Exploring the molecular mechanism of Artemisia rupestris L. for the treatment of hepatocellular carcinoma via PI3K/AKT pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Hepatocellular carcinoma (HCC) is a common gastrointestinal malignancy in China. Most tumors develop from chronic inflammation. Artemisia rupestris L. (ARL) has been found to have a significant effect on viral influenza and hepatitis, but the mechanism of action of ARL against liver cancer is unclear. AIM OF THE STUDY: The study objective was to explore the mechanism of action of ARL for the treatment of hepatocellular carcinoma (HCC) by ethanol extract and in vitro experimental design. MATERIALS AND METHODS: Interactions between ARL and cellular target proteins against HCC were analyzed through network pharmacology and network topology with the utilization of the DAVID database. The rate of HepG2 cells' growth inhibition was assessed using the MTT assay in vitro cellular assay; hoechst33342 detects apoptosis of cells; the ability of HepG2 cells to migrate and invade was assessed using the transwell assay and the cell scratch experiment; and the levels of protein expression in HepG2 cells were assessed using the western blot assay. RESULTS: Network pharmacology prediction results demonstrated that 22 active ingredients were tested, 176 possible action targets were discovered, and the PI3K/Akt signaling pathway was found to be the most pertinent action pathway for the treatment of hepatocellular carcinoma. In vitro results showed that it can effectively restrict HepG2 cell proliferation, apoptosis, migration, and invasion as well as the regulation of protein expressions. CONCLUSION: Conclusively, Quercetin, Linarin, and Kaempferol were found most essential active ingredients from ARL that regulate the antitumor effects against HCC through the PI3K/Akt signaling pathway. The study provides a fundamental basis for further comprehensive evaluation of ARL to treat tumor diseases in general and its therapeutic potential against hepatocellular carcinoma in particular.

Uncovering quality markers of Yiqi-Tongluo capsule against myocardial ischemia and optimization of its extraction process.

Myocardial ischemia (MI), a condition in which the heart is unable to function due to insufficient blood and oxygen supply, is a major cause of death from coronary heart disease (CHD). Yiqi Tongluo capsule (YTC) is a Chinese patent drug which commonly used for treatment of MI in clinic. However, the related active components of YTC for treatment of MI were still uncovered. This paper is aimed to study the quality markers (Q-markers) of YTC and further optimize the extraction process of YTC based on Q-markers, providing research foundation for the further modern pharmaceutical preparations of YTC. We firstly used UPLC-QTOF-MS to analyze the constituents of YTC absorbed in blood, then isoprenaline (ISO) induced H9c2 cell model was used further screen the active constituents with protective effects on cardiomyocytes. After that, the orthogonal table (L9 (34)) was used to optimize the extraction process with three levels of 4 factors (water addition, immersion time, extraction time and decoction times). Finally, the HPLC fingerprint of 15 batches of optimized YTC was established. In our present study, a total of 33 components were identified in YTC, of which 10 components were absorbed in blood. Among the 10 components, 8 compounds had significant protective effects on ISO stimulated H9c2 cells, including Paeoniflorin, Ferulic acid, Calycosin, Senkyunolide A, N-butylphthalide, Z-ligustilide, LevistilideA, and Astragaloside IV, which were considered as the Q-markers of YTC. The optimized extraction process based on Q-marker as follows: soaking 1 h, then adding 8 times water to extract 3 times by decoction, each extraction lasts 1.5 h. The HPLC fingerprint of optimized YTC was established with 15 batches of YTC samples, and the optimized YTC samples has no significant toxicity to the heart, liver, spleen, lungs, and brain tissues of rats.

Active substances and molecular mechanisms of the anti-myocardial ischemia effects of Carthami flos by network pharmacology and in vitro experiments.

Myocardial ischemia is a predominant cardiovascular disorder that can result in a series of life-threatening cardiovascular diseases. Carthami flos (CF), the flower of Carthamus tinctorius L., is a commonly used herbal medicine in Chinese medicine for treating coronary atherosclerotic heart diseases based on its anti-myocardial ischemia (MI) effects. This paper aimed to investigate the active substances and mechanisms of the anti-MI effects of CF by network pharmacology and in vitro experiments. The results indicated that 9 constituents showed high degree of association with multiple targets of MI, including quercetin, kaempferol, ß-sitosterol, luteolin, baicalein, safflomin A, safflomin C, safflower-yellow-B and hydroxysafflor yellow A. In addition, AKT1, EGFR, CASP3, MYC, JUN, ALB, CTNNB1, VEGFA, ESR1, and IL1B were screened as the leading targets with a degree number ≥50. Bioinformatic annotation of GO-MF and KEGG showed that the anti-MI effects of CF are related to apoptosis and response to antioxidative stress pathways. The in vitro results showed that CF reduced LDH and CK levels, alleviated cell cycle arrest, and decreased ROS levels in H2O2-treated H9c2 cells. In addition, CF also promoted the nuclear shift of Nrf2 and the mRNA expressions of Akt, Nrf2 and Bcl-2 but decreased the expression of caspase-3 in H2O2-treated H9c2 cells. Collectively, the anti-MI effects of CF involve inhibiting apoptosis and antioxidative stress in cardiomyoblasts by regulating Akt/Nrf2/Caspase-3/Bcl-2, and the possible active substances of CF are quercetin, kaempferol, ß-sitosterol, luteolin, baicalein, safflomin C, safflower-yellow-B, and hydroxysafflor yellow A. The results of this study will be helpful for further drug development of CF and its active monomers.

MNX1 promotes cell proliferation and activates Wnt/ß-catenin signaling in colorectal cancer.

Aberrant Wnt/ß-catenin signaling is a characteristic feature of colorectal cancer (CRC), therefore, understanding the underlying mechanisms of aberrant Wnt/ß-catenin signaling will improve the treatment outcome of CRC. Expression of MNX1 in paired fresh CRC tissues and corresponding adjacent normal tissues were examined by qPCR and Western blotting. The levels of MNX1 in paraffin-embedded CRC specimens were detected by immunohistochemistry (IHC). The role of MNX1 in growth and proliferation of CRC cells was evaluated by MTT and colony formation assay. Luciferase reporter analysis and western blotting were carried out to explore the influence of MNX1 on Wnt/ß-catenin signaling. The results showed that expression of MNX1 is markedly upregulated in CRC tissues and positively correlated with level of Ki67, and overexpression of MNX1 significantly promotes the proliferation of CRC cells. Further study showed that ectopic expression of MNX1 activates the Wnt/ß-catenin signaling and upregulates the expression of c-Myc and CCND1, the downstream genes of Wnt/ß-catenin signaling. Therefore, MNX1 plays an indispensable role in promoting of human CRC progression and may represent a novel therapeutic target for CRC.

Molecular and phenotypic characterization of Colletotrichum species associated with anthracnose disease in peppers from Sichuan Province, China.

The anthracnose caused by Colletotrichum species is an important disease that primarily causes fruit rot in pepper. Eighty-eight strains representing seven species of Colletotrichum were obtained from rotten pepper fruits in Sichuan Province, China, and characterized according to morphology and the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sequence. Fifty-two strains were chosen for identification by phylogenetic analyses of multi-locus sequences, including the nuclear ribosomal internal transcribed spacer (ITS) region and the ß-tubulin (TUB2), actin (ACT), calmodulin (CAL) and GAPDH genes. Based on the combined datasets, the 88 strains were identified as Colletotrichum gloeosporioides, C. siamense, C. fructicola, C. truncatum, C. scovillei, and C. brevisporum, and one new species was detected, described as Colletotrichum sichuanensis. Notably, C. siamense and C. scovillei were recorded for the first time as the causes of anthracnose in peppers in China. In addition, with the exception of C. truncatum, this is the first report of all of the other Colletotrichum species studied in pepper from Sichuan. The fungal species were all non-host-specific, as the isolates were able to infect not only Capsicum spp. but also Pyrus pyrifolia in pathogenicity tests. These findings suggest that the fungal species associated with anthracnose in pepper may inoculate other hosts as initial inoculum.

[Problems and solutions in minimally invasive process of posterior discectomy for lumbar disc herniation].

OBJECTIVE: To analyze the problems and complications of posterior discectomy for lumbar disc herniation. METHODS: From January 2005 to June 2010, 497 patients with lumbar disc herniation were treated by posterior discectomy, which data were analyzed retrospectively. There were 395 males and 102 females,ranging in age from 20 to 78 years with an average of 43.7 years. Among them, 405 cases were in single gap, 86 cases were in double gaps and 6 cases were in three gaps. The complication of operation and solution was analyzed. RESULTS: The mean operative time was 70 min (from 45 to 210 min), and the mean hospitalization was 10 d (from 5 to 20 d). Forty-seven cases suffered operative complications. There were 16 cases of wrong location of segments (14 cases occurred in operation and 2 occurred after operation), 15 cases of less alleviation or aggravation of nerve symptoms (12 cases were poor alleviation and 3 cases were aggravation), 10 cases of urinary retention, 5 cases of cerebrospinal fluid leakage and 1 case of infection. CONCLUSION: Minimally invasive process of posterior discectomy in treating lumbar disc herniation may complicate with many problems. The operative effects can be improved and the complication can be decreased if the ability of location is improved,surgical indications is correct and the operation is exactly performed.

[The first exploration of a minimally invasive lysis subcutaneouly for the treatment of gluteal muscle contracture based on relatively safe region around standard injection point of gluteal muscle].

OBJECTIVE: To explore the solution of choosing the minimally invasive incision site for gluteal muscle contracture patient based on standard injection point of gluteal muscle. METHODS: from September 2008 to August 2010, 25 patients (14 males and 11 females with an average of 16.5 years, ranging from 12 to 26 years) with injected gluteal muscle contracture were prospectively studied. The course of disease was from 6 to 12 years. Firstly, the connective skin Surface line from anterior superior iliac spine to coccyx (line AD) was delineated and the point (point O) was marked out as the standard gluteal muscle injection site which was on the one-third of the distance from the anterior superior iliac spine(point A) to the coccyx (point D). Secondly, the anterior and posterior edge lines of surface projection of the gluteal muscle contracture banding (line a, line p) were delineated. Thirdly, the distance from B to O and C to O (B is the point of intersection of line a and line AD,C is the point of intersection of line P and line AD)were measured which was the intersection of line a,p and line AD to point O. Lastly, the minimally invasive surgery was operformed via the skin entry of point C. RESULTS: OB = (0 +/- 0.76) cm, OC = (2.86 +/- 0.78) cm, BC = (2.86 +/- 1.01) cm,the mean postoperative drainage was less than 10 ml,there was no nerve damage,hematoma and other complications. All patients achieved the function of squatting in 4 to 6 days. CONCLUSION: The solution of choosing the minimally invasive incision site based on standard injection point of gluteal muscle has advantages of positioning precisely,handling easily, recoverying quickly, less trauma and safety, etc.

DIFFEOMORPHIC SURFACE FLOWS: A NOVEL METHOD OF SURFACE EVOLUTION.

We describe a new class of surface flows, diffeomorphic surface flows, induced by restricting diffeomorphic flows of the ambient Euclidean space to a surface. Different from classical surface PDE flows such as mean curvature flow, diffeomorphic surface flows are solutions of integro-differential equations in a group of diffeomorphisms. They have the potential advantage of being both topology-invariant and singularity free, which can be useful in computational anatomy and computer graphics. We first derive the Euler-Lagrange equation of the elastic energy for general diffeomorphic surface flows, which can be regarded as a smoothed version of the corresponding classical surface flows. Then we focus on diffeomorphic mean curvature flow. We prove the short-time existence and uniqueness of the flow, and study the long-time existence of the flow for surfaces of revolution. We present numerical experiments on synthetic and cortical surfaces from neuroimaging studies in schizophrenia and auditory disorders. Finally we discuss unresolved issues and potential applications.