Calycosin ameliorates spinal cord injury by targeting Hsp90 to inhibit oxidative stress and apoptosis of nerve cells.

BACKGROUND: Zhenbao pill is effective in protecting against spinal cord injury (SCI). We attempt to explore the characteristics of calycosin (a main monomer of Zhenbao pill) in SCI and its relative mechanism. METHODS: The target of calycosin was screened using pharmacological network analysis. The SCI cell model was constructed using hydrogen peroxide (H2O2), and the animal model was developed by compressing spinal cord with a vascular clamp. Flow cytometry was conducted to test reactive oxygen species (ROS) levels and cell apoptosis. Detection of malondialdehyde (MDA) activity and Superoxide dismutase (SOD) activity were performed using relative kits. Heat shock protein 90 (HSP90) was examined using western blot and quantitative real-time PCR. Motor function tests were carried out. The hematoxylin-eosin and Nissl staining were conducted. RESULTS: In SCI models, ROS, MDA, and cell apoptosis were elevated, SOD and HSP90 levels were restrained, while calycosin addition reversed the above results. Besides, calycosin application or HSP90 overexpression enhanced phosphorylation of protein kinase B (Akt) but weakened that of apoptosis signal-regulating kinase 1 (ASK1) and p38, while HSP90 inhibitor 17-AAG treatment restrained the above results. Meanwhile, the injection of calycosin improved the motor function in SCI model rats. Furthermore, the pathologic results also clarified the positive effect of calycosin on SCI. CONCLUSION: HSP90 was lowly expressed in SCI models. Calycosin alleviated SCI by promoting HSP90 up-regulation and inhibiting oxidative stress and apoptosis of nerve cells.

Identification of Hub Genes to Regulate Breast Cancer Spinal Metastases by Bioinformatics Analyses.

Breast cancer (BC) had been one of the deadliest types of cancers in women worldwide. More than 65% of advanced-stage BC patients were identified to have bone metastasis. However, the molecular mechanisms involved in the BC spinal metastases remained largely unclear. This study screened dysregulated genes in the progression of BC spinal metastases by analyzing GSE22358. Moreover, we constructed PPI networks to identify key regulators in this progression. Bioinformatics analysis showed that these key regulators were involved in regulating the metabolic process, cell proliferation, Toll-like receptor and RIG-I-like receptor signaling, and mRNA surveillance. Furthermore, our analysis revealed that key regulators, including C1QB, CEP55, HIST1H2BO, IFI6, KIAA0101, PBK, SPAG5, SPP1, DCN, FZD7, KRT5, and TGFBR3, were correlated to the OS time in BC patients. In addition, we analyzed TCGA database to further confirm the expression levels of these hub genes in breast cancer. Our results showed that these regulators were significantly differentially expressed in breast cancer, which were consistent with GSE22358 dataset analysis. Furthermore, our analysis demonstrated that CEP55 was remarkably upregulated in the advanced stage of breast cancer compared to the stage I breast cancer sample and was significantly upregulated in triple-negative breast cancers (TNBC) compared to other types of breast cancers, including luminal and HER2-positive cancers, demonstrating CEP55 may have a regulatory role in TNBC. Finally, our results showed that CEP55 was the most highly expressed in Basal-like 1 TNBC and Basal-like 2 TNBC samples but the most lowly expressed in mesenchymal stem-like TNBC samples. Although more studies are still needed to understand the functions of key regulators in BC, this study provides useful information to understand the mechanisms underlying BC spinal metastases.

Comparison of Nephrotoxicity Induced By MongolianMeng-Gen-Wu-Su (Mercury) Processed Products, MongolianMeng-Gen-Wu-Su(Mercury)-18-Composition Pill and Mercuric Sulfide, Mercuric Chloride and Mercurous Chloride / 世界科学技术-中医药现代化

The renal toxicity of rats after a single dose ofMeng-Gen-Wu-Su (mercury) processed products,Meng-Gen-Wu-Su (mercury)-18-composition pill, mercuric sulfide, mercuric chloride, and mercurous chloride was studied. Fifty-four male Wistar rats were randomly divided into nine groups according to body weights (6 rats in each group): normal control group, low and high dose groups (0.033, 0.33 g·kg-1·d-1) ofMeng-Gen-Wu-Su (mercury) processed products, low and high dose groups (0.29, 2.9 g·kg-1·d-1) ofMeng-Gen-Wu-Su (mercury)-18-composition pill, simplified prescription ofMeng-Gen-Wu-Su (mercury)-18-composition pill group (0.26 g·kg-1·d-1), mercuric sulfide group (17.39 mg·kg-1·d-1), mercuric chloride group (4.06 mg·kg-1·d-1) and mercurous chloride group (35.3 mg·kg-1·d-1). After acclimation for one week, once oral administration was given to each group of rats. After 24 h, function and morphological changes of liver and kidney were detected. Mercury accumulation in kidney was determined by inductively coupled plasma optical emission spectroscopy (ICP-OES) and inductively coupled plasma source mass spectrometer (ICP-MS). Apoptosis of renal cell was determined by terminal-deoxynucleoitidyl transferase mediated Nick End Labeling (TUNEL). Renal typeⅢ collagen protein's expression was determined by immunohistochemical (HIC) method and expression changes of MT-1, MT-2 mRNA in kidney were also determined by real-time fluorescence quantitative PCR (real-time-PCR). There was no significant difference of ALT, AST in serum between normal control group and other groups (P>0.05). CREA and UREA in mercurous chloride group were apparently higher than normal control group and low dose group of Meng-Gen-Wu-Su processed products (P<0.01). Hepatic and renal pathologic examination results showed that liver cell of low dose groups ofMeng-Gen-Wu-Su processed products andMeng-Gen-Wu-Su-18-composition pill swelled to a low degree and glomerular disease was not obvious. In high-dose groups ofMeng-Gen-Wu-Su processed products,Meng-Gen-Wu-Su-18-composition pill and mercuric sulfide group, liver and kidney appeared some pathological changes and such changes were more significant in mercuric chloride and mercurous chloride groups. Compared with normal control group and low dose group ofMeng-Gen-Wu-Su processed products, the mercury kidney volume in mercuric chloride and mercurous chloride groups increased significantly (P<0.01). The apoptosis rate of renal cell and expression of typeⅢ collagen protein increased significantly in the groups of mercuric sulfide, mercuric chloride and mercurous chloride (P<0.01). MT-1and MT-2 mRNA gene expression rised significantly in the groups of mercuric chloride and mercurous chloride (P<0.05 orP<0.01). In summary, the rats renal toxicity after a single dose ofMeng-Gen-Wu-Su (Mercury) processed products or MongolianMeng-Gen-Wu-Su (Mercury)-18-composition pill were both far less than that of mercuric chloride or mercurous chloride.

Research on Composition of Mongolian Betel Shi-San-Wei Ingredients Pill (Gao-You-13) / 世界科学技术-中医药现代化

Betel Shi-San-Wei Ingredients Pill(BSSWIP) was first recorded in the 19th century writings Meng-Yi Jin-Gui with the name of Gao-Y ou-13. The name of BSSWIP was first recorded in the book of the 1977 edition of the Drug Standard of the Jilin Province, which was formerly known as Tai De Hu Ran Gu Lu Ge Qi Nai Ran Ta, Se Me Ji De Ji De, and etc. Although in the book of Tong-Wa-Ga-Ji-De, Se Me Ji De Ji De was documented, it was the same name of different compositions. It had no original relation with BSSWIP. In different periods, the BSSWIP was consisted of 13, 14 or 15 kinds of herbs. There were at least five different types of herbs appeared in the Gao-Y ou-13. The evolution of prescription was mainly from the 19th century to the first half of the 20th century. There was no major change on prescription composition and proportion since 1971. Among them, 10 kinds of herbs, which were Bing-Lang, Guang-Zao, Mu-Xiang, Ding-Xiang, Rou-Dou-Kou, Zi-Nao-Sha, Gan-Jiang, Bi-Ba, Hu-Jiao, and Chen-Xiang were fixed. The ratio of single herbal medicine changed the most was Zhi-Cao-W u, which was followed by Mu-Xiang, Ding-Xiang and Chen-Xiang. There were no marked sources of BSSWIP in the recording of the Drug Standard of the Jilin Province and the Mongolian Medicine V olume·Ministry of Health of the People's Republic of China. The composition and proportion were considered to be from the book Meng-Y i Jin-Gui according to notes of Standards on Mongolian Patent Medicine in Inner Mongolia. Recordings of three standards are in consistence with the Meng-Y i Jin-Gui on Gao-Y ou-13 except for Y e-Mao-Niu Xin and the different ratio of Zhi-Cao-W u. In the appendix of the Standards on Mongolian Patent Medicine in Inner Mongolia, it marked the differences from the original prescription. Therefore, the other two criteria should also mark the similarities and differences compared with the original prescription properly.

Effects of Betel shisanwei ingredients pill on AC-cAMP-PKA signal transduction pathways in hippocampus and prefrontal cortex of depressive rats / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effects of Mongolian pharmaceutical Betel shisanwei ingredients pill on AC-cAMP-PKA signal transduction pathways in hippocampus and prefrontal cortex of depressive rats.</p><p><b>METHOD</b>Sixty male Wistar rats were randomly divided into six groups according to the sugar consumption test (10 rats in each group), normal control group,model group,fluoxetine group (3.3 mg x kg(-1)) and low dose, medium dose and high dose group (0.25, 0.5, 1 g x kg(-1)) of Betel shisanwei ingredients pill. Except the normal control,the other groups were treated with the chronic unpredictable mild stress stimulation combined with lonely raising for 28 days. 10 mL x kg(-1) of drugs were given to each rat once daily,continuously for 28 days. The AC activity of the hippocampus and prefrontal cortex were determined by radiation immunity analysis (RIA), while cAMP and PKA quantity were determinated by Enzyme-linked immunosorbent (ELISA).</p><p><b>RESULT</b>The AC activity, cAMP and PKA quantity of hippocampus and prefrontal of mouse model of Chronic stress depression decreased significantly than those of control group (P < 0.05 or P < 0.01). However, the AC activity, cAMP and PKA quantity of rat hippocampus and prefrontal cortex in the fluoxetine group and the Mongolian pharmaceutical Betel shisanwei ingredients pill group indecreased significantly than those of model group (P < 0.01 or P < 0.05). Especially for the high dose group of Mongolian pharmaceutical Betel shisanwei ingredients pill.</p><p><b>CONCLUSION</b>The AC-cAMP-PKA signal transduction pathways in hippocampus and prefrontal cortex of depression model of rats is down-regulated, whereas Mongolian pharmaceutical Betel shisanwei ingredients pill could up-regulated it to resist depression.</p>

Mongolian Medicine Meng-Gen-Wu-Su (Mercury)-18-composition Pill Research / 世界科学技术-中医药现代化

Meng-Gen-Wu-Su (mercury)-18-composition pill was firstly recorded in the book of Gan-Lu-Si-Bu with the name of Jin-18 pill . The name of Me ng-G e n-W u-Su ( mercury )-18-composition pill was firstly recorded in the book of Me ng-Y i-Jin-G ui . Its composition and dosage had always been adjusted in the later dynasties . Until the issue of the book Ne i-Me ng G u-Me ng Che ng-Y ao Biao-Zhun , the composition and dosage of Meng-Gen-Wu-Su was promulgated. In different periods, the Meng-Gen-Wu-Su consisted of 17, 18 or 19 kinds of herbs. There are at least five different types of herbs appeared in the Meng-Gen-Wu-Su-18-composition pill. But 16 kinds of medicinals such as mercury, He-Zi, Cao-Wu, Liu-Huang, Qing-Ma-Zi, Jue-Ming-Zi, Bai-Yun-Xiang, Mu-Xiang, Shi-Chang-Pu, Su-Ge-Mu-Le, Shi-Gao, Rou-Dou-Kou, Ding-Xiang, Cao-Guo, Hong-Hua, Hei-Yun-Xiang are fixed in the composition. The proportion of Meng-Gen-Wu-Su-18-composition pill is inconsistent in different periods. The significant difference of drug dosage proportion are Liu-Huang and Bai-Yun-Xiang, followed by Qing-Ma-Zi, Jue-Ming-Zi, Cao-Guo, Wen-Guan-Mu. The Meng-Gen-Wu-Su-18-composition pill and Jin-18 pill from the book of Gan-Lu-Si-Bu are same prescription with the same composition but different names. The composition and dosage proportion of Meng-Gen-Wu-Su-18-composition pill from the book of Meng-Yi-Jin-Gui and He-Li-Le Jing-Zhu Jie-Y i Nan-Jing are the same with the same prescription name .