ObjectiveTo establish a quantitative analysis multi-components by single marker method （QAMS） for five main components （aucubin， geniposidic acid， chlorogenic acid， asperuloside and rutin） in Eucommiae Folium， to verify its feasibility and applicability in the determination of Eucommiae Folium， so as to provide a scientific basis for the development of quality standard of this herb. MethodHigh performance liquid chromatography was performed on a Welch Boltmatetm™ C₁₈ column （4.6 mm×100 mm， 2.7 μm） with methanol （A）-0.2% phosphoric acid aqueous solution （B） as the mobile phase for gradient elution （0-8 min， 3%A； 8-10 min， 3%-11%A； 10-26 min， 11%A； 26-27 min， 11%-25%A； 27-60 min， 25%-32%A）， the column temperature was set at 30 ℃， the flow rate was 0.6 mL·min^-1， the detection wavelengths were at 210 nm and 254 nm. Chlorogenic acid was used as an internal reference to establish the relative correction factors （f） between it and the other four components， and the contents of the five components in 14 batches of Eucommiae Folium were determined by QAMS and external standard method （ESM）， respectively. ResultThe f values of chlorogenic acid to aucubin， geniposidic acid， asperuloside and rutin were 3.13， 1.45， 2.64 and 0.56， respectively. Repeatability was good under different experimental conditions， relative standard deviation （RSD） was <5.0%. The contents of aucubin， geniposidic acid， chlorogenic acid， asperuloside and rutin in 14 batches of Eucommiae Folium were 1.340-28.975， 0.252-36.086， 10.016-27.443， 1.396-8.646， 0.533-1.766 mg·g^-1， respectively. There were no significant difference between content results of QAMS and that of ESM （RSD<5.0%）. ConclusionQAMS established with chlorogenic acid as the internal reference can be used to determine the contents of five components in Eucommiae Folium， and this method is simple and accurate. After comprehensive evaluation， the quality standard of Eucommiae Folium in subsequent editions of Chinese Pharmacopoeia is suggested that three main active components， chlorogenic acid， aucubin and geniposidic acid， are selected as quality markers， and their content limits are recommended not less than 1.5%， 1.0% and 1.0%， respectively. This quality standard draft can avoid the potential quality risk due to poor specificity and low content limit of the index component （chlorogenic acid） in the previous editions of Chinese Pharmacopoeia.

ObjectiveTo prepare matrine lipid-based cubic liquid crystalline nanoparticle （MAT-LLCN） gels and investigate its in vitro release and transdermal absorption behavior. MethodTaking entrapment efficiency as the index， the optimal formulation of MAT-LLCN was screened by extreme vertex mixture method based on the optimal ratio of glycerol monooleate （GMO） to poloxamer 407 （P407）， and its drug loading was investigated. MAT-LLCN gels was prepared by mixing MAT-LLCN with pre-swelled carbomer 940 as the gel matrix. The structure of MAT-lipid-based cubic liquid crystalline （LLC） was characterized by polarized light microscopy （PLM） and small angle X-ray scattering （SAXS）. The in vitro release and transdermal absorption properties of MAT-LLCN gels and MAT ordinary gels were compared by modified Franz diffusion cell method， skin structure changes caused by them were observed by hematoxylin-eosin （HE） staining. ResultThe optimal formulation of MAT-LLCN gels was 5.5% of GMO-P407 （9∶1）， 1%-6% of MAT， 0.6% of carbomer 940， adding water to sufficient amount. The prepared MAT-LLC was confirmed as body-centered （Im3m） LLC. The in vitro release behavior of MAT-LLCN gels was in accordance with the Weibull equation （R²=0.954 0）， and the release mechanism was the Fick diffusion. In vitro transdermal test showed that all the parameters of MAT-LLCN gels were higher than those of MAT ordinary gels （P<0.05）， including cumulative release rate， steady-state release rate and the amount of drug retention in skin. HE staining results showed that MAT-LLCN gels could loose the cellular arrangement of skin stratum corneum， and maintain the stability of the cell structure of the dermis. ConclusionThe prepared MAT-LLCN gels can accelerate the transdermal drug transport and form drug storage in the dermis by rapidly opening the skin stratum corneum barrier， suggesting that LLC has good application prospects in the field of transdermal drug delivery.

Objective:To externally validated the intra-aortic balloon pump (IABP) shockⅡ score and CardShock score for predicting in-hospital mortality in patients with acute myocardial infarction complicated by cardiogenic shock (AMI-CS) and compared them with the Acute Physiology and Chronic Health EvaluationⅡ (APACHEⅡ) score.Methods:According to the inclusion and exclusion criteria, patients admitted to the cardiac care center (CCU) of our center from December 2010 to May 2019 were enrolled in this study. Patients’ baseline characteristics, in-hospital interventions, and outcomes were collected. The APACHEⅡ score was calculated during hospitalization by clinicians and collected by researchers. Two researchers independently calculated the IABP-ShockⅡ score and CardShock score; any disagreement was discussed with the third researcher. The performance of risk scores was evaluated by discrimination and calibration. The discriminative ability of risk scores was evaluated using the area under the receiver operating characteristic curve (AUC) and compared by the Delong method. The calibration of these risk scores was examined by the Hosmer-Lemeshow goodness-of-fit test. The calibration plot was also built.Results:A total of 150 patients enrolled in our study, and the in-hospital mortality was 60%. According to the IABP-ShockⅡ score, patients scored as low risk (0-2), moderate risk (3-4), and high risk (5-9) had in-hospital mortality of 29%, 68%, and 80%, respectively. According to the CardShock score, patients scored as low risk (0-3), moderate risk (4-5), and high risk (6-9)had in-hospital mortality of 21%, 57%, and 82%, respectively. According to the APACHEⅡ score, patients scored<20, 20-30, and >30 had in-hospital mortality of 19%, 69%, and 93%, respectively. For predicting the in-hospital mortality, the APACHEⅡ score demonstrated excellent discrimination (AUC=0.90, 95% CI: 0.84-0.95). The IABP-ShockⅡ score and CardShock score showed good discrimination (AUC=0.76, 95% CI: 0.68-0.83 and AUC=0.79, 95% CI: 0.72-0.85). The discriminative ability did not significantly differ between the IABP-ShockⅡ score and the CardShock score (0.76 vs 0.79, P>0.05), but both of which were significantly lower than the APACHEⅡ score (0.76 vs 0.90, P<0.05, and 0.79 vs 0.90, P<0.05). At the same time, it was not significantly different between the IABP-ShockⅡ score and the CardShock score (0.76 vs 0.79, P>0.05). All of these three scores were adequately calibrated according to the Hosmer-Lemeshow goodness-of-fit test ( P>0.05).The calibration plot showed accurate calibration of these three scores. Conclusions:Although less accurate than the APACHEⅡ score, the IABP-ShockⅡ score and CardShock score can show accurate prediction for in-hospital mortality of AMI-CS patients.

Positive end-expiratory pressure (PEEP) is a common method to maintain alveolar patency in patients undergoing mechanical ventilation. However, in patients undergoing tracheotomy, alveolar collapse often occurs due to bedridden, aspiration, and other factors. There is currently no effective means to provide PEEP support for such patients. The application of a high-flow oxygen inhalation device with a PEEP valve was designed to provide patients with continuously adjustable PEEP, which helps to improve the patient's oxygenation and maintain the lung's physiological functions.

Objective To design a novel electronic device for measuring the pressure in the cuff of the artificial airway; and to study the advantage of this device on continuous and intermittent cuff pressure monitoring. Methods ① a portable electronic device for cuff pressure measurement was invented, which could turn pressure signal into electrical signal through a pressure transducer. Meantime, it was possible to avoid pressure leak from the joint and the inside of the apparatus by modified Luer taper and sophisticated design. If the cuff pressure was out of the normal range, the apparatus could release a sound and light alarm. ② Six traditional mechanical manometers were used to determine the cuff pressure in 6 tracheal tubes. The cuff pressure was maintain at 30 cmH2O (1 cmH2O =0.098 kPa) by the manometer first, and repeated every 30 seconds for 4 times. ③ Study of continuous cuff pressure monitoring: We used a random number generator to randomize 6 tracheal tubes, 6 mechanical manometers and 6 our products by number 1-6, which has the same number of a group. Every group was further randomized into two balanced groups, one group used the mechanical manometer first, and the other used our product first. The baseline pressure was 30 cmH2O, measurement was performed every 4 hours for 6 times. Results When traditional mechanical manometer was used for cuff pressure monitoring, cuff pressure was decreased by an average of 2.9 cmH2O for each measurement (F = 728.2, P = 0.000). In study of continually monitoring, at each monitoring point, the pressure measured by electronic manometer was higher than the mechanical manometer. All the pressures measured by mechanical manometer were dropped below 20 cmH2O at 8th hour, and there was no pressure decrease below 20 cmH2O measured by electronic manometer in 24 hours by contrast. In study of intermittent monitoring, the same result was found. The pressure was dropped significantly with time when measured by mechanical manometer (F = 61.795, P = 0.000), the drops below 20 cmH2O began at 8th hour; but when measured by electronic manometer, all the value stayed unchanged around the baseline in 24 hours (F = 0.511, P = 0.796). Conclusions Compared with traditional mechanical manometer, cuff pressures monitored by our novel electronic manometer were steadier in both continuous and intermittent monitoring. The device is compact and convenient, and can provide a good solution for continuously monitor of the tracheal cuff pressure.

Obejective To explore the diagnostic and curative evaluation value of gastrin-releasing pep-tide precursor (ProGRP) and neuron specific enolization enzyme (NSE) in small cell lung cancer (SCLC). Methods Sixty SCLC patients, sixty non-small cell lung cancer (NSCLC) patients and forty patients with be-nign pulmonary disease were collected fromJanuary 2014 to October 2015. The levels of serum ProGRP and NSE in all patients were determined by ELISA method and radioimmunoassay respectively then the clinical value of ProGRP and NSE on SCLC was evaluated. Results The levels of ProGRP and NSE in SCLC group were signif-icantly higher than those in NSCLC group and those in lung benign disease group (P < 0.05). The levels of Pro-GRP and NSE in extensive stage were higher than those in limited stage in SCLC group (P < 0.05). The bound-ary value of SCLC through ProGRP identified through ROC curve was 64.68 pg/mL. The diagnostic sensitivity , specific degree and Youdenindex of ProGRP in SCLC were 86.7%, 97.5% and 0.842 respectively, which were significantly higher than NSE (P < 0.05). After 2 cycles of chemotherapy, serum ProGRP in SCLC disease con-trol groupwere significantly decreased(P < 0.05) but on difference of serum ProGRP was found in SCLC progres-siongroup (P > 0.05). Conclusion ProGRP and NSE can be used as markers for the diagnostic and curative evaluation of SCLC. And ProGRP has higher sensitivity and specificity than NSE and can be promoted in clinic.

Amphibian skin antimicrobial peptides exhibit a broad spectrum of antimicrobial activity against Gram-positive and Gram-negative bacterium and cytotoxic activity responsible for inhibiting the growth of cancer cells. In this present study, six cDNAs encoding antimicrobial peptide precursors were cloned from the skin of Chinese brown frog, Rana chensinensis by RT-PCR and 3'-RACE procedure and identified as preprotemporin-1CEa, preprotemporin-1CEb, preprotemporin-1CEc, preprobrevinin-1CEa, preprobrevinin-1CEb, and preprochensinin-1, respectively. The nucleotide sequences of cDNA encoding 59-65 amino acid composed of 289-315 bp. Preprotemporin-1CEa, preprotemporin-1CEb and preprotemporin-1CEc are members of temporin family, which usually are short, hydrophobic, and C-terminally alpha-amidated antimicrobial peptides. Preprobrevinin-1CEa and preprobrevinin-1CEb were identified as the members of the brevinin-1 family of antimicrobial peptides since both peptides contain "RANA box" that it's responsible for forming Cys-bridged cyclic heptapeptides at the C-terminal region of peptide. The nucleotide acid sequence and the deduced amino acid Sequence of preprochensinin-1 were not found to be identity with any known amphibian skin defensive peptides, so, preprochensinin-1 was identified as a novel peptide precursor. Four of bioactive peptides: temporin-1CEa, temporin-1CEb, brevinin-1CEa and chensinin-1 were synthesized to investigate their antimicrobial, anticancer and haemolysis activities. The results showed that all of the synthesized antimicrobial peptides in this study inhibited the growth of the Gram-positive bacterium, and exhibited the anticancer activity against the growth of MCF-7 cells and HeLa cells. Analysis of the R. chensinensis bioactive peptides and their gene expression will be beneficial for preservation of this species.
Amino Acid Sequence ; Animals ; Anti-Infective Agents ; pharmacology ; Antimicrobial Cationic Peptides ; genetics ; pharmacology ; Antineoplastic Agents ; pharmacology ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Hemolysis ; drug effects ; Molecular Sequence Data ; Protein Precursors ; genetics ; pharmacology ; Proteins ; genetics ; pharmacology ; Ranidae ; genetics ; metabolism ; Skin ; metabolism

OBJECTIVETo assess the local immune response in patients with hepatocellular carcinoma after percutaneous microwave coagulation therapy (PMCT).

METHODSSeventy-eight patients with hepatocellular carcinoma underwent PMCT. Both cancerous and adjacent liver tissue were taken before, and 3, 17 and 30 days after PMCT using ultrasound-guided liver biopsy. Specimens were stained by immunohistochemical techniques for detecting CD3, CD45RO, CD56, CD68, and CD20 positive cells.

RESULTSA few CD3, CD45RO, CD56, CD68 and CD20 positive cells were observed in the cancer stroma and surrounding sinusoids in liver tissue pre-PMCT. The number of immunocytes, except for CD20 positive cells, was significantly increased both within the cancer and the adjacent liver tissue, with a larger increase in tumor tissue at day 3 post-PMCT compared with pre-PMCT. These immunocytes were enlarged in size. The number of CD3, CD45RO and CD56 positive cells peaked at day 17 and CD68 positive cells peaked at days 3 post-PMCT. At day 30 post-PMCT, this increase still existed. These infiltrating immunocytes distributed in the parenchyma of the tumor, and within the lumen of small blood vessels after PMCT. In addition, more infiltrated immune cells were seen in cancer cell spaces.

CONCLUSIONSA change in immunocyte infiltration takes place in number, configuration and location after patients with HCC are treated with percutaneous microwave coagulation, suggesting that local immune function is enhanced post-PMCT.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; analysis ; Carcinoma, Hepatocellular ; immunology ; pathology ; therapy ; Electrocoagulation ; Female ; Humans ; Immunohistochemistry ; Liver Neoplasms ; immunology ; pathology ; therapy ; Male ; Microwaves ; therapeutic use ; Middle Aged ; T-Lymphocytes ; immunology

Objective To investigate the anticoagulant effect of polysaccharides from the heads of Rana temporaria chensinensis(Chinese Forest Frogs). Methods In-vitro experiments were applied to test the effect of polysaccharides on recalcification time, prothrombin time, thrombase time , the activities of thrombin, hydrolyzing fibrin content, activated fibrin dissolve zymogen and blood clotting.. In the in-vivo experiment, bleeding time was measured by cutting mouse tail and clotting time was measured by glass sheet method. Results The polysaccharides from the heads of Chinese Forest Frogs had an obvious anticoagulant effect in a dose-effect manner, but had on thrombolytic action. It could prolong the bleeding time and the clotting time in vivo. Condusion The polysaccharides from the heads of Chinese Forest Frogs have the anticoagulatant activities in vitro and in vivo.