This study was aimed to investigate the relationship between the quality ofRheum palmatum L. and ecological factors, in order to provide the basis for nurture and cultivation ofR. palmatumL. The ultra performance liquid chromatography (UPLC) was used in the determination of 8 effective components in samples from 3 majorR. palmatumL. producing provinces, which wereGansu,SichuanandQinghai province. The SPSS software was applied to analyze the effective components by the one-way analysis of variance. And the correlation analysis between the content and ecological factors was also conducted. The results showed that the pressure, relative humidity and temperature had close relationship with the content of bioactive compounds inR. palmatumL. The content of effective components inR. palmatumL. was positively related to pressure, relative humidity, and temperature. It was concluded that the quality ofR. palmatumL. in Gansu province was better. And the ecological factors affected the accumulation of effective components. This research provided experimental basis for the quality ofR. palmatumL. in different regions and ecological adaptation. The results demonstrated critical meaning forR. palmatumL. quality improvement, appropriate ecological division, and industrialized development promotion ofR. palmatumL.

Objective To establish a fast and accurate technique of identifying the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation.Methods Using STA-PUT velocity sedimentation method to isolate the pachytene spermatocytes, round spermatids and elongating and condensing spermatids from mouse testes.To determine the cell populations` distribution,each tube of cell fraction was then partially transfered to the 96 plate well,and each well was added with acridine orange dye.Then each well was analyzed using fluorescence microscopy.Results Three types of spermatogenic cells can be identified quickly and accurately by it`s specific cytoplasm/nucleus character using the acridine orange dye staining under fluorescence detection.Conclusions A rubust method to quickly and accurately determine the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation is successfully developed.

AIM To investigate whether dipfluzine (Dip) possesses antiarrhythmic effect on experimental arrhythmias and effect on cytosolic calcium in ventricular myocytes of guinea-pig. METHODS Experimental arrhythmias were induced by strophanthin G infusion through jugular vein in guinea-pigs and by myocardial ischemia-reperfusion (I-R) in rats respectively. Cytosolic calcium concentration ([Ca2+]i) of isolated guinea-pig ventricular myocytes was examined with laser confocal scanning microscope. RESULTSIn guinea-pigs pretreatment with Dip 20 mg·kg-1 increased the dosages of strophanthin G required to induce ventricular premature contraction (VP), ventricular tachycardia (VT), ventricular fibrillation (VF) and cardiac arrest (CA), pretreatment with Dip 10 mg·kg-1 increased the dosages of strophanthin G required to induce VP. In the I-R-induced arrhythmic model of rats, Dip 20 mg·kg-1 decreased the number of rats exhibiting VT, VF and CA, and the number of rats exhibiting VF and CA was decreased by Dip 10 mg·kg-1. Both Dip and verapamil (Ver) decreased [Ca2+]i of the ventricular myocytes in normal Tyrode′s solution. The Ca2+ overload evoked by high extracellular Ca2+ levels was inhibited by Dip and Ver, and the prophylactic effect of Dip was less than that of Ver, while the curative effect of Dip was more obvious than that of Ver. CONCLUSION Dip has antiarrhythmic effect, which is likely related to the modulation on the intracellular calcium homeostasis.

Objective To investigate the combined biological effects of low dose radiation,carbon monoxide,benzene and noise on rats.Methods Sixteen male SD rats were randomly divided into experiment group and control group.The experiment group was exposed to carbon monoxide,benzene,low dose radiation and noise daily,the control group was in common environment.Peripheral blood,organ index,and marrow DNA content were detected.Two-dimensional electrophoresis (2-DE) was performed on serum protein analysis.Differential expressed proteins were identified by a matrix assisted laser desorption/ionization time of flight mass spectrometry (MAIDI-TOF-MS).Results Compared to control group,the liver index,spleen index,thymus index,leukocytes,platelets count,and marrow DNA content of the experiment group were decreased significantly (t =2.732,4.141,3.053,2.211,2.668,11.592,P ＜0.05).12 altered proteins were detected and through identification,3 proteins were definite in terms of serum amyloid A-4 protein (SAA4),trichoplein keratin filament-binding protein (TCHP) and tubulin alpha-4A chain (TUBA4A).Conclusions The hematopoietic system and immune system of rats are damaged significantly with the changes of several serum protein expressions by the combined exposure of low dose radiation,carbon monoxide,benzene and noise.This study may provide new information for the mechanism of the combination effects.

BACKGROUND:It has been found that central nervous system is involved in Guillain-Barre syndrome and Miller-Fisher syndrome, and the involved sites include optic nerve, brain stem and cerebellum. Abnormal signal of MRI can be observed in the brainstem and spinocerebellar tract of patients with Miller-Fisher syndrome. To establish an animal model of encephalitis after infection of Campylobacter jejuni, and investigate the mechanism of formation by means of imaging, immunology and pathology.OBJECTIVE: To construct an animal model of lesion of central nervous system after infection of Campylobacter jejuni Penner 4.DESIGN: A randomized grouping designed, controlled animal experiment.SETTING: Department of Imaging and Department of Neurology, Second Hospital of Hebei Medical University.MATERIALS: The experiment was carried out in the Laboratory of Molecular Imaging, the Second Hospital of Hebei Medical University between August and December 2003. Fifteen healthy flap-eared rabbits were randomly divided into experimental group (n=10) and control group (n=5).METHODS: In the experimental group, Campylobacter jejuni inactivated bacteria liquor was completely emulsified with complete Freund adjuvant (CFA) of the same volume in week 1, and then the rabbits were immunized with subcutaneous injection at multiple points of bilateral axilla, bilateral groins and side of back spine, 1 mL for each site, and 5 mL for each rabbit; The rabbits were further immunized with intraperitoneal injection of simple Campylobacterjejuni inactivated bacteria liquor in the following every two weeks, 5 mL for each time in each rabbit for 5 times. In the control group, the Campylobacter jejuni inactivated bacteria liquor was replaced by saline of the same volume, the injected method and time were all the same as those in the experimental group. Evaluative methods: ①Symptoms and physical signs: their mental status, conditions of diet, urine and excrement, and activities of limbs were observed; ② Serological examination: the contents of anti-Campylobacterjejuni antibody, anti-IgG GM1 antibody and myelin basic protein (MBP) were detected with enzyme-linked immunoadsorbent assay (ELISA); ③ MRI examination was applied to the randomly selected rabbits before every immunization with Toshiba 1.5 T MRI instrument. The scanning sequence included spin-echo T1-weighted image with the scanning parameter of 500/15 ms (TR/TE); rapid spin-echo T2-weighted image, 4 000/108 ms (TR/TE); fluid attented inversion recovery (Flair) sequence, the parameter was 10 000/120 ms (TR/TE), inversion angle was 90°. The thickness of scanning layer was 4.0 mm, and the layer space was 0.8 mm. ④ Histological examination: At 4 weeks after the first immunization, the attacked animals were induced to death by cardiac perfusion, and the skull was opened immediately to remove optic nerve, part white matter, hippocampus, brainstem, cerebellum and spinal cords of neck, chest and waist, which were fixed with formaldehyde solution (40 g/L),and hematoxylin-eosin (HE) staining, fast blue staining and MBP immunohistochemical staining were performed respectively. At 10 weeks after immunization, 5 randomly selected rabbits in the experimental group and the 5 rabbits in the control group were treated with the same methods to obtain the histological samples.MAIN OUTCOME MEASURES: The symptoms and physical signs,contents of anti-Campylobacterjejuni antibody, anti-IgG GM1 antibody and MBP, imaging observation and histological examination were mainly observed.RESULTS: Fifteen animals were enrolled, 14 were involved in the analysis of results, 1 rabbit in the experimental group died at 4 weeks after immunization. ① Mental symptoms and disorder of limb's activity occurred in 1 rabbit in the experimental group at 2 weeks after immunization. ② In the experimental group, titre of anti-Campylobacterjejuni-IgG antibody in serum reach the peak at 2-4 weeks. From week 2, the serum A value was significantly higher in the experimental group than in the control group (1.923±0.403, 0.973±0.633, P ＜ 0.05). The IgG type GM1 (A value) was obviously elevated at week 8, but insignificantly different from that in the control group (0.115±0.042, 0.097±0.039, P ＞ 0.05). The MBP content (Avalue) in serum was significantly elevated at the 8th week (0.134±0.041).③ The imaging examination showed that abnormal MRI signal of different degree occurred at 2-4 weeks after immunization in the experimental group. ④ The histological changes showed that there was swelling of myelin sheath at the sites of brainstem, medulla oblongata, cervical spinal cord, thoracic spinal cord and lumbar spinal cord in the experimental group, no inflammatory cell infiltration and deletion of myelin sheath were observed. No obvious changes at the above site were observed in the contro1 group.CONCLUSION: Campylobacterjejuni Penner 4 can induce lesion of central nervous system.

OBJECTIVE: To investigate the expression and significance of hypoxia inducible factor-2α (HIF-2α) and transcription factor OCT-4 in non-small cell lung cancer (NSCLC), and evaluate their roles in the prognosis of NSCLC. METHODS: Tissues from 51 cases of NSCLC were collected and immunohistochemistry (SP method) was used to detect the expression of HIF-2α and OCT-4 proteins. The correlation between the protein expression and the prognosis of NSCLC was analyzed. RESULTS: The positive rates of HIF-2α and OCT-4 expression in the NSCLC were 52.9% and 72.5%, respectively. There was significant relation between the expression of HIF-2α and OCT-4 (r=0.514,P<0.01). High expression of them revealed poor prognosis for NSCLC patients characterized with a bad overall survival(P<0.05). CONCLUSION There is a negative corelation between the expression of HIF-2α and OCT-4 and the prognosis of NSCLC. Combined examination of HIF-2α and OCT-4 expression might be an important biomarker for NSCLC prognosis.
Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; Female ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; Male ; Octamer Transcription Factor-3 ; metabolism ; Prognosis

Objective:To analyze the effects of proton pump inhibitors(PPIs)on the prevention of stress ulcers(SU)in elderly patients with acute respiratory distress syndrome(ARDS), and to analyze related factors for the risk of short-term death.Methods:This study was a multicenter retrospective cohort study.Two hundred elderly ARDS patients diagnosed and treated at Peking University International Hospital, Anzhen Hospital and Ezhou Central Hospital from November 2017 to December 2019 were continuously included.These patients were treated with PPIs(omeprazole, pantoprazole, rabeprazole, lansoprazole and esomeprazole)within 48 hours after ICU admission to prevent SU and were considered as the PPI group.According to the propensity score matching method, 200 elderly ARDS patients admitted to the hospitals with similar ages, medical history and sequential organ failure assessment(SOFA)scores who did not use PPIs were selected as the control group.All patients were followed up for 30 days.Kaplan-Meier survival analysis and the log-rank test were used to compare the 30-day mortality risk between the two groups.Cox regression analysis was used to analyze the relevant factors affecting the 30-day mortality.The 30-day mortality risk and the incidence of clinically significant gastrointestinal bleeding were evaluated among patients using different PPIs.Results:The average time of PPI use was 8.4±4.4 d in the PPI group.In the control group, 38.0% of patients were treated with H 2 receptor antagonists, and the average time of use was 8.1±5.2 days.There was no significant difference in the 30-day all-cause mortality risk between the two groups(20.5% or 41 cases vs.23.5% or 47 cases, P>0.05). The incidences of clinically significant upper gastrointestinal tract bleeding(2.5% or 5 cases vs.7.0% or 14, P<0.05), gastrointestinal bleeding(5.5% or 11 cases vs.12.5% or 25 cases, P<0.05)and hospital-acquired pneumonia(9.0% or 18 vs.4.0% or 8 cases, P<0.05)had significant differences between the PPI group and the control group.Multivariate Cox regression analysis showed that age>70 years( HR=1.845, 95% CI: 1.131-3.010, P<0.05), arterial oxygen partial pressure <78.0 mmHg(1 mmHg=0.133 kPa, HR=2.143, 95% CI: 1.317-3.487, P<0.01), SOFA score>14( HR=3.603, 95% CI: 1.741-7.456, P<0.01)and blood lactic acid>3.8 mmol/L( HR=2.725, 95% CI: 1.437-5.167, P<0.01)were related factors for the 30-day mortality.Compared with the control group, there was no significant difference in 30-day mortality between the five subgroups taking different PPIs including omeprazole, pantoprazole, rabeprazole, lansoprazole and esomeprazole( P>0.05), and the incidence of clinically significant gastrointestinal bleeding was significantly reduced( P<0.05), but there was no significant difference between the five PPIs subgroups( P>0.05). Conclusions:Although PPIs have no effect on short-term death in elderly ARDS patients, it can increase the risk of hospital acquired pneumonia while reducing the occurrence of gastrointestinal bleeding.With PPI use, advanced age, low arterial oxygen partial pressure, high SOFA score and high blood lactate are risk factors for the 30-day mortality.

OBJECTIVETo develop a high performance liquid chromatography coupled with a photodiode array detector (HPLC-DAD) method for simultaneous determination of 7 nucleosides and nucleobases in Fritillaria taipaiensis.

METHODThe analyses were performed on an Agilent Zorbax-SB-Aq-C18 column (4.6 mm x 250 mm, 5 microm) eluted with water and methanol in gradient mode. The flow rate was 1.0 mL x min(-1). The detection wavelength was 260 nm. The temperature of sample manager was set at 25 degrees C, and the injection volume was 20 microL.

RESULTThe investigated compounds including uracil, cytidine, uridine, guanosine, thymidine, adenosine and adenine were shown good linearity (r > or = 0.999 8) over the tested ranges. The average recoveries were within 96.96% - 103.5% with RSD < or = 3.8%.

CONCLUSIONThe accuracy, stability, repeatability and average recovery of the method are satisfying, and the seven nucleosides and nucleobases components in F. taipaiensis can be rapidly and accurately quantified by HPLC-DAD. This work provided helpful information for comprehensive quality evaluation of F. taipaiensis.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Fritillaria ; chemistry ; Nucleosides ; analysis ; Reproducibility of Results ; Water ; chemistry

OBJECTIVETo establish a HPLC fingerprint of water-soluble constituents of Fritillaria unibracteata.

METHODZorbax SB Aq C18 chromatographic column (4. 6 mm x 250 mm, 5 microm) was adopted for gradient elute with the mobile phase consisting of methanol and water. The flow rate was 1.0 mL x min(-1); the detection wavelength was 260 nm, and the temperature of sample manager was set at 25 degrees C. Similarity Evaluation System for Chromatographic Fingerprint of traditional Chinese medicine (version 2.0) published by the State Pharmacopeia Committee of China was adopted for the fingerprint analysis on the 11 batches of F. unibracteata herbs.

RESULTThe 11 batches of F. unibracteata herbs had 14 common peaks, nine of which were identified with good separating degrees. The similarities of the 11 batches were more than 0. 970, with good quality homogeneity.

CONCLUSIONThe method is so accurate, highly reproducible and stable that it is suitable for the comprehensive quality evaluation of F. unibracteata herbs.

Chromatography, High Pressure Liquid ; methods ; Dermatoglyphics ; Drugs, Chinese Herbal ; chemistry ; Fritillaria ; chemistry ; Medicine, Chinese Traditional ; Solubility ; Water ; chemistry

Acetaminophen (APAP) overdose is a major cause of liver injury. Neural precursor cell expressed developmentally downregulated 4-1 (NEDD4-1) is an E3 ubiquitin ligase that has been implicated in the pathogenesis of numerous liver diseases; however, its role in APAP-induced liver injury (AILI) is unclear. Thus, this study aimed to investigate the role of NEDD4-1 in the pathogenesis of AILI. We found that NEDD4-1 was dramatically downregulated in response to APAP treatment in mouse livers and isolated mouse hepatocytes. Hepatocyte-specific NEDD4-1 knockout exacerbated APAP-induced mitochondrial damage and the resultant hepatocyte necrosis and liver injury, while hepatocyte-specific NEDD4-1 overexpression mitigated these pathological events both in vivo and in vitro. Additionally, hepatocyte NEDD4-1 deficiency led to marked accumulation of voltage-dependent anion channel 1 (VDAC1) and increased VDAC1 oligomerization. Furthermore, VDAC1 knockdown alleviated AILI and weakened the exacerbation of AILI caused by hepatocyte NEDD4-1 deficiency. Mechanistically, NEDD4-1 was found to interact with the PPTY motif of VDAC1 through its WW domain and regulate K48-linked ubiquitination and degradation of VDAC1. Our present study indicates that NEDD4-1 is a suppressor of AILI and functions by regulating the degradation of VDAC1.