Objective To establish a method for simultaneous determination of contents of schisandrin B, schisantherin A, lobetyolin and ruscogenin in Huangqi Shengmai Decoction with HPLC method. Methods The chromatographic column was an Agilent Eclipse C18 column (4.6 mm × 250 mm, 5 μm) that was maintained at a constant temperature of 30 ℃. The mobile phase was 0.1% acetic acid solution-acetonitrile, with gradient elution. The flow rate was 1.0 mL/min. The detection was at wavelength of 280 nm. Results The standard curves of schisandrin B, schisantherin A, lobetyolin and ruscogenin had good linearity in the ranges of 0.862 5-21.562 5μg (r 2=0.999 1), 0.737 5-18.437 5μg (r 2=0.999 4), 0.095 2-2.380 0 μg (r 2=0.999 6), and 0.810 0-20.250 0 μg (r 2=0.999 0), respectively. The average recoveries of the four components were 98.06%(RSD=1.64%), 99.61%(RSD=2.72%), 97.98%(RSD=1.45%), and 99.30%(RSD=1.25%), respectively. Conclusion This method is accurate, rapid and specific, achieving the stable results with high reproducibility.

This study was aimed to observe the protective effect of extract from Rhizoma A nemones Raddeanae (RAR) on hepatic fibrosis induced by porcine serum in rats. A total of 68 SD rats were randomly divided into 5 groups, which were the normal group, model group, RAR group, extraction of RAR (EXRAR) group, Fu-Zheng Hua-Y u(FZHY) group. Each rat was intraperitoneally injected with 0.5~0.6 ml of porcine serum twice a week for 15 suc-cessive weeks to establish liver fibrosis model. Intragastric administration was given after the model was successfully established. The FZHY group was given FZHY capsule (0.525 g·kg-1). The RAR group was given RAR decoction (0.7 g·kg-1). The EXRAR group was given EXRAR (0.071 g·kg-1). The model group and normal group were given e-qual amount of physiological saline. The medication was given once a day. And the treatment course was 8 weeks. At the end of the 23th week, rats were sacrificed. Contents of SOD and MDA in blood serum were assayed. The protein expressions of α-SMA and TGF-β1 in liver tissues were detected by SABC. The results showed that compared with the model group, content of MDA decreased in the EXRAR group, RAR group and FZHY group (P<0.05), and content of SOD increased obviously (P<0.05). In the model group, expression of α-SMA and TGF-β1 increased, with dark brown dyeing and diffusion area. Expression area and strength of the FZHY group, RAR group, and EXRAR group were ob-viously weak with tasteless interval dyeing and no formation of typical pseudolobule in comparison with the model group. The color rendering index showed that compared with the model group, the protein expression of α-SMA and TGF-β1 decreased obviously in liver tissues of the FZHY group, EXRAR group, and RAR group (P< 0.05). It was concluded that RAR and its extract had a good antifibrosis effect. And the EXRAR had basically the same antifibrosis effect as RAR. It was assumed that the possible mechanism was related with the inhibiting of hepatic stellate cell (HSC) activation and the expression of TGF-β1 as well as the resisting of lipid peroxidation.

Objective To investigate the changes and clinical significance of the frequency of circulating CD4+ CD25+ regulatory T cells (Treg) in nonalcoholic fatty liver (NAFL) patients.Methods CD4+ CD25+ Treg in the peripheral blood from 50 NAFL patients and 50 healthy subjects were quantitatively analyzed using flow cytometry. Group t test or Mann-Whitney U test and Spearman's rank correlation test were used for statistical analysis.Results The proportion of circulating CD4+CD25+ Treg in NAFL patients was (5.39 ± 1.94)％,which was significantly higher than that in healthy controls [(4.21±1.52)％](t=3.385,P＜0.01).Further analysis revealed that the frequency of Treg was positively correlated with triglyceride (TG) level and body mass index (r=0.307 and 0.251,respectively; P=0.002 and 0.012,respectively),and negatively correlated with high density lipoproteincholesterol (HDL-C) (r=-0.306,P=0.002).Meanwhile,Treg in patients with high body mass index,high TG,low HDL-C,hypertension and metabolic syndrome (MS) were all higher than those in controls (t=2.294,2.533,3.154,2.010 and 4.454,respectively; all P＜0.05).But there was no significant difference between patients with high fasting blood glucose and controls (U=1143.500,P=0.471).Conclusion The increased frequency of peripheral Treg in NAFL patients may have some relations with the imbalance of proinflammation and anti-inflammation in NAFL patients who coexisting with MS.

Objective To observe the effects of modifiedDanshen Decoction on spermidine/spermine acetyltransferase (SSAT) /polyamine pathways of SD rats with IRI; To investigate its protective mechanism. Methods The model of IRI was established by ligating left anterior descending coronary artery for 30 min followed by reperfusion for 90 min. The SD rats were randomly divided into the control group, sham-operation group, model group and modifiedDanshen Decoction group, with 10 rats in each group. The myocardial infarction size was measured by using TTC staining. The contents of SSAT were measured by ELISA. The SSAT mRNA and SSAT protein expression level were detected with real-time fluorescent quantitative PCR method and Western blot, respectively. The contents of polyamines (putrescine, spermidine, spermine) in cardiac tissue were detected by HPLC. Results Compared with sham-operation group, the myocardial infarction size, the SSAT content, the SSAT mRNA and SSAT protein expression levels of model group increased significantly, the contents of polyamines decreased significantly, with statistical significance (P<0.01); Compared with model group, the myocardial infarction size of modifiedDanshen Decoction group was significantly reduced, while the SSAT content and SSAT mRNA and protein expression level decreased significantly, the contents of polyamines increased, with statistical significance (P<0.05, P<0.01).ConclusionModifiedDanshen Decoction can adjust the SSAT polyamine pathways and increase polyamine content in cardiomyocytes, and thus play a role of protection of myocardial ischemia-reperfusion injury.

Objective:To study the influence of triptolide on neutrophils asthmatic mice of WBC and EOS in bronchoalveolar lavage fluid.Methods:Using ovalbumin ( OVA) combined with lipopolysaccharide ( LPS) method to establish sensitized asthmatic mice,BALB/c mice were randomly divided into 32 neutrophilic asthma group ( NA group ) , neutrophil triptolide intervention group (TLN group),neutrophil dexamethasone group (DXN group) and normal control group (NC group),n=8,hemocytometer calculated for each group of mice bronchoalveolar lavage fluid ( BALF) of the total number of WBC and EOS;smears stained Switzerland View in-flammatory cell infiltration.Results: In bronchioalveolar lavage fluid, numbers and infiltrations of WBC and EOS were significantly decreased in the DXN,TLN group than those in the NA group(P<0.05);but were significantly higher than the NC group (P<0.05), the DXN group above parameters were significantly higher than the TLN group ( all P<0.05 ) .Conclusion: Triptolide can reduce the total number of BALF WBC and EOS,inhibit lung WBC,EOS infiltration,ease neutrophilic airway inflammation.

OBJECTIVE: To compare the hemodynamics and post-anesthetic recovery of total intravenous anesthesia (TIVA) with remifentanil or fentanyl combined with propofol administered by target controlled infusion (TCI) in neurosurgery. METHODS: A total of 80 patients undergoing selective neurosurgery were randomly divided into a remifentanil group (Group R, n=40) and a fentanyl group (Group F, n=40). In Group R,remifentanil and propofol was administered by TCI and the blood concentration were 3 approximately 5 microg/L and 3 approximately 5 mg/L each. In Group F, fentanyl was continuously infused at 2 approximately 3 microg/(kg.h) and propofol was administered by TCI with the same blood concentration as that in Group R.Vecuronium was injected at intervals to maintain muscle relaxant.Mean arterial pressure and heart rate during the anesthesia and post-anesthetic recovery were recorded. RESULTS: Mean arterial pressure of all the patients was decreased significantly from induction of anesthesia to termination of operation compared with that before the induction( P<0.01). The heart rate of Group R was increased obviously from recovery of respiration to extubation and heart rate of Group F was decreased obviously from fixed headframe to termination of operation compared with that before the induction (P<0.01). But there was no significant difference between the 2 groups (P>0.05). The eyes opened and extubed time of Group R were decurtated obviously and the scores of pain were increased significantly (P<0.01). CONCLUSION TIVA with remifentanil or fentanyl combined with propofol administered by TCI in neurosurgical operation can provided steadible hemodynamics. Resuscitation of remifentanil with propofol administered by target controlled infusion were more quickly but the scores of pain were more higher than that of fentanyl.
Adolescent ; Adult ; Aged ; Anesthesia Recovery Period ; Anesthesia, Intravenous ; methods ; Anesthetics, Intravenous ; administration & dosage ; Craniotomy ; Female ; Fentanyl ; Humans ; Infusion Pumps ; Male ; Middle Aged ; Neurosurgical Procedures ; Piperidines ; Propofol ; Remifentanil ; Vecuronium Bromide ; administration & dosage ; Young Adult

OBJECTIVE: To construct the recombinant lentivirus vector containing short hairpin RNA (shRNA) inhibited DREAM expression and to investigate the gene therapy of neuropathic pain by inhibiting the expression of DREAM gene by RNA interference. METHODS: An effective short hairpin RNA targeting to rat DREAM was cloned into the plasmids on the base of Lentivirous vectors, pKCSHR-Puro/GFP, and both of the pKCSHR-Puro/GFP-DREAM and Lentivector package plasmids mix were transferred into the 293T cells. The culture supernatant was harvested, and the virus titer was detected 48 hours after transferring. Thirty-six sheer breed pathogen free adult Sprague Dawley rats were randomly divided into 6 groups (6 in each group): normal control group (N); sham-operated group (S); CCI group (C0 group):CCI model without any intervention; Saline control group (C1 group); empty vector control group (C2 group); and LV-shRNADREAM lentiviral vector treatment group (C3 group). The rats in the last 3 groups respectively accepted injection of normal saline, blank vector, LV-shRNADREAM lentiviral vector in the subarachnoid on the 7th day after CCI, and the pain behavior was observed after 3, 7, 10, 14, 21 d after CCI. Green fluorescent protein (GFP) expression was detected by fluorescence microscope and the contents of DREAM mRNA and DREAM protein were detected by Realtime PCR and Western blot respectively in the rat lumbar spinal cord. RESULTS: The short hairpin RNA sequences targeting at rat DREAM were cloned into the vectors, and an entry clone and an expression clone were constructed successfully confirmed by sequence analysis. Lentiviral packaging was successful in 293 T cell line and the transfection titer of the lentivirus was 1 x 10(8) IFU/mL. LV-shRNADREAM lentivirus vector was transfected successfully in the rat spine with Intrathecal injection of LV-shRNADREAM. Compared with the other 3 groups, heat pain threshold and mechanical pain threshold in Group C3 improved significantly (P<0.01), and the expression of DREAM mRNA and DREAM protein in the lumbar spinal cord in Group C3 were lowered significantly (P<0.01). CONCLUSION Lentivirus vectors containing rat DREAM gene are constructed successfully, and lentivirus mediated shRNA can inhibit the DREAM expression in the rat spine, which may prove to be an effective method for neuropathic pain.
Analgesia ; methods ; Animals ; Base Sequence ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Kv Channel-Interacting Proteins ; biosynthesis ; genetics ; Lentivirus ; genetics ; metabolism ; Male ; Molecular Sequence Data ; Pain ; etiology ; Pain Management ; RNA Interference ; RNA, Small Interfering ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Repressor Proteins ; biosynthesis ; genetics ; Sciatic Nerve ; injuries ; Transfection

Objective:To implement an intervention with community-dwelling middle-aged and elderly people through a new science promotion approach, and to evaluate changes in the level of bone health awareness.Methods:From April 2017 to June 2017, 200 community-dwelling middle-aged and elderly people aged 50 years and over were randomly included.They were divided into the intervention group receiving a new science promotion intervention(n=100, with 50 males and 50 females)and the control group(n=100, with 50 males and 50 females). Baseline and follow-up assessments of bone health awareness were performed before and after the one-year intervention.A questionnaire survey on preferences for different types of science promotion articles in WeChat accounts was conducted.Differences in scores for intervention effects before and after intervention were compared between the two groups.Potential influencing factors for the scale score were examined by a multi-factor analysis.Results:After one year of intervention, scores for intervention effects were higher in the intervention group than in the control group( P<0.001). In the intervention group, scores were higher after intervention than before intervention( P<0.001). In the control group, scores after follow-up were slightly higher than those at baseline(29.4±11.4 vs.27.9±10.1, P<0.001). The increase in scores before and after intervention was greater in the intervention group than in the control group( P<0.001). Baseline score, group designation, history of drinking and diabetes impacted the scores in middle-aged and elderly women, while other factors did not.Most middle-aged and older people preferred texts illustrated with pictures, and older people were more receptive to videos. Conclusions:The new science promotion method is beneficial to bone health management, can improve bone health awareness in middle-aged and elderly people.

Objective:To explore the feasibility in evaluating segmental extracellular volume (ECV) using dual-layer spectral detector CT in patients with heart failure with various percentages of delayed hyper-enhanced volume, using MRI as a reference.Methods:Twenty-seven patients with heart failure in Peking Union Medical College Hospital were prospectively enrolled in this study from July 2019 to January 2021. All the patients underwent both CT late iodine enhancement and MR late gadolinium enhancement imaging within a week. According to percentages of hyper-enhanced volume on cardiac MR image, myocardial segments were classified into 3 groups: 0-4% (group A), 5%-49% (group B) and 50%-100% (group C). ECV images were independently observed by 2 experienced radiologists. The correlation and agreement between CT-ECV and MRI-ECV for the whole myocardial assessment and for the segmental assessment were evaluated using Spearman correlation and Bland-Altman analysis. Inter-observer agreement was assessed using the ICC.Results:Of the 27 patients, 432 segments were analyzed, with 199 segments in group A, 151 segments in group B and 82 segments in group C. In the whole myocardial analysis, the CT-ECV and MRI-ECV were 32.25% (28.04%, 34.28%) and 32.27% (28.35%, 34.77%), respectively. A good correlation was found between CT-ECV and MRI-ECV ( r=0.969, P<0.001). Bland-Altman analysis between CT-ECV and MRI-ECV showed a small bias (0.4%), with 95% limits of agreement of -6.9% to 7.8%. as for the segmental myocardial assessment, the CT-ECV was 28.23% (26.18%, 29.92%) for group A, 33.29% (30.16%, 35.96%) for group B and 40.22% (34.06%, 46.70%) for group C. There was statistically significant difference in ECV among three groups (all P<0.001). Good correlations between CT-ECV and MRI-ECV were found in group A ( r=0.614, P<0.001), group B ( r=0.852, P<0.001) and group C ( r=0.953, P<0.001). Bland-Altman analysis between CT-ECV and MRI-ECV showed a small bias 2.0% (95% limits of agreement:-17.6%-21.6%) in group A, a small bias -0.4% (95% limits of agreement: -13.8%-12.9%) in group B and a small bias -1.8% (95% limits of agreement: -17.4%-13.9%) in group C. The ICC values between 2 observers were 0.877 and 0.945 on CT-ECV and MRI-ECV, respectively. Conclusions:The larger percentage of hyper-enhanced volume on cardiac MR image, the stronger correlation are between the CT-ECV and MRI-ECV. CT-ECV may be taken as an alternative tool for quantitative evaluation of myocardial tissue in patients with heart failure.

Objective:To evaluate the feasibility of high concentrated contrast media combined with monochromatic images to improve image quality with double low scanning in dual-layer spectral coronary CT angiography.Methods:Fifty-six patients with suspected coronary artery disease were enrolled and randomly separated into two groups. All patients were scanned at 120 kVp in step-and-shoot mode using a dual-layer detector CT (IQon spectral CT). Patients were either injected with 18 ml high concentration contrast medium(400 mgl/ml) at 2 ml/s (group A) or 45 ml contrast medium (370 mgl/ml) at 4 ml/s (group B). Forty to 80 keV monoenergetic images with 10 keV increment and conventional image (group A2) were reconstructed for group A, conventional poly-energetic image was reconstructed for group B. Region of interest was placed on aorta root (AO), middle segment of left anterior descending (LAD), left circumflex (LCX) and right coronary artery (RCA) to measure the attenuation and noise, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR). Image quality was assessed by two reviewers independently with a 4-point scale on image quality (1-undiagnostic,4-excellent). The objective and subjective image evaluation were compared using the Kruskal-Wallis test. The Steel Dwass was used for multiple comparisons between monoenergetic images in Group A and conventional images in Group B, after the Kruskal-Wallis test.Results:There were significant differences among the attenuation, noise, SNR and CNR of group B, group A2, and monoenergetic images(all P<0.001).The CT value in the aortic root was significantly higher at 40-50 keV monoenergetic images than that in group B(all P<0.05), and similarly, the CT value in LAD,LCX and RCA at 40 keV were superior than that in group B(all P<0.001). There was no significant difference in the noise of the aortic root among 40 keV image, group A2 and group B(all P>0.05),while the noise in the 50-80 keV monoenergetic images were significantly lower than that in Group B(all P<0.001). The SNR and CNR in the aortic root were significantly higher at 40-60 keV monoenergetic images than that in group B(all P<0.001).The SNR in LAD and RCA at 40，50 keV and the CNR in LAD and RCA at 40-60 keV were higher than that in group B(all P<0.01).The SNR at 40-60 keV and CNR at 40-70 keV in LCX were superior than that in group B(all P<0.05). There were significant differences among the subjective image quality score of group B, group A2, and monoenergetic images(all P<0.001).The subjective image quality score of 40，50 keV images were not significantly different from that in group B (all P>0.05),while the score in 60-80 keV image and group A2 were lower than that of Group B(all P<0.001). Conclusion:40，50 keV low monochromatic images derived from dual-layer spectral detect CT combined with high concentrated contrast media can provide comparable or superior image quality with double low scanning in CCTA study.