes of HepG-2 cells more significantly than A549 cells.The number of cells in the oxymatrine group was significantly reduced;And the cell villi was decreased,the nucleus was small and round,and the cell apoptosis morphological characteristics were obvious.Conclusion Oxymatrine can inhibit the proliferation cells and promote the apoptosis of tumor cells.It affected HepG-2 cells more obviously,which suggests that it may have different sensitivity to different tumor cells.