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OBJECTIVE To establish the method for determination of bile acid active constituents （cholic acid， hyodeoxycholic acid and sodium taurocholate） in Babao jingfeng powder， and to compare the differences of 3 constituents in Babao jingfeng powder from different manufacturers. METHODS UPLC-MS method was adopted to determine the contents of cholic acid， hyodeoxycholic acid and sodium taurocholate in 9 batches of Babao jingfeng powder from 3 manufacturers. The separation was performed on a Phenomenex Luna® C18 column with mobile phase consisted of acetonitrile-water （gradient elution） at a flow rate of 0.5 mL/min. The column temperature was 40 ℃ and the sample size was 2 μL. MS system was operated in a negative ion single quadrupole mode， and the m/z of cholic acid， hyodeoxycholic acid and sodium taurocholate were 407.2， 392.2 and 514.2， respectively. With the contents of three components as the index， cluster analysis and principle component analysis were performed on 9 batches of samples. RESULTS The linear ranges of cholic acid， hyodeoxycholic acid and sodium taurocholate were 5.48- 548.40， 5.38-538.40， 4.74-474.05 ng/mL， respectively （r≥ 0.999 3）. RSDs of precision， repeatability and stability tests（24 h） were all lower than or equal to 3.7% （n＝6）， and the average recoveries were 103.3%， 103.3% and 101.6% with RSDs of 3.3%， 3.4%， 4.2% （n＝6）， respectively. The contentsof cholic acid ranged 0.702-1.711 mg/g， those of deoxycholic acid ranged 0.599-2.049 mg/g， and those of sodium taurocholate ranged 0.664-1.752 mg/g in 9 batches of samples. The average content of 3 components in samples from manufacturer A was high， and the difference between batches was the smallest； the average content of 3 components from manufacturer C was the lowest， and the difference between batches was large. Cluster analysis could basically distinguish samples from different manufacturers； cluster analysis was conducted by using the comprehensive score of principal components as an indicator， the results of which were basically consistent with those of cluster analysis for content. CONCLUSIONS The method is established successfully for the content determination of the 3 bile acid active constituents of artificial bezoar in Babao jingfeng powder. The contents of 3 components in Babao jingfeng powder from 3 manufacturers are significantly different.
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OBJECTIVE To establish the met hod for the purific ation of test sample of Banxialu granules ，and to determine the contents of 4 ephedrine components such as ephedrine ，pseudoephedrine，norpseudoephedrine and methylephedrine. METHODS Three batches of Banxialu granules were extracted with methanol（containing 1％formic acid ）and pretreated with QuEChERS method of N-propyl ethylenediamine adsorbent （PSA）and octadecyl bonded silica gel adsorbent （C18）. Ultra high performance liquid chromatography tandem triple quadrupole mass spectrometry （UHPLC-MS/MS）was adopted. The separation was performed on an Agilent XDB-C 18 column with 5 mmol/L ammonium acetate solution （containing 0.1％ formic acid ）-acetonitrile as mobile phase（gradient elution ）at the flow rate of 0.40 mL/min. The column temperature was set at 30 ℃，and sample size was 2 μL. The electrospray ionization source was adopted ，and positive ion scanning was performed in multiple reaction monitoring mode. The ion pairs used for quantitative analysis were m/z 166.2→148.1（ephedrine，pseudoephedrine），m/z 152.2→134.1（norpseudoephedrine）， m/z 180.2→162.2（methylephedrine）. RESULTS The solution obtained by QuEChERS purification method was clear and nearly colorless. The linear ranges of ephedrine ， pseudoephedrine， norpseudoephedrine and methylephedrine were 1.38-206.82， 1.41-212.13，1.29-19.34，1.99-59.83 ng/mL（r＞0.99）. The limits of detection were 0.41，0.42，0.39 and 0.60 ng/mL. The limits of quantitation were 1.38 ，1.41 ，1.29 and 1.99 ng/mL，respectively. RSDs of precision ，stability（48 h）and repeatability tests were all lower than 2％. The average recoveries were 95.75％-100.87％（RSD＜2％，n＝9）. The contents of above 4 ephedrine components were 20.62-26.02，20.96-24.90，2.26-2.63，5.36-6.32 μg/g， respectively. CONCLUSIONS Established method is simple ，rapid，sensitive and suitable for simultaneous determination of 4 ephedrine components in Banxialu granules.
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DNA methylation as an important aspect of epigenetics plays an important role in spermatogenesis and embryonic development. In recent years, researchers have found that male infertility, in particular abnormal semen quality, is related to abnormal DNA methylation. To further delineate the pathogenesis of male infertility and inspire new ideas for the treatment of male infertility, a comprehensive review over the correlation between abnormal methylation of imprinted genes, repetitive DNA elements and non-imprinted genes, semen quality (including sperm count, morphology, and vitality) and male infertility is provided.
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Humans , Male , DNA Methylation , Infertility, Male/genetics , Semen Analysis , Sperm Count , Spermatogenesis , Spermatozoa/pathologyABSTRACT
OBJECTIVE：To establish a method for the determination of plasma protein binding rate of rosmarinic acid ，caffeic acid and chlorogenic acid from Glechoma longituba . METHODS ：UHPLC method combined with ultrafiltration method was adopted to determine the plasma protein binding rate of rosmarinic acid ，caffeic acid and chlorogenic acid from G. longituba in the plasma of New Zealand rabbits. The determination was performed on a Phenomenex Luna ® C18 column with mobile phase consisted of acetonitrile （A）-0.1％ formic acid solution （B）（gradient elution ）at the flow rate of 0.5 mL/min. The column temperature was set at 45 ℃，and the detection wavelength was 327 nm. The sample size was 3 μL. RESULTS：At low ，medium and high concentrations，the plasma binding rates of rosmarinic acid were （97.78 ± 1.67）％ ，（94.32 ± 1.42）％ ，（95.12 ± 1.51）％ ， respectively（n＝3）；those of caffeic acid were （90.12±2.33）％，（89.53±1.98）％，（90.23±1.56）％，respectively（n＝3）；those of chlorogenic acid were （63.23 ± 2.12）％ ，（67.87 ± 1.06）％ ，（62.34 ± 1.34）％ ，respectively （n＝3）. CONCLUSIONS ： Established method is easy to operate and shorter time for analysis. It can be used to determine the plasma protein binding rate of rosmarinic acid ，caffeic acid and chlorogenic acid in G. longituba .
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Objective@#To investigate the reversibility of ischemic core defined by CT perfusion imaging in acute ischemic stroke (AIS) patients receiving intravenous thrombolysis within different time windows and influencing factors.@*Methods@#The data of AIS patients who received intravenous thrombolysis in the Department of Neurology of Lishui People′s Hospital from May 2016 to December 2018 were retrospectively reviewed. All patients had finished multi-model CT imaging before thrombolysis and multi-model MRI examination 24-48 hours after thrombolysis. The baseline ischemic core volume (hypoperfusion area with relative cerebral blood flow (rCBF)<30%) was quantitatively assessed based on CT perfusion images using MIStar software, and the final ischemic core volume was assessed based on diffusion weighted imaging acquired 24-48 hours after thrombolysis. The reversibility of ischemic core was defined as baseline ischemic core volume-the final infarct volume ≥5 ml. Then the clinical and imaging features of the patients between reversible group and irreversible group were compared, and the predictors of ischemic core reversibility were analyzed by binary Logistic regression analysis.@*Results@#Finally, 97 patients were enrolled in the present study, of which 64 (66%) patients achieved successful recanalization, 51 (53%) patients with reversible baseline ischemic core. For patients with recanalization, the incidence of reversibility was 76% (26/34), 71% (17/24), 2/5 and 0 (0/1) in patients with time window from onset to thrombolysis (ONT) <3.0 h, 3.0-4.5 h, 4.6-6.0 h, and >6.0 h, respectively. In the non-recanalization group, six patients were also showed with ischemic core reversibility, including 4 (4/12) in the ONT<3.0 h group and 2 (2/12) in the ONT 3.0-4.5 h group. It was found that the reversible volume was positively correlated with baseline ischemic core volume (r=0.805, P<0.001) by Spearman correlation analysis. Finally, binary Logistic regression analysis revealed that the history of hypertension, ONT and recanalization were independent predictors of reversible changes of baseline ischemic core.@*Conclusions@#The ischemic core defined by CT perfusion imaging (rCBF<30%) was considerably inaccurate for patients with ONT<6.0 h. If recanalization could be achieved within this time window, most of the patients would manifest with ischemic core reversibility, the predictors of which also included hypertension history and ONT.
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Objective To investigate the reversibility of ischemic core defined by CT perfusion imaging in acute ischemic stroke (AIS) patients receiving intravenous thrombolysis within different time windows and influencing factors.Methods The data of AIS patients who received intravenous thrombolysis in the Department of Neurology of Lishui People's Hospital from May 2016 to December 2018 were retrospectively reviewed.All patients had finished multi-model CT imaging before thrombolysis and multi-model MRI examination 24-48 hours after thrombolysis.The baseline ischemic core volume (hypoperfusion area with relative cerebral blood flow (rCBF)＜30％) was quantitatively assessed based on CT perfusion images using MIStar software,and the final ischemic core volume was assessed based on diffusion weighted imaging acquired 24-48 hours after thrombolysis.The reversibility of ischemic core was defined as baseline ischemic core volume-the final infarct volume ≥5 ml.Then the clinical and imaging features of the patients between reversible group and irreversible group were compared,and the predictors of ischemic core reversibility were analyzed by binary Logistic regression analysis.Results Finally,97 patients were enrolled in the present study,of which 64 (66％) patients achieved successful recanalization,51 (53％) patients with reversible baseline ischemic core.For patients with recanalization,the incidence of reversibility was 76％ (26/34),71％ (17/24),2/5 and 0 (0/1) in patients with time window from onset to thrombolysis (ONT) ＜3.0 h,3.0-4.5 h,4.6-6.0 h,and ＞6.0 h,respectively.In the non-recanalization group,six patients were also showed with ischemic core reversibility,including 4 (4/12) in the ONT＜3.0 h group and 2 (2/12) in the ONT 3.0-4.5 h group.It was found that the reversible volume was positively correlated with baseline ischemic core volume (r=0.805,P＜0.001) by Spearman correlation analysis.Finally,binary Logistic regression analysis revealed that the history of hypertension,ONT and recanalization were independent predictors of reversible changes of baseline ischemic core.Conclusions The ischemic core defined by CT perfusion imaging (rCBF＜30％) was considerably inaccurate for patients with ONT＜6.0 h.If recanalization could be achieved within this time window,most of the patients would manifest with ischemic core reversibility,the predictors of which also included hypertension history and ONT.
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Objective To compare the clinical features between radiological isolated syndrome (RIS) and classical multiple sclerosis (CMS), in order to improve the understanding of the RIS. Methods All 35 patients with RIS and 32 patients with CMS were selected. The epidemiological and clinical findings, cerebrospinal fluid, neural electrophysiological examination and magnetic resonance imaging (MRI) data were analyzed. Results There were no statistical differences in sex ratio and onset age between RIS patients and CMS patients (P>0.05). The main symptoms of in patients with RIS were headache (45.7%, 16/35), dizziness (40.0%, 14/35), hypomnesis (20.0%, 7/35) and psychiatric disorders (11.4%, 4/35). But the main symptoms of in patients with CMS were limb weakness (75.0%, 24/32), sensory abnormalities (68.8%, 22/32) and ocular symptoms (34.4%,11/32). The incidences of limb weakness, sensory abnormalities and ocular symptoms in patients with CMS were significantly higher than those in patients with RIS:75.0%(24/32) vs. 0, 68.8%(22/32) vs. 0 and 34.4%(11/32) vs. 0, and there were statistical differences (P0.05). The cerebrospinal fluid protein and the incidences of IgG index>0.7 in patients with RIS were significantly lower than those in patients with CMS:0.175 (0.03-0.69) g/L vs. 0.440 (0.04-1.09) g/L and 3/18 vs. 47.6%(10/21), and there were statistical differences (P0.05). On MRI, the demyelinating lesions of RIS and CMS were both mainly distributed in the periventricular, semi-oval center, infratentorial white matter, partly involving corpus callosum or cortical. The rates of demyelinating lesions in brainstem and cerebellum in patients with RIS were significantly lower than those in patients with CMS:5.7%(2/35) vs. 34.4% (11/32) and 2.9% (1/35) vs. 25.0% (8/32), and there were statistical differences (P<0.01 or <0.05). Comparison with CMS lesions, RIS lesions mainly showed patching and stippled, and there were statistical differences (P<0.01 or <0.05). The rates of lesions enhancement and spinal cord injury in patients with RIS were significantly lower than those in patients with CMS: 2/17 vs. 45.0% (9/20) and 1/14 vs. 43.5% (10/23), and there were statistical differences (P<0.05). Conclusions There are differences in clinical findings, cerebrospinal fluid, neural electrophysiological examination and MRI appearances between RIS and CMS.
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Objective To study the default mode network (DMN) of mild cognitive impairment (MCI) related to leukoaraiosis (LA) with resting-state functional magnetic resonance imaging (rs-fMRI). Methods 31 LA patients (Clinical Dementia Rating of 0.5) and 27 nor-mal controls (Clinical Dementia Rating of 0, and matched in age, gender and lever of education) were scanned with the rs-fMRI. The data was analyzed with SPM5 software, using independent component analysis. The differences between the both groups were compared with two-sample t-test. Results The DMN during resting-state of normal controls was posterior cingulate cortex/precuneus, bilateral medial fron-tal cortex, bilateral middle temporal gyrus, bilateral inferior parietal gyrus, angular gyrus, bilateral hippocampus. The DMN of MCI group was consistent with the normal controls, but the activation decreased in anterior cingulate cortex/left medial frontal lobe, right parahippcam-pus/uncus, right inferior temporal gyrus, left deep frontal white matter/head of caudate nucleus;and increased in the left caudate nucleus/an-terior cingulate cortex, left frontal lobe, and left superior temporal gyrus/inferior parietal gyrus. Conclusion Activation of resting-state func-tional network is disorder in LA, which may relate to cognitive impairment.