Expression and significance of CDK1 based on bioinformatics in lung adenocarcinoma / 中国胸心血管外科临床杂志

@#Objective    To analyze the expression and clinical significance of cyclin-dependent kinase 1 (CDK1) in lung adenocarcinoma by bioinformatics. Methods    Based on the gene expression data of lung adenocarcinoma patients in The Cancer Genome Atlas (TCGA), the differential expression of CDK1 in lung adenocarcinoma tissues and normal lung tissues was analyzed. The expression of CDK1 gene in lung adenocarcinoma was analyzed by UALCAN at different angles. Survival analysis of different levels of CDK1 gene expression in lung adenocarcinoma was performed using Kaplan-Meier Plotter. Correlation Cox analysis of CDK1 expression and overall survival was based on clinical data of lung adenocarcinoma in TCGA. Gene set enrichment analysis was performed on gene sequences related to CDK1 expression in clinical cases. The protein interaction network of CDK1 from Homo sapiens was obtained by STRING. CDK1-related gene proteins were obtained and analyzed by the web server Gene Expression Profiling Interactive Analysis (GEPIA). Results    Based on the analysis of TCGA gene expression data, CDK1 expression in lung adenocarcinoma was higher than that in normal lung tissues. UALCAN analysis showed that high CDK1 expression may be associated with smoking. Survival analysis indicated that when CDK1 gene was highly expressed, patients with lung adenocarcinoma had a poor prognosis. Univariate and multivariate Cox regression analysis of CDK1 expression and overall survival showed that high CDK1 expression was an independent risk factor for survival of patients with lung adenocarcinoma. Gene set enrichment analysis revealed that high CDK1 expression was closely related to DNA replication, cell cycle, cancer pathway and p53 signaling pathway. Conclusion    CDK1 may be a potential molecular marker for prognosis of lung adenocarcinoma. In addition, CDK1 regulation may play an important role in DNA replication, cell cycle, cancer pathway and p53 signaling pathway in lung adenocarcinoma.

Study on Quality Standard for Fushiming Capsule / 中国药房

OBJECTIVE:To establish quality standard of Fushiming capsule. METHODS:TLC was used to qualitatively identify the ligustilide,aurantio obtusin,chrysophanol,fruit of Chinese wolfberry and Whitmania pigra,respectively. HPLC method was used to determine the contents of puerarin and ginsenosides Rb1. The determination was performed on Intersil C18 column with mobile phase consisted of acetonitrile-0.3% phosphoric acid(gradient elution)at flow rate of 1.0 mL/min;the detection wavlength was set at 203 nm,and column temperature was 25 ℃;the sample size was 10 μL. RESULTS:TLC spots of ligustilide,aurantio obtusin,chrysophanol,the fruit of C. wolfberry and W. pigra were clear and well separated without negative interference. The linear range of puerarin and ginsenoside Rb1were 10.56-337.92 μg/mL(r=0.999 7)and 17.80-569.70 μg/mL(r=0.999 6). The limits of quantitation were 2.20,1.86 μg/mL,and the limits of detection were 0.12,0.13 μg/mL,respectively. RSDs of precision,stability and reproducibility tests were lower than 2.0%. The recoveries were 95.65%-99.66%(RSD=1.45%,n=6) and 96.95%-98.52%(RSD=0.77%,n=6),respectively. CONCLUSIONS:Established quality standard can be used for the quality control of Fushiming capsule.

Cinnamaldehyde Derivatives Inhibit Coxsackievirus B3-Induced Viral Myocarditis

The chemical property of cinnamaldehyde is unstable in vivo, although early experiments have shown its obvious therapeutic effects on viral myocarditis (VMC). To overcome this problem, we used cinnamaldehyde as a leading compound to synthesize derivatives. Five derivatives of cinnamaldehyde were synthesized: 4-methylcinnamaldehyde (1), 4-chlorocinnamaldehyde (2), 4-methoxycinnamaldehyde (3), α-bromo-4-methylcinnamaldehyde (4), and α-bromo-4-chlorocinnamaldehyde (5). Neonatal rat cardiomyocytes and HeLa cells infected by coxsackievirus B3 (CVB3) were used to evaluate their antiviral and cytotoxic effects. In vivo BALB/c mice were infected with CVB3 for establishing VMC models. Among the derivatives, compound 4 and 5 inhibited the CVB3 in HeLa cells with the half-maximal inhibitory concentrations values of 11.38 ± 2.22 μM and 2.12 ± 0.37 μM, respectively. The 50% toxic concentrations of compound 4 and 5-treated cells were 39-fold and 87-fold higher than in the cinnamaldehyde group. Compound 4 and 5 effectively reduced the viral titers and cardiac pathological changes in a dose-dependent manner. In addition, compound 4 and 5 significantly inhibited the secretion, mRNA and protein expressions of inflammatory cytokines TNF-α, IL-1β and IL-6 in CVB3-infected cardiomyocytes, indicating that brominated cinnamaldehyde not only improved the anti-vital activities for VMC, but also had potent anti-inflammatory effects in cardiomyocytes induced by CVB3.

Determination of seven components in Pifubingxuedu tablet by HPLC method / 药学实践杂志

Objective To establish a HPLC method for simultaneous determination of seven constituents in Pifubingxuedu tablet.Methods The determination was performed on Kromasil-C18 column (4.6 mm × 250 mm, 5 μm).The mobile phase was acetonitrile-0.5％ phosphoric acid aqueous solution with a flow rate of 0.8ml/min.The detector was PDA and detection wavelength was set at 245,325,403 nm, respectively.Results A method has been established for the determination of chlorogenic acid, hydroxy Safflower Pigment A, paeoniflorin, forsythiaside A, ferulic acid, berberine and glycyrrhizin acid in one run by HPLC.Their average contents and RSD in each Pifubingxuedu tablet were 0.299 5 mg/tablet (2.25％);0.700 0 mg/tablet(1.33％);0.429 2 mg/tablet (1.21％);0.039 1 mg/tablet (2.34％);0.014 8 mg/tablet(2.23％);0.209 0 mg/tablet (2.06％);0.272 7 mg/tablet (2.68％), respectively.Conclusion The established method is simple, convenient, accurate and reliable.It is suitable for the quality control of Pifubingxuedu tablet.

Effect of Siji Kangbingdu Mixture on Inhibition of the Proliferation of Coxsackievirus A16 / 医药导报

Objective To verify the antiviral effects of Siji Kangbingdu mixture (SJKBDM) against coxsackievirus A16 (CoxA16).Methods Vero cells and 5-day-old suckling mice, injected with 75/50 and 1×106 TCID50 CoxA16, were used as evaluation models.The preventive influences of SJKBDM against CoxA16 in Vero cells were assessed in the models.The effects of SJKBDM on the mortality, survival time, change rate of body weight, and clinical symptom scores of suckling mice were observed.Results ①The half maximal inhibitory concentration of SJKBDM on Vero cells was 9.59 mg·mL-1.②Toxic effects were not observed from 32.3 g·kg-1 single dose or continuous intraperitoneal injectin of SJKBDM in suckling BALB/c mice.③The SJKBDM had significant inhibitory effect against CoxA16 virus.Doses higher than 1.22 mg·mL-1 could significantly improve the Vero cell survival rate, and the SJKBDM inhibition of 75/50 TCID50 CoxA16 induced pathological changes in Vero cells.④The SJKBDM significantly improved clinical symptoms of mice with CoxA16 viral infection, especially with crude drug doses of higher than 1.62 g·kg-1.The survival rate and other indicators were comparable or slightly higher compared with ribavirin, and the clinical score was higher than that of ribavirin.Conclusion The SJKBDM has significant inhibitory effect on CoxA16 cell proliferation, significantly decreases death rate, and improves clinical symptoms of mice infected with CoxA16 virus.

Improved anti-tumor efficacy and pharmacokinetics of bufalin via PEGylated liposomes / 中国药理学与毒理学杂志

OBJECTIVE To determine the characterization, anti-tumor efficacy and pharmacokinetics of bufalin- loaded PEGylated liposomes compared with bufalin entity. METHODS Bufalin- loaded PEGylated liposomes and bufalin- loaded liposomes were prepared reproducibly with homogeneous particle size by the combination of thin film evaporation method and high pressure homogenization method. The particle size and zeta potential of the liposomes were determined by dynamic light scattering technique. The direct imaging of morphology of liposomes was charactered by transmission electron microscope. The content of bufalin in liposomes was analysed by HPLC method. The entrapment efficiency and the particle size was applied to assess the stability profile, after storage at 4℃ on day 0, 7, 15, 30 and 90. The in-vitro release behaviours of bufalin from liposomes were conducted using dialysis bag technique at 37℃. In-vitro cytotoxicity studies were carried out using MTT〔3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide〕assay on several kinds of tumor cell lines including SW620, PC-3, MDA-MB-231, A549, U251, U87 and HepG2. In-vivo pharmacokinetic study of bufalin liposomes was evaluated by HPLC method. RESULTS Their mean particle sizes were 127.6 nm and 155.0 nm, mean zeta potentials were 2.24 mV and - 18.5 mV, entrapment efficiencies were 76.31% and 78.40% , respectively. In- vitro release profile revealed that the release of bufalin in bufalin- loaded PEGylated liposomes was slower than that of bufalin-loaded liposomes. The cytotoxicity of blank liposomes has been found within acceptable range, whereas bufalin-loaded PEGylated liposomes showed enhanced cytotoxicity to U251 cells compared with bufalin entity. In-vivo pharmacokinetics indicated that bufalin-loaded PEGylated liposomes could extend eliminate half-life time of bufalin in plasma in rats. CONCLUSION The results suggested that bufalin-loaded PEGylated liposomes improved the solubility and increased the drug concentration in plasma.

Evaluating Pharmacological Effects of Two Major Components of Shuangdan Oral Liquid: Role of Danshensu and Paeonol in Diabetic Nephropathy Rat

Shuangdan oral liquid (SDO) containing radix Salviae miltiorrhizae (Chinese name Danshen) and cortex moutan (Chinese name Mudanpi) is a traditional Chinese medicine using for treating vascular diseases. Danshensu (DSS) is a main effective monomer composition derived from radix Salviae miltiorrhizae and paeonol (Pae) from cortex moutan. Although the two herbs are widely used in traditional Chinese medicine, the pharmacological functions of their active compositions were not reported. Therefore, the research of DSS and Pae in mechanisms and pharmacodynamics interaction can provide scientific evidence to support clinical application. The diabetic nephropathy (DN) rats which were induced by streptozotocin (STZ) were treated with SDO, DSS, Pae, and DSS+Pae for eight weeks. The positive effects on DN animal models were investigated by detection of physiological and biochemical indexes and oxidative stress markers, within five treatments: SDO, DSS, Pae, DSS+Pae and insulin group. Compared with the model group, the DSS+Pae group improved the renal function, blood lipid metabolism and blood viscosity, increased the vitality of T-SOD or T-AOC and decreased the level of MDA or NO after the treatment. The study was successfully showed that the DSS+Pae group could delay the process of DN, especially in the renal injury part of histopathology changes. Our results suggest that the co-administration of DSS and Pae significantly may play a protective role in DN rats through decreasing the oxidative stress and improving the blood lipid metabolism mechanisms.

Analysis of rhythm features of EEG for driving fatigue / 生物医学工程学杂志

With extracting separately delta, theta, alpha and beta rhythms of electroencephalogram (EEG), we studied the characters of EEG for fatigued drivers by analyzing relative power spectrum, power spectral entropy and brain electrical activity mapping. The experimental results showed that with the average relative power spectrum in delta and theta rhythms of EEG increasing, the average relative power spectrum in alpha and beta rhythms decreased, while the average relative power spectrum in delta, theta and alpha rhythms increased in deep fatigue. The average power spectral entropy of EEG decreases with the increasing fatigue level. The average relative power spectrum and the average power spectral entropy of EEG could be expected to serve as the index for detecting fatigue level of drivers.

Effect of Rhizoma curcumae oil on the learning and memory in rats exposed to chronic hypoxia and the possible mechanisms / 生理学报

The effect of Rhizoma curcumae oil on the learning and memory in rats exposed to chronic hypoxia and the possible mechanisms were investigated. The rats were divided randomly into 5 groups (14 animals in each group): control, chronic hypoxia, chronic hypoxia with low (5 mg/kg body weight), middle (10 mg/kg body weight) and high (20 mg/kg body weight) concentrations of Rhizoma curcumae oil injection. The animals undergoing chronic hypoxia were exposed to hypoxia in a hypoxic chamber containing 10% O(2) and 5% CO(2) for 10 h/d, lasting 28 d. Morris water maze (MWM) test was used to obtain the scores of leaning and memory. The superoxide dismutase (SOD) activity and malonaldehyde (MDA) content were determined in the serum and hippocampus as well as [Ca(2+)](i) in the hippocampus. The expression of phosphorylated Ca(2+)/calmodulin-dependent protein kinase II (p-CaMKII) in the hippocampus was evaluated by using immunohistochemistry and Western blot. Compared with the control group, the chronic hypoxia group showed the following changes: (1) The escape latency to the hidden platform was remarkably prolonged (P<0.05); (2) The content of MDA and [Ca(2+)](i) were obviously higher, but the activity of SOD and the expression of p-CaMKII were significantly lower (P<0.05, P<0.01). Compared with the chronic hypoxia group, groups with Rhizoma curcumae oil injection had the following changes: (1) The escape latency to the hidden platform was remarkably shorter in 10, 20 mg/kg body weight groups (P<0.05); (2) The content of MDA and [Ca(2+)](i) were markedly decreased in 5, 10, 20 mg/kg body weight groups (P<0.05, P<0.01), but the activity of SOD in the serum and the expression of p-CaMKII were significantly higher in 10, 20 mg/kg body weight groups (P<0.05, P<0.01). The results showed that the capacity of learning and memory was degraded following chronic hypoxia. The decrease in MDA content and [Ca(2+)](i) and (or) the increase in SOD activity and p-CaMKII expression might participate in the enhancing effect on learning and memory induced by Rhizoma curcumae oil.

Changes of learning, memory and levels of CaMKII, CaM mRNA, CREB mRNA in the hippocampus of chronic multiple-stressed rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not simulate complicated living circumstances that people confronted with. The aim of this study was to investigate the effects of chronic multiple-stress on learning and memory as well as on the levels of calcium/calmodulin-dependent protein kinase II (CaMKII), calmodulin (CaM) mRNA, and cAMP-response element binding protein (CREB) mRNA in the hippocampus of rats.</p><p><b>METHODS</b>The rats were divided randomly into stressed and control groups. The stressed group was given chronic multiple-stress for 6 weeks to set up a chronic multiple-stressed model. The rats' performance of spatial learning and memory was tested using Morris Water Maze (MWM) and Y-maze. Meanwhile, the expressions of CaMKII, CaM mRNA and CREB mRNA of rats' hippocampus were detected by immunohistochemistry, Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. In addition, the width of synaptic cleft and the thickness of post-synaptic densities (PSD) were observed in the hippocampal CA3 region of rats by electron microscopy.</p><p><b>RESULTS</b>After exposure to chronic multiple-stress for 6 weeks, the ability of learning and memory of the stressed group was higher than that of the control group (P < 0.05, P < 0.01). The width of synaptic cleft was smaller and the thickness of PSD was larger in the hippocampal CA3 region of the stressed group than in that of the control group (P < 0.01). The CaMK II immunostaining of the stressed group was stronger than that of the control group in the stratum radiatum and oriens of the hippocampal CA1 and CA3, especially in the stratum oriens. Quantitative analysis indicated that the expression of CaMK II, CaM mRNA, and CREB mRNA in the hippocampus of the stressed group was higher than that of the control group (P < 0.05, P < 0.01).</p><p><b>CONCLUSIONS</b>The capacity of learning and memory can be enhanced after chronic multiple-stress. The increased levels of CaMK II, CaM mRNA, and CREB mRNA may contribute to the enhancing effect of chronic multiple-stress on learning and memory.</p>

Determination of Oleanolic Acid and Ursolic Acid in the Fruits of Ligustrum Iucidum Ait.of Shanxi by HPLC / 中国中医药信息杂志

Objective To determine oleanolic acid(OA)and ursolic acid(UA)from Fructus Ligustri Lucidi in diffrernt habitat and various growing stages.Method We picked the fruits of Fructus Ligustri Lucidi in Otober in five cities of Shanxi,and in August,September,October,November and December in Xi'an.Removed impurities and storaged the fruits under room temperature.By HPLC with Waters 600 as its chromatographic system,and Lichrospher C_(18)(4.6mmx250mm,5?m) column was applied with CH_3CN-CH_3OH-H_2O-H_3PO_4-(C_2H_5)_3N(50:30:20:0.02:0.04)as its mobile phase,the flow rate was 1 mL/min.The standard working curve was made to determine the contents of OA and UA at different habitat and different time spot from samples.Result The contents of OA and UA were highest in Ankang city.During prolonging growing stages,the highest contents of OA and UA were October and August,respectively.They both reduced to the lowest point in December.Conclusion The contents of OA and UA changed in different habitat and diffrernt growing stages of Fructus Ligustri Lucidi. It was suggested that we should mainly base on the highest contents to select the harvest time according to our demands.

Dose-dependent pharmacokinetic study of genistein in Beagle dogs / 药学学报

<p><b>AIM</b>To study the pharmacokinetics of genistein at different doses in Beagle dogs.</p><p><b>METHODS</b>Suspended in 0.5% CMC-Na solution, genistein was orally administered to Beagle dogs at doses of 2.67, 5.34 and 10.68 mg.kg(-1). At various time intervals, 1.5 mL of blood was drawn from the femoral vein of dogs in their front legs. The plasma was treated with beta-glucuronidase. The genistein in plasma was extracted twice by vortexing with 2.0 mL mixture of methyl tert-tubtyl ether and pentane (v/v = 8:2). The organic phase was removed into the tubes and then evaporated in ventilation cabinet. The residue was dissolved in 50 microL of methanol. 20 microL solution was drawn and detected by high-performance liquid chromatography. The pharmacokinetic parameters were calculated by 3P97 software.</p><p><b>RESULTS</b>The plasma drug concentration-time data were fitted to the two-compartment model. When the dose was 2.67 mg.kg(-1), the MRT and AUC of parent compound were 52.9 min and 6.7 mg.min. L(-1), respectively. When the dose rose to 5.34 mg.kg(-1), the MRT and AUC of parent compound became 224.8 min and 26.1 mg.min.L(-1), respectively. However, when the dose increased to 10.68 mg .kg(-1), the MRT and AUC of parent compound increased to 267.7 min and 33.2 mg.min L(-1), respectively. The AUC of glucuronidated genistein was 33.9, 70.1 and 140.5 mg.min.L(-1) at the dose of 2.67, 5.34 and 10.68 mg.kg(-1), respectively.</p><p><b>CONCLUSION</b>Due to significant first pass metabolism, the drug was mainly existed in the form of glucuronidated genistein in the plasma. With the increase of dose, the absorption of genistein became saturated and the half life prolonged.</p>

Research of mechanisms of swainsonine-induced apoptosis in the human gastric carcinoma cell line SGC-7901 / 中国临床药理学与治疗学

AIM: To investigate the effects and mechanisms of swainsonine-induced apoptosis on SGC-7901 cells. METHODES: After being treated with swainsonine, effective dose and median inhibition concentration (IC50) of swainsonine to SGC-7901 cells were examined by MTT assay. Cell cycle distribution and apoptotic rates were analyzed by flow cytometry. Expression of p53, c-myc and Bcl-2 were determined by immunocyto- chemical method, and the concentration of Ca2+ intra-cellular ([Ca2+]i ) was measured by the laser scanning confocal microscope (LSCM). RESULTS: Swainsonine inhibited cell growth of SGC-7901 in vitro, IC50 of 24 h was 0.84 μg·ml-l, and complete inhibition concentration of swainsonine was 6.2 μg·ml-l. Treated with swainsonine at the concentrations of 0.5, 1.5 and 4.5 μg·ml-l for 24 h, the expression of apoptosis inhibiting gene p53 and bcl-2 decreased, and apoptotic trigger gene c-myc increased (P<0.05), as well as [Ca2+]i overloading, SGC-7901 cell was induced to apoptosis in the end. The percentage of S phase were 38.8%, 39.7% and 29.6%, respectively (20.0% in control group and 23.2% in 5-Fu group), the percentage of G2/M phase were 4.5%, 1.7% and 5.3%, respectively (5.5% in control group and 9.0% in 5-Fu group), and the percentage of G1/M phase was not altered. SGC-7901 cells were treated by swainsonine at the concentrations of 0.5, 1.5 and 4.5 μg·ml-l for 24 h. Compared with the control group, the percentage of S phase were increased and that of G2/M cells were decreased significantly in treatment groups (P<0.01). CONCLUSION: Swainsonine can inhibit the cell proliferation and induce apoptosis of SGC-7901 cells, the mechanisms of swainsonine-induced apoptosis may related with [Ca2+]i overloading and expression of apoptosis-related genes.

Study on Extraction Technique for Xiangjiang Granula Essential Oil / 中国药房

OBJECTIVE: To study the optimum supercritical- fluid extraction technique for Xiangjiang Granula essential oil. METHODS: The orthogonal test was adopted to optimize the extraction process using the content of Ligustilide and the yield rate of essential oil as indicators, and 95% ethanol as co- solvents. The content of Ligustilide was determined by HPLC, using Phenomenex Luna C18( 250nm? 4. 6nm, 5? m) as column and methanol- 0. 5% glacial acetic acid( 30∶ 70) as mobile phase, with the detection wavelength set at 280 nm. RESULTS: The optimal extraction conditions were: temperature at 50℃ , pressure at 45MPa, extraction time for 3h, and 95% ethanol as co- solvents. The Ligustilide had a good linearty relationship between 5. 1～ 25. 5? g? mL- 1( r=0. 999 8) . CONCLUSIONS: This technique is easy, convenient and workable, and can provide theoretical support for production.

Preparation and Appraising of Eucalyptus Oil?-cyclodextrin Inclusion Compound / 中国药房

OBJECTIVE:To prepare and appraise eucalyptus oil?-cyclodextrin inclusion compound,and to testify the feasibility of transforming the dosage form of eucalyptus oil by clathration techniques.METHODS:The physicochemical properties of inclusion compound was identified by thin-layer chromatography(TLC),infrared spectroscopy(IR),ultra-violet spectroscopy(UV)and gas chromatograph mass spectrometer(GC-MS),respectively.Meanwhile,the changes in constituents and clathration outcomes of eucalyptus oil before and after clathration were also investigated.RESULTS:The analytic result of TLC,IR and UV showed that stable inclusion compound has been formed from eucalyptus oil and?-cy?clodextrin.The result of GC-MS demonstrated that there was no significant change in the essential components and the percentage composition of eucalyptus oil before and after inclusion.CONCLUSION:Stable inclusion compound can be made from eucalyptus oil and?-cyclodextrin meanwhile without changes in main components and the percentage composition of eucalyptus oil.

Induction of apoptosis by Tryptanthrin on K562 cells / 中国药理学通报

Aim To study the effect of Tryptanthrin(Try) on proliferation and apoptosis of erythroleukemia K562 cells.Methods The cell proliferation effect of Try(1.56～50 mg?L-1) on K562 cells was assessed by MTT assay.The morphologic change was observed by Hoechst 33258 fluore-scent stain.The flow cytometer was used to detect cell apoptosis and cell cycle.Results MTT showed that in the range of 3.12～50 mg?L-1 Try obviously inhibited the proliferation of K562 cells in a dose and time-dependent manner.Typical apoptosis changes were observed in K562 cells treated with Try for 48 h by flourescence inverted microscope.With Annexin V-FITC and PI double staining,folw cytometer result showed that the apoptosis state was obvious in K562 cells treated with 25,50 mg?L-1 Try for 48 h.The cell cycle distribution of K562 was changed.The G0/G1 phase was blocked and the DNA synthesis was inhibited,accompanied with subdiploid apoptotic peak.Conclusion Try has an effect on inhibiting the cell proliferation and inducing the apoptosis of K562.

Research progress of tryptanthrin / 中国药理学通报

Tryptanthrin is a kind of indole quinazoline alkaloid.In this review,recently reported research progresses of tryptanthrin on extraction,systhesis and pharmalogical action have been mainly summarized.The effect of tryptanthrin on antimicrobial,antiinflammatory and antitumor activities has been found,showing its good application and development perspective.