Synthesis and antitumor activity of α,ß-unsaturated carbonyl moiety- containing oleanolic acid derivatives targeting PI3K/AKT/mTOR signaling pathway.

Oleanolic acid (OA) and its semi-synthetic derivatives have been reported to have a wide range of biological activities. The introduction of electrophilic Michael acceptor group can increase the reactivity of OA to cellular targets and thus improve the anti-tumor activity. In this work, a series of novel α,ß-unsaturated carbonyl derivatives of OA were designed and synthesized. Their in vitro cytotoxic activity against MCF-7, HepG2 and HeLa cells were tested. Most derivatives exhibited improved cell growth inhibitory activity, especially for 3d with an IC50 of 0.77 µM in MCF-7 cells. Moreover, 3d inhibited the migration of MCF-7 and HeLa cells at the concentration of 4 µM. Flow cytometric analysis revealed that 3d induced cell apoptosis and S phase arrest in a concentration-dependent manner. Western blotting experiment demonstrated that 3d inhibited the phosphorylation of AKT and mTOR. These results suggest that this series of OA derivatives bearing exocyclic methylene ketone pharmacophore are promising anticancer agents as potential PI3K/AKT/mTOR pathway inhibitors.

Oxidative Radical Addition/Cyclization Cascade for the Construction of Carbonyl-Containing Quinoline-2,4(1H,3H)-Diones.

Oxidative radical addition/cyclization cascade of o-cyanoarylacrylamides with α-keto acids as well as aldehydes is reported. This transformation exhibits a wide substrate scope and significant functional group tolerance and provides a convenient and highly efficient access to carbonyl-containing quinoline-2,4(1H,3H)-diones. A possible mechanism for the transformation is proposed.

Copper-catalyzed cascade addition/cyclization: highly efficient access to phosphonylated quinoline-2,4(1H,3H)-diones.

A novel and facile Cu-catalyzed addition/cyclization cascade of o-cyanoarylacrylamide was developed. The process exhibits significant functional group tolerance, and provides an efficient and straightforward pathway for the synthesis of various phosphonylated quinoline-2,4(1H,3H)-diones.

[Study on antithrombotic activities and structure-activity relationships of Veratrum nigrum Alkaloids].

OBJECTIVE: To explore antithrombotic effects and mechanisms of total alkaloids (VnA) and single alkaloids (Al - A6) from Veratrum nigrum. METHODS: The effects of VnA and Al - A6 on thrombosis and coagulation parameters (APTT, PT and TT) were examined in rat in vivo, while their effects on platelet aggregation induced by ADP and thrombin were determined in rabbit in vitro. RESULTS: VnA and Al ~ A5 decreased thrombus wet weight and platelet aggregation, increased TT, while they had no influence on APTT or PT. A6 had no obvious effect on thrombus wet weight, platelet aggregation or coagulation parameters. CONCLUSIONS: VnA and Al ~ A5 have antithrombotic activities on venous thrombosis. The effective component in VnA is A2 probably. The antithrombotic effects are possibly related to the inhibition of platelet aggregation and the extension of TT. The cevanine type skeleton of steroidal alkaloids is necessary for antithrombotic activities and the ester substitute at C3 can enhance antithrombotic activities.

[Quantitative models for Baicalin content using NIR technology for the study of Shang Jie plaster].

A dynamic prediction model for the content of Baicalin in Shang Jie plasters extract solutions was developed using near-infrared spectroscopy in transmission mode. Sixty five spectra were obtained through near-infrared transmission mode during extracting process. Refering to the content of Baicalin performed by reversed-phase high performance liquid chromatography (HPLC), the calibration model was developed with the application of partial least squares regression algorithm (PLSR). The constructed model was validated by 30 samples; some parameters of the calibration model were optimized by cross-validation. The root mean square error (RMSECV) of Baicalin was 0.006 8 mg x g(-1), the correlation coefficient (R) was 0.9991, and the optimal dimension factor was 8; After predicted by test set, the root mean square error (RMSEP) and correlation coefficient (R) of prediction obtained were 0.009 2 mg x g(-1) and 0.998 7 respectively. This work demonstrated that NIR spectroscopy combined with PLS could be used for the determination of Baicalin in Shang Jie plasters extract.

Correlation and mechanism between cardiac magnetic resonance imaging and oral streptococcus count in patients with primary microvascular angina pectoris.

BACKGROUND: Although primary microvascular angina (PMVA) can be diagnosed clinically, the etiology and pathophysiology of PMVA remain unclear. The effects of conventional clinical medications (aspirin, statins, and nitrates) are unsatisfactory, and PMVA can lead to serious cardiovascular events. The present study was designed to analyze the correlation between the load perfusion cardiovascular magnetic resonance imaging (CMR) results and the Streptococcus sanguinis(S sanguinis) count and the correlations between the S sanguinis count in oral cavity subgingival plaque and changes in the plasma levels of platelet alpha-granule membrane glycoprotein 140 (GMP-140), fibrinopeptide A (FPA), von Willebrand factor (vWF), and homocysteine (Hcy) in patients with PMVA after increased anti-infective treatment of the oral cavity. This study also discusses the pathogenesis of PMVA from this perspective. The differences in the S sanguinis count in oral cavity subgingival plaque and oral health status between healthy people and PMVA patients will be compared, and the correlation between the oral cavity health status and disease in PMVA patients will be analyzed. METHODS: The present randomized controlled trial with a parallel control group will be conducted in 68 PMVA patients diagnosed by the in-patient cardiology department. The selected patients will be randomly divided into 2 groups, one receiving routine drug treatment and the other a combination of anti-infective treatments. The normal control group will comprise 30 healthy people with no infectious oral cavity disease matched by age and sex. We will conduct CMR, and the presence of S sanguinis in subgingival plaques will be used to determine the bacterial count in PMVA patients. Blood samples will also be collected to determine the levels of GMP-140, FPA, vWF, and Hcy. S sanguinis in the subgingival plaque of PMVA patients will be further analyzed after increasing the oral cavity anti-infective treatment; the resulting changes and their correlations with changes in GMP-140, FPA, vWF, and Hcy levels will be assessed. Additionally, the differences in the S sanguinis count and the oral cavity health status of oral cavity dental plaque between healthy people and PMVA patients will be determined, and the correlation between the oral cavity conditions and PMVA will be analyzed. The relationship between the perfusion CMR results and the oral cavity S sanguinis count of PMVA patients, and the potential pathogenesis, will be explored. We will use the SPSS19.0 statistical software package to analyze the data. The measurements will be expressed as means±standard deviation. Student t test will be used for intergroup comparisons, a relative number description will be used for the count data, and the chi-square test will be used for intergroup comparisons. Multivariate logistic regression will be performed to identify associations. A P value < .05 will be considered significant. DISCUSSION: In this study, the correlation between the perfusion CMR results and the S sanguinis count in oral cavity subgingival plaque of PMVA patients will be analyzed. Changes in the levels of GMP-140, FPA, vWF, and Hcy of PMVA patients after receiving increased oral cavity anti-infective treatment will be explored, and the difference in the S sanguinis count in oral cavity subgingival plaque and the oral cavity health status between healthy people and PMVA patients will be compared. ATRIAL REGISTRATION: Chinese Clinical Trial Registry, (http://www.chictr.org.cn/showprojen.aspx?proj=45091).

NLRC4 gene silencing-dependent blockade of NOD-like receptor pathway inhibits inflammation, reduces proliferation and increases apoptosis of dendritic cells in mice with septic shock.

Septic shock is one of the most significant health concerns across the world, involving hypo-perfusion and defects in tissue energy. The current study investigates the role of NLR family CARD domain containing protein 4 (NLRC4) in septic shock-induced inflammatory reactions, lung tissue injuries, and dendritic cell (DC) apoptosis. Septic shock mice models were established by modified cecal ligation and puncture and injected with retroviral vector expressing siRNA-NLRC4. DCs were then isolated and transfected with siRNA-NLRC4. The degree of lung tissue injury, cell cycle distribution, cell apoptosis and cell viability of DCs were assessed. NLRC4 was found to be expressed at high levels in mice with septic shock. NLRC4 silencing inhibited the activation of the NOD-like receptor (NLR) pathway as evidenced by the decreased levels of NOD1, NOD2, RIP2, and NF-κB. In addition, NLRC4 silencing reduced the inflammatory reaction as attributed by reduced levels of IL-1ß, TNF-α and IL-6. Suppressed NLRC4 levels inhibited cell viability and promoted cell apoptosis evidenced by inhibited induction of DC surface markers (CD80, CD86, and MHC II), along with alleviated lung tissue injury. In conclusion, NLRC4 silencing ameliorates lung injury and inflammation induced by septic shock by negatively regulating the NLR pathway.

Cardiovascular magnetic resonance image analysis and mechanism study for the changes after treatments for primary microvascular angina pectoris.

ABSTRACT: Most cases of primary microvascular angina pectoris (PMVA) are diagnosed clinically, but the etiology and pathological mechanisms are unknown. The effect of routine clinical medications is minimal, and PMVA can progress to serious cardiovascular events. To improve the diagnosis and effective treatment of this disease, this study was designed to diagnose PMVA via cardiovascular magnetic resonance (CMR) and the coronary angiography thrombolysis in myocardial infarction (TIMI) blood flow grade, as well as to analyze vascular endothelial function to elucidate the pathogenesis of PMVA and compare the effects of routine clinical medications.The present randomized controlled trial including a parallel control group will be conducted on 63 PMVA patients in our cardiovascular department. The patients will be selected and randomly divided into the control, diltiazem, and nicorandil groups. The control group will be administered routine drug treatments (aspirin, atorvastatin, betaloc ZOK, perindopril, and isosorbidemononitrate sustained-release tablets). The diltiazem group will be additionally treated with 90 mg qd diltiazem sustained-release capsules. The nicorandil group was additionally given 5 mg tid nicorandil tablets. Coronary angiography will be performed before treatment, the severity and frequency of chest pain will be evaluated before and after 9 months of treatment, and homocysteine and von Willebrand factor levels will be measured. Electrocardiography, echocardiography, dynamic electrocardiography, a treadmill exercise test, and CMR will be performed. Sex, age, body mass index, complications, smoking, and family history will also be recorded. The SPSS19.0 statistical software package will be used to analyze the data. The measurements will be expressed as the mean ±â€Šstandard deviation. Measurement data will be compared between the groups using Student's t-test. A relative number description will be used for the counting data, and the chi-squaretest will be used to compare the groups. A multivariate logistic regression analysis will be performed A P-value < .05 will be considered significant.The direct indices (CMR and coronary angiographic TIMI blood flow grade) may improve after adding diltiazem or nicorandil during routine drug treatments (such as aspirin, statins, and nitrates) in PMVA patients, and indirect indices (homocysteine and von Willebrand factor levels) may be reduced. TRIAL REGISTRATION: Chinese Clinical Trial Registry (http://www.chictr.org.cn/showprojen.aspx?proj=41894), No. CHiCTR1900025319, Registered on August 23, 2019; pre initiation.

Mineralocorticoid receptor, CYP11B2 mRNA expression, and atrial matrix remodelling in patients with atrial fibrillation.

OBJECTIVE: Atrial fibrillation is associated with the activation of the renin-angiotensin-aldosterone system in the atria. It is not clear whether the expression of mineralocorticoid receptor (MR) and aldosterone synthase CYPII B2 in patients with atrial fibrillation is altered. This study aimed to investigate the mRNA expression of MR and CYPIIB2 and to reveal the correlation between CYPIIB2 mRNA and matrix remodelling in patients with atrial fibrillation. METHODS: Twenty-five patients with rheumatic heart valve disease, 12 in sinus rhythm and 13 in atrial fibrillation (> or = 6 months), underwent a valve replacement operation and right and left atrial lateral wall tissue samples were obtained. The MR and CYPI IB2 expressions were analysed at the mRNA level and collagen volume fraction was determined by Van Gieson's staining. Results - Collagen volume fraction was found to be increased significantly in atrial fibrillation groups compared with sinus rhythm groups (P < 0.001). Both the mRNA of MR and CYPIIB2 were significantly increased in the fibrillation group compared with the group in sinus rhythm (P < 0.01). Collagen volume fraction significantly and positively correlated with left atrial dimension (r = 0.845, P < 0.001).There was a positive correlation between CYPI I B2 mRNA and collagen volume fraction (r = 0.757, P < 0.001). CONCLUSION: Increased expression of MR and CYPIIB2 in the atria is one of the molecular mechanisms for the development of atrial interstitial fibrosis in patients with atrial fibrillation.

A new eudesmane sesquiterpene lactone from Curcuma wenyujin.

The aim of the study was to investigate the sesquiterpene constituents from the rhizomes of Curcuma wenyujin Y. H. Chen et C. Ling. The isolation and purification of the constituents from the 50% EtOH extracts of the rhizomes were performed with repeated column chromatography over sillica gel and macroporous resin. Eight sesquiterpenes were obtained and identified as wenyujinlactone A (1), neolitamone A (2), zedoarondiol (3), isozedoarondiol (4), aerugidiol (5), curcumol (6), curdione (7) and (1R, 10R)-epoxy-(-)-1, 10-dihydrocurdine (8) by means of spectral analysis. Among them, compound 1 was found to be a new eudesmane sesquiterpene lactone, whilst compounds 2-5 were obtained from this plant for the first time.

Effect of matrine and carvedilol on collagen and MMPs activity of hypertrophy myocardium induced by pressure overload.

OBJECTIVE: To explore the effect and mechanism of matrine (Mt.) on myocardial interstitial fibrosis induced by pressure overload. METHODS: Pressure overloaded myocardial hypertrophy was produced by banding of aorta abdominalis in 67 male Sprague-Dawley rats weighing (200+/-15) g. The rats were assigned into one of the following groups: sham-operation control, operation control, operation group treated with matrine (15 mg/(kg.d)) and treated with carvedilol (Car.) (3.6 mg/(kg.d)) group. The rats were given drugs one day after operation. Five weeks after treatment, the left ventricular weight (LVW) was measured and the volume of myocardial cells was detected with Hematoxylin-Eosin (H-E) stain and Masson stain was used to assess the level of fibrosis of the myocardial matrix. Myocardial metalloproteinase activity was quantified with zymography, and survival rate was calculated. RESULTS: Survival rate significantly decreased (P<0.05), LVW/BW (body weight), MMP-2 (matrix metalloproteinase-2) activity (P<0.05), size of cardiomyocytes and interstitial fibrosis obviously increased in the operation group compared with sham control group. Mt. and Car. treatment can significantly increase survival rate (P<0.05), decrease LVW/BW (P<0.05) and MMP-2 activity (P<0.05), decrease size of cardiomyocytes and interstitial fibrosis compared with operation group. But there was difference compared with sham group. CONCLUSION: Matrine was shown to be able to prevent cardiac remodelling of hypertrophy cardium induced by pressure overload including myocardial hypertrophy and fibrosis which may be associated with the decrease in MMP-2 activity of heart.

Determination of urea content in urea cream by centrifugal partition chromatography.

The objective of this study is to establish a centrifugal partition chromatography (CPC) method for determination of the urea ingredient in urea cream. The mechanism of this method is that urea is determined by UV detector at 430 nm after being extracted from the cream and derivatized on line via Ehrlich reaction in rotor of CPC, where the reaction products dissolve in the mobile phase and the cream matrix retains in the stationary phase. The mixed solvent consisting of n-hexane, methanol, hydrochloric acid and p-dimethylaminobenzaldehyde with a ratio of 1000 mL:1000 mL:18 mL:2.0 g is used for solvent system of CPC. The CPC method proposed offers good precision and convenience without complex sample pretreatment processes.

Rapid discrimination and quantification of alkaloids in Corydalis Tuber by near-infrared spectroscopy.

With the application of near-infrared spectroscopy (NIRS), a convenient and rapid method for determination of alkaloids in Corydalis Tuber extract and classification for samples from different locations have been developed. Five different samples were collected according to their geographical origin, 2-Der with smoothing point of 17 was applied as the spectral pre-treatment, and the 1st to scaling range algorithm was adjusted to be optimal approach, classification model was constructed over the wavelength range of 4582-4270 cm⁻¹, 5562-4976 cm⁻¹ and 7000-7467 cm⁻¹ with a great recognition rate. For prediction model, partial least squares (PLS) algorithm was utilized referring to HPLC-UV reference method, the optimum models were obtained after adjustment. Pre-processing methods of calibration models were COE for protopine and min-max normalization for palmatine and MSC for tetrahydropalmatine, respectively. The root mean square errors of cross-validation (RMSECV) for protopine, palmatine, tetrahydropalmatine were 0.884, 1.83, 3.23 mg/g. The correlation coefficients (R²) were 99.75, 98.41 and 97.34%. T test was applied, in the model of tetrahydropalmatine; there is no significant difference between NIR prediction and HPLC reference method at 95% confidence interval with t=0.746

[Effect of long-term fertilization on dissipation of pentachlorophenol in red paddy field].

Dissipation of pentachlorophenol (PCP) with initial concentration of 85 mg/kg in red paddy field with rice cultivation and non-rice cultivation were investigated under the conditions of four long-term fertilization treatments, including the ones without fertilizer (CK), application of urea (N), application of organic fertilizer (OM), and N + OM. The results showed that the extractable PCP residues in surface soil in the treatments CK, N, OM and N + OM under non-rice cultivation condition were 28.3, 34.2, 19.3, 18.7 mg/kg after harvesting rice plant, and the extractable PCP residues in subsurface soil were 6.3, 9.1, 5.1, 4.1 mg/kg,respectively. Under rice cultivation condition, the extractable PCP residues in surface soil were 19.4, 30.9, 16.7, 8.7 mg/kg, and the extractable PCP residues in subsurface soil were 3.7, 6.1, 2.6, 2.8 mg/kg, respectively. The results indicated that the long-term application of organic fertilizer or inorganic plus organic fertilizer significantly accelerated the dissipation of PCP in surface soil, and decreased the transportation of PCP down the soil profile. However, long-term application of urea inhibited the dissipation of PCP in surface soil, and increased the transportation of PCP down the soil profile. When compared with non-rice cultivation, rice cultivation significantly accelerated the dissipation of PCP in surface soil other than the treatment N. In addition,the rice cultivation significantly decreased the transportation of PCP down the soil profile. No matter rice was cultivated or not, the dechlorinated metabolites of PCP detected in paddy soil were 2,3,4,5-TeCP and 3,4,5-TCP. 2,3,4,5-TeCP was the major dechlorinated metabolite of PCP under non-rice cultivation condition,whereas 3,4,5-TCP was the major dechlorinated metabolite of PCP under rice cultivation condition.

Two new isoflavone glycosides from Pueraria lobata.

Two new isoflavone diglycosides, formononetin 8-C-[beta-D-apiofuranosyl-(1 --> 6)]-beta-D-glucopyranoside (1) and formononetin 8-C-[beta-D-xylopyranosyl-(1 --> 6)]-beta-D-glucopyranoside (2), were isolated from the roots of Pueraria lobata, together with four known compounds, 4'-methoxypuerarin (3), daidzin (4), genistin (5), and daidzein (6). The structures of these compounds were elucidated by the spectroscopic methods.

Two new iridoid glycosides from the Tibetan folk medicine Swertia franchetiana.

Two new iridoid glycosides designated as senburiside III (2) and senburiside IV (3), together with one known iridoid glycoside senburiside I (1) and three known secoiridoid glucosides swertiamarin (4), gentiopicroside (5) and sweroside (6), were isolated from the whole plant of Swertia franchetiana. The structures of the two new compounds were elucidated by spectroscopic methods.

Xanthone glycosides from Swertia franchetiana.

A new xanthone glycoside (1) has been isolated from Swertia franchetiana together with five known xanthone glycosides. Their structures were elucidated as 7-O-[beta-D-xylopyranosyl-(1-->2)-beta-D-xylopyranosyl]-1,7,8-trihydroxy-3-methoxyxanthone (1), 7-O-[alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl]-1,7,8-trihydroxy-3-methoxyxanthone (2), 8-O-beta-D-glucopyranosyl-1,3,5,8-tetrahydroxyxanthone (3), 1-O-beta-D-glucopyranosyl-1-hydroxy-3,7,8-trimethoxyxanthone (4), 1-O-[beta-D-xylopyranosyl-(1-->6)-beta-D-glucopyranosyl]-1-hydroxy-2,3,5-trimethoxyxanthone (5) and 1-O-[beta-D-xylopyranosyl-(1-->6)-beta-D-glucopyranosyl]-1-hydroxy-3,5-dimethoxyxanthone (6) on the basis of spectroscopic evidence.