Aquatic plant duckweed can enrich high concentration of arsenic, it is thus used as the representative of phytofiltration.The mechanism of arsenic tolerance in duckweeds has received much concern.In this study, synchrotron radiation X-ray fluorescence (SRXRF) and X-ray absorption near edge structure (XANES) techniques were used to study the micro-distribution and speciation of arsenic in natural As-rich duckweed from lead-zinc mine.Two monolithic duckweeds, FP1 and FP2, were analyzed by micro SRXRF, setting single point scan time and spot size were 5 s, 70 μm×80 μm and 2 s, 100 μm×100 μm respectively.Six points of FP2 were selected and analyzed by micro XANES in energy range of 11.81-11.96 keV.Pressed-pellet duckweed was analyzed by bulk XANES in energy range of 11.67-12.27 keV.The result showed that As(Ⅲ) was the major speciation of duckweed from bulk XANES and micro-XANES data.SRXRF micro analysis showed that arsenic had significant vein distribution in duckweed, and was not spread into the photosynthetic mesophyll within certain concentration, which may reduce the leaf toxicity triggered by arsenic.This vein distribution may play a role in arsenic tolerance in duckweed.

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.

Objective: To investigate the effects of developmental exposure to DEHP on learning and memory of mice. Methods: Male littermates of ICR mice randomly assigned to five experimental groups (n=14 for each condition) on PND4 to receive distilled water, vehicle and 10, 50 and 200 mg/ (kg·d) DEHP from PND5 to PND38 by gavage, weighing and recording body weight of mice. Open field task were conducted on PND 26 and Morris water maze task were begun from PND30 to PND 37 to evaluate spontaneous exploration activity and emotion, spatial learning and memory performance of pubertal mice, respectively. On PND39, all animals were killed and hippocampi were isolated on ice, then total proteins of hippocampus were extracted, followed by determining the expression of PSD95 and synapsin I by western blotting. Results: 200 mg/ (kg·d) DEHP significantly reduced the growth of body weight of mice and the time staying in the central area in open field, prolonged the time searching the hidden platform in Morris water maze (P<0.05) . 50 mg/ (kg·d) DEHP didn’t change the growth of body weight and the emotion (P>0.05) , but reduced the percent of time and distance in the target quadrant during the probe trial of mice in Morris water maze (P<0.05) . The results of western blotting showed that DEHP significantly reduced the expression of PSD95 in hippocampus of mice with all dose groups (P<0.01) , but only 200 mg/ (kg·d) DEHP reduced the expression of synapsin I (P<0.05) . Conclusion Developmental exposure to DEHP can damage the development of synapse in hippocampus, adversely impacting spatial memory performance of mice at a dose that are insufficient to significantly influence the general development and result in anxiety.

Objective: To investigate the effects of herbal cake-partitioned moxibustion on the plasma levels of trimethylamine (TMA), trimethylamine-N-oxide (TMAO), and flavin-containing monooxygenase 3 (FMO3) in rabbits with atherosclerosis (AS), as well as to explore the possible mechanism of herbal cake-partitioned moxibustion in treating AS. Methods: After 1-week adaptive feeding, 28 male New Zealand rabbits were divided into a blank group, a model group, an antibiotic group, and a herbal cake-partitioned moxibustion group according to the random number table method, with 7 rabbits in each group. Rabbits were fed with a basic diet in the blank group, while with a basic diet plus 1％ choline in the remaining groups to prepare the AS model. Rabbits were given drinking water with broad-spectrum antibiotics in the antibiotic group, and herbal cake-partitioned moxibustion in the herbal cake-partitioned moxibustion group for 12 weeks. The atherosclerotic plaques by hematoxylin-eosin (HE) staining, the blood lipid levels, the plasma TMA and TMAO levels by liquid chromatography-mass spectrometry were detected for rabbits in each group at the end of interventions. Liver FMO3 protein expression was detected by Western blotting. Liver FMO3 mRNA expression was detected by real-time fluorescence quantitative polymerase chain reaction. Results: HE staining showed that the arterial wall was rough, the intima was significantly thickened, and more foam cells and lipid deposits were seen in rabbits of the model group. Arterial wall thickening was not obvious with a few foam cells and lipid deposits in the herbal cake-partitioned moxibustion group. Compared with the blank group, the serum levels of low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC) were increased (P<0.01), the plasma levels of TMA and TMAO were increased (P<0.05, P<0.01), and the expression levels of liver FMO3 protein and mRNA were all increased (P<0.05, P<0.01); while the serum high-density lipoprotein cholesterol (HDL-C) level was decreased in the model group (P<0.05). Compared with the model group, the LDL-C and TC levels were decreased (P<0.01 or P<0.05), the HDL-C levels were increased (P<0.01), the TMA and TMAO levels were decreased (P<0.05), while the protein and mRNA expression levels of FMO3 were decreased without statistical significance in the herbal cake-partitioned moxibustion group and the antibiotic group. Conclusion: Herbal cake-partitioned moxibustion can slow atherosclerotic plaque formation and regulate lipid levels in AS rabbits, and the mechanism may be related to the down-regulation of TMA and TMAO expression in the plasma.

Objective : To investigate the epidemiological and clinical characteristics of cases with pertussis in Yiwu City, Zhejiang Province, so as to provide insights into pertussis diagnosis and control. Methods: A total of 186 patients with definitive diagnosis of pertussis in medical institutions in Yiwu City from November 1, 2020 to August 31, 2022 were recruited, and subjects' demographic, clinical symptoms and history of pertussis vaccination were collected using questionnaire surveys. The temporal, population, and spatial distribution and clinical symptoms of pertussis were analyzed using a descriptive epidemiological method, and the clinical characteristics of pertussis patients with different doses of pertussis vaccination were compared. Results: Pertussis was found to predominantly occur during the period between July and November (101 cases, 54.30%), and the three highest-incidence regions included Jiangdong Street, Beiyuan Street and Choujiang Street (87 cases, 46.77%). The 186 pertussis cases included 105 males (56.45%) and 81 females (43.55%), and included 144 cases with age of onset under 7 year (77.42%). Preschool and diaspora children were predominant among all pertussis cases, and the main clinical symptoms included spasmodic cough (97 cases, 52.15%), post-tussive vomiting (82 cases, 44.09%) and aggravated cough at night (77 cases, 41.40%). Routine blood tests measured 119 cases with abnormal white blood cell counts (63.98%), 137 cases with abnormal lymphocyte counts (73.66%), 39 cases with abnormal neutrophil counts (20.97%), 21 cases with abnormal platelet counts (11.29%) and 111 cases with abnormal hemoglobin concentrations (59.68%). There were 55 cases that were unvaccinated (29.57%), 23 cases that were not fully vaccinated (12.37%), and 108 cases that were fully vaccinated (58.06%). There were significant differences among pertussis cases with different doses of vaccination in terms of age, incidence of post-tussive vomiting, percentage of abnormal platelet counts and percentage of hemoglobin concentrations (all P<0.05). Conclusions The majority of pertussis cases are preschool and diaspora children in Yiwu City from November 1, 2020 to August 31, 2022, and the clinical symptoms mainly include spasmodic cough, post-tussive vomiting and aggravated cough at night, with atypical symptoms. The capability for differential diagnosis of pertussis is required to be improved in medical institutions.

Objective: The present study was represented by di-(2-ethylhexyl) phthalate (DEHP), to explore the role of thyroid hormones (THs) disruption in the connection of placenta and neurodevelopmental toxicity. Methods: During fetal mice neural tube closed (pregnancy 9.5 days, E9.5d) to begin synthesis of THs (E15.5 d), all pregnant mice were administered with different concentration of DEHP (0、10、50、200 mg/kg) by gavage once a day(10 mice per group). All pregnant mice were conducted with BrdU administration in E14d by subcutaneous injection. Seven pregnant mice from each group were scarified after anesthesia in E15.5 d, serum and amniotic fluid were collected to determinate the levels of THs(T₃, T₄, FT₃ and FT₄) by the automatic biochemical analyzer, detecting fetal mice placental protein expression of monocarboxylate transporter 8 (MCT8), organic anion transporting polypeptide 1C1 (OATP1C1) and deiodinaseⅡ&Ⅲ (DIO₂, DIO₃) by Western blot. Each group of the remaining three pregnant mices were killed after anesthesia in E18d, take the male fetal brain, BrdU immunohistochemistry was used to detect the proliferation and migration of fetal brain cortical neurons. Results: There was no abnormalities in diet, water intake, body weight and general activity of pregnant mice in each treatment group, and there were no difference in the general physiolo. Results There was no abnormalities in diet, water intake, body weight and general activity of pregnant mice in each treatment group, and there were no difference in the general physiological development status of body weight, brain weight, brain body ratio between the mice of each group. There was no statistically significant differences in serum T₃, T₄, FT₃, FT₄ and amniotic fluid FT₄ in pregnant mice of each group (P>0.05), Compared with the control group, the FT₃ levels in the amniotic fluid of the DEHP 50 and 200 mg/kg groups were significantly decreased(P<0.05). Compared with the control group, the placental MCT8 and DIO₂ protein levels of male fetal mice in the DEHP 50 and 200 mg/kg group decreased, and the level of OATP1C1 protein in 200 mg/kg group decreased(P<0.05), and there was no statistically significant difference in DIO₃ protein levels among all groups (P>0.05). Compared with the control group, the number of BrdU positive cells in the cerebral cortex of male mice in DEHP 200 mg/kg group decreased, 56.5% was distributed in VZ-SVZ layer, and the percentage of BrdU positive cells in the IZ layer of 50 mg/kg group increased (P<0.05). Conclusion DEHP 50, 200 mg/kg may affect the proliferation and migration of neural cells in the developing brain, which may be related to its interference with thyroid hormone by placental transport.

Objective:Flotillin-2(FLOT2)is a prototypical oncogenic and a potential target for cancer therapy.However,strategies for targeting FLOT2 remain undefined.Post-translational modifications are crucial for regulating protein stability,function,and localization.Understanding the mechanisms and roles of post-translational modifications is key to developing targeted therapies.This study aims to investigate the regulation and function of lysine acetylation of FLOT2 in nasopharyngeal carcinoma,providing new insights for targeting FLOT2 in cancer intervention. Methods:The PhosphoSitePlus database was used to analyze the lysine acetylation sites of FLOT2,and a lysine acetylation site mutation of FLOT2[FLOT2(K211R)]was constructed.Nasopharyngeal carcinoma cells were treated with histone deacetylase(HDAC)inhibitor trichostatin A(TSA)and Sirt family deacetylase inhibitor nicotinamide(NAM).TSA-treated human embryonic kidney(HEK)-293T were transfected with FLOT2 mutant plasmids.Co-immunoprecipitation(Co-IP)was used to detect total acetylation levels of FLOT2 and the effects of specific lysine(K)site mutations on FLOT2 acetylation.Western blotting was used to detect FLOT2/FLAG-FLOT2 protein expression in TSA-treated nasopharyngeal carcinoma cells transfected with FLOT mutant plasmids,and real-time reverse transcription PCR(real-time RT-PCR)was used to detect FLOT2 mRNA expression.Nasopharyngeal carcinoma cells were treated with TSA combined with MG132 or chloroquine(CQ)to analyze FLOT2 protein expression.Cycloheximide(CHX)was used to treat HEK-293T cells transfected with FLAG-FLOT2(WT)or FLAG-FLOT2(K211R)plasmids to assess protein degradation rates.The BioGrid database was used to identify potential interactions between FLOT2 and HDAC6,which were validated by Co-IP.HEK-293T cells were co-transfected with FLAG-FLOT2(WT)/FLAG-FLOT2(K211R)and Vector/HDAC6 plasmids,and grouped into FLAG-FLOT2(WT)+Vector,FLAG-FLOT2(WT)+HDAC6,FLAG-FLOT2(K211R)+Vector,and FLAG-FLOT2(K211R)+HDAC6 to analyze the impact of K211R mutation on total lysine acetylation levels.In 6-0B cells,overexpression of FLOT2(WT)and FLOT2(K211R)was performed,and the biological functions of FLOT2 acetylation site mutants were assessed using cell counting kit-8(CCK-8),colony formation,and Transwell invasion assays. Results:The PhosphoSitePlus database indicated that FLOT2 has an acetylation modification at the K211 site.Co-IP confirmed significant acetylation of FLOT2,with TSA significantly increasing overall FLOT2 acetylation levels,while NAM had no effect.Mutation at the K211 site significantly reduced overall FLOT2 acetylation,unaffected by TSA.TSA decreased FLOT2 protein expression in nasopharyngeal carcinoma cells without affecting FLOT2 mRNA levels or FLOT2(K211R)protein expression in transfected cells.The degradation rate of FLOT2(K211R)protein was significantly slower than that of FLOT2(WT).The proteasome inhibitor MG132 prevented TSA-induced FLOT2 degradation,while the lysosome inhibitor CQ did not.BioGrid data suggested a potential interaction between FLOT2 and HDAC6,confirmed by Co-IP.Knockdown of HDAC6 in nasopharyngeal carcinoma cells significantly increased FLOT2 acetylation;co-transfection of HDAC6 and FLAG-FLOT2(WT)significantly decreased total lysine acetylation levels,whereas co-transfection of HDAC6 and FLAG-FLOT2(K211R)had no effect.Knockdown of HDAC6 significantly reduced FLOT2 protein levels without affecting mRNA levels.MG132 prevented HDAC6-knockdown-induced FLOT2 degradation.Knockdown of HDAC6 significantly accelerated FLOT2 degradation.Nasopharyngeal carcinoma cells transfected with FLOT2(K211R)showed significantly higher proliferation and invasion than those transfected with FLOT2(WT). Conclusion:The K211 site of FLOT2 undergoes acetylation modification,and HDAC6 mediates deacetylation at this site,inhibiting proteasomal degradation of FLOT2 and maintaining its stability and tumor-promoting function in nasopharyngeal carcinoma.

Three-dimensional finite element models of the large defect of proximal femur were reconstructed with allograft prosthesis composite in clinical bone-healing phase; current model was under the given conditions of 138mm-intramedullary stem-length of host bone and 135mm defect-length of proximal femur. The femur was constructed with efilm software from CT data, then three-dimensional concrete models were created by using Proe-Wildfire software; the three-dimensional finite element models of allograft prosthesis composite were made in ANSYS11 software. Loads were simulated using the peaking values during stance walking. The stress on femur-cement-callus-prostheses and the influence of stress on the clinical bone-healing phase were analysed. The highest stress value of femur is on the medial side of the tip of the prostheses. The highest stress value of cement mantle is on the medical side of the cement mantle at the tip of the stem. The highest stress value of the prostheses is on the medial side near the upper 4cm of the stem tip. The highest stress value on the callus is at the medial side of the callus layer. The highest stress value on every part is under the corresponding fatigue strength. Clinical bone-healing phase model is well enough for stance walking.
Biomechanical Phenomena ; Female ; Femoral Neoplasms ; diagnostic imaging ; surgery ; Finite Element Analysis ; Hip Prosthesis ; Humans ; Imaging, Three-Dimensional ; Male ; Osteosarcoma ; diagnostic imaging ; surgery ; Tomography, Spiral Computed ; Weight-Bearing ; physiology ; Wound Healing

Objective:To observe the effects of alprostadil at different doses on C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in patients with angina pectoris.Methods:From August 2017 to April 2018, 120 patients with angina pectoris admitted to China Coast Guard Hospital of the People's Armed Police Force were selected and divided into control group (60 cases) and observation group (60 cases) by random number table method.Both two groups received routine anti-angina treatment, while the control group received 10 μg alprostadil, the observation group received 20 μg alprostadil for 2 weeks.The changes of CRP, TNF-α, IL-6 and hemorheological indicators before and after treatment were compared between the two groups.Results:Before treatment, there were no statistically significant differences in the CRP, TNF-α and IL-6 between the two groups (all P>0.05). After treatment, the CRP, TNF-α and IL-6 levels in the observation group were (4.63±0.62)mg/L, (0.46±0.08)μg/L, (46.59±4.72)ng/L, respectively, which were significantly lower than those in the control group [(6.18±0.74)mg/L, (1.19±0.28)μg/L, (58.62±5.07)ng/L]( t=12.437, 19.418, 13.452, all P<0.05). The high shear viscosity, low shear viscosity, plasma viscosity and platelet aggregation rates in the observation group were (4.27±0.46)mPa/s, (8.07±0.18)mPa/s, (1.03±0.25)mPa/s, (37.42±1.05)%, respectively, which were lower than those in the control group [(5.14±0.588)mPa/s, (10.43±0.42)mPa/s, (2.01±0.46)mPa/s, (40.19±1.86)%] ( t=9.103, 40.006, 14.499, 10.046, all P<0.05). Conclusion:The conventional dose of alprostadil (20 μg) is effective in the treatment of angina pectoris.It can improve CRP, TNF-α, IL-6 and hemorheological parameters of patients.

Objective:To observe the efficacy of meglumine adenosine cyclophosphate (MAC) in the adjuvant treatment of chronic congestive heart failure(CHF) in the elderly, and its effects on cystatin C(Cys-C) and plasma B-type natriuretic peptide(BNP).Methods:From August 2017 to August 2018, 110 elderly patients with chronic CHF admitted to Zhejiang Provincial Armed Police Corps Hospital were selected and randomly divided into control group(55 cases) and observation group(55 cases) by random number table method.The control group was treated with routine treatment, while the observation group was treated with MAC on the basis of routine treatment.Both two groups were treated for 2 weeks.The clinical efficacy, left ventricular ejection fraction (LVEF), cardiac output, Cys-C and plasma BNP were compared between the two groups.Results:The total improvement rate of the observation group was 94.55%(52/55), which was higher than 78.18%(43/55) of the control group, the difference was statistically significant(χ 2=6.253, P<0.05). Before treatment, there were no statistically significant differences in LVEF, cardiac output, BNP and Cys-C between the two groups(all P>0.05). After treatment, the LVEF[(44.16±6.82)%], cardiac output[(8.07±1.45)L/min] of the observation group were significantly higher than those of the control group[(40.71±5.38)%, (6.44±1.37)L/min], and BNP[(1.65±0.24)mg/L], Cys-C[(552.79±10.46)mg/L] of the observation group were significantly lower than those of the control group[(2.31±0.48)mg/L, (681.73±12.71)mg/L], the differences were statistically significant( t=2.945, 6.060, 9.121, 0.551, all P<0.05). Conclusion:MAC has significant effect in the treatment of elderly patients with chronic CHF.It can improve the cardiac function of patients and reduce the plasma concentrations of Cys-C and BNP.