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.

Light is the main source for plants to obtain energy.Asarum forbesii is a typical shade medicinal plant, which generally grows in the shady and wet place under the bushes or beside the ditches.It can grow and develop without too much light intensity.This experiment explores the effects of shading on the growth, physiological characteristics and energy metabolism of A.forbesii, which can provide reference and guidance for its artificial planting.In this experiment, A.forbesii was planted under 80%, 60%, 40%, 20% and no shade.During the vigorous growth period, the photosynthetic physiological characteristics such as fluorescence parameters, photosynthetic parameters, photosynthetic pigment content and ultrastructure, as well as the content of mitochondrial electron transport chain(ETC) synthase and nutrients were measured.The results showed that the photosynthetic pigment content, chlorophyll fluorescence parameters and net photosynthesis rate(P_n) decreased with the decrease of shading.Under 20%-40% shading treatment, the plants had damaged ultrastructure, expanded and disintegrated chloroplast, disordered stroma lamella and grana lamella, and increased osmiophi-lic granules and starch granules.The activities of nicotinamide adenine dinucleotide dehydrogenase(NADH), succinate dehydrogenase(SDH), cytochrome C oxidoreductase(CCO) and adenosine triphosphate(ATP) synthasewere positively related to light intensity.With the reduction of shading, the content of total sugar and protein in nutrients increased first and then decreased, and the content was the highest under 60% shade.In conclusion, under 60%-80% shading treatment, the chloroplast and mitochondria had more complete structure, faster energy metabolism, higher light energy-conversion efficiency, better absorption and utilization of light energy and more nutrient synthesis, which was more suitable for the growth and development of A.forbesii.
Asarum ; Chlorophyll/metabolism* ; Chloroplasts ; Energy Metabolism ; Photosynthesis/physiology* ; Plant Leaves/metabolism*

Objective: To explore the safety and feasibility of stereotactic radiation therapy (SBRT) strategy for irradiating porcine ventricular septum, see if can provide a preliminary experimental evidence for clinical SBRT in patients with hypertrophic obstructive cardiomyopathy (HOCM). Methods: Five male pigs (39-49 kg, 6 months old) were used in this study. Pigs were irradiated at doses of 25 Gy (n=2) or 40 Gy (n=3). Delineation of the target volume was achieved under the guidance of 3-dimensional CT image reconstruction, and SBRT was then performed on defined target volume of porcine ventricular septum. Blood biomarkers, electrocardiogram and echocardiography parameters were monitored before and after SBRT. Pathological examination (HE staining, Masson staining) was performed on the target and non-target myocardium at 6 months post SBRT. Results: SBRT was successful and all animals survived to the designed study endpoint (6 months) after SBRT. Serum cardiac troponin T (cTnT) level was significantly higher than the baseline level at 1 day post SBRT, and reduced at 1 week after SBRT, but was still higher than the baseline level(P<0.05). Serum N-terminal pro-B type natriuretic peptide (NT-proBNP) was also significantly increased at 1 day post SBRT (P<0.05) and returned to baseline level at 1 week post SBRT. The serum NT-proBNP level was (249±78), (594±37) and (234±46) pg/ml, respectively, and the cTnT was (14±7), (240±40) and (46±34) pg/ml, respectively at baseline, 1 day and 1 week after SBRT in the 40 Gy dose group. The serum NT-proBNP level was (184±20), (451±49) and (209±36) pg/ml, respectively, the cTnT values ​​were (9±1), (176±29) and (89±27) pg/ml, respectively at baseline, 1 day and 1 week after SBRT in the 25 Gy dose group. Both NT-proBNP and cTnT values tended to be higher post SBRT in the 40 Gy dose group as compared with the 25 Gy dose group, but the difference was not statistically significant (P>0.05). The left ventricular ejection fraction and the left ventricular end-diastolic diameter remained unchanged before and after SBRT (P>0.05). The interventricular septum thickness showed a decreasing trend at 6 months after SBRT, but the difference was not statistically significant ((9.54±0.24) mm vs. (9.82±8.00) mm, P>0.05). The flow velocity of the left ventricular outflow tract, and the valve function and morphology were not affected by SBRT. At 6 months after SBRT, HE staining revealed necrosis in the irradiated target area of ​​the myocardium in the 40 Gy dose group and the 25 Gy dose group, and the degree of necrosis in the irradiated interventricular septum was more obvious in the 40 Gy dose group as compared with the 25 Gy group. The combined histological analysis of the two groups showed that the necrotic area of ​​the irradiated target area accounted for (26±9)% of the entire interventricular septum area, which was higher than that of the non-irradiated area (0) (P<0.05). There was no damage or necrosis of myocardial tissue outside the target irradiation area in both groups. The results of Masson staining showed that the percentage area of myocardial fibrosis was significantly higher in the irradiated target area than non-irradiated area ((12.6±5.3)% vs. (2.5±0.8)%, P<0.05). Conclusion: SBRT is safe and feasible for irradiating porcine ventricular septum.
Animals ; Feasibility Studies ; Male ; Necrosis ; Radiosurgery/methods* ; Stroke Volume ; Swine ; Ventricular Function, Left ; Ventricular Septum

OBJECTIVE: To evaluate the effects of a 48-week course of adefovir dipivoxil (ADV) plus Chinese medicine (CM) therapy, namely Tiaogan Jianpi Hexue () and Tiaogan Jiedu Huashi () fomulae, in hepatitis B e antigen (HBeAg)-positive Chinese patients. METHODS: A total of 605 HBeAg-positive Chinese CHB patients were screened and 590 eligible participants were randomly assigned to 2 groups in 1:1 ratio including experimental group (EG, received ADV plus CM) and control group (CG, received ADV plus CM-placebo) for 48 weeks. The major study outcomes were the rates of HBeAg and HBV-DNA loss on week 12, 24, 36, 48, respectively. Secondary endpoints including liver functions (enzymes and bilirubin readings) were evaluated every 4 weeks at the beginning of week 24, 36, and 48. Routine blood, urine, and stool analyses in addition to electrocardiogram and abdominal B scan were monitored as safety evaluations. Adverse events (AEs) were documented. RESULTS: The combination therapy demonstrated superior HBeAg loss at 48 weeks, without additional AEs. The full analysis population was 560 and 280 in each group. In the EG, population achieved HBeAg loss on week 12, 24, 36, and 48 were 25 (8.90%), 34 (12.14%), 52 (18.57%), and 83 (29.64%), respectively; the equivalent numbers in the CG were 20 (7.14%), 41 (14.64%), 54 (19.29%), and 50 (17.86%), respectively. There was a statistically significant difference between these group values on week 48 (P<0.01). No additional AEs were found in EG. Subgroup analysis suggested different outcomes among treatment patterns. CONCLUSION Combination of CM and ADV therapy demonstrated superior HBeAg clearance compared with ADV monotherapy. The finding indicates that this combination therapy may provide an improved therapeutic effect and safety profile (ChiCTR-TRC-11001263).
Adenine ; analogs & derivatives ; therapeutic use ; Adult ; Antiviral Agents ; therapeutic use ; Double-Blind Method ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hepatitis B e Antigens ; immunology ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Male ; Medicine, Chinese Traditional ; Organophosphonates ; therapeutic use ; Young Adult

【Objective】 To study the function and mechanism of INI1 in human epithelioid sarcoma cells. 【Methods】 Western blotting was used to test the expressionlevel of INI1in epithelioid sarcoma cells. The expression of INI1 in clinical specimens of epithelioid sarcoma was detected by immunohistochemistry. Tet-on system was used to establish the expression of INI1 in epithelioid sarcoma cells. Following the induction of INI1 expression, the cell morphology, proliferation and the molecular markers of adipocytic differentiation were detected. Then western blot was employed to detect the transcription factors of adipocytic differentiation, and oil red O staining was also tested. 【Results】 The expression of INI1 was absent in both epithelioid sarcoma cells and clinical tissues(P<0.05). The INI1 expression in the epithelioid sarcoma cells was successfully established by Tet-on system. After induction by doxycycline, the expression of INI1 was up-regulated. With continuous expression of INI1, the morphology of VA-ES-BJ cells was changed, with the appearance of abundant vacuoles in the cytoplasm. The cell proliferation was obviously inhibited(P<0.05). Epithelial cell marker Cytokeratin was found significantly reduced(P<0.05). In addition, the adipocyte markers, leptin, adiponectin and lipoprotein lipase were significantly up-regulated(P<0.05). The adipogenic transcriptional factors PPAR-γ and CEBP-α were found upregulated(P<0.05) and the oil red staining was significantly positive. 【Conclusions】 The expression of INI1 was absent in epithelioid sarcoma. Reconstruction of the expression of INI1 could induce adipocytic differentiation via upregulation of transcriptional factors PPAR-γand CEBP-αin epithelioid sarcoma cells.

Objective:To observe the effect of external phrenic nerve electrical stimulation on respiratory function for convalescent stroke patients. Methods:From October, 2017 to July, 2018, 27 convalescent stroke patients were randomly divided into control group (n = 14) and observation group (n = 13). Both groups received routine rehabilitation, while the observation group accepted external phrenic nerve electrical stimulation additionaly. Their ventilation and respiratory muscle strength were measured before and four weeks after treatment. Results:The forced vital capacity (FVC), forced expiratory volume in 1st second (FEV1), peak expiratory flow (PEF), maximal inspiratory pressure (MIP) and maximal expiratory pressure (MEP) improved in both groups after treatment (t > 5.075, Z = -3.366, P < 0.001), and improved more in FVC, FEV1, PEF and MIP in the observation group than in the control group (t > 0.921, Z = -2.746, P < 0.05). Conclusion:External phrenic nerve electrical stimulation can improve ventilation and inspiratory muscle strength for convalescent stroke patients.

Objective To observe the clinical efficacy and safety of ozagrel sodium injection combined with mild hypothermia in the treatment of craniocerebral injury acute cerebral infarction.Methods Sixty-two patients of craniocerebral injury acute cerebral infarction were randomly divided into control group (n =31 cases) and treatment group (n =31 cases).Control group was given Xueshuantong injection 10 mL qd,intravenous drip.Treatment group was taken with ozagrel sodium 100 mL bid,intravenous drip,and mild hypothermia treatment.Two groups were treated for 14 d.The clinical efficacy,national institutes of health stroke scale (NIHSS) scores,Barthel index (BI),changes of hemodynamic indexes and adverse drug reactions were compared between two groups.Results After treatment,the total effective rates in treatment and control groups were 90.32％ (28 cases/31 cases) and 61.29％ (19 cases/ 31 cases) with significant difference (P ＜0.05).After treatment,the main indexes in treatment and control groups were compared:NIHSS scores were (8.24 ± 0.76),(15.42 ± 1.65) points;BI were (75.23 ± 3.48),(52.32 ± 3.33) points;whole blood high shear viscosity respectively were (3.46 ±0.21),(4.63 ±0.26) mPa · s;whole blood low shear viscosity were (7.69 ±0.32),(8.38 ±0.31) mPa · s;plasma viscosity were (1.24 ±0.11),(1.76 ±0.16) mPa · s;fibrinogen were (3.07 ±0.41),(4.16 ±0.18)g · L-1,there were statistically significant differences (all P ＜0.05).No adverse drug reactions were observed during the treatment in two groups.Conclusion Ozagrel sodium injection combined with mild hypothermia has a definitive clinical efficacy and safety in the treatment of craniocerebral injury acute cerebral infarction,which can improve the patient's hemodynamic parameters,neurological function and activities of daily living.

A new benzaldehyde, 3-hydroxy-4-(4-(2-hydroxyethyl) phenoxy) henzaldehyde(1), together with six known compounds, including isovanillic acid(2), pyrocatechol(3), glutinosalactone A(4), chrysoeriol(5), apigenin(6) and luteolin(7) were isolated from aerial part of Rehmannia glutinosa. The compounds were isolated by macroporous resin, silica gel, Sephadex LH-20 and HPLC chromatographies. The chemical structures of 1-7 were elucidated on the basis of spectral analysis (MS, 1D NMR and 2D NMR).
Benzaldehydes ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Rehmannia ; chemistry ; Spectrometry, Mass, Electrospray Ionization

To investigate the chemical compounds from aerial part of Rehmannia glutinosa, six compounds were isolated and deter- mined by extensive spectroscopic analysis as(+)-(7S, 8S, 8'S)-9-O-[β-D-glucopyranoyl] asarininone(1), 2α,3β,19α,23-tetrahydroxy-olean-12-en-28-oic acid(2),7,3'-dihydroxyl-5'-methoxyisoflavone (3), aeginetic acid (4), corchorifattty acid B (5), pinellic acid (6). Among them, compound 1 was a new natural product. Compounds 2, 3 and 5 were obtained from the Rehmannia genus for the first time. In vitro study showed that none of the six compounds exhibited obvious activities to BEL-7402 and HCT-8 at the concentration of 10 mg x L(-1).
Antineoplastic Agents, Phytogenic ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Rehmannia ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Triterpenes ; chemistry ; pharmacology